| 1. |
- de la Cochetiére, Marie France, et al.
(author)
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3-Hydroxy fatty acids in faeces of patients with Crohn's disease or ulcerative colitis as a non-invasive diagnostic approach
- 2007
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In: Microbial Ecology in Health and Disease. - : Informa UK Limited. - 0891-060X .- 1651-2235. ; 19, s. 1-6
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Journal article (peer-reviewed)abstract
- The microbiota is part of the gastrointestinal ecosystem. A more detailed understanding should provide insight into multiple human disease states. This study investigated inflammatory bowel disease (Crohn's disease and ulcerative colitis). Previous analyses have suggested a role of gram-negative bacteria including Escherichia coli. An integrated procedure is presented where gas chromatography-mass spectrometry is used to determine chemical markers of gram-negative bacterial lipopolysaccharide (3-hydroxy fatty acids with 10-18 carbon atoms) in faecal samples. Six patients with Crohn's disease (CD), five with ulcerative colitis (UC) and six healthy adults were chosen as groups of interest. Nine saturated straight-chain 3-OH fatty acids of 10-18 carbon chain lengths and six iso- and anteiso-branched-chain 3-OH fatty acids of 15-18 carbon chain lengths were detected. Significant differences were found in the 3-OH n-C17:0, 3-OH i-C18:0 and 3-OH n-C18:0 composition of faeces. The present study therefore confirms that alteration of the composition of the endogenous gram-negative microbiota may be of importance in inflammatory bowel disease and those alterations could be detected with a non-invasive chemical-analytical approach.
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| 2. |
- Ferrando, R, et al.
(author)
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3-Hydroxy fatty acids in saliva as diagnostic markers in chronic periodontitis
- 2005
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In: Journal of Microbiological Methods. - : Elsevier BV. - 1872-8359 .- 0167-7012. ; 62:3, s. 285-291
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Journal article (peer-reviewed)abstract
- Saturated straight- and branched-chain 3-hydroxy fatty acids (3-OH FAs) of 10-18 carbon chain lengths were determined in saliva from 27 individuals with chronic periodontitis and 18 healthy individuals by using gas chromatography-tandem mass spectrometry. Of the 14 different 3-OH FAs detected, 3-OH-C-i17:0 was the most abundant in the periodontitis samples while 3-OH-C-14:0 was the most abundant in the healthy individuals. Considering the relative percentages of 3-OH-C-12:0, 3-OH-C-14:0, 3OH-C-i17:0, and 3-OH-C-17:0, 95.6% of all cases were correctly classified as healthy individuals or periodontitis patients by means of discriminant analysis. The sensitivity, specificity, positive predictive value and negative predictive value of 3-OH FA analysis in diagnosing peridontitis were, respectively, 0.92, 1.00, 1.00, and 0.90. The results indicate that 3-OH FA analysis of saliva samples is a useful diagnostic method in chronic periodontitis.
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| 3. |
- Hynes, Sean, et al.
(author)
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Comparative chemical and biological characterization of the lipopolysaccharides of gastric and enterohepatic helicobacters
- 2004
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In: Helicobacter. - : Wiley. - 1083-4389 .- 1523-5378. ; 9:4, s. 313-323
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Journal article (peer-reviewed)abstract
- Background. The lipopolysaccharide of Helicobacter pylori plays an important role in colonization and pathogenicity. The present study sought to compare structural and biological features of lipopolysaccharides from gastric and enterohepatic Helicobacter spp. not previously characterized.Materials and methods. Purified lipopolysaccharides from four gastric Helicobacter spp. (H. pylori, Helicobacter felis, Helicobacter bizzozeronii and Helicobacter mustelae) and four enterohepatic Helicobacter spp. (Helicobacter hepaticus, Helicobacter bilis, Helicobacter sp. flexispira and Helicobacter pullorum) were structurally characterized using electrophoretic, serological and chemical methods.Results. Structural insights into all three moieties of the lipopolysaccharides, i.e. lipid A, core and O-polysaccharide chains, were gained. All species expressed lipopolysaccharides bearing an O-polysaccharide chain, but H. mustelae and H. hepaticus produced truncated semirough lipopolysaccharides. However, in contrast to lipopolysaccharides of H. pylori and H. mustelae, no blood group mimicry was detected in the other Helicobacter spp. examined. Intra-species, but not interspecies, fatty acid profiles of lipopolysaccharides were identical within the genus. Although shared lipopolysaccharide-core epitopes with H. pylori occurred, differing structural characteristics were noted in this lipopolysaccharide region of some Helicobacter spp. The lipopolysaccharides of the gastric helicobacters, H. bizzozeronii and H. mustelae, had relative Limulus amoebocyte lysate activities which clustered around that of H. pylori lipopolysaccharide, whereas H. bilis, Helicobacter sp. flexispira and H. hepaticus formed a cluster with approximately 100010,000-fold lower activities. H. pullorum lipopolysaccharide had the highest relative Limulus amoebocyte lysate activity of all the helicobacter lipopolysaccharides (10-fold higher than that of H. pylori lipopolysaccharide), and all the lipopolysaccharides of enterohepatic Helicobacter spp. were capable of inducing nuclear factor-Kappa B(NF-B) activation.Conclusions. The collective results demonstrate the structural heterogeneity and pathogenic potential of lipopolysaccharides of the Helicobacter genus as a group and these differences in lipopolysaccharides may be indicative of adaptation of the bacteria to different ecological niches.
