Use of gas chromatography-ion trap tandem mass spectrometry for the detection and characterization of microorganisms in complex samples

Larsson, Lennart; Saraf, Anita

doi:10.1007/bf02740818

Cited by 22 publications

(13 citation statements)

References 30 publications

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“…In brief, samples were heated in 2 M methanolic HCl at 80°C 18 hours. The liberated fatty acid methyl esters were extracted with heptane and the hydroxylated fatty acid methyl esters were purified by solid phase extraction as previously described [21]. Prior to analysis, the hydroxy fatty acid methyl esters were transformed to trimethylsilyl ethers.…”

Section: Methodsmentioning

confidence: 99%

Endotoxin markers in bronchoalveolar lavage fluid of patients with interstitial lung diseases

Szponar

Larsson

Domagała‐Kulawik

2012

Multidiscip Respir Med

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BackgroundExposure 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).MethodsBALf 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.ResultsThe 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.ConclusionsA 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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Section: Methodsmentioning

confidence: 99%

Endotoxin markers in bronchoalveolar lavage fluid of patients with interstitial lung diseases

Szponar

Larsson

Domagała‐Kulawik

2012

Multidiscip Respir Med

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“…Two laboratories have recently begun assessing airborne PG levels. 21,27,28 Muramic acid (MA), present in PG but not elsewhere in nature, was used as the unique marker for PG. MA levels have been assayed in dust collected from floors and air conditioning systems of homes, a hospital, 16,29 and from the air of agricultural environments.…”

Section: Introductionmentioning

confidence: 99%

Investigation of the Concentration of Bacteria and Their Cell Envelope Components in Indoor Air in Two Elementary Schools

Liu

Krahmer

Fox

et al. 2000

Journal of the Air & Waste Management Association

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Bacterial cell envelope components are widely distributed in airborne dust, where they act as inflammatory agents causing respiratory symptoms. Measurements of these agents and other environmental factors are assessed in two elementary schools in a southeastern city in the United States. Muramic acid (MA) was used as a marker for bacterial peptidoglycan (PG), and 3-hydroxy fatty acids (3-OH FAs) were used as markers for Gram-negative bacterial lipopolysaccharide (LPS). Culturable bacteria were collected using an Andersen sampler with three different culture IMPLICATIONS Levels of bacteria and their constituents (PG and LPS) are correlated with total surface dust and TSP levels in school environments. This suggests an important environmental health guideline for school administrators: decreasing both surface dust and TSP levels will likely reduce potential exposure to harmful biocontaminants. The high correlations between CO 2 and levels of bacteria and their constituents also indicate that an appropriate air-exchange rate may help minimize biocontamination. The analytical MA and 3-OH FA methods measure culturable and nonculturable bacteria as well as bacterial cell wall remnants, whereas the culture method only assesses the culturable portion of the bacterial population. Together, these three different measures provide complementary information for characterizing biocontamination of indoor air, which is more informative than information provided by culture alone. Additional evaluation of other environmental factors, including CO 2 and air-exchange rate, might suggest recommendations for lowering biocontamination in buildings.media. In addition, temperature (T), relative humidity (RH), and CO 2 were continuously monitored.Concentrations of airborne MA and 3-OH FAs were correlated with total suspended particulate (TSP) levels. Outdoor MA (mean = 0.78-1.15 ng/m 3 ) and 3-OH FA levels (mean = 2.19-2.18 ng/m 3 ) were similar at the two schools. Indoor concentrations of airborne MA and 3-OH FAs differed significantly between schools (MA: 1.44 vs. 2.84 ng/m 3 ; 3-OH FAs: 2.96 vs. 4.57 ng/m 3 ). Although indoor MA levels were low, they were significantly related to teachers' perception of the severity of indoor air quality (IAQ) problems in their classrooms. Concentrations of CO 2 correlated significantly with all bacteria measurements. Because CO 2 levels were related to the number of occupants and the ventilation rates, these findings are consistent with the hypothesis that the children and teachers are sources of bacterial contamination. Many culturable bacteria present in indoor air are opportunistic organisms that can be infectious for compromised individuals, while both culturable and nonculturable bacterial remnants act as environmental toxins for both healthy and compromised individuals. Measuring the "total bacteria load" would be most accurate in assessing the biotoxicity of indoor air. Chemical analysis of MA and 3-OH FAs, when coupled with the conventional culture method, provides complementary infor...

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“…Saturated straight-and branched-chain 3-OH fatty acid profiles were determined in saliva, essentially as previously described (Larsson and Saraf, 1997;Sebastian and Larsson, 2003;Ferrando et al, 2005). Briefly, pellets of centrifuged saliva samples (1500 x g, 15 min) were subjected to methanolysis by the dry methanolhydrochloric-acid method (2-M, 80°C, 18 hrs) (Sebastian and Larsson, 2003;Ferrando et al, 2005).…”

Section: Establishment Of Salivary 3-oh Fatty Acid Profilesmentioning

confidence: 99%

“…The injector, transfer line, and ion trap were held at, respectively, 280°C, 290°C, and 180°C (Ferrando et al, 2005). Ions of m/z 131 representing product ions of m/z 175 (resulting from cleavage of C3/C4 of the 3-OTMS methyl ester derivative) were monitored (Larsson and Saraf, 1997).…”

Section: Establishment Of Salivary 3-oh Fatty Acid Profilesmentioning

confidence: 99%

Fatty Acid Profiles in Smokers with Chronic Periodontitis

et al. 2010

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We hypothesized that tobacco smoke induces alterations to the 3-OH fatty acids present in lipid A in a manner consistent with a microflora of reduced inflammatory potential. Whole saliva samples and full-mouth clinical periodontal recordings were obtained from persons with (22 smokers; 15 non-smokers) and without (14 smokers; 15 non-smokers) chronic periodontitis. Clear differences in the contributions of multiple saturated 3-OH fatty acid species were noted in the group with disease compared with healthy individuals. Increases in the long-chain fatty acids associated with anaerobic bacterial periodontopathogens, particularly 3-OH-C(i17.0) (146.7%, relative to controls), were apparent. Significant reductions in the 3-OH fatty acids associated with the consensus (high potency) enteric LPS structure (3-OH-C(12.0) and 3-OH-C(14.0); 33.3% and 15.8% reduction, respectively) were noted in smokers compared with non-smokers with chronic periodontitis. Thus, smoking is associated with specific structural alterations to the lipid-A-derived 3-OH fatty acid profile in saliva that are consistent with an oral microflora of reduced inflammatory potential. These findings provide much-needed mechanistic insight into the established clinical conundrum of increased infection with periodontal pathogens but reduced clinical inflammation in smokers.

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Use of gas chromatography-ion trap tandem mass spectrometry for the detection and characterization of microorganisms in complex samples

Cited by 22 publications

References 30 publications

Endotoxin markers in bronchoalveolar lavage fluid of patients with interstitial lung diseases

Endotoxin markers in bronchoalveolar lavage fluid of patients with interstitial lung diseases

Investigation of the Concentration of Bacteria and Their Cell Envelope Components in Indoor Air in Two Elementary Schools

Fatty Acid Profiles in Smokers with Chronic Periodontitis

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