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The cyclin-dependent kinase inhibitor 3 (CDKN3) gene, involved in mitosis, is upregulated in cervical cancer (CC). We investigated CDKN3 mRNA as a survival biomarker and potential therapeutic target for CC. CDKN3 mRNA was measured in 134 CC and 25 controls by quantitative PCR. A 5-year survival study was conducted in 121 of these CC patients. Furthermore, CDKN3-specific siRNAs were used to investigate whether CDKN3 is involved in proliferation, migration, and invasion in CC-derived cell lines (SiHa, CaSki, HeLa). CDKN3 mRNA was on average 6.4-fold higher in tumors than in controls (p = 8 x 10-6, Mann-Whitney). A total of 68.2% of CC patients over expressing CDKN3 gene (fold change ≥ 17) died within two years of diagnosis, independent of the clinical stage and HPV type (Hazard Ratio = 5.0, 95% CI: 2.5-10, p = 3.3 x 10-6, Cox proportional-hazards regression). In contrast, only 19.2% of the patients with lower CDKN3 expression died in the same period. In vitro inactivation of CDKN3 decreased cell proliferation on average 67%, although it had no effect on cell migration and invasion. CDKN3 mRNA may be a good survival biomarker and potential therapeutic target in CC.

Autophagy is a process that maintains homeostasis during stress, although it also contributes to cell death under specific contexts. Ceramides have emerged as important effectors in the regulation of autophagy, mediating the crosstalk with apoptosis. Melatonin induces apoptosis of cancer cells; however, its role in autophagy and ceramide metabolism has yet to be clearly elucidated. This study was aimed to evaluate the effect of melatonin administration on autophagy and ceramide metabolism and its possible link with melatonin-induced apoptotic cell death in hepatocarcinoma (HCC) cells. Melatonin (2 mm) transiently induced autophagy in HepG2 cells through JNK phosphorylation, characterized by increased Beclin-1 expression, p62 degradation, and LC3II and LAMP-2 colocalization, which translated in decreased cell viability. Moreover, ATG5 silencing sensitized HepG2 cells to melatonin-induced apoptosis, suggesting a dual role of autophagy in cell death. Melatonin enhanced ceramide levels through both de novo synthesis and acid sphingomyelinase (ASMase) stimulation. Serine palmitoyltransferase (SPT) inhibition with myriocin prevented melatonin-induced autophagy and ASMase inhibition with imipramine-impaired autophagy flux. However, ASMase inhibition partially protected HepG2 cells against melatonin, while SPT inhibition significantly enhanced cell death. Findings suggest a crosstalk between SPT-mediated ceramide generation and autophagy in protecting against melatonin, while specific ASMase-induced ceramide production participates in melatonin-mediated cell death. Thus, dual blocking of SPT and autophagy emerges as a potential strategy to potentiate the apoptotic effects of melatonin in liver cancer cells., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The detection and identification of ignitable liquid residues in fire debris can be meaningful in fire investigations. However, background pyrolysis products and weathering hinder the identification and classification steps. In addition to those processes, the acidification of the ignitable liquids before the combustion process could make those tasks even more difficult. Nevertheless, there are no systematic studies assessing the extraction, analysis, and composition of acidified ignitable liquid residues obtained from fire debris. In this work, a method for the study of acidified ignitable liquid residues in fire debris by solid-phase microextraction with gas chromatography and mass spectrometry is proposed. This methodology has been evaluated, first with simulated solutions (gasoline/sulfuric acid mixtures set on fire under controlled conditions), and then with analysis of samples from real fire debris obtained from 18 chemical ignition Molotov cocktails made with sulfuric acid and three different ignitable liquids (two types of gasoline and diesel fuel). In addition, the extensive modifications observed in chromatograms of acidified ignitable liquid residues regarding neat and weathered samples were studied. These alterations were produced by the combustion and acidification processes. As a consequence, tert-butylated compounds are proposed as diagnostic indicators for the identification