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| 4. |
- Knasnik, L, et al.
(author)
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Routine clinical laboratory tests correspond to increased serum levels of 3-hydroxy fatty acids, markers of endotoxins, in cardiosurgery patients
- 2006
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In: Archivum Immunologiae et Therapiae Experimentalis. - : Springer Science and Business Media LLC. - 0004-069X .- 1661-4917. ; 54:1, s. 55-60
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Journal article (peer-reviewed)abstract
- Introduction: Endotoxemia developing during cardiosurgery as elevated endotoxin concentrations in patient's serum may prevail over 24 h after operation. A major reason is thought to be increased gut permeability resulting in endotoxin and bacterial leakage. In this study we aimed to measure endotoxin levels on samples obtained during and after cardiovascular procedures and compare them with clinical observations and laboratory test results. Materials and Methods: 3-Hydroxy fatty acids (3-OH FAs) of 10-18 carbon chain length, chemical markers of endotoxin (lipopolysaccharide), were determined in patient sera by gas chromatography-mass spectrometry-based analysis. Results were compared with routine laboratory tests: blood morphology, urine, ALT, AST, bilirubin, kidney parameters, clotting parameters, and gasometry. Results: Of a total of 16 patients, 5 patients (group I) showed increased serum 3-OH FA levels and 11 patients (group II) did not show any change in 3-OH FA levels 24 h after operation. All group I patients revealed leukocytosis, two developed post-operative anemia. Significantly different changes were observed: the initial, pre-operative 3-OH FA levels were similar for both groups, while group I patients showed increased levels of all the studied 3-OH FAs during the operation (p <= 0.05), and 3-OH C14 and 3-OH C16 remained elevated 24 h after the operation. Conclusions: Cardiosurgery may strongly promote gut endotoxin translocation to the blood in some patients. Prolonged leukocytosis, deep anemia, and increased liver dysfunction markers may indicate the need for observation of possible endotoxemia development. It is recommended to monitor the endotoxin level and/or endotoxemia markers in cardiosurgery patients.
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| 5. |
- Larsson, Lennart, et al.
(author)
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Identification of bacterial and fungal components in tobacco and tobacco smoke.
- 2008
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In: Tobacco Induced Diseases. - : E.U. European Publishing. - 1617-9625. ; 4:1, s. 4-4
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Journal article (peer-reviewed)abstract
- ABSTRACT: The microbiological composition of tobacco products was studied using culture and chemical analysis (of tobacco leaves) or chemical analysis only (tobacco and tobacco smoke). The chemical analyses utilized gas chromatography-tandem mass spectrometry for determining 3-hydroxy fatty acids, muramic acid, and ergosterol as markers of respectively lipopolysaccharide (LPS), peptidoglycan, and fungal biomass. Mesophilic bacteria dominated in both fresh and cured tobacco leaves; a range of additional bacteria and fungi were also found albeit in minor amounts. The peptidoglycan and LPS concentrations were approximately the same in tobacco leaves as in cigarette tobacco. The concentrations of the measured microbial components were much lower in some cigarettes locally produced in China, Korea, and Vietnam than in cigarettes of international brands purchased in the same countries, and the concentrations in the smoke were in general agreement with the concentrations in cigarette tobacco. No differences in microbial load in tobacco of "light" and "full flavor" cigarettes were seen. Storing cigarettes at high humidity resulted in elevated levels of fungi in the cigarette tobacco leading to increased ergosterol concentrations in the smoke. The fact that tobacco smoke is a bioaerosol may help to explain the high prevalence of respiratory disorders among smokers and non-smokers exposed to second hand smoke since the same symptoms are also commonly associated with exposure to bioaerosols.
