The aim of this study was to characterise the microbial and metabolite diversity in samples of biodeteriorated 18th century silk from the burial crypts of the Church of St Francis of Assisi in Kraków (Poland). Highthroughput Illumina sequencing, surface-assisted laser desorption/ionisation mass spectrometry on a silver nanoparticle enhanced target (109Ag SALDI), and laser ablation-remote-electrospray ionisation-selected reaction monitoring-ambient mass spectrometry imaging (LARESI MSI) were employed. The use of high throughput Illumina sequencing helped obtain a broader picture of microbiocenosis compared to earlier studies. The 109Ag SALDI method enabled the qualitative analysis of the entire chemical compound profile of silk. A few hundred metabolites, including peptides, amino acids, urea and organic acids dominated by N-(3-oxododecanoyl)-L-homoserine, decanoylcholine, formiminoalanine and hexacosatrienoic acid were detected in archaeological silk, whereas about 100 metabolites, mainly represented by polymers, were detected in contemporary and restored silk. The novel ambient MS imaging method LARESI MSI, used for the first time for the archaeological silk biodeterioration analysis, allowed the direct detection and mapping of selected amino acids, historical dyes and dihydroxybenzoic acid (decomposition product of tannins) on the silk samples. Based on our results, the suitability of the tested methods should also be considered for other historical objects.

Infrared (IR) laser ablation-remote-electrospray ionization (LARESI) platform coupled to a tandem mass spectrometer (MS/MS) operated in selected reaction monitoring (SRM) or multiple reaction monitoring (MRM) modes was developed and employed for imaging of target metabolites in human kidney cancer tissue. SRM or MRM modes were employed to avoid artifacts that are present in full scan MS mode. Four tissue samples containing both cancerous and noncancerous regions, obtained from three patients with renal cell carcinoma (RCC), were imaged. Sixteen endogenous metabolites that were reported in the literature as varying in abundance between cancerous and noncancerous areas in various human tissues were selected for analysis. Target metabolites comprised ten amino acids, four nucleosides and nucleobases, lactate, and vitamin E. For comparison purposes, images of the same metabolites were obtained with ultraviolet (UV) desorption/ionization mass spectrometry imaging (UV-LDI-MSI) using monoisotopic silver-109 nanoparticle-enhanced target (109AgNPET) in full-scan MS mode. The acquired MS images revealed differences in abundances of selected metabolites between cancerous and noncancerous regions of the kidney tissue. Importantly, the two imaging methods offered similar results. This study demonstrates the applicability of the novel ambient LARESI SRM/MRM MSI method to both investigating and discovering cancer biomarkers in human tissue.

Climatic factors, mainly rainfall, temperature and relative humidity, air pollution and biological activity influence the processes of stone degradation. This article analyses results of studies on microbial (mainly cyanobacterial) colonization of 32 cultural heritage stone monuments in Argentina, Belize, Brazil, Cambodia, Laos and Mexico, performed by four of the six authors over several years. Principal components analysis (PCA) and canonical analysis of principal coordinates (CAP) revealed that stone type, calcareous or siliceous, affected cyanobacterial community structure in biofilms. Köppen climate class was important, more temperate climates grouping separately from more tropical sites. Highly polluted cities showed reduced cyanobacterial diversity, with centroid completely separate from the rest; Chroococcus appeared to be relatively pollution-resistant. Buildings at marine sites showed a different and somewhat reduced cyanobacterial diversity. The genus Gloeocapsa was the most frequent genus detected overall, present at all but two sites. Various cases are discussed in more detail, along with the accompanying biodeterioration, and special mention is made of black crusts seen on some buildings. Although some next generation sequencing analyses are discussed, the statistical analyses are based on results of standard microbiological techniques that have been in use for many years and were employed similarly in all the studies.