of acidified gasoline in fire debris, even in strongly weathered samples., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background & Aims: Liver fibrosis, an important health concern associated to chronic liver injury that provides a permissive environment for cancer development, is characterized by accumulation of extracellular matrix components mainly derived from activated hepatic stellate cells (HSCs). Axl, a receptor tyrosine kinase and its ligand Gas6, are involved in cell differentiation, immune response and carcinogenesis., Methods: HSCs were obtained from WT and Axl(-/-) mice, treated with recombinant Gas6 protein (rGas6), Axl siRNAs or the Axl inhibitor BGB324, and analyzed by western blot and real-time PCR. Experimental fibrosis was studied in CCl4-treated WT and Axl(-/-) mice, and in combination with Axl inhibitor. Gas6 and Axl serum levels were measured in alcoholic liver disease (ALD) and hepatitis C virus (HCV) patients., Results: In primary mouse HSCs, Gas6 and Axl levels paralleled HSC activation. rGas6 phosphorylated Axl and AKT prior to HSC phenotypic changes, while Axl siRNA silencing reduced HSC activation. Moreover, BGB324 blocked Axl/AKT phosphorylation and diminished HSC activation. In addition, Axl(-/-) mice displayed decreased HSC activation in vitro and liver fibrogenesis after chronic damage by CCl4 administration. Similarly, BGB324 reduced collagen deposition and CCl4-induced liver fibrosis in mice. Importantly, Gas6 and Axl serum levels increased in ALD and HCV patients, inversely correlating with liver functionality., Conclusions: The Gas6/Axl axis is required for full HSC activation. Gas6 and Axl serum levels increase in parallel to chronic liver disease progression. Axl targeting may be a therapeutic strategy for liver fibrosis management., (Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

A novel micro-injector for capillary electrophoresis for the handling of samples with volumes down to as little as 300 nL was designed and built in our laboratory for analyses in which the available volume is a limitation. The sample is placed into a small cavity located directly in front of the separation capillary, and the injection is then carried out automatically by controlled pressurization of the chamber with compressed air. The system also allows automated flushing of the injection chamber as well as of the capillary. In a trial with a capillary electrophoresis system with contactless conductivity detector, employing a capillary of 25 μm diameter, the results showed good stability of migration times and peak areas. To illustrate the technique, the fast separation of five inorganic cations (Na(+) , K(+) , NH4 (+) , Ca(2+) , and Mg(2+) ) was set up. This could be achieved in less than 3 min, with good limits of detection (10 μM) and linear ranges (between about 10 and 1000 μM). The system was demonstrated for the determination of the inorganic cations in porewater samples of a lake sediment core., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Raman spectroscopy has emerged as a viable technique for the organic analysis of gunshot residues (GSRs), offering additional information to the well-established analysis using scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy (SEM-EDX). In this article, a Raman imaging system with an electron-multiplying coupled-charged device (EMCCD) camera was used to analyze complete GSR particles from both conventional and nontoxic ammunition fired at different cloth targets. The same cloths were then stained with blood to mimic real evidence and measured. The direct analysis using Raman imaging of the GSR particles collected with the stubs used for SEM-EDX analysis (the frequent method used for GSR collection) was evaluated. Multivariate curve-resolution and chemical-mapping methods were applied to the spectroscopic data to identify and highlight the signal corresponding to the GSR particles and differentiate them from the substrates. It was confirmed that both measurement approaches (on the targets and the stubs) could be used for the identification of GSR particles, even under unfavorable conditions such as the presence of blood. The results obtained demonstrate the huge potential of Raman imaging for the fast analysis of complete GSR particles and prove its complementary usefulness in the analysis of the stubs used by the well-established SEM-EDX technique.

