Competition for growth substrates in river water between Escherichia coli and indigenous bacteria illustrated by high-resolution mass spectrometry.

Escherichia coli normally cannot grow in the environment. One environmental stress that prevents E. coli growth may be the competition for growth substrates with co-existing micro-organisms. In this study, the growth substrates of E. coli were screened by high-resolution mass spectrometry and compared with those of indigenous bacteria in river water. In an incubation experiment, E. coli multiplied in sterilized river water, but did not multiply when indigenous micro-organisms were present in the water. By analysing dissolved organic matter in the river water before and after E. coli growth, 35 compounds were identified as putative growth substrates of E. coli. Among them, 33 compounds were also identified as putative growth substrates of indigenous bacteria. These results indicate that E. coli and indigenous bacteria compete for organic substrates in river water, which could suppress the growth of E. coli.

Association of the colistin resistance gene mcr-1 with faecal pollution in water environments in Hanoi, Vietnam.

Colistin is one of the antibiotics of last resort for human health. However, the dissemination of the plasmid-mediated colistin resistance gene mcr-1 is of great concern globally. In the One Health framework, the environment is an important component for managing antimicrobial resistance. However, little information is available concerning the prevalence of mcr-1 in water environments. We aimed to reveal the prevalence of mcr-1 in different water environments in Hanoi, Vietnam. Quantitative PCR was applied to detect mcr-1 in four urban drainages receiving untreated domestic wastewater, three rivers, five lakes and two groundwater samples. Urban drainages contained higher concentrations of mcr-1, suggesting that urban residents carry the gene. The class 1 integron-integrase gene was identified as a good surrogate of antibiotic resistance genes including mcr-1. A significant correlation was found between the levels of mcr-1 and the human-specific cross-assembly phage, which is an indicator of human faecal pollution. These results indicated that the primary source of mcr-1 in urban water environments is human faeces, which is consistent with the fact that most domestic wastewater is untreated in Hanoi. The control of untreated wastewater is critical for alleviating the spread of mcr-1 in water environments in Vietnam.

Identification of estrone-degrading Betaproteobacteria in activated sludge by microautoradiography fluorescent in situ hybridization.

UNLABELLED: Natural oestrogens, which are degraded but not completely removed in wastewater treatment plants, are suspected of causing the endocrine disruption of aquatic organisms in the receiving water body. While several bacterial isolates were reported to be oestrogen-degrading bacteria, our previous study implied that only the unidentified rod-shaped Betaproteobacteria in chains were responsible for estrone (E1) degradation by activated sludge especially at the sub-milligram per litre level. The Betaproteobacteria were suspected to be related to genera Sphaerotilus and Leptothrix according to morphological observations. Probe Spha823 was newly developed to target 16S rRNA gene clones obtained from activated sludge and closely related to the above genera. [(3) H]E1-incubated sludge samples showed that most of the (3) H-labelled cells hybridized with probe Spha823 by microautoradiography (MAR) fluorescent in situ hybridization. Spha823-defined cells were present in all three activated sludge samples tested, where they accounted for up to 3% of the total microbial biomass. Spha823-defined cells comprised 59·5-80·1% of the total MAR-positive cells, which suggested that the Sphaerotilus-Leptothrix-related bacteria were the most abundant micro-organisms involved in E1 degradation (at 200 µg l(-1) ) in the activated sludge samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Estrone (E1) is one of the natural estrogens, which can be degraded but is not always completely removed in wastewater treatment plants. E1 is suspected of causing the endocrine disruption of aquatic organisms in the receiving water body. We identified dominant E1-incorporating bacteria, which should include E1-degrading bacteria, in activated sludge treating domestic wastewater. Sphaerotilus-Leptothrix-related bacteria, which had never been reported in the previous attempts based on culture-dependent approach, occupied 60-80% of the E1-incorporating bacteria. This study demonstrates the identification of functionally active bacteria to degrade micro-pollutants at sub-milligram per litre level.

Application of whole sediment toxicity identification evaluation procedures to road dust using a benthic ostracod Heterocypris incongruens.

