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Isolates of Mycobacterium avium-intracellulare complex (MAI) from AIDS and non-AIDS patients were compared by pyrolysis mass spectrometry (PyMS). Those from AIDS patients were more closely related to each other than those from non-AIDS patients which were significantly more disparate.
Clinical (66: dental 53; vaginal 4; wound 9) and reference (5*) strains of pigmented Gram-negative anaerobic bacilli were examined in pyrolysis mass spectrometry (PMS) and conventional tests (CTs). The strains were identified in CTs as: Prevotella intermedia (48*); Pr. melaninogenica (1); Pr. corporis (7); Porphyromonas asaccharolytica (12*); P. endodontalis (1*) and P. gingivalis (2*). Numerical classification based on CTs resolved five clusters comprising strains identified as (I) Pr. corporis, (II) Pr. melaninogenica, (III) Pr. intermedia, (IV) P. gingivalis and (V) P. asaccharolytica and P. endodontalis. Numerical classification based on PMS showed a similar division, with decreasing homogeneity in the order Pr. intermedia, Pr. corporis, P. asaccharolytica, in agreement with the ordering of homogeneity for these species in CTs. PMS clusters corresponding the Porphyromonas spp. were clearly distinct from those of Prevotella spp. PMS and CT classifications disagreed on cluster membership for only six of the strains. PMS identification from blind challenge sets agreed with conventional identification for 64 of 67 strains.
Pyrolysis mass spectrometry was used to produce complex biochemical fingerprints of Eubacterium exiguum, E. infirmum, E. tardum and E. timidum. To examine the relationship between these organisms the spectra were clustered by canonical variates analysis, and four clusters, one for each species, were observed. In an earlier study we trained artificial neural networks to identify these clinical isolates successfully; however, the information used by the neural network was not accessible from this so-called 'black box' technique. To allow the deconvolution of such complex spectra (in terms of which masses were important for discrimination) it was necessary to develop a system that itself produces 'rules' that are readily comprehensible. We here exploit the evolutionary computational technique of genetic programming; this rapidly and automatically produced simple mathematical functions that were also able to classify organisms to each of the four bacterial groups correctly and unambiguously. Since the rules used only a very limited set of masses, from a search space some 50 orders of magnitude greater than the dimensionality actually necessary, visual discrimination of the organisms on the basis of these spectral masses alone was also then possible.
Near infrared (NIR) spectroscopy and pyrolysis-molecular beam mass spectrometry (py-MBMS) analysis can be used in conjunction with multivariate regression and principal components analysis to differentiate brown-rot-degraded wood from non-degraded spruce and to follow the temporal changes in wood undergoing brown-rot degradation. Regression of NIR test results vs. percent weight loss for Postia placenta- and Gloeophyllum trabeum-infected spruce wood blocks yielded a correlation coefficient of 0.96. Regression of MBMS test results for the same samples yielded a correlation coefficient of 0.96. Principle components analysis was used to differentiate non-infected wood and P. placenta- and G. trabeum-infected wood. These techniques may be used to detect different types of biodegradation and to develop a better understanding of the chemical changes that the wood undergoes when it is subjected to brown-rot biodegradation.
Diamond powder was synthesized through a metallic reduction-pyrolysis-catalysis route with the reaction of carbon tetrachloride and sodium at 700 degreesC, in which the sodium was used as reductant and flux. This temperature is much lower than that of traditional methods. The x-ray powder diffraction patterns showed three strong peaks of diamond. The Raman spectrum showed a sharp peak at 1332 inverse centimeters, which is characteristic of diamond. Although the yield was only 2 percent, this method is a simple means of forming diamond.
Bacterial samples were prepared for pyrolysis gas-liquid chromatography using cells grown on membrane filters. Pyrochromatograms were reproducible when cells harvested from the filters were pyrolyzed without being washed.
Pyrolysis gas-liquid chromatography was performed on dried Bacillus microorganisms to evaluate the effects of growth media. Six cultures of Bacillus and six lot numbers of Trypticase soy agar (BBL) were used to test the hypothesis that a microorganism grown on various lot numbers of the same chromatogram. Also tested was the effect of three different media on chromatogram reproduction using the same six cultures. Results show little or no differences observed between the chromatograms of the individual Bacillus spp. grown on the six lot numbers of Trypticase soy agar. When chromatograms of the three different media were compared, several differences were observed, particularly in the areas most characteristic of individual species. Pryolysis gas-liquid chromatography can be a useful tool for the characterization or identification of the genus Bacillus if the chromatographic and cultural conditions are maintained.