Nail polishes are cosmetic paints that may be susceptible of forensic analysis offering useful information to assist in a crime reconstruction. Although the nail polish appearance could allow a quick visual identification of the sample, this analysis is subjected to the perception and subjective interpretation of the forensic examiner. The chemical analysis of the nail polishes offers great deal of information not subjected to analyst interpretation. Confocal Raman spectroscopy is a well-suited technique for the analysis of paints due to its non-invasive and non-destructive nature and its ability to supply information about the organic and inorganic components of the sample. In this work, 77 regular and gel nail polishes were analyzed with confocal Raman spectroscopy using two laser wavelengths (532 and 780 nm). The sample behavior under the two laser wavelengths and the differences in the spectra taken at different points of the sample were studied for each nail polish. Additionally, the spectra obtained for all the nail polishes were visually compared. The results concluded that the longer laser wavelength prevents sample burning and fluorescence effects; the similarity among the spectra collected within the sample is not directly related with the presence of glitter particles; and 64% of the samples analyzed showed a characteristic spectrum. Additionally, the use of confocal Raman spectroscopy for the forensic analysis of nail polishes evidence in the form of flakes or smudges on different surfaces were studied. The results showed that both types of evidence can be analyzed by the technique. Also, two non-invasive sampling methods for the collection of the evidence from the nails of the suspect or the victim were proposed: (i) to use acetone-soaked cotton swabs to remove the nail varnishes and (ii) to scrape the nail polish from the nail with a blade. Both approaches, each exhibiting advantages and drawbacks in terms of transport and handling were appropriate for a further chemical analysis of the samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background & Aims: Sustained c-Jun N-terminal kinase (JNK) activation by saturated fatty acids plays a role in lipotoxicity and the pathogenesis of non-alcoholic steatohepatitis (NASH). We have reported that the interaction of JNK with mitochondrial Sab leads to inhibition of respiration, increased reactive oxygen species (ROS), cell death and hepatotoxicity. We tested whether this pathway underlies palmitic acid (PA)-induced lipotoxicity in hepatocytes., Methods: Primary mouse hepatocytes (PMH) from adeno-shlacZ or adeno-shSab treated mice and HuH7 cells were used., Results: In PMH, PA dose-dependently up to 1mM stimulated oxygen consumption rate (OCR) due to mitochondrial β-oxidation. At ⩾1.5mM, PA gradually reduced OCR, followed by cell death. Inhibition of JNK, caspases or treatment with antioxidant butylated hydroxyanisole (BHA) protected PMH against cell death. Sab knockdown or a membrane permeable Sab blocking peptide prevented PA-induced mitochondrial impairment, but inhibited only the late phase of both JNK activation (beyond 4h) and cell death. In PMH, PA increased p-PERK and its downstream target CHOP, but failed to activate the IRE-1α arm of the UPR. However, Sab silencing did not affect PA-induced PERK activation. Conversely, specific inhibition of PERK prevented JNK activation and cell death, indicating a major role upstream of JNK activation., Conclusions: The effect of p-JNK on mitochondria plays a key role in PA-mediated lipotoxicity. The interplay of p-JNK with mitochondrial Sab leads to impaired respiration, ROS production, sustained JNK activation, and apoptosis., (Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

This manuscript presents a preliminary investigation on the applicability of Raman imaging for non-destructive and rapid analysis of blue crossing ink lines. The MCR method was used to facilitate visualization of the distribution of inks of the same colour and the most predominant Raman signature at the crossing was used to interpret the order of application of inks. Different pen ink types, different times separating the application of the two ink lines and different paper substrates were used. From the 90 Raman images examined, the correct order of application was determined in more than 60% by direct observation. The remainder cases were not as clear due to the uneven distribution of inks and the empty spaces similar to a net-like pattern observed at the crossing. This pattern was possibly caused by physical impediments (the first ink applied acting as a physical barrier) or chemical impediments (the two inks did not stick). Such impediments were more strongly observed in the crossings involving the U. Eye pen ink, causing the complete skipping of this ink line. Moreover, most crossings showed some mixing between the two inks and it was more accentuated when the times separating the application of the inks were shorter, since the ink was fresher. The use of white or certificate papers did not seem to influence in the inks distribution nor determining the order of the inks. Although this study provided useful insights regarding crossing ink lines, future statistic studies may be