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| 6. |
- Larsson, L, et al.
(author)
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Tobacco smoke as a major source of endotoxin in indoor air
- 2005
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In: Indoor Air 2005: Proceedings of the 10th International Conference on Indoor Air Quality and Climate, Vols 1-5. ; , s. 1655-1658
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Conference paper (peer-reviewed)abstract
- Mass spectrometry was used to determine the endotoxin (lipopolysaccharide, LPS) marker 3-hydroxytetradecanoic acid and 3-deoxy -D-manno-2-octulosonic acid in cigarette tobacco and smoke particles. Tobacco from cigarettes of different brands purchased in four different countries in Europe and Asia contained similar amounts of LPS. Indoor exposure to environmental tobacco smoke (ETS) in rooms of smoking students entailed inhalation of LPS in amounts that were 6-60 times higher than in student rooms without ongoing smoking. The fact that cigarette smoke contains large amounts of endotoxin may partly explain the high prevalence of respiratory disorders among smokers and may also draw attention to a hitherto unknown or neglected risk factor of ETS.
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| 7. |
- Larsson, Lennart, et al.
(author)
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Tobacco smoking increases dramatically air concentrations of endotoxin.
- 2004
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In: Indoor Air. - : Hindawi Limited. - 0905-6947 .- 1600-0668. ; 14:6, s. 421-424
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Journal article (peer-reviewed)abstract
- We used a mass spectrometry-based assay for identifying the endotoxin (lipopolysaccharide, LPS) marker (R)-3-hydroxytetradecanoic acid in cigarette smoke particles and found that smoking involved inhalation of 17.4 pmol of endotoxin per each smoked cigarette. Indoor exposure to environmental tobacco smoke (ETS) entailed inhalation of 12.1 pmol of LPS/m3 air, an amount that was 120 times higher than the levels found in smoke-free indoor air. Endotoxin is one of the most potent inflammatory agents known, hence our results may help to explain the high prevalence of respiratory disorders among smokers, and they may also draw attention to a hitherto unknown or neglected risk factor of ETS.
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| 8. |
- Mackiewicz, Barbara, et al.
(author)
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Respiratory Disorders In Two Workers Of Customs Depositories Occupationally Exposed To Mouldy Tobacco
- 2008
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In: Annals of Agricultural and Environmental Medicine. - 1898-2263. ; 15:2, s. 317-322
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Journal article (peer-reviewed)abstract
- Work-related respiratory symptoms, including dyspnoea, cough, fever, tiredness and malaise, were recorded in two customs officers employed in 2 depositories of confiscated cigarettes, of which one showed signs of dampness. Microbiological sampling of the air and the cigarettes stored in a damp depository revealed the presence of potentially pathogenic fungi and bacteria and the biochemical markers of bacterial lipopolysaccharide and fungal biomass. The Penicillium species (P. simplicissimum, P. inflatum, P commune) dominated in the damp depository, while in the other one Aspergillus fumigatus was prevalent. The patients under study did not show a specific sensitization to microbial allergens in the precipitin test, the test for inhibition of leukocyte migration and the bronchial provocation challenge, except for a weak reaction to fungal allergens in the test for inhibition of leukocyte migration. Moreover, one patient responded with subjective symptoms after exposure to inhalation of increased doses of Penicillium simplicissimum antigen. Both cases were diagnosed as a specific form of organic dust toxic syndrome (ODTS). It is hypothesized that the symptoms were evoked most probably by the non-specific action of low molecular fungal metabolites, such as mycotoxins or VOCs (volatile organic compounds), with the possible contribution of bacterial endotoxin. However, as there is no a direct proof to support this presumption, and the effects of nicotine and other tobacco constituents cannot be excluded, further studies are needed to elucidate etiopathogenesis of the disorders associated with the exposure to stored tobacco.