Road dust is considered to be an important source of sediment contamination in receiving water bodies; however, few studies have evaluated the toxicity of road dust to benthic organisms. This study evaluated the toxicity of road dust to a benthic ostracod, Heterocypris incongruens, using a six-day direct exposure experiment. We applied whole sediment toxicity identification evaluation (TIE) methods to identify the primary group of toxicants in road dust. Three road dust samples from Tokyo caused high ostracod mortality. The addition of hydrophobic adsorbents, Ambersorb and XAD, eliminated toxicity in all samples, suggesting that hydrophobic compounds were the main toxicants in road dust samples. A cation exchange resin, Chelex, also reduced the toxicity of two samples, although the measured concentrations of dissolved heavy metals in the test solution did not exceed the LC(50) values in the literature. In addition, the sum of toxic unit (TU=measured concentration/LC(50)) of each individual metal which predicts the toxicity of the metal mixtures did not exceed 1.0 in all samples, suggesting that heavy metal mixtures did not have additive effects. We hypothesized that the toxicity reduction by XAD and Chelex was due to the removal of hydrophobic compounds, rather than heavy metals. Thus, a toxicity test was conducted on fractions eluted with organic solvents from the XAD and Chelex recovered from one of the road dust samples. Methanol-eluted fractions of XAD and Chelex showed 100 percent ostracod mortality, indicating that the hydrophobic organic compounds removed by these adsorbents were the principal toxicants in the road dust sample.

Influence of backwashing on the microbial community in a biofilm developed on biological activated carbon used in a drinking water treatment plant.

The influence of backwashing on the biofilm community developed on biological activated carbon (BAC) used in a drinking water treatment plant was investigated by means of bacterial cell enumeration and terminal-restriction fragment length polymorphism (T-RFLP) fingerprinting analysis of bacterial and eukaryotic ribosomal RNA genes (rDNA). After backwashing, the attached bacterial abundance in the top layer of the BAC bed decreased to 64% of that before backwashing. The community level changes caused by backwashing were examined through the T-RFLP profiles. In the bacterial 16S rDNA analysis, the relative abundances of some terminal-restriction fragments (T-RFs) including the Planctomycetes-derived fragment increased; however, the relative abundances of some T-RFs including the Betaproteobacteria-derived fragments decreased. In the eukaryotic 18S rDNA analysis, the relative abundances of some T-RFs including the protozoan Cercozoa-derived fragments increased; however, the relative abundances of some T-RFs including the metazoan Chaetonotus- and Paratripyla-derived fragments decreased. The T-RFLP analysis suggests that backwashing can cause changes in the relative compositions of microorganisms in a BAC biofilm in the top layer of the bed.

Lack of association of group specific component haplotypes with lung function in smokers.

BACKGROUND: Airway inflammation may affect the decrease in lung function that occurs in response to cigarette smoke, and is an important pathological feature in chronic obstructive pulmonary disease (COPD). Group specific component (GC) can act as an inflammatory mediator and may therefore have important influences on the inflammatory reaction in the airway. Several reports have described associations between GC haplotypes and COPD but these remain controversial. In addition, most of these studies were based on a small number of subjects. METHODS: We have studied the contribution of GC haplotypes to the level of lung function in a large cohort of smokers with high or low lung function (mean FEV(1) % predicted 91.8 and 62.6, respectively). The frequency of the three major GC haplotypes (1S, 1F and 2) was investigated in 537 individuals with high lung function and 533 with low lung function. RESULTS: No significant difference was found in the frequency of any GC haplotype between the high and low lung function groups. There was also no significant difference between the groups in genotype frequency of the two single nucleotide polymorphisms that underlie the haplotypes. CONCLUSION: The GC haplotype does not contribute to reduced lung function in this cohort of smokers.

Tumor-related leukocytosis is linked with poor prognosis in patients with lung carcinoma.

BACKGROUND: Tumor-related leukocytosis is a paraneoplastic syndrome that is encountered occasionally in the clinical course of patients with lung carcinoma. Recently, autonomous production of hematopoietic cytokines (granulocyte-colony stimulating factor [G-CSF], granulocyte-macrophage-colony stimulating factor [GM-CSF], and interleukin-6 [IL-6]) were identified in some of these patients. However, the incidence and clinical characterization of this phenomenon have not been clarified. METHODS: During a 7-year period, 227 patients with carcinoma of the lung were investigated, and 33 patients were diagnosed with tumor-related leukocytosis. Except for one patient with small cell lung carcinoma, the other 32 patients had nonsmall cell lung carcinoma, and the highest incidence is recognized in large cell carcinoma. These 33 patients were examined for serum G-CSF, GM-CSF, and IL-6 levels using enzyme immunoassays and enzyme-linked immunosorbent assays. Tumor specimens were stained for antihuman cytokine (G-CSF, GM-CSF, and IL-6) monoclonal antibodies. RESULTS: Sixteen patients showed high serum G-CSF levels, 4 patients showed high serum GM-CSF levels, and 18 patients showed high serum IL-6 levels. Twelve specimens stained positively against anti-G-CSF antibody. Two specimens stained positively against anti-GM-CSF antibody, and three specimens were stained positively against anti-IL-6 antibody, including one double positive specimen for both G-CSF and IL-6. All specimens that were positive for monoclonal antibodies were from patients with nonsmall cell lung carcinoma. These patients had a poor outcome compared with the other patients. CONCLUSIONS: Tumor-related leukocytosis and cytokine production frequently occur in the clinical course of lung carcinoma, and this phenomenon is related mainly to nonsmall cell lung carcinoma. Its occurrence appears to be an ominous prognostic sign in patients with lung carcinoma.