helpful for more objective examinations., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Two prototypes of microstructured capillaries (MSCs) were designed, manufactured, and used to carry out different experiments. MSC-1 consisted of six holes of ≈28 μm id whereas MSC-2 consisted of 85 holes of ≈7.7 μm id. A fundamental study on the hydrodynamic injection through a commercial CE equipment was conducted. Experimental times to flush specific volumes were approximately three times larger than the theoretical values. Then, the detection of starch was carried out by using the MSCs and conventional capillaries, and the electropherograms were compared on the basis of analytical parameters employed in CE. An improvement in peak asymmetry was obtained for the MSC-1 compared to the conventional capillaries. S/N was one order of magnitude increased with the MSC, improving ten times the sensitivity. Considering this advantage, the separation and detection of nitrostarch was performed as a first application of the MSC-1. Minimal sample amounts of nitrostarch (1.7 μg) were detected. Results present a real interest in forensics since this substance had not been previously detected through CE, leading to new investigations in the design of new capillaries capable of enhancing CE performance., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Hepatocellular carcinoma (HCC) frequently develops in a pro-inflammatory and pro-fibrogenic environment with hepatic stellate cells (HSCs) remodeling the extracellular matrix composition. Molecules secreted by liver tumors contributing to HSC activation and peritumoral stromal transformation remain to be fully identified. Here we show that conditioned medium from HCC cell lines, Hep3B and HepG2, induced primary mouse HSCs transdifferentiation, characterized by profibrotic properties and collagen modification, with similar results seen in the human HSC cell line LX2. Moreover, tumor growth was enhanced by coinjection of HepG2/LX2 cells in a xenograft murine model, supporting a HCC-HSC crosstalk in liver tumor progression. Protein microarray secretome analyses revealed angiogenin as the most robust and selective protein released by HCC compared to LX2 secreted molecules. In fact, recombinant angiogenin induced in vitro HSC activation requiring its nuclear translocation and rRNA transcriptional stimulation. Moreover, angiogenin antagonism by blocking antibodies or angiogenin inhibitor neomycin decreased in vitro HSC activation by conditioned media or recombinant angiogenin. Finally, neomycin administration reduced tumor growth of HepG2-LX2 cells coinjected in mice. In conclusion, angiogenin secretion by HCCs favors tumor development by inducing HSC activation and ECM remodeling. These findings indicate that targeting angiogenin signaling may be of potential relevance in HCC management.

During the last years (2000-2014), many publications concerning the forensic analysis of questioned documents have been published, and new techniques and methodologies are nowadays employed to overcome forensic caseworks. This article reviews a comprehensive collection of the works focused on this issue, including dating studies, the analysis of inks from pens and printers, the analysis of paper, the analysis of other samples related to questioned documents and studies on intersecting lines. These sections highlight the most relevant analytical studies by a wide range of analytical techniques. Separation and spectrometric techniques are critically discussed and compared, emphasizing the advantages and disadvantages of each one. Finally, concluding remarks on the research published are included., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A new automated portable dual-channel capillary electrophoresis instrument was built and applied to the concurrent determination of cations and anions. The system uses a single buffer and hydrodynamic injection of the sample is performed autonomously. A novel engraved flow-cell interface is used at the injection ends of the capillaries allowing the autonomous operation of the system. The engraved flow-cell replaces traditionally used split injectors in purpose made capillary electrophoresis systems and makes the system design easier. A new software package with graphical user interface was employed to control the system, making its operation simple and increasing its versatility. The electrophoretic method was optimized to allow the baseline separation of 12 cations and anions commonly found in fireworks. The system was proven to be useful for the analysis of consumer fireworks, saving time and expenses compared to separate analyses for anions and cations. This is the first time that cationic and anionic compositions of fireworks are investigated together. The analysis of samples revealed several inaccuracies between the declared compositions for the fireworks and the obtained results, which could be attributed to cross-contamination during their manufacture or to a transfer between other components of the pyrotechnic item. The presence of certain unexpected peaks, however, had no apparent reason and might represent an irregularity in the manufacture of some devices., (Copyright © 2014 Elsevier B.V. All rights reserved.)

