Emission Fingerprints Research Articles

Variation in biogenic volatile organic compound emission pattern of Fagus sylvatica L. due to aphid infection

Volatile organic compounds (VOCs) have been the focus of interest to understand atmospheric processes and their consequences in formation of ozone or aerosol particles; therefore, VOCs contribute to climate change. In this study, biogenic VOCs (BVOCs) emitted from Fagus sylvatica L. trees were measured in a dynamic enclosure system. In total 18 compounds were identified: 11 monoterpenes (MT), an oxygenated MT, a homoterpene (C 14H 18), 3 sesquiterpenes (SQT), isoprene and methyl salicylate. The frequency distribution of the compounds was tested to determine a relation with the presence of the aphid Phyllaphis fagi L. It was found that linalool, (E)-β-ocimene, α-farnesene and a homoterpene identified as (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), were present in significantly more samples when infection was present on the trees. The observed emission spectrum from F. sylvatica L. shifted from MT to linalool, α-farnesene, (E)-β-ocimene and DMNT due to the aphid infection. Sabinene was quantitatively the most prevalent compound in both, non-infected and infected samples. In the presence of aphids α-farnesene and linalool became the second and third most important BVOC emitted. According to our investigation, the emission fingerprint is expected to be more complex than commonly presumed.

Wood smoke as a source of particle-phase organic compounds in residential areas

The objective of this study was to investigate the organic composition of wood smoke emissions and ambient air samples in order to determine the wood smoke contribution to the ambient air pollution in the residential areas. From November 2005 to March 2006 particle-phase PM 10 samples were collected in the residential town Dettenhausen surrounded by forests near Stuttgart in southern Germany. Samples collected on pre-baked glass fibre filters were extracted using toluene with ultrasonic bath and analysed by gas chromatography mass spectrometry (GC-MS). 21 polycyclic aromatic hydrocarbons (PAH) including 16 USEPA priority pollutants, different organic wood smoke tracers, primarily 21 species of syringol and guaiacol derivatives, levoglucosan and its isomers mannosan, galactosan and dehydroabietic acid were detected and quantified in this study. The concentrations of these compounds were compared with the fingerprints of emissions from hardwood and softwood combustion carried out in test facilities at Universitaet Stuttgart and field investigations at a wood stove during real operation in Dettenhausen. It was observed that the combustion derived PAH was detected in higher concentrations than other PAH in the ambient air PM 10 samples. Syringol and its derivatives were found in large amounts in hardwood burning but were not detected in softwood burning emissions. On the other hand, guaiacol and its derivatives were found in both softwood and hardwood burning emissions, but the concentrations were higher in the softwood smoke compared to hardwood smoke. So, these compounds can be used as typical tracer compounds for the different types of wood burning emissions. In ambient air samples both syringol and guaiacol derivatives were found which indicates the wood combustion contribution to the PM load in such residential areas. Levoglucosan was detected in high concentrations in all ambient PM 10 samples. A source apportionment modelling, Positive Matrix Factorization (PMF) was implemented to quantify the wood smoke contribution to the ambient PM 10 bound organic compounds in the residential area.

Detection of fungal development in a closed environment through the identification of specific VOC: Demonstration of a specific VOC fingerprint for fungal development

The occurrence of disease amongst the occupants of “mouldy” environments has been widely described in the literature. However, the detection of such moulds in closed environments remains difficult, particularly in the event of recent (before the first deterioration) or masked contamination (behind a material). In this context, the present study aimed to determine a specific chemical fingerprint for fungal development detectable in closed environments (dwellings, office, museum…). To achieve this, chemical emissions from sterile and artificially contaminated by moulds materials were analyzed and compared using a descriptive statistical method. Principal Component Analysis is thus chosen to analyze the results. PCA generated optimum and similar graphical representations of the scatterplot representing the data matrix. This statistical approach made it possible to identify an emission fingerprint without applying any preconception as to the type of emitted compound. Statistical analysis of the data then enabled confirmation of the impact of moulds on total VOC emissions. This emission of specific compounds resulted in obtaining a signature for the presence of fungal development in an environment, defined by specific ions. This analysis, and use of these ions applied to dwellings, made it possible to distinguish those with proven fungal development from those with no sign of mould or with a context favorable to fungal development, thus demonstrating that a chemical fingerprint specific to fungal development could be detected in indoor environments.

