John V. Goodpaster

Proteomic Study of Low-Temperature Responses in Strawberry Cultivars (Fragaria × ananassa) That Differ in Cold Tolerance

To gain insight into the molecular basis contributing to overwintering hardiness, a comprehensive proteomic analysis comparing crowns of octoploid strawberry (Fragaria × ananassa) cultivars that differ in freezing tolerance was conducted. Four cultivars were examined for freeze tolerance and the most cold-tolerant cultivar (‘Jonsok’) and least-tolerant cultivar (‘Frida’) were compared with a goal to reveal how freezing tolerance is achieved in this distinctive overwintering structure and to identify potential cold-tolerance-associated biomarkers. Supported by univariate and multivariate analysis, a total of 63 spots from two-dimensional electrophoresis analysis and 135 proteins from label-free quantitative proteomics were identified as significantly differentially expressed in crown tissue from the two strawberry cultivars exposed to 0-, 2-, and 42-d cold treatment. Proteins identified as cold-tolerance-associated included molecular chaperones, antioxidants/detoxifying enzymes, metabolic enzymes, pathogenesis-related proteins, and flavonoid pathway proteins. A number of proteins were newly identified as associated with cold tolerance. Distinctive mechanisms for cold tolerance were characterized for two cultivars. In particular, the ‘Frida’ cold response emphasized proteins specific to flavonoid biosynthesis, while the more freezing-tolerant ‘Jonsok’ had a more comprehensive suite of known stress-responsive proteins including those involved in antioxidation, detoxification, and disease resistance. The molecular basis for ‘Jonsok’-enhanced cold tolerance can be explained by the constitutive level of a number of proteins that provide a physiological stress-tolerant poise.

Comparing the Effects of Weathering and Microbial Degradation on Gasoline Using Principal Components Analysis

Abstract:  Ignitable liquid residues recovered from a fire scene will often show signs of weathering as a result of exposure to the heat of the fire. In addition, when the substrate is rich in organic matter, both weathering and microbial degradation may be observed. In this study, 20 μL aliquots of fresh gasoline samples were intentionally weathered and also subjected to microbial degradation in potting soil. These samples were then analyzed using a passive adsorption–elution recovery method and gas chromatography/mass spectrometry. Peak areas from compounds of interest were normalized and autoscaled and then subjected to principal components analysis. This analysis showed that while lower boiling compounds are subject to weathering, a different set of compounds are subject to microbial degradation. Of the compounds studied, heptane, octane, toluene, and ethylbenzene were the most vulnerable to both weathering and microbial degradation. In contrast, 1,3,5‐trimethylbenzene and 2‐ethyltoluene were the most resistant to both phenomena.

Forensic analysis of dyed textile fibers

AbstractTextile fibers are a key form of trace evidence, and the ability to reliably associate or discriminate them is crucial for forensic scientists worldwide. While microscopic and instrumental analysis can be used to determine the composition of the fiber itself, additional specificity is gained by examining fiber color. This is particularly important when the bulk composition of the fiber is relatively uninformative, as it is with cotton, wool, or other natural fibers. Such analyses pose several problems, including extremely small sample sizes, the desire for nondestructive techniques, and the vast complexity of modern dye compositions. This review will focus on more recent methods for comparing fiber color by using chromatography, spectroscopy, and mass spectrometry. The increasing use of multivariate statistics and other data analysis techniques for the differentiation of spectra from dyed fibers will also be discussed. FigureMIP image of red cotton fiber

The effects of microbial degradation on ignitable liquids.

AbstractThe identification of ignitable liquid residues in fire debris is a key finding for determining the cause and origin of a suspicious fire. However, the complex mixtures of organic compounds that comprise ignitable liquids are susceptible to microbiological attack following collection of the sample. Biodegradation can result in selective removal of many of the compounds required for identification of an ignitable liquid. Such degradation has been found to occur rapidly in substrates such as soil, rotting wood, or other organic matter. Furthermore, fire debris evidence must often be stored for extended periods at room temperature prior to analysis due to case backlogs and available evidence storage. Hence, extensive damage to ignitable liquid residues by microbes poses a significant threat to subsequent laboratory work. In this work, the effects of microbial degradation of ignitable liquids in soil have been evaluated as a function of time. Key findings include the loss of n-alkanes, particularly C9–C16, which showed the most dramatic decrease in gasoline as well as the petroleum distillates, while branched alkanes remained unchanged. Monosubstituted benzenes also showed the most dramatic loss in gasoline. In the heavy petroleum distillates, n-alkanes with even carbon numbers were degraded more than n-alkanes with odd carbon numbers. FigureA “fully involved” house fire in Indianapolis, IN

Chemical Characterization of Dissolvable Tobacco Products Promoted To Reduce Harm

In 2009, the R. J. Reynolds Tobacco Co. released a line of dissolvable tobacco products that are marketed as an alternative to smoking in places where smoking is prohibited. These products are currently available in Indianapolis, IN, Columbus, OH, and Portland, OR. This paper describes the chemical characterization of four such products by gas chromatography-mass spectrometry (GC-MS). The dissolvable tobacco products were extracted and prepared by ultrasonic extraction using acetone, trimethylsilyl derivatization, and headspace solid phase microextraction (SPME). The following compounds were identified in the dissolvables using either ultrasonic extractions or trimethylsilyl derivatization: nicotine, ethyl citrate, palmitic acid, stearic acid, sorbitol, glycerol, and xylitol. The following compounds were identified in the dissolvables using headspace SPME: nicotine, ethyl citrate, cinnamaldehyde, coumarin, vanillin, and carvone. With the exception of nicotine, the compounds identified thus far in the dissolvables are either flavoring compounds or binders. The concentration of free nicotine in the dissolvables was determined from the Henderson-Hasselbalch equation and by measuring the pH and nicotine concentration by GC-MS. The results presented here are the first to reveal the complexity of dissolvable tobacco products and may be used to assess potential oral health effects.