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| 9. |
- Nilsson, Anders, et al.
(author)
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Microorganisms and volatile organic compounds in airborne dust from damp residences
- 2004
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In: Indoor Air. International Journal of Indoor Environment and Health. - : Hindawi Limited. - 0905-6947 .- 1600-0668. ; 14:2, s. 74-82
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Journal article (peer-reviewed)abstract
- Airborne dust samples from damp (n = 9) and control (n = 9) residences were analyzed for microorganisms (molds and bacteria), bacterial markers (3-hydroxy fatty acids and muramic acid), and adsorbed volatile organic compounds (VOCs). The number of mold species was greater in the damp residences than in the controls (23 vs.18) and nine mold species were found only in damp residences. The levels of 3-hydroxy fatty acids and muramic acid correlated better in damp residences than in controls, indicating that damp conditions affect the bacterial flora of airborne dust. Identifications made by culture and microscopy of the major molds found, i.e. Aspergillus, Cladosporium, and Penicillum, coincided with the identification of VOCs known to be produced by these species. A number of additional VOCs irritating to the skin, eyes, or respiratory tract were also found. The results from this pilot study illustrate the diversity of microorganisms and VOCs present in the indoor environment and suggest that analysis of airborne dust may help to assess human exposure to microorganisms and chemical compounds.
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| 10. |
- Park, J H, et al.
(author)
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Characterization of lipopolysaccharides present in settled house dust
- 2004
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In: Applied and Environmental Microbiology. - 0099-2240. ; 70:1, s. 262-267
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Journal article (peer-reviewed)abstract
- The 3-hydroxy fatty acids (3-OHFAs) in lipopolysaccharides (LPS) play an important role in determining endotoxin activity, and childhood exposure to endotoxin has recently been associated with reduced risk of atopic diseases. To characterize the 3-OHFAs in house dust (HD), we used gas chromatography-mass spectrometry to assay 190 HD samples. Dust from beds, bedroom floors, family rooms, and kitchen floors was collected as part of a birth cohort study of childhood asthma (study 1) and a longitudinal study of home allergen and endotoxin (study 2). We also measured endotoxin activity with a Limulus assay and computed specific activity (endotoxin activity per nanomole of LPS). Longer-chain (C16:0 and C18:0) 3-OHFAs were predominant in HD compared with short-chain (C10:0, C12:0, and C14:0) acids. Endotoxin activity was positively correlated with short-chain 3-OHFAs in both studies. In study 2, 3-OH C16:0 was negatively correlated and 3-OH C18:0 was not correlated with endotoxin activity, consistent with previous findings that the Limulus assay responds preferentially to LPS containing short-chain 3-OHFAs. Kitchen dust contained the highest concentrations of 3-OH C10:0, the highest endotoxin activities, and the highest specific activities (P < 0.03). Bed dust contained the largest amounts of long-chain 3-OHFAs, the highest concentrations of LPS, and the lowest specific activities. Apartments had significantly different types of LPS (P = 0.03) compared with single-family homes in study 2. These data suggest that the Limulus assay may underestimate exposure to certain types of LPS. Because nontoxic LPS may have immune modulating effects, analysis of 3-OHFAs may be useful in epidemiologic studies.
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| 11. |
- Sebastian, Aleksandra, et al.
(author)
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Characterization of the microbial community in indoor environments by chemical marker analysis: an update and critical evaluation.
- 2005
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In: Indoor Air. - : Hindawi Limited. - 0905-6947 .- 1600-0668. ; 15 Suppl 9:9, s. 20-26
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Journal article (peer-reviewed)abstract
- We published recently an integrated procedure for applying chemical marker analysis to characterize the microbiology of indoor environments comprising a scheme for extraction and analysis of markers of endotoxin, peptidoglycan/bacterial biomass, and fungal biomass. In the present paper, we report some significant improvements and also new possibilities of the described approach. We found that while 3-hydroxy fatty acids (3-OH FAs) of 10-14 carbon chain lengths are useful endotoxin markers, longer 3-OH FAs (i.e. with 16 carbon atoms and more) may rather serve as markers of Actinobacteria. We introduced C-13-labeled 3-hydroxytridecanoic acid, from labeled Pectinatus cerevisiiphilus, as an internal standard to improve quantification of the 3-OH FAs in the gas chromatography-mass spectrometry analysis. Finally, in experiments aiming to identify a suitable method for collection of house dust for chemical marker analysis, we found that the marker compositions of dusts sedimented on plexiglass plates that were spatially well-distributed in a studied room at different heights above floor level, were undistinguishable. This type of sampling thus appears to be well suited for use, e.g. in epidemiological studies. In summary, the presented work describes important new capabilities of chemical marker analysis in defining human exposure to microorganisms in indoor environments.