This manuscript aims to study the inter and intra brand, model and batch variability in the Raman spectral signature among modern pen inks that will help forensic document examiners during the interpretation process. Results showed that most oil-based samples have similar Raman signatures that are characteristic of the Crystal Violet dye, independently of the brand. Exception was the Pilot samples that use Victoria Pure Blue BO instead. This small inter-brand variability makes oil-based pens difficult to discriminate by brand. On the contrary, gel and liquid-based samples use different colorants such as Rhodamine B, Copper Phthalocyanine, Ethyl Violet and Victoria Blue B. No particular pattern was observed regarding the colorants used by each brand, except the Pilot samples that were the only brand using the Victoria Blue B dye, which is a clear distinct feature. Additionally, the intra-brand variability was also large among gel-based Pilot samples. The small spectral differences observed among several batches of Bic Crystal Medium samples demonstrated that changes were introduced in their chemical formula over the years. The intra-batch variability was small and no spectral differences were observed within batches. This manuscript demonstrates the potential of Raman spectroscopy for discriminating pens inks from different brands and models and even, batches. Additionally, the main colorants used in modern pens were also identified., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

This study examines the utility of near infrared hyperspectral imaging (NIR-HSI) combined with chemometrics for the detection of dynamite residues on human handprints. Polyvinyl sheets containing dynamite residues were then analysed with the NIR-HSI system. A spectral library was developed by using partial least squares-discriminant analysis model (PLS-DA) to detect and classify the pixels contaminated with the dynamite residues. Values of sensitivity and specificity of 100% were obtained for both calibration and cross validation of dynamite and ammonium nitrate. The results were tested in real human handprints. Seven volunteers deposited their handprints into polyvinyl transparent sheets after the manipulation of a common type of dynamite which was mainly composed by ammonium nitrate. These results highlight the extremely high potential and capability of NIR-HSI combined with chemometrics for the fast and easy identification of explosive residues and additionally, its potential competence to detect the explosive manipulation., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Over the last few years, there has been an increase in the reports of drug-facilitated crimes. The list of drugs associated with these crimes is extensive and benzodiazepines constitute one of the groups of substances more commonly used. The sedative properties, which characterize benzodiazepines, are enhanced when such drugs are combined with alcohol, being more attractive for committing these types of crimes. In this work, a capillary electrophoresis method was applied to the analysis of 63 different samples of club drinks spiked with benzodiazepine tablets. The resulting electropherograms were processed and analyzed with the chemometric multivariate techniques: principal component analysis (PCA) and soft independent modeling of class analogies (SIMCA) classification. The PCA results allowed a clear differentiation of each drug class in a 3D plot. In addition, the SIMCA classification model (5% significance level) showed that eight out of nine test samples were automatically assigned by software to their proper sample class. The conflicting sample was correctly classified in the Coomans' plot (95% confidence). This novel approach based on the comparison of electrophoretic profiles of spiked drinks by chemometric tools allows determining the benzodiazepine used for drink spiking without the use of drug standards. Moreover, it provides an opportunity for the forensic laboratories to incorporate the identification capability provided by the electrophoretic fingerprinting of benzodiazepine solutions in existing or new databases., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Consumer fireworks are a heterogeneous group of pyrotechnic items widely used by citizens around the world. There are a wide number of forensic cases related to consumer fireworks that require knowing their chemical composition and variety of designs to conduct accurate and comprehensive analyses. In this research paper, a selection of six consumer firework types (firecracker, rocket, pyrotechnic fountain, pyrotechnic battery, sparkler, and smoke bomb) is physically described and their anionic compositions are determined. Preblast (fuses and charges) samples and postblast residues of the different consumer fireworks were analyzed by CE in order to determine their anionic composition. Different types of chemical compositions in fuses and pyrotechnic charges were determined, although they were not related to any type of item. Additionally, several discrepancies were found between the analytical results and the declared item compositions. Regarding postblast residues, a huge variety of anions were identified and attributed to some unconsumed starting materials and different chemical reactions occurring during combustion., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This work reports an investigation for the analysis of different paper samples using CE with laser-induced detection. Papers from four different manufactures (white-copy paper) and four different paper sources (white and recycled-copy papers, adhesive yellow paper notes and restaurant serviettes) were pulverized by scratching with a surgical scalpel prior