Renal cell carcinoma (RCC) is the most prevalent and lethal malignancy of the kidney. Despite all the efforts made, no tissue biomarker is currently used in the clinical management of patients with kidney cancer. A search for possible biomarkers in urine for clear cell renal cell carcinoma (ccRCC) has been conducted. Non-targeted metabolomic analyses were performed on paired samples of surgically removed renal cancer and normal tissue, as well as on urine samples. Extracts were analyzed by liquid chromatography/high-resolution mass spectrometry (LC-HRMS). Hydroxybutyrylcarnitine, decanoylcarnitine, propanoylcarnitine, carnitine, dodecanoylcarnitine, and norepinephrine sulfate were found in much higher concentrations in both cancer tissues (compared with the paired normal tissue) and in urine of cancer patients (compared with control urine). In contrast, riboflavin and acetylaspartylglutamate (NAAG) were present at significantly higher concentrations both in normal kidney tissue as well as in urine samples of healthy persons. This preliminary study resulted in the identification of several compounds that may be considered potential clear cell renal carcinoma biomarkers.

A novel method to investigate penetration, distribution and metabolic effects of biocides in microbial biofilms is described. LASCA (laser ablation solvent capture by air flow) mass spectrometry imaging (MSI) allowed for the visualization of distributions of specific ion masses, including the signature of a quaternary ammonium salt biocide, benzyl dimethyl dodecyl ammonium chloride (BDDAC), throughout the biofilm. The biocide penetrated, the upper layers of a Marinobacter sp. biofilm on a carbon steel coupon in seawater after 30 min. The concentration initially decreased with depth; however, it again increased close to the metal surface. Penetration of the biocide primarily occurred close to the center of the circular coupon, where ions with negative mass defects were preferentially detected, indicating that this is the anodic region of the corroding coupon. Future work using high performance liquid chromatography/mass spectrometry (HPLC/MS) will allow the identification within the biofilm of bacterial metabolites directly affected by biocide action. (C) 2017 Elsevier Ltd. All rights reserved.

The aim of the study was to explore the halophile metabolome in building materials using untargeted metabolomics which allows for broad metabolome coverage. For this reason, we used high-performance liquid chromatography interfaced to high-resolution mass spectrometry (HPLC/HRMS). As an alternative to standard microscopy techniques, we introduced pioneering Coherent Anti-stokes Raman Scattering Microscopy (CARS) to non-invasively visualize microbial cells. Brick samples saturated with salt solution (KCl, NaCl (two salinity levels), MgSO4, Mg(NO3)2), were inoculated with the mixture of preselected halophilic microorganisms, i.e., bacteria: Halobacillus styriensis, Halobacillus naozhouensis, Halobacillus hunanensis, Staphylococcus succinus, Marinococcus halophilus, Virgibacillus halodenitryficans, and yeast: Sterigmatomyces halophilus and stored at 28°C and 80% relative humidity for a year. Metabolites were extracted directly from the brick samples and measured via HPLC/HRMS in both positive and negative ion modes. Overall, untargeted metabolomics allowed for discovering the interactions of halophilic microorganisms with buildings materials which together with CARS microscopy enabled us to elucidate the biodeterioration process caused by halophiles. We observed that halophile metabolome was differently affected by different salt solutions. Furthermore, we found indications for haloadaptive strategies and degradation of brick samples due to microbial pigment production as a salt stress response. Finally, we detected changes in lipid content related to changes in the structure of phospholipid bilayers and membrane fluidity.