Age‐dependent accumulation of lipofuscin in perivascular and subretinal microglia in experimental mice

Fundus autofluorescence (AF) imaging by confocal scanning laser ophthalmoscopy has been widely used by ophthalmologists in the diagnosis/monitoring of various retinal disorders. It is believed that fundus AF is derived from lipofuscin in retinal pigment epithelial (RPE) cells; however, direct clinicopathological correlation has not been possible in humans. We examined fundus AF by confocal scanning laser ophthalmoscopy and confocal microscopy in normal C57BL/6 mice of different ages. Increasingly strong AF signals were observed with age in the neuroretina and subretinal/RPE layer by confocal scanning laser ophthalmoscopy. Unlike fundus AF detected in normal human subjects, mouse fundus AF appeared as discrete foci distributed throughout the retina. Most of the AF signals in the neuroretina were distributed around retinal vessels. Confocal microscopy of retinal and choroid/RPE flat mounts demonstrated that most of the AF signals were derived from Iba-1+ perivascular and subretinal microglia. An age-dependent accumulation of Iba-1+ microglia at the subretinal space was observed. Lipofuscin granules were detected in large numbers in subretinal microglia by electron microscopy. The number of AF+ microglia and the amount of AF granules/cell increased with age. AF granules/lipofuscin were also observed in RPE cells in mice older than 12 months, but the number of AF+ RPE cells was very low (1.48 mm(-2) and 5.02 mm(-2) for 12 and 24 months, respectively) compared to the number of AF+ microglial cells (20.63 mm(-2) and 76.36 mm(-2) for 6 and 24 months, respectively). The fluorescence emission fingerprints of AF granules in subretinal microglia were the same as those in RPE cells. Our observation suggests that perivascular and subretinal microglia are the main cells producing lipofuscin in normal aged mouse retina and are responsible for in vivo fundus AF. Microglia may play an important role in retinal aging and age-related retinal diseases.

Identification of novel dendritic cell populations in normal mouse retina.

Whether tissue resident or infiltrating antigen-presenting cells (APCs) are involved in modulating immune responses in the retina and initiating inflammation is controversial. In this histologic study, the authors examine the retinas of mice strains with different susceptibility to experimental autoimmune uveoretinitis (EAU) for tissue resident APC. Retinal wholemounts from normal and inflamed eyes of B10R III, C57BL/6, BALB/c, and ABH Biozii mice were immunostained for APC markers (33D1, CD11c, CD11b, major histocompatibility complex [MHC] class II, F4/80, CD80, CD86, CD205, mPDCA, B220, and GR1) and analyzed by confocal fluorescence microscopy using emission fingerprinting and three-dimensional reconstruction techniques. Hematoxylin and eosin-stained histologic sections were used to evaluate EAU disease scores and to assess outer blood retina barrier (retinal pigment epithelium [RPE]) structures. A population of 33D1(+) cells was identified exclusively in the peripheral margins and juxtapapillary areas of the retina in normal, nonimmunized C57BL/6 adult mice. These cells were also MHC class II(high), and their location corresponded to sites of earliest inflammation in EAU. Numbers in the papillary area were very low (less than 10), but this region marked the predominant anatomic site for initiation of inflammation in this moderately susceptible strain. The distribution and phenotype of these cells within the retinas differed between mouse strains exhibiting different disease susceptibility. In EAU-resistant BALB/c mice, many more 33D1(+) dendritic cells were present in the normal retina but were MHC class II(low/-). Conversely, no 33D1(+) or MHC class II (+) dendriform cells could be found in the normal retinas of highly EAU-susceptible B10.RIII mice. A novel population of 33D1(+) DCs was identified in normal mouse retina. The function of these cells remains to be defined, but increased numbers correlate positively with structural abnormalities in the RPE and increased resistance of the strain to EAU.