to their derivatization with a fluorescent labeling agent, 8-aminopyrene-1,3,6-trisulfonic acid. Methodological conditions were evaluated, specifically the derivatization conditions with the aim to achieve the best S/N signals and the separation conditions in order to obtain optimum values of sensitivity and reproducibility. The best conditions, in terms of fastest, and easiest sample preparation procedure, minimal sample consumption, as well as the use of the simplest and fastest CE-procedure for obtaining the best analytical parameters, were applied to the analysis of the different paper samples. The registered electropherograms were pretreated (normalized and aligned) and subjected to multivariate analysis (principal component analysis). A successful discrimination among paper samples without entanglements was achieved. To the best of our knowledge, this work presents the first approach to achieve a successful differentiation among visually similar white-copy paper samples produced by different manufactures and paper from different paper sources through their direct analysis by CE-LIF and subsequent comparative study of the complete cellulose electropherogram by chemometric tools., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background & Aims: Acid sphingomyelinase (ASMase) is activated in non-alcoholic steatohepatitis (NASH). However, the contribution of ASMase to NASH is poorly understood and limited to hepatic steatosis and glucose metabolism. Here we examined the role of ASMase in high fat diet (HFD)-induced NASH., Methods: Autophagy, endoplasmic reticulum (ER) stress and lysosomal membrane permeabilization (LMP) were determined in ASMase(-/-) mice fed a HFD. The impact of pharmacological ASMase inhibition on NASH was analyzed in wild type mice fed a HFD., Results: ASMase deficiency determined resistance to hepatic steatosis mediated by a HFD or methionine-choline deficient diet. ASMase(-/-) mice were resistant to HFD-induced hepatic ER stress, but sensitive to tunicamycin-mediated ER stress, indicating selectivity in the resistance of ASMase(-/-) mice to ER stress and steatosis. Autophagic flux, determined in the presence of rapamycin and/or chloroquine, was lower in primary mouse hepatocytes (PMH) from ASMase(-/-) mice and accompanied by increased p62 levels, suggesting autophagic impairment. Moreover, autophagy suppression by chloroquine and brefeldin A caused ER stress in PMH from ASMase(+/+) mice but not in ASMase(-/-) mice. ASMase(-/-) PMH exhibited increased lysosomal cholesterol loading, decreased LMP and apoptosis resistance induced by O-methyl-serine dodecylamide hydrochloride or palmitic acid, effects that were reversed by decreasing cholesterol levels by oxysterol 25-hydroxycholesterol. In vivo pharmacological ASMase inhibition by amitriptyline, a widely used tricyclic antidepressant, protected wild type mice against HFD-induced hepatic steatosis, fibrosis, and liver damage, effects indicative of early-stage NASH., Conclusions: These findings underscore a critical role for ASMase in diet-induced NASH and suggest the potential of amitriptyline as a treatment for patients with NASH., (Copyright © 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

In this study near infrared hyperspectral imaging (NIR-HSI) is used to provide a fast, non-contact, non-invasive and non-destructive method for the analysis of explosive residues on human handprints. Volunteers manipulated individually each of these explosives and after deposited their handprints on plastic sheets. For this purpose, classical explosives, potentially used as part of improvised explosive devices (IEDs) as ammonium nitrate, blackpowder, single- and double-base smokeless gunpowders and dynamite were studied. A partial-least squares discriminant analysis (PLS-DA) model was built to detect and classify the presence of explosive residues in handprints. High levels of sensitivity and specificity for the PLS-DA classification model created to identify ammonium nitrate, blackpowder, single- and double-base smokeless gunpowders and dynamite residues were obtained, allowing the development of a preliminary library and facilitating the direct and in situ detection of explosives by NIR-HSI. Consequently, this technique is showed as a promising forensic tool for the detection of explosive residues and other related samples., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Glutathione (GSH) is the main non-protein thiol in cells whose functions are dependent on the redox-active thiol of its cysteine moiety that serves as a cofactor for a number of antioxidant and detoxifying enzymes. While synthesized exclusively in the cytosol from its constituent amino acids, GSH is distributed in different compartments, including mitochondria where its concentration in the matrix equals that of the cytosol. This feature and its negative charge at physiological pH imply the existence of specific carriers to import GSH from the cytosol to the mitochondrial matrix, where it plays a key role in defense against respiration-induced reactive oxygen species and in the detoxification of lipid hydroperoxides and electrophiles. Moreover, as mitochondria play a central strategic role in the activation and mode of cell death, mitochondrial GSH has been shown to critically regulate the level of sensitization to secondary hits that induce mitochondrial membrane permeabilization and release of proteins confined in the intermembrane space that once in the cytosol engage the molecular machinery of cell death. In this review, we summarize recent data on the regulation of mitochondrial GSH and its role in cell death and prevalent human diseases, such as cancer, fatty liver disease, and Alzheimer's disease.