Corrosion processes in two North Sea oil production pipelines were studied by analyzing pig envelope samples via metagenomic and metabolomic techniques. Both production systems have similar physico-chemical properties and injection waters are treated with nitrate, but one pipeline experiences severe corrosion and the other does not. Early and late pigging material was collected to gain insight into the potential causes for differential corrosion rates. Metabolites were extracted and analyzed via ultra-high performance liquid chromatography/high-resolution mass spectrometry with electrospray ionization (ESI) in both positive and negative ion modes. Metabolites were analyzed by comparison with standards indicative of aerobic and anaerobic hydrocarbon metabolism and by comparison to predicted masses for KEGG metabolites. Microbial community structure was analyzed via 16S rRNA gene qPCR, sequencing of 16S PCR products, and MySeq Illumina shotgun sequencing of community DNA. Metagenomic data were used to reconstruct the full length 16S rRNA genes and genomes of dominant microorganisms. Sequence data were also interrogated via KEGG annotation and for the presence of genes related to terminal electron accepting (TEA) processes as well as aerobic and anaerobic hydrocarbon degradation. Significant and distinct differences were observed when comparing the 'high corrosion' (HC) and the 'low corrosion' (LC) pipeline systems, especially with respect to the TEA utilization potential. The HC samples were dominated by sulfate-reducing bacteria (SRB) and archaea known for their ability to utilize simple carbon substrates, whereas LC samples were dominated by pseudomonads with the genetic potential for denitrification and aerobic hydrocarbon degradation. The frequency of aerobic hydrocarbon degradation genes was low in the HC system, and anaerobic hydrocarbon degradation genes were not detected in either pipeline. This is in contrast with metabolite analysis, which demonstrated the presence of several succinic acids in HC samples that are diagnostic of anaerobic hydrocarbon metabolism. Identifiable aerobic metabolites were confined to the LC samples, consistent with the metagenomic data. Overall, these data suggest that corrosion management might benefit from a more refined understanding of microbial community resilience in the face of disturbances such as nitrate treatment or pigging, which frequently prove insufficient to alter community structure toward a stable, less-corrosive assemblage.

Fuel biodegradation linked to sulfate reduction can lead to corrosion of the metallic infrastructure in a variety of marine environments. However, the biological stability of emerging biofuels and their potential impact on copper-nickel alloys commonly used in marine systems has not been well documented. Two potential naval biofuels (Camelina-JP5 and Fisher-Tropsch-F76) and their petroleum-derived counterparts (JP5 and F76) were critically assessed in seawater/sediment incubations containing a metal coupon (70/30 Cu-Ni alloy). Relative to a fuel-unamended control (1.2 +/- 0.4 mu M/d), Camelina-JP5 (86.4 +/- 1.6 mu M/ d) and JP5 (77.6 +/- 8.3 M/d) stimulated much higher rates of sulfate reduction than either FT-F76 (11.4 +/- 2.7 mu M/d) or F76 (38.4 +/- 3.7 mu M/d). The general corrosion rate (r(2) = 0.91) and pitting corrosion (r(2) = 0.92) correlated with sulfate loss in these incubations. Despite differences in microbial community structure on the metal or in the aqueous or sediment phases, sulfate reducing bacteria affiliated with Desulfarculaceae and Desulfobacteraceae became predominant upon fuel amendment. The identification of alkylsuccinates and alkylbenzylsuccinates attested to anaerobic metabolism of fuel hydrocarbons. Sequences related to Desulfobulbaceae were highly enriched (34.2-64.8%) on the Cu-Ni metal surface, regardless of whether the incubation received a fuel amendment. These results demonstrate that the anaerobic metabolism of biofuel linked to sulfate reduction can exacerbate the corrosion of Cu-Ni alloys. Given the relative lability of Camelina-JP5, particular precaution should be taken when incorporating this hydroprocessed biofuel into marine environments serviced by a Cu-Ni metallic infrastructure. (C) 2017 Elsevier Ltd. All rights reserved.

Environmental legislation has driven the reduction of sulfur levels in automotive fuels worldwide (<= 10 ppm). We evaluate the behavior of microbial biomass in terms of community composition, metabolite production, and degradation of the Brazilian blend B10, made with ultra-low-sulfur diesel (ULSD, 6.3 ppm), high-sulfur diesel (HSD, 327 ppm), and ultra-high sulfur diesel (UHSD, 861 ppm) during simulated storage. The microcosm was assembled in glass flasks containing an aqueous phase (mineral medium) and an oil phase (fuels) at two conditions of microbial contamination: natural;(similar to 10(3) colony-forming units (CFU) per liter) and inoculated(similar to 10(6) bacterial cells and fungal spores per milliliter), evaluated each 10 days for 40 days. The results showed that biomass production was more pronounced in inoculated treatment and could be described, at T-40 as UHSD < ULSD < HSD B10. The higher degradation of terminal methyl ester fraction (50 1%), and aromatic compounds (46 2%) was in HSD B10, and ULSD B10 suffered the lowest degradation (23 +/- 3%; 26 +/- 7%, respectively) (p < 0.05). Pseudomonas,(Proteobacteria) was the predominant bacterial genus at the interface (similar to 91%), but in the water phase, changes in relative abundance and development of Pandoraea (Proteobacteria) and Propionispora (Firmicutes) were observed (p < 0.05). Ascomycota and Basidiomycota were the most abundant fungal phyla (similar to,78%). Putative fatty acids myristic, palmitic, stearic, oleic, linoleic, and alpha-linolenic acid were detected in the water phase with high relative abundance at all sulfur levels compared to controls (p < 0.05), indicating the degradation products of the fatty acid methyl esters present in soybean biodiesel. The data set suggests that the reduction of sulfur content may have favored microbial growth; however, the addition of fatty acid methyl ester (FAIVLE), additives, and the origin of petroleum-based fuels may be a more -relevant factor in the B10-blend, aerobic biodegradation.