Stromal cells associated with early invasive foci in human mammary ductal carcinoma in situ coexpress urokinase and urokinase receptor

The transition from ductal carcinoma in situ (DCIS) of the breast to invasive ductal carcinoma is facilitated by proteolytic degradation of basement membrane. The transition can be identified as microinvasive foci in a small proportion of DCIS lesions. We have previously found that MMP-13 is frequently expressed in such foci. To establish whether plasmin-directed proteolysis is likely to be involved in early invasion, we have here studied the expression of urokinase plasminogen activator (uPA) and its receptor (uPAR) in human DCIS lesions with and without microinvasion. uPA mRNA was detected in periductal stromal cells in all of 9 DCIS lesions with microinvasion and in 2 of 9 DCIS lesions without microinvasion by in situ hybridization. The uPA mRNA signal was seen in numerous stromal cells in microinvasive areas together with MMP-13 mRNA expressing cells. Double immunofluorescence analyses, using emission fingerprinting, showed that the uPA expressing stromal cells included both myofibroblasts and macrophages. The early invasive carcinoma cells were negative for uPA. uPAR immunoreactivity was focally upregulated in periductal stromal cells in all of the 9 DCIS lesions with microinvasion and in only 2 of the 9 DCIS lesions without microinvasion. uPAR was seen in both macrophages and myofibroblasts in microinvasive areas, and it was evident that uPA and uPAR colocalized in both fibroblast-like cells and macrophage-like cells. We conclude that periductal macrophages and myofibroblasts are strongly involved in the initial steps of breast cancer invasion by focally upregulating the expression of the plasminogen activation system and MMP-13.

Quantification and Localization of Bacteria in Plant Tissues Using Quantitative Real-Time PCR and Online Emission Fingerprinting

In order to quantify and localize specific bacterial target genes in plant tissue, this project has generated relevant new insights in the combined application of quantitative real-time PCR in parallel with the in situ PCR + probe-hybridization and online emission fingerprinting using LSM 510 META. After designing an Enterobacter radicincitans species-specific probe, introduced bacterial cells were monitored in growing plant parts and their colonization behaviour was examined in relation to the native bacterial community. For this purpose, the plant growth-promoting rhizobacterial (PGPR) strain Enterobacter radicincitans was applied to Brassica oleracea plants in increasing inoculum concentrations 107, 108 and 109 cells per plant. Inoculation of 109E. radicincitans cells per plant to Brassica oleracea leaves and roots resulted in significant increases of root, leaf and tuber growth. Total bacterial cell numbers were estimated using quantitative real-time PCR to be between 107 and 109 cells g−1 fresh leaf weight and about 108 cells g−1 fresh root weight of Brassica oleracea plants. Using quantitative real-time PCR, a significant colonization of Brassica oleracea leaves and roots with E. radicincitans cells was measured. Roots were colonized with a density of 107 cells g−1 fresh root weight up to at least 14 days after inoculation. That is equivalent to a proportion of E. radicincitans 16S rDNA-gene copy numbers compared to the total bacterial communities of about 10–16%. Online emission fingerprinting established that the introduced bacteria proliferated on and inside the root and that they colonized the intercellular spaces of the root cortex layer. Hence, E. radicincitans was able to successfully compete with the native bacterial population.

Amino Acid-stimulated Ca2+ Oscillations Produced by the Ca2+-sensing Receptor Are Mediated by a Phospholipase C/Inositol 1,4,5-Trisphosphate-independent Pathway That Requires G12, Rho, Filamin-A, and the Actin Cytoskeleton

The G protein-coupled Ca(2+)-sensing receptor (CaR) is an allosteric protein that responds to two different agonists, Ca(2+) and aromatic amino acids, with the production of sinusoidal or transient oscillations in intracellular Ca(2+) concentration ([Ca(2+)](i)). Here, we examined whether these differing patterns of [Ca(2+)](i) oscillations produced by the CaR are mediated by separate signal transduction pathways. Using real time imaging of changes in phosphatidylinositol 4,5-biphosphate hydrolysis and generation of inositol 1,4,5-trisphosphate in single cells, we found that stimulation of CaR by an increase in the extracellular Ca(2+) concentration ([Ca(2+)](o)) leads to periodic synthesis of inositol 1,4,5-trisphosphate, whereas l-phenylalanine stimulation of the CaR does not induce any detectable change in the level this second messenger. Furthermore, we identified a novel pathway that mediates transient [Ca(2+)](i) oscillations produced by the CaR in response to l-phenylalanine, which requires the organization of the actin cytoskeleton and involves the small GTPase Rho, heterotrimeric proteins of the G(12) subfamily, the C-terminal region of the CaR, and the scaffolding protein filamin-A. Our model envisages that Ca(2+) or amino acids stabilize unique CaR conformations that favor coupling to different G proteins and subsequent activation of distinct downstream signaling pathways.