Detection and quantification of human herpesvirus 6 genomes using bronchoalveolar lavage fluid in immunocompromised patients with interstitial pneumonia.

Human herpesviruses have been recognized as a pathogen involved in interstitial pneumonia (IP), especially in immunocompromised patients. So far, little is known about involvement of human herpesvirus 6 (HHV-6) in systemic respiratory tract disease. Currently, routine diagnostic tests for HHV-6 are inefficient for HHV-6 reactivation, therefore, we established a rapid quantification system of HHV-6 using real-time PCR in order to determine the possible role of human HHV-6 reactivation in immunocompromised patients showing IP. Bronchoalveolar lavage fluid (BALF) specimens were obtained from 84 consecutively treated patients with interstitial lung diseases (ILD) including various types of IP. First, we determined the viral burden in BALF and peripheral blood obtained from healthy volunteers. In healthy volunteers, the prevalence of HHV-6 in BALF was higher (4/12, 33.3%) than in peripheral blood (8/53, 15.1%), ranging from 0 to 101.65 HHV-6 genome copies per 1 microg of DNA. Among 84 patients with ILD analyzed, the prevalence of HHV-6 in BALF was 27.4% (23/84), ranging from 0 to 103.87 copies per 1 microg of DNA. Three specimens obtained from patients with collagen vascular disease, 2 from Hodgkin's disease, and 1 with sarcoidosis had high level of HHV-6 viral DNA, while none of the patients with idiopathic IP showed elevation of HHV-6 (more than 102) in BALF. Our results suggest that measurement of HHV-6 genomes in BALF using real-time PCR may be useful in management of the care of respiratory complications in immunocompromised patients.

Interstitial pneumonia possibly due to a novel anticancer drug, TS-1: first case report.

A newly approved oral fluoropyrimidine, TS-1, is a dihydropyrimide dehydrogenase (DPD)-inhibiting fluoropyrimidine (DIF) drug. We describe a case of interstitial pneumonia probably caused by TS-1. A peripheral blood lymphocytes stimulating test (DLST) with TS-1 demonstrated a substantial positive reaction. So far only three cases of TS-1-induced interstitial pneumonia have been reported but the relationship between interstitial pneumonia and TS-1 was demonstrated only in this case. Considering that interstitial pneumonia has also been reported with 5-FU, it is necessary in the future to clarify which component of this drug is directly related to interstitial pneumonia.

A new macrocyclic trichothecene, 12,13-deoxyroridin E, produced by the marine-derived fungus Myrothecium roridum collected in Palau.

A new macrocyclic trichothecene, 12,13-deoxyroridin E (1), and three known compounds, roridin E (2), verrucarin A (3), and verrucarin J (4), were obtained as cytotoxic components from the marine-derived fungus Myrothecium roridum, isolated in Palau. 12,13-Deoxyroridin E is the second example of a macrocyclic trichothecene possessing a double bond at C-12-C-13 and was about 80-fold less cytotoxic than roridin E, the epoxide variant.

Malignant pleural mesothelioma produces functional granulocyte-colony stimulating factor.

We report a patient with diffuse malignant pleural mesothelioma showing marked elevation of neutrophils. The level of serum granulocyte-colony stimulating factor (G-CSF) was elevated (138 pg/mL; normal range, < 20 pg/mL). The patient died 6 weeks after disease progression had been noted, and immunohistochemistry using a specific monoclonal antibody against recombinant G-CSF at autopsy demonstrated that the malignant mesothelioma cells actually produced G-CSF. Only three cases of malignant pleural mesothelioma, including the current patient, have been reported to produce G-CSF. We demonstrated an elevated serum level of G-CSF and G-CSF-bearing tumor cells by immunochemistry.