Lipsticks are very popular cosmetic products that can be transferred by contact to different surfaces, being important forensic evidence with an intricate analysis if they are found in a crime scene. This study evaluates the use of confocal Raman microscopy at 780 nm excitation wavelength for the nondestructive identification of 49 lipsticks of different brands and colors, overcoming the lipstick fluorescence problem reported by previous works using other laser wavelengths. Although the lipsticks samples showed some fluorescence, this effect was not so intense to completely overwhelm the Raman spectra. Lipsticks smudges on twelve different surfaces commonly stained with these samples were also analyzed. In the case of the surfaces, some of them provided several bands to the smudge spectra compromising the identification of the lipstick. For these samples spectral subtraction of the interfering bands from the surface was performed. Finally, five different red lipsticks with very similar color were measured on different surfaces to evaluate the lipstick traceability with their smudges even on interfering surfaces. Although previous spectral subtraction was needed in some cases, all the smudged were linked to their corresponding lipsticks even when they are smeared on the interfering surfaces. As a consequence, confocal Raman microscopy using the 780 nm excitation laser is presented as a nondestructive powerful tool for the identification of these tricky samples., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Shooting distance estimation offers useful information for the reconstruction of firearm related incidents. The muzzle to target distance is usually estimated by examining the bullet entrance hole and the gunshot residue pattern. To visualize the pattern the forensic analyst usually uses presumptive tests based on color chemical reactions that are applied using long and tedious proceedings. Due to the drawbacks of the chemical tests recent developments for shooting distance estimation not based on color chemical tests were described in the literature. The present review covers the approaches for shooting distance estimation published in the last 10 years considering two types of target, clothing and skin., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Sponges corresponding to the Jaspidae family have proved to be a prolific source of bioactive natural products. Among these, the bengamides and the bengazoles stand out by virtue of their unprecedented molecular architectures and impressive biological profiles, including antitumor, antibiotic and anthelmintic properties. As a consequence, intense research activity has been devoted to these compounds from both chemical and biological standpoints. This review describes in detail the research into these classes of natural products and the benefits they offer in chemistry and biology.

Raman microscopy is a useful tool for the analysis of drug particles collected with adhesive tapes. In this work, first, the spectra of thirty drugs of abuse, degradation products, metabolites, and common cutting agent standards were recorded and the Raman bands observed were summarized providing the forensic analyst useful information for the identification of drug evidence. Then, the collection of different drug particles by a fingerprint lifting tape commonly used to remove and store fingerprints and fibers, and a white and green packaging tape, followed by the subsequent identification of the drugs by confocal Raman spectroscopy was performed. The particles were analyzed on top of the tapes, trapped between glass slides and the tapes, trapped in the tape folded over itself in the case of the transparent tape, and after folding and unfolding the tape in the case of the colored tape. The results obtained by the different approaches show that both tapes did not compromise the drugs spectra. However, the use of transparent tape is preferred because this tape allows the previous visual detection of the particles. Finally, several drug and sugar particles were spread over a clean table and inside a pocket, and the particles were collected with transparent tape and then properly identified. Although good results were obtained in both cases, the amount of fibers and other substances present in the collection area made the previous detection of the particles difficult and increases the analysis time., (Copyright © 2013 Forensic Science Society. Published by Elsevier Ireland Ltd. All rights reserved.)

Determining the sequencing of intersecting ink lines is one of the current problems for forensic document examiners. The way two inks will distribute and interact with each other and the paper at the crossing is a dynamic process that can be affected by many variables. Thus, the main purpose of this manuscript is to visualize and have a more comprehensive understanding of this process as well as study a methodology for determining the correct order of intersecting ink lines. For this, overlapping layers of different types of inks from writing instruments and printers were cross-sectioned and examined with a microscope. Results from pen/pen crossings showed that liquid-liquid and gel-gel intersections tended to form a double layer but oil-oil intersections usually formed mixtures. Additionally, oil-liquid and oil-gel intersections tended to form a double layer whenever the oil ink was on top and liquid-gel intersections tended to form a double layer for almost all crossings with exception of the ones involving a gel pen ink from one manufacturer. Results from pen/printer crossings showed the formation of a double layer only when the printer ink was on top of the pen ink. On the other permutation, the pen ink tended to penetrate through the printer ink producing the mixture of both inks. The inks drying time was found to be an important factor affecting the interaction between two inks in a crossing, particularly crossings involving gel pen inks. On the contrary, the type of paper and the writing pressure showed no significant influence on the inks distribution at the crossing. The methodology developed was reproducible with overlapping layers but there were many experimental difficulties during the validation process of intersections representing real crossings. Moreover, interpretation was dependent on the operator's eye which was a limiting factor., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Alcoholic liver disease (ALD) is a major cause of chronic liver disease and a growing health concern in theworld. While the pathogenesis of ALD is poorly characterized key players identified in experimental models and patients, such as perturbations in mitochondrial structure and function, selective loss of antioxidant defense and susceptibility to inflammatory cytokines, contribute