In aquatic environments, the presence of microorganisms often leads to severe deterioration of metallic materials. While sessile cell (biofilm)-driven microbiologically-influenced corrosion (MIC), or biocorrosion, is recognized as a contributor to metal failures in marine systems, uncertainty still exists whether and to what extent products resulting from metabolic activity of the planktonic cells impact corrosion. Electrochemical measurements, molecular ecology techniques, comparative untargeted metabolomics, light and electron microscopy and surface analyses were employed to determine whether there is a difference in corrosion of 1020 carbon steel electrodes exposed to a native North Pacific seawater bacterial population that was either allowed to or prevented from developing a biofilm on the electrode surface. Laboratory studies were carried out using batch reactors. The investigation re-addressed the following issues (a) whether physical contact between cells and the metal surface is essential to enhance corrosion of 1020 carbon steel; (b) what, if any, is the impact of metabolic products secreted by cells kept in the planktonic phase, through confinement within a dialysis tube, on the severity of 1020 carbon steel deterioration; (c) are there any differences between the mineralogy of corrosion products formed on biofilm–free and biofilmed electrodes and (d) whether and to what extent does the biofilm population differ from that of the initial planktonic inoculum and the implication of this difference for diagnosing and mitigating MIC. Increased corrosion rates and extensive pitting damage were observed on the surface of biofilmed electrode (BE) compared to the biofilm-free (BFE) electrode which served as a “sterile” control. Corrosion products recovered from the BE surface had a significantly higher content (% w/w) of CaCO3 (aragonite) than those recovered from BFE surface (24.6% and 1.3% respectively). A reverse trend was observed for goethite (α-FeOOH); the latter dominated the BFE surface (43.8 wt %) and was relatively scarce (6.4 wt %) on BE surface. A vast disparity, mainly in putative lipids content, was found between metabolomes of the bacterial population allowed to form a biofilm and the same population kept in planktonic phase. DNA profiles of planktonic and biofilm bacterial populations, obtained through Illumina sequencing, were also considerably different. Extracellular DNA (e-DNA) comprising 16S rRNA sequences (up to 590 OTU), some of which were characteristic of bacteria implicated in MIC, was observed in the corrosion products recovered from the BFE surface. These results confirmed other reports that, in an oxygenated marine environment, the physical presence of an actively metabolizing bacterial biofilm leads to a pronounced pitting damage and increases corrosion rates of carbon steel. The study revealed that when cells are prevented from interacting with the surface, i.e. confined within a planktonic phase, their metabolic products released into the planktonic phase do not cause significant pitting corrosion. The corrosion in this case, is similar to that measured in filter sterilized seawater. Importantly, the investigation demonstrated that DNA profile of a biofilm differed significantly from that of planktonic seawater population, thus confirming that MIC risk assessment should not be carried out using sampling of planktonic cells alone. The presence of eDNA within corrosion products recovered from the biofilm-free electrode showed that identifying biocorrosion exclusively based on DNA profiling can be misleading and should not be used as a sole MIC diagnostic method.