Visualization of glucocorticoid receptor in the brain of green fluorescent protein–glucocorticoid receptor knockin mice

Glucocorticoids exert various neuroendocrinological effects, including stress response, in the central nervous system via glucocorticoid receptor (GR). GRs are transported from the cytoplasm to the nucleus upon ligand binding, and then exert the transcriptional activity. Although it is important for unraveling the actual property of the GR in vivo, subcellular dynamics of the GR are still unclear within the brain tissue in which the neuronal circuitry is maintained. To address this issue, we generated green fluorescent protein (GFP)–GR knockin mice, whose GR has been replaced by a GFP–GR fusion protein that is functionally indistinguishable from endogenous GR. In fixed brain sections of the GFP–GR knockin mice, the distribution of the green fluorescence was similar to that of GR immunoreactivity. By subtracting autofluorescence using fluorescent emission fingerprinting method with confocal laser scanning microscope, nuclear localization of GFP–GR was identifiable in the hippocampal CA3 subregion, where subcellular localization of the GR has been unsolved compared with other areas. To examine the subcellular trafficking of GFP–GR in vivo, we performed adrenalectomy on the GFP–GR knockin mice. GFP–GR was translocated from the nucleus to the cytoplasm and neurites two days after adrenalectomy. Furthermore, laser scanning cytometry by which fluorescence intensity in situ can be quantitatively measured revealed the entire GFP–GR expression level was increased. We then examined the dynamic changes in the subcellular localization of GFP–GR in living hippocampal neurons both in dissociated culture and in tissue slices. GFP–GR was localized in not only the perikarya but also neurites in the absence of ligand, and nuclear translocation following ligand treatment was observed. This is the first report visualizing subcellular trafficking of the GR in the mouse brain in more physiological condition. The present results propose new avenues for the research of the GR dynamics both in vitro and in vivo.

The relationship between volatile organic profiles and emission sources in ozone episode region—a case study in Southern Taiwan

This study investigates the relationship between volatile organic profiles in the atmosphere and emission sources in an ozone non-attainment in region Southern Taiwan. Dynamometer test of vehicles and stack sampling from industrial facilities were conducted to obtain the fingerprints of emissions from on-road mobile sources and stationary sources, respectively. In addition, field sampling of non-methane organic compounds (NMOC) concentration at monitoring stations during episode seasons were also collected by canisters. The influences of different emissions sources on airborne concentrations were estimated by back-trojectory analysis and chemical mass balance model (CMB 8.0) calculation. Field measurement data indicated that the daily average concentration of NMOC ranges between 26.4 and 69.8 ppb at different sites. The mass fraction for paraffins, oleffins and aromatics in airborne samples at these sites were 28–47%, 7–12% and 41–52%, respectively. Toluene was the dominant species among these species, followed by isopentane, n-butane and 1,2,4-trimethylbenzene. The source apportionment of airborne NMOC in the ozone non-attainment region, based on CMB simulation, is passenger cars (28–51%), motorcycles (9–24%), industrial sources (14–33%), solvent application (13–46%) and biogenic emissions (<1–2.4%), respectively. Both field measurement and model analysis showed that the vehicle exhaust and industrial emission are the dominant contributors of NMOC in the region.

Evaluation of modeled spatially and temporarily highly resolved emission inventories of photosmog precursors for the city of Augsburg: The experiment EVA and its major results

Emission inventories of NOx, CO, and individual volatile organic compounds (VOC), highly resolved in space and time, belong to the most important input parameters for chemistry and transport models (CTM) used for ozone prediction. Because of the decisive influence of the input quality on the outcome of CTM simulations, the quality of emission inventories has to be assessed. This paper presents an experimental evaluation of the highly resolved emission inventories for the city of Augsburg. The emissions of the city, determined in March and October 1998 using mass balance and tracer techniques, and derived from the measured receptor concentration ratios, were compared with emissions modeled from an emission inventory. The modeled CO emissions were in agreement with the measured ones within the combined experimental and model uncertainties. More detailed CO emission model simulations suggest that the tendency of calculated CO emissions being smaller than the measured ones may be due to higher traffic activity in Augsburg. Modeled NOx emissions were in agreement with the measured ones within the combined experimental and model uncertainties. Large deviations between modeled and measured values have been found for some individual NMHC compounds. The measured NMHC emission fingerprints were dominated by mobile sources. Substantial model predicted NMHC emissions from the solvent use could not be detected by measurements suggesting that they may not be correctly represented by the emission model.