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| 12. |
- Szponar, Bogumila, et al.
(author)
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Bacterial and fungal markers in tobacco smoke.
- 2012
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In: Science of the Total Environment. - : Elsevier BV. - 1879-1026 .- 0048-9697. ; 438:Nov.,1, s. 447-451
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Journal article (peer-reviewed)abstract
- Previous research has demonstrated that cigarette smoke contains bacterial and fungal components including lipopolysaccharide (LPS) and ergosterol. In the present study we used gas chromatography-mass spectrometry to analyze tobacco as well as mainstream and second hand smoke for 3-hydroxy fatty acids (3-OH FAs) of 10 to 18 carbon chain lengths, used as LPS markers, and ergosterol, used as a marker of fungal biomass. The air concentrations of LPS were 0.0017nmol/m(3) (N=5) and 0.0007/m(3) (N=6) in the smoking vs. non-smoking rooms (p=0.0559) of the studied private houses, and 0.0231nmol/m(3) (N=5) vs. 0.0006nmol/m(3) (N=5) (p=0.0173), respectively, at the worksite. The air concentrations of ergosterol were also significantly higher in rooms with ongoing smoking than in rooms without smoking. A positive correlation was found between LPS and ergosterol in rooms with smoking but not in rooms without smoking. 3-OH C14:0 was the main 3-OH FA, followed by 3-OH C12:0, both in mainstream and second hand smoke and in phenol:water smoke extracts prepared in order to purify the LPS. The Limulus activity of the phenolic phase of tobacco was 3900endotoxin units (EU)/cigarette; the corresponding amount of the smoke, collected on filters from 8 puffs, was 4EU/cigarette. Tobacco smoking has been associated with a range of inflammatory airway conditions including COPD, asthma, bronchitis, alveolar hypersensitivity etc. Significant levels of LPS and ergosterol were identified in tobacco smoke and these observations support the hypothesis that microbial components of tobacco smoke contribute to inflammation and airway disease.
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| 13. |
- Szponar, Bogumila, et al.
(author)
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Endotoxin markers in bronchoalveolar lavage fluid of patients with interstitial lung diseases
- 2012
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In: Multidisciplinary Respiratory Medicine. - : PAGEPress Publications. - 2049-6958. ; 7
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Journal article (peer-reviewed)abstract
- Background: Exposure to inhaled endotoxins (lipopolysaccharides, LPS) of Gram-negative bacteria commonly found in indoor environments and assessed in secondary tobacco smoke, has been associated with airway inflammation and asthma exacerbation. The bronchoalveolar lavage fluid (BALf) from patients with interstitial lung diseases (sarcoidosis, lung fibrosis, smoking-related ILD, eosinophilic disorders) was analyzed for the markers of lipopolysaccharide (LPS, endotoxin). Methods: BALf was obtained from patients with diffuse lung diseases: idiopathic pulmonary fibrosis (n = 42), sarcoidosis (n = 22), smoking-related-ILD (n = 11) and eosinophilic disorders (n = 8). Total cell count and differential cell count were performed. In addition, samples were analyzed for 3-hydroxy fatty acids (3-OHFAs) of 10-18 carbon chain lengths, as markers of LPS, by gas chromatography-tandem mass spectrometry. Results: The highest LPS concentration was found in patients with eosinophilic disorders and the lowest in patients with sarcoidosis (p< 0.05) followed by the lung fibrosis and the sr-ILD patients. The difference between LPS in BALf with extremely high eosinophil proportion (> 25%) and those with lower proportion was also significant (p = 0.014). A significant correlation was found between LPS and eosinophils, but not between LPS and lymphocytes, neutrophils, or macrophages count. Conclusions: A positive relationship of LPS and eosinophilic pulmonary disorders may be linked to a persistent eosinophil activation mediated by Th2 pathway: chronic endotoxin exposure would intensify Th2 pathway resulting in fibrosis and, at the same time, eosinophil stimulation, and hence in eosinophilic pulmonary disorders.