Elevated serum soluble CD44 level in sarcoidosis.

Sarcoidosis is a chronic multi-organ granulomatous disease of unknown etiology. Several studies have suggested an involvement of immunologic background in sarcoidosis. The lymphocyte surface marker CD44 is a multifunctional molecule which mediates the adhesion of lymphocytes to the extracellular matrix. Recently, we developed a system to quantitate soluble CD44 (sCD44) which we employed to determine serum and bronchoalveolar lavage fluid (BALF) levels of sCD44 to obtain further insights into immunologic aspects of sarcoidosis. Serum sCD44 levels were measured in 13 consecutive patients with sarcoidosis and 56 normal healthy controls using enzyme-linked immunoabsorbent assay. BALF sCD44 levels were also measured in 11 patients with sarcoidosis and 10 normal healthy controls. In patients with sarcoidosis, the serum sCD44 level was significantly higher than that of normal controls (348.5+/-164.2 ng/ml vs 145.4+/-22.9 ng/ml; p<0.001). Also BALF sCD44 levels tended to be higher in sarcoidosis than in normal controls (23.7+/-13.4 ng/ml vs 18.1+/-8.4 ng/ml), but no statistically significant difference was recognized. We also found that there was a positive correlation between the serum sCD44 and angiotensin converting enzyme (r=0.78). Our data indicate that sCD44 may be related to immunologic background and may be a useful new marker of sarcoidosis.

Detection of nocturnal wheezing in bronchial asthma using intermittent sleep tracheal sounds recording.

Common clinical features of bronchial asthma include bronchoconstriction during the night, particularly while asleep. Although bronchoconstriction reduces the quality of life and can cause life-threatening events, a clinical technique for evaluating bronchoconstriction during sleep has not been widely applied. In this study, we measured nocturnal wheezing by intermittent sleep tracheal sounds recording (ISTSR) to detect bronchoconstriction during the hours of sleep. Using ISTSR, we studied the number and duration of nocturnal wheezing episodes in 27 adult patients with bronchial asthma. Nocturnal wheezing was detected in 36 of 39 recordings. Although the pattern of hourly nocturnal wheezing count (hourly NWC pattern) varied among subjects, there appeared to be a reproducible pattern within individuals. When wheezing alternated between long and short duration, bronchoconstriction tended to be more severe. The NWC in 1 h (NWC/H) was positively correlated with subjective symptoms and inversely correlated with the morning per cent peak expiratory flow. The hourly NWC was significantly greater at 05:00 than that at midnight. Intermittent sleep tracheal sounds recording has potential to be a non-invasive clinical tool for detecting nocturnal bronchoconstriction during hours of sleep in patients with asthma.

Sarcoidosis induced by interferon therapy for chronic myelogenous leukaemia.

A 31-year-old male was diagnosed as having chronic myelogenous leukaemia and has been treated with hydroxyurea and interferon-alpha since February 1995. After 16 months, he complained of low-grade fever and a cough. Bilateral hilar lymph node enlargement was detected on the chest X-ray film and multiple subcutaneous erythematous nodules appeared. A skin biopsy revealed subcutaneous sarcoid granuloma. Two months after the cessation of interferon therapy, the subcutaneous nodules and the hilar lymph node enlargement resolved. It is possible that continuous interferon administration can promote granuloma formation in sarcoidosis by activating T cells and macrophages.

Alpha-fetoprotein producing pulmonary blastoma in a patient with systemic sclerosis: pathogenetic analysis.

We describe a rare case of pulmonary blastoma complicated with systemic sclerosis. The serum alpha-fetoprotein level was elevated at presentation and the biopsied tumour stained positively against anti-alpha-fetoprotein antibody. The alpha-fetoprotein produced autonomously by tumour cells was of yolk-sac origin. Although the pathogenesis of pulmonary blastoma has not been clarified, we suggest that this pulmonary blastoma is a type of yolk-sac tumour.

Elevation of antibodies to cytomegalovirus and other herpes viruses in pulmonary fibrosis.