Fuels that biodegrade too easily can exacerbate through-wall pitting corrosion of pipelines and tanks and result in unintentional environmental releases. We tested the biological stability of two emerging naval biofuels (camelina-JP5 and Fischer-Tropsch-F76) and their potential to exacerbate carbon steel corrosion in seawater incubations with and without a hydrocarbon-degrading sulfate-reducing bacterium. The inclusion of sediment or the positive control bacterium in the incubations stimulated a similar pattern of sulfate reduction with different inocula. However, the highest rates of sulfate reduction were found in incubations amended with camelina-JP5 [(57.2 ± 2.2)-(80.8 ± 8.1) μM/day] or its blend with petroleum-JP5 (76.7 ± 2.4 μM/day). The detection of a suite of metabolites only in the fuel-amended incubations confirmed that alkylated benzene hydrocarbons were metabolized via known anaerobic mechanisms. Most importantly, general (r(2) = 0.73) and pitting (r(2) = 0.69) corrosion were positively correlated with sulfate loss in the incubations. Thus, the anaerobic biodegradation of labile fuel components coupled with sulfate respiration greatly contributed to the biocorrosion of carbon steel. While all fuels were susceptible to anaerobic metabolism, special attention should be given to camelina-JP5 biofuel due to its relatively rapid biodegradation. We recommend that this biofuel be used with caution and that whenever possible extended storage periods should be avoided.

This study aimed to assess the interaction between bacteria and food processing surfaces using novel methods. Microbial cross contamination between stainless steel, a common food processing material, and raw chicken was studied using microbiological culture, specialized microscope and molecular techniques. Confocal laser scanning microscopy (CLSM) allowed the visualization of biofilms containing single or dual species of Escherichia coli O157:H7, Salmonella typhimurium, Bacillus cereus, Staphylococcus aureus and Pseudomonas aeruginosa, formed after 6 days’ incubation on stainless steel or 4h on raw chicken. The results provided information on intra-biofilm location and stratification of species within dual species biofilms. Top-to-bottom Z-stack images revealed that, on both materials, S. typhimurium and E. coli attached concurrently, the former in greater numbers. E. coli and B. cereus segregated on steel, E. coli more frequent near the metal surface, B. cereus almost the only species in outer layers. Few cells of S. aureus, found at all depths, were seen in the 2.9 µm thick biofilm on steel with E. coli. Greatest attachment was shown by P. aeruginosa, followed by S. typhimurium, E. coli and finally Gram positive species. Large amounts of EPS in P. aeruginosa biofilms made visualization difficult on both materials, but especially on chicken meat, a limitation of this technique. Nevertheless, CLSM was useful for determining time sequence of adhesion and species makeup of thin biofilms. The technique showed that five min contact between bacterially-contaminated chicken and sterile steel resulted in greatest transfer of P. aeruginosa, followed by S. typhimurium. This was confirmed using DGGE. Gram positive bacteria transferred poorly. A biofilm containing 2.3 × 105 cfu·cm−2 B. cereus on steel transferred an undetectable number of cells to chicken after 5 min contact. This species was unable to form biofilm on chicken when incubated for 4 h in growth medium. S. typhimurium and P.aeruginosas were most efficiently transferred from contaminated steel to raw chicken within 5 min contact, with 20–30% transfer from single species biofilms. All other species, and all cells in dual species biofilms, showed less than 2% transfer. CLSM and DGGE were shown to be useful techniques for the study of bacterial adhesion to stainless steel.

The use of gas or liquid chromatography coupled with mass spectrometry are underexplored, but extremely powerful, tools to address questions on the metabolic fate of hydrocarbons in systems that range in complexity from individual bacterial cells to natural or man-made environmental compartments. Such studies can be divided into targeted analysis, traditionally performed using gas chromatography/mass spectrometry, and untargeted analysis, for which high-performance liquid chromatography (HPLC)/mass spectrometry is preferred. The general utility of the approach is predicated on the recognition that phylogenetically diverse life forms often have somewhat similar patterns for the metabolism of individual substrates. We make the distinction between metabolic profiling and metabolomics and illustrate the use of the aforementioned tools, in combination with the requisite experimental designs, laboratory procedures, and data analysis methods, to understand the fate of hydrocarbons under both aerobic and anaerobic conditions and in both freshwater and marine environments.