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| 14. |
- Szponar, Bogumila
(author)
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Endotoxins in Environmental and Clinical Samples Assessed by GC-Tandem MS
- 2010
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In: Detection of Biological Agents for the Prevention of Bioterrorism. - Dordrecht : Springer Netherlands. - 1874-6489. ; , s. 245-265
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Conference paper (peer-reviewed)abstract
- Bacteria appeared on the Earth millions years before us and human evolution was triggered by the constant presence of pathogenic and symbiotic microorganisms in our surroundings. Interplay occurred between higher organism and microbial consortia residing in the host organs and on the epithelial surfaces; another natural space of bacteria human interaction is the indoor environment where we spend the majority of our lifetime. Indoor microbial exposure affects our well-being and can result in respiratory symptoms, such as allergies and asthma, since both dead and live microorganisms and their cell constituents, including lipopolysaccharides (LPS, endotoxins), interact with our immune system. Thus, there is a demand for robust tools for qualitative and quantitative determination of the microbial communities that we are exposed to. This work described the reproducible approach of the Gram-negative bacteria and endotoxins assessment by their specific chemical markers, 3-hydroxy fatty acids. Gas chromatography-tandem mass spectrometry proved to be an excellent means for specific and selective detection of bacteria/endotoxin markers in the complex matrices like indoor bioaerosol and clinical samples: blood, saliva or feces. Using this method, epidemiological studies were conducted in the field of indoor air quality research, as well as in clinical investigations when bacterial consortia were involved: in Crohn's disease, periodontitis, and newborn gut microbial colonisation in association with allergy development.
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| 15. |
- Szponar, Bogumila, et al.
(author)
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Limitations in the use of 3-hydroxy fatty acid analysis to determine endotoxin in mammalian samples
- 2002
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In: Journal of Microbiological Methods. - 1872-8359 .- 0167-7012. ; 50:3, s. 283-289
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Journal article (peer-reviewed)abstract
- 3-Hydroxy fatty acids (3-OH FAs) of 10-18-carbon chain lengths are constituents of the lipopolysaccharide of Gram-negative bacteria. These acids are used as chemical markers for determining endotoxin in environmental samples. The present communication addresses the question whether this type of analysis also would be applicable to mammalian samples. Low levels (6.1 +/- 1.6-94.0 +/- 23.2 pmol/ml) of the studied 3-OH FAs were detected in blood from both conventional and germ-fine rats. The levels were considerably higher (0.0-1.06 +/- 0.17 nmol/mg) in livers. The amounts of the 3-OH FAs did not differ between the two groups of rats. All analyses were made by gas chromatography-tandem mass spectrometry (GC-MSMS) for unequivocal identification. The results illustrate a limitation in using 3-OH FA analysis to determine endotoxin in mammalian samples since these acids may represent not only endotoxin but also products from mammalian mitochondrial fatty acid beta-oxidation. (C) 2002 Elsevier Science B.V. All rights reserved.
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| 16. |
- Vanitha, JD, et al.
(author)
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Identification of a group of nontuberculous mycobacteria isolated from the South Indian BCG trial area by HPLC
- 2002
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In: Current Science. - 0011-3891. ; 82:2, s. 189-191
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Journal article (peer-reviewed)abstract
- Twenty-five isolates of nontuberculous mycobacteria isolated from the South Indian BCG trial area were analysed by high performance liquid chromatography (HPLC) for mycolic acid pattern. The chromatograms differentiated the isolates into four species, namely M. terrae complex, M. intracellulare, M. parafortuitum and M. fortuitum. Three strains were unidentified, one of which did not show any mycolic acid peaks. All isolates had been identified as M. diernhoferi by biochemical methods in a previous study. Nineteen of the isolates were analysed by gas chromatography-mass spectrometry (GC-MS) for the presence of tuberculostearic acid, 2-eicosanol and mycolic acid cleavage products, and were classified as nonchromogens or rapidly growing mycobacteria. The results show that HPLC can discriminate the described mycobacterial species better than biochemical methods and GC-MS.
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| 17. |
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