Washed organisms, including strains of Streptococcus mitior, S. mutans, and S. sanguis, were examined by curie-point pyrolysis gas-liquid chromatography. A linear discriminant function based upon three items from the output data was adequate for segregating the strains according to species. Strains with intermediate properties were also encountered. Sources of variability in cultures were evaluated, chromatographic performance was maintained throughout the investigation, and matching performance from a duplicate pair of chromatographic columns was demonstrated.
Pyrolysis-gas-liquid chromatography (PGLC) of whole cells and cell fragments was used to differentiate 10 Salmonella serotypes. Lyophilized samples (200 microgram) of whole cells, cell walls, flagella, and deoxyribonucleic acid from each serotype were analyzed in duplicate by PGLC. Pyrochromatograms recorded as pyrolytic elution patterns represented thermal fragmentation products of the samples. Mathematical expressions of percent similarity and percent conformity were calculated for all possible pair combinations of the 10 serotypes. Stepwise discriminant analysis of the PGLC data showed that 100 percent correct classification of the 10 serotypes was possible from the flagella or deoxyribonucleic acid pyrochromatograms. The classification matrix of the whole-cell data showed a 90 percent correct classification. PGLC of cell fragments may provide useful information for taxonomic studies of Salmonella and other microorganisms.
High-resolution pyrolysis gas-liquid chromatography was applied to three bacteria (Escherichia coli NCTC 9001, Pseudomonas putida (NCIB 9494, and Staphylococcus aureus NCTC 8532) grown under a variety of conditions. Changing the culture medium drastically altered the quantitative aspects of the pyrograms of all three organisms, but the effects of culture time and incubation temperature were less severe. Mathematical analysis of the relative peak heights showed that four peaks could be used to discriminate the three bacteria however they were cultured.
Curie-point pyrolysis (Py)-mass spectrometry has been used to differentiate 19 microorganisms by Gram type on the basis of the methyl esters of their fatty acid distribution. The mass spectra of gram-negative microorganisms were characterized by the presence of palmitoleic acid (C(inf16:1)) and oleic acid (C(inf18:1)), as well as a higher abundance of palmitic acid (C(inf16:0)) than pentadecanoic acid (C(inf15:0)). For gram-positive microorganisms, a signal of branched C(inf15:0) (isoC(inf15:0) and/or anteisoC(inf15:0)) more intense than that of palmitic acid was observed in the mass spectra. Principal components analysis of these mass spectral data segregated the microorganisms investigated in this study into three discrete clusters that correlated to their gram reactions and pathogenicities. Further tandem mass spectrometric analysis demonstrated that the nature of the C(inf15:0) fatty acid isomer (branched or normal) present in the mass spectrum of each microorganism was important for achieving the classification into three clusters.
Vegetative cells and spores of 10 strains of Clostridium botulinum representing types A, B, and E were grown in Trypticase-peptone-sucrose-yeast extract (TPSY) medium. Five type E strains were also grown in Multipeptone-sucrose-Nutramino acids (MSN) medium. Lyophilized samples were subjected to pyrolysis-gas-liquid chromatography (PGLC) analysis, and the resulting pyrograms were examined for variations in elution patterns between spores and vegetative cells of types A, B, and E grown in the TPSY medium and spores and vegetative cells of type E grown in the TPSY medium and spores and vegetative cells of type E grown in TPSY and MSN media. Growth and toxin production of all 10 strains of C. botulinum were investigated by using a modified dialysis sac culture technique. The dialysate supernatant fluid (DSF) obtained after centrifugation of the 5-day-old cultures from the dialysate was also subjected to PGLC analysis. Control samples consisting of (i) noninoculated DSF, (ii) noninoculated DSF plus partially purified toxin, and (iii) 1.0 mg of partially purified toxin were also analyzed by PGLC. Differences between pyrograms of cultures were suitable for positive identification at the type level but not at the strain level. Pyrograms permitting differentiation were also obtained between spores and vegetative cells as well as between the same cultures grown in different media. The dialysis sac technique was useful in detecting growth but not toxin production of C. botulinum.