Multi-Spectral Imaging and Linear Unmixing Add a Whole New Dimension to Laser Scanning Fluorescence Microscopy

Multicolor fluorescence microscopy has become a popular way to discriminate between multiple proteins, organelles, or functions in a single cell or animal and can be used to approximate the physical relationships between individual proteins within the cell, for instance, by using fluorescence resonance energy transfer (FRET). These techniques have become invaluable to all biologists from cytogeneticists to zoologists. Resolving multiple, co-localized chromosome probes and individual fluorescent species in corals share the common problem of spectral overlap. Even in simple systems where two or three fluorochromes are used, spectral overlap or cross-talk can be difficult to eliminate, limiting the ability to distinguish one signal from another with any confidence. However, this problem is not unique to the biologist; geologists who use remote sensing techniques have been struggling with the problem for a long time. In fact, their problems are actually worse because many geological spectra are very similar and the number of possible spectral classes is much larger than the number of fluorescent probes in a biology experiment. Nonetheless, these issues have been tackled successfully, and biologists have recently begun to employ these methods to decipher multiple spectral signatures within cells and organisms, a process known as emission fingerprinting. Here, we review some approaches that have been used to enhance spectral resolution in fluorescence microscopy.

Mineralogical fingerprints of industrial emissions — an example from Ni mining and smelting on the Kola Peninsula, NW Russia

Fifteen large (>20 kg) snowpack samples, representing the atmospheric deposition of one winter (1995/1996), were collected in the vicinity (1.5–8 km) of the copper–nickel processing plants on the Kola Peninsula, NW Russia. Microscopical (reflected light) and electron microprobe investigations were carried out on polished sections of filter residues (>0.45 μm) of these samples to identify the mineralogical composition of the particulate deposition. The technique allows us to directly distinguish between particles originating from geogenic sources (e.g. windblown rock dust) and additional anthropogenic input from industrial sources. Furthermore, it was possible to gain information on the metallurgical processes used in the different factories (one roasting plant and two smelters using different technology). Each industry emits a very characteristic spectrum of particulates. The mineralogy of the particulate inpute can thus be directly used to unravel the source of observed heavy metal inputs at any one place even in complex situations with several industries located at close distance.

Apportionment of sulfur oxides at canyonlands during the winter of 1990 — II. fingerprints of emissions from point and regional sources impacting canyonlands

During January-March of 1990 a study was conducted to determine the sources of sulfur oxides present at Canyonlands and Green River, Utah. Samples were collected at these two receptor sites and at several sites intended to characterize the chemical composition of air masses reaching the receptor sites from various geographical regions. The results of the sampling program have been given in the first paper in a series of three papers. In this paper, the concentrations of spherical aluminosilicate (SAS) particles, total fluoride, and particulate selenium, arsenic and lead are combined with meteorological data to obtain source fingerprints for the ratios of these species to SO x , from the various regional sources that can influence the Canyonlands sampling site. The results indicate that the variability in the ratios of these various species to SO x , from the various regional sources is large enough to provide useful input to a receptor based, chemical mass balance source apportionment of SO, at the receptor sites. The chemical mass balance analysis is given in the final paper of this series. The ratio of SAS: SO x , varies by a factor of 25 in emissions from various coal-fired power plants. The concentrations of lead and arsenic relative to sulfur oxides is also variable in emissions from different coal-fired power plants. The ratio of particulate selenium and total fluoride to SO x , in emissions from these coal-fired sources is relatively constant. Emissions from source regions containing smelters are high in Se, As and Pb. Emissions from source regions containing fluoride ore processing industries have a high F Tots;: SO x , ratio.