Owing to their low CO2 emissions the use of bio-fuels and their blends in the Navy has become not only increasingly attractive but also mandatory. Onboard Navy ships, fuel is stored in seawater compensated fuel ballast tanks (SWCFBT) that are connected in series using copper alloy sluice pipes. Majority of Navy ships have several groups of such tanks which are typically made of carbon steel. Seawater is drawn into the tanks to compensate for fuel reduction during operations. While petro-fuels in contact with seawater are known to suffer from microbial contamination problems, bio-fuels, in particular methyl-ester (ME)-based fuels, are susceptible to biodegradation, which compromises the fuel quality, equipment performance and can lead to microbially-influenced corrosion (MIC) of SWCFBT. ME-based biofuels are, therefore, not recommended for use during naval operations. Of concern to the Navy is the possible impact of marine microbial proliferation in the presence of a second generation, methyl ester-free biofuels on corrosion of SWCFBT. Commonly, preliminary risk assessment of MIC is undertaken through laboratory investigations. The difficulty in designing and performing studies that can accurately simulate real life operational conditions in SWCFBT onboard Navy ships is one of the major drawbacks to diagnostics and mitigation of MIC in these systems. This problem is further compounded by the fact that military ship’s transit data and operational conditions are mostly classified and not easily accessible. As a result, corrosion rates observed in most laboratory experiments seldom reflect those observed in real service life environments. Here presented laboratory investigation attempted to simulate the operational condition of a SWCFBT that is adjoining to the overflow/expansion tank onboard a Navy ship and to compare marine corrosion of 1018 carbon steel in the presence of a conventional and a 50/50 blend of an alternative Navy fuel in fully oxygenated and oxygen-limited North-Pacific seawater. Studies were carried out employing six independently operated batch reactors containing as-received and 0.1 µm filter-sterilised San Diego Bay seawater (SDBSW). These reactors were augmented with either a conventional Navy fuel (petro-F76), a 50/50 blend of petro-F76 fuel with an alternative algal FT-F76 biofuel and with corresponding fuel-free controls. Linear polarization resistance (LPR) and corrosion potential (Ecorr) measurements were conducted using 312 mm2 cylindrical electrodes manufactured from 1018 carbon steel. For each electrode, LPR data were recorded every 20 s by scanning through a narrow potential range within ±5 mV of the corrosion potential (Ecorr) at a scan rate of 0.125 mV/s for 8 weeks at 23oC. Reactors were sampled at regular time intervals. Aliquots of bulk fluids were collected to (i) confirm the sterility of the bulk phase in the control reactor and (ii) for genomic and metabolomic analyses. Upon reactor decommissioning, corrosion deposits were aseptically recovered from the electrode surfaces and subjected to comprehensive chemical and microbiological characterization. Metabolomics analysis of recovered deposits and of liquid samples collected from the planktonic phase was performed using Agilent 1290 binary HPLC coupled with Agilent 6538 UHD Accurate Mass Q-ToF mass spectrometer. Characterization of crystalline phases in the corrosion deposits, imaging and elemental analysis were carried out using powder X-ray diffraction (XRD) and field emission scanning electron microscopy coupled with energy dispersive X-ray analysis (FEM/EDX), respectively. Biofilm and planktonic prokaryotic community structures were determined through DNA extraction from corrosion deposits and from liquid samples retrieved from reactors bulk phases, followed with lllumina sequencing of 16S rDNA. Electrochemical measurements, metabolomics and mineralogical analyses demonstrated marked differences between the corrosion of 1018 carbon steel exposed in the reactors. Corrosion rates recorded in the reactors varied with fuel type and oxygen concentration. The highest level of corrosion was recorded for 1018 carbon steel exposed to as-received SDB seawater augmented with conventional petro-F76 fuel. Extensive surface pitting damage was observed on all carbon steel electrodes exposed in the reactors. The severity of pitting damage decreased in the following order: conventional Navy fuel > 50/50 blend Navy fuel > SDB seawater. Metabolomics data analysis, revealed considerable differences in chemical signatures between deposits recovered from the surfaces of 1018 carbon steel electrodes exposed in the conventional Navy fuel compared to the 50/50 fuel blend. XRD patterns demonstrated that magnetite (Fe3O4) and Goethite (αFeOOH) were the dominant crystalline phases detected in the corrosion deposits recovered from the surfaces of all carbon steel electrodes. DNA profiling of biofilm communities in fuel augmented and fuel-free reactors is in progress. It is anticipated that the outcome of this multidisciplinary study will aid in elucidating the risk of MIC of carbon steel in compensated fuel tanks containing blends of a second generation naval petro- and bio-fuels.