The aim of this study was to determine whether latent viral infection is associated with idiopathic pulmonary fibrosis (IPF), an interstitial lung disease whose aetiology remains to be elucidated. Cytomegalovirus (CMV) immunoglobulin G (IgG) and complement fixation (CF), Epstein-Barr (EB) viral capsid antigen (VCA) IgG, herpes simplex virus (HSV) IgG, adenovirus CF, and parainfluenza 3 virus haemagglutinin inhibition (HI) titres were measured in the serum from patients with pulmonary diseases. The study included five subject groups: 35 normal controls (aged (mean +/- SD) 38 +/- 17 yrs); 43 IPF (63 +/- 10 yrs), seven collagen vascular disease-related interstitial pneumonitis (CVD-IP) (62 +/- 12 yrs); 22 sarcoidosis (36 +/- 14 yrs); and 17 emphysema (66 +/- 11 yrs). Levels of CMV IgG in IPF (87.6 +/- 51.7) and CVD-IP (101.2 +/- 69.9) were significantly elevated compared to those in the control (30.9 +/- 24.1), sarcoidosis (34.4 +/- 38.3) and emphysema groups (40.3 +/- 24.6), whereas CMV immunoglobulin M (IgM) was generally below the limit of detection. Similarly, CMV CF titres in IPF and CVD-IP were elevated compared to those in other diseases. EB VCA IgG titres in IPF, CVD-IP and emphysema and HSV IgG in IPF were also elevated. In contrast, adenovirus CF and parainfluenza 3 HI titres demonstrated no significant difference among all of the groups investigated. Increases in cytomegalovirus immunoglobulin G and complement fixation titres with negative cytomegalovirus immunoglobulin M suggest that latent cytomegalovirus infection may be more prominent in idiopathic pulmonary fibrosis or collagen vascular disease-related interstitial pneumonitis. Together with the elevation of Epstein-Barr virus viral capsid antigen and herpes simplex virus immunoglobulin G in idiopathic pulmonary fibrosis and/or collagen vascular disease-related interstitial pneumonitis, it is rational to assume that these viruses may be implicated in the development of pulmonary fibrosis. Further study is necessary to investigate the relationship between latent viral infection and pulmonary fibrosis.

Multiple pulmonary nodules in association with pyoderma gangrenosum.

This report describes a patient with extensive pyoderma gangrenosum in whom there were co-existent lung abnormalities. The patient's X-ray showed peripherally sited multiple pulmonary lesions bilaterally. A lung biopsy showed chronic non-specific inflammatory changes with neutrophil and lymphocyte infiltration which were similar to the skin lesions. This case was diagnosed as multiple aseptic nodules in pyoderma gangrenosum. The pulmonary infiltrative shadows were controlled only with prednisolone treatment. Steroid therapy is considered to be the first choice to control pulmonary lesions of this disease.

[Use of a bronchoscope for thoracoscopic observation and diagnosis of pleural plaques].

A 62-year-old man was admitted to our hospital for evaluation of bilateral tumor-like shadows along the ribs. He had been a diamond-grinder for twenty years. Thoracoscopic examination with a flexible bronchoscope revealed many well-circumscribed tumors with shiny, smooth, convex surfaces on the parietal pleura. The biopsy specimen had hyaline collagen with no cellular components. Based on these findings the tumors were diagnosed as pleural plaques. Cases of pleural plaques are usually followed up with chest radiography only, but the clinical course may be complicated by malignant mesothelioma and other malignant disease. To differentiate these conditions we performed biopsy during intrathoracic observation. The flexible bronchoscope can be useful in such procedures because it is relatively easy to operate and the procedure is relatively non invasive.

Protein kinase inhibitor attenuates an increase in endothelial monolayer permeability induced by tumour necrosis factor-alpha.

We questioned the mechanism of the increase in pulmonary endothelial permeability induced by tumour necrosis factor-alpha (TNF-alpha), a cytokine implicated in the pathogenesis of adult respiratory distress syndrome. As a measure of permeability, we determined the albumin transferred across cultured pulmonary endothelial monolayers prepared on a porous filter. The agents evaluated included protein kinase inhibitors H-7 and H-8, a calmodulin antagonist W-7, and protein kinase C (PKC) activators, phorbol myristate acetate (PMA) and SC-9. H-7, more potent in inhibiting PKC than H-8, failed to attenuate the increase in permeability induced by TNF-alpha. Neither PMA nor SC-9 increased permeability. However, H-8, which is a potent inhibitor of cyclic nucleotide-dependent protein kinases, prevented the increase in permeability induced by TNF-alpha. These results suggest that protein kinase other than PKC are involved in the signal transduction in endothelial permeability increase induced by TNF-alpha. Calmodulin pathway may not be implicated in the increase in permeability induced by TNF-alpha.