The effect of microbial degradation on the chromatographic profiles of tiki torch fuel, lamp oil, and turpentine.

Abstract:  Biodegradation can result in selective removal of many of the compounds required for the identification of an ignitable liquid. In this study, the effects of microbial degradation on tiki torch fuel, lamp oil, and turpentine are reported. Samples of soil spiked with 20 μL of the liquids were stored at room temperature for up to 7 days. The ignitable liquids were then recovered using passive headspace concentration onto charcoal strips followed by solvent elution using pentane. Microbial degradation of tiki torch fuel resulted in the loss of the n‐alkanes relative to the branched alkanes. Changes in the profile of the lamp oil were minor due to the highly branched nature of its alkanes. Microbial degradation of turpentine resulted in the selective loss of limonene and o‐cymene. Overall, significant degradation by microbial action could result in the inability to identify the presence of an ignitable liquid or misclassify the ignitable liquid found.

Characterization of Automotive Paint Clear Coats by Ultraviolet Absorption Microspectrophotometry with Subsequent Chemometric Analysis

Clear coats have been a staple in automobile paints for almost thirty years and are of forensic interest when comparing transferred and native paints. However, the ultraviolet (UV) absorbers in these paint layers are not typically characterized using UV microspectrophotometry, nor are the results studied using multivariate statistical methods. In this study, measurements were carried out by UV microspectrophotometry on 71 samples from American and Australian automobiles, with subsequent chemometric analysis of the absorbance spectra. Sample preparation proved to be vital in obtaining accurate absorbance spectra and a method involving peeling the clear coat layer and not using a mounting medium was preferred. Agglomerative hierarchical clustering indicated three main groups of spectra, corresponding to spectra with one, two, and three maxima. Principal components analysis confirmed this clustering and the factor loadings indicated that a substantial proportion of the variance in the data set originated from specific spectral regions (230–265 nm, 275–285 nm, and 300–370 nm). The three classes were well differentiated using discriminant analysis, where the cross-validation accuracy was 91.6% and the external validation accuracy was 81.1%. However, results showed no correlation between the make, model, and year of the automobiles.

Design and Optimization of a Total Vaporization Technique Coupled to Solid-Phase Microextraction

Solid-phase microextraction (SPME) is a popular sampling technique in which chemical compounds are collected with a sorbent-coated fiber and then desorbed into an analytical instrument such as a liquid or gas chromatograph. Typically, this technique is used to sample the headspace above a solid or liquid sample (headspace SPME), or to directly sample a liquid (immersion SPME). However, this work demonstrates an alternative approach where the sample is totally vaporized (total vaporization SPME or TV-SPME) so that analytes partition directly between the vapor phase and the SPME fiber. The implementation of this technique is demonstrated with polydimethylsiloxane-divinylbenzene (PDMS-DVB) and polyacrylate (PA) coated SPME fibers for the collection of nicotine and its metabolite cotinine in chloroform extracts. The most important method parameters were optimized using a central composite design, and this resulted in an optimal extraction temperature (96 °C), extraction time (60 min), and sample volume (120 μL). In this application, large sample volumes up to 210 μL were analyzed using a volatile solvent such as chloroform at elevated temperatures. The sensitivity of TV-SPME is nearly twice that of liquid injection for cotinine and nearly 6 times higher for nicotine. In addition, increased sampling selectivity of TV-SPME permits detection of both nicotine and cotinine in hair as biomarkers of tobacco use where in the past the detection of cotinine has not been achieved by conventional SPME.

Identification of Ascorbic Acid and Its Degradation Products in Black Powder Substitutes

Low explosives such as smokeless powder, black powder, and black powder substitutes have been used in illicit pipe bombings throughout the United States. Some of the newer black powder substitutes are formulated with ascorbic acid, which gradually decomposes as the powder ages, making it difficult if not impossible for the forensic chemist to identify it by traditional bulk techniques. A sensitive method for the identification of residual levels of ascorbic acid in black powder substitutes is presented. Powder samples are extracted with a mixture of acetonitrile and bis(trimethylsilyl)acetamide (BSA), which converts carboxylic acid and alcohol functional groups to trimethylsilyl esters and ethers, respectively. Samples are then analyzed by gas chromatography-mass spectrometry (GC-MS). Results have shown that trace amounts of ascorbic acid can be identified at detection limits that are well below those for traditional bulk techniques. Degradation products for ascorbic acid (hydroxylated carboxylic acids, furanones, and lactones) can also be detected.

Headspace concentrations of explosive vapors in containers designed for canine testing and training: Theory, experiment, and canine trials

It is a common misconception that the amount of explosive is the chief contributor to the quantity of vapor that is available to trained canines. In fact, this quantity (known as odor availability) depends not only on the amount of explosive material, but also the container volume, explosive vapor pressure and temperature. In order to better understand odor availability, headspace experiments were conducted and the results were compared to theory. The vapor-phase concentrations of three liquid explosives (nitromethane, nitroethane and nitropropane) were predicted using the Ideal Gas Law for containers of various volumes that are in use for canine testing. These predictions were verified through experiments that varied the amount of sample, the container size, and the temperature. These results demonstrated that the amount of sample that is needed to saturate different sized containers is small, predictable and agrees well with theory. In general, and as expected, once the headspace of a container is saturated, any subsequent increase in sample volume will not result in the release of more vapors. The ability of canines to recognize and alert to differing amounts of nitromethane has also been studied. In particular, it was found that the response of trained canines is independent of the amount of nitromethane present, provided it is a sufficient quantity to saturate the container in which it is held.