Renal cell carcinoma (RCC) accounts for several percent of all adult malignant tumor cases and is directly associated with over 120 thousand death cases worldwide annually. Therefore, there is a need for cancer biomarker tests and methods capable of discriminating between normal and malignant tissue. It is demonstrated that gold nanoparticle enhanced target (AuNPET), a nanoparticle-based, surface-assisted laser desorption/ionization (SALDI)-type mass spectrometric method for analysis and imaging, can differentiate between normal and cancerous renal tissue. Diglyceride DG(18:1/20:0)-sodium adduct and protonated octadecanamide ions were found to have greatly elevated intensities in cancerous part of analyzed tissue specimen. Compounds responsible for mentioned ions formation were pointed out as a potential clear cell RCC biomarkers. Their biological properties and localization on the tissue surface are also discussed. Potential application of presented results may also facilitate clinical decision making during surgery for large renal masses.

Preservation of cultural heritage is of paramount importance worldwide. Microbial colonization of construction materials, such as wood, brick, mortar, and stone in historic buildings can lead to severe deterioration. The aim of the present study was to give modern insight into the phylogenetic diversity and activated metabolic pathways of microbial communities colonized historic objects located in the former Auschwitz II-Birkenau concentration and extermination camp in Oświecim, Poland. For this purpose we combined molecular, microscopic and chemical methods. Selected specimens were examined using Field Emission Scanning Electron Microscopy (FESEM), metabolomic analysis and high-throughput Illumina sequencing. FESEM imaging revealed the presence of complex microbial communities comprising diatoms, fungi and bacteria, mainly cyanobacteria and actinobacteria, on sample surfaces. Microbial diversity of brick specimens appeared higher than that of the wood and was dominated by algae and cyanobacteria, while wood was mainly colonized by fungi. DNA sequences documented the presence of 15 bacterial phyla representing 99 genera including Halomonas, Halorhodospira, Salinisphaera, Salinibacterium, Rubrobacter, Streptomyces, Arthrobacter and nine fungal classes represented by 113 genera including Cladosporium, Acremonium, Alternaria, Engyodontium, Penicillium, Rhizopus, and Aureobasidium. Most of the identified sequences were characteristic of organisms implicated in deterioration of wood and brick. Metabolomic data indicated the activation of numerous metabolic pathways, including those regulating the production of primary and secondary metabolites, for example, metabolites associated with the production of antibiotics, organic acids and deterioration of organic compounds. The study demonstrated that a combination of electron microscopy imaging with metabolomic and genomic techniques allows to link the phylogenetic information and metabolic profiles of microbial communities and to shed new light on biodeterioration processes.

A novel interface for ambient, laser ablation-based mass spectrometric imaging (MSI) referred to as laser ablation and solvent capture by aspiration (LASCA) is presented and its performance demonstrated using selected, unaltered biological materials. LASCA employs a pulsed 2.94 μm laser beam for specimen ablation. Ablated materials in the laser plumes are collected on a hanging solvent droplet with electric field-enhanced trapping, followed by aspiration of droplets and remaining plume material in the form of a coarse aerosol into a collection capillary. The gas and liquid phases are subsequently separated in a 10 μL-volume separatory funnel, and the solution is analyzed with electrospray ionization in a high mass resolution Q-ToF mass spectrometer. The LASCA system separates the sampling and ionization steps in MSI and combines high efficiencies of laser plume sampling and of electrospray ionization (ESI) with high mass resolution MS. Up to 2000 different compounds are detected from a single ablation spot (pixel). Using the LASCA platform, rapid (6 s per pixel), high sensitivity, high mass-resolution ambient imaging of “as-received” biological material is achieved routinely and reproducibly.