Cholera-like vibrios examined by pyrolysis gas-liquid chromatography could be distinguished from other common aerobic gram-negative bacilli, including oxidase-positive organisms, e.g., Aeromonas. Vibrios in Heiberg group I were subdivided into three types on the basis of differences in one complex in the chromatogram, and these closely corresponded with the identification as classical, El Tor, or "intermediate" biotypes of Vibrio cholerae by conventional methods.
Nine coded duplicate strains of capsular nontypable Klebsiella pneumoniae were analyzed by pyrolysis-gas-liquid chromatography-mass spectrometry. All duplicate strains were correctly matched, and individual strains, including seven nontypable strains, were clearly distinguishable from one another. The addition of mass spectrometry to the analysis has aided the process of identification and has provided chemical structural information on K. pneumoniae strains. Application of this technique to the identification of some disease outbreaks or nosocomial problems could be of epidemiological importance, especially when conventional methods do not identify the epidemic strain.
Two rapid spectroscopic approaches for whole-organism fingerprinting of pyrolysis-mass spectrometry (PyMS) and Fourier transform-infrared spectroscopy (FT-IR) were used to analyze a group of 29 clinical and reference Candida isolates. These strains had been identified by conventional means as belonging to one of the three species Candida albicans, C. dubliniensis (previously reported as atypical C. albicans), and C. stellatoidea (which is also closely related to C. albicans). To observe the relationships of the 29 isolates as judged by PyMS and FT-IR, the spectral data were clustered by discriminant analysis. On visual inspection of the cluster analyses from both methods, three distinct clusters, which were discrete for each of the Candida species, could be seen. Moreover, these phenetic classifications were found to be very similar to those obtained by genotypic studies which examined the HinfI restriction enzyme digestion patterns of genomic DNA and by use of the 27A C. albicans-specific probe. Both spectroscopic techniques are rapid (typically, 2 min for PyMS and 10 s for FT-IR) and were shown to be capable of successfully discriminating between closely related isolates of C. albicans, C. dubliniensis, and C. stellatoidea. We believe that these whole-organism fingerprinting methods could provide opportunities for automation in clinical microbial laboratories, improving turnaround times and the use of resources.
Carbon nanocoils were prepared by the chemical vapor deposition process of the catalytic pyrolysis of acetylene at 700-800 degrees C with various catalysts. Twisting or coiling-formed carbon nanocoils with changing coiling-chirality and zigzag-formed carbon nanofibers were obtained with SUS 304, WS2, Pt-Pd, TiN, and Ni as the catalysts. Their morphologies and microstructures were examined in detail, and then the changing mechanism of the coiling-chirality was discussed. No apparent difference in the microstructure between the part of a nanocoil with changing coiling-chirality and the part of a zigzag nanofiber with changing zigzag-chirality, or between a bulk right-clockwise coil and a bulk left-clockwise coil was observed. It was supposed that changing coiling-chirality was mainly caused by the gradual or successive change in chemical composition on the thin layers present on the surface of catalyst grains during the chemical vapor deposition process.
Nano-crystalline Co3O4 and CoO powders have been prepared by a spray pyrolysis approach. The effects of the reaction temperature and initial salts on the crystallinity and phase composition have been studied. Based on the TEM and XRD results, the crystal sizes were in the range of 1-10 nm. SEM and TEM observations also reveal that the nano-powders easily create micron-scale spherical agglomerates. The Co3O4 powders obtained by spraying nitrate solution at 500 degrees C show high specific surface area, which according to the BET method is 82.37 m2/g. The time/temperature phase diagram of cobalt oxides developed from XRD and DTA/TGA analyses shows the existence of a CoO phase at low and high temperature ranges when some specific preparation conditions are applied.
The cytotoxicity of heterocyclic amines, dietary carcinogens derived from cooked foods, to primary cultured rat hepatocytes was studied. A tryptophan pyrolysis product, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) was the most cytotoxic of 11 compounds tested. Trp-P-1 was found to induce apoptosis as measured by morphological changes in nuclear chromatin and internucleosomal DNA fragmentation. 3-Amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) showed a moderate apoptotic effect, and other compounds had a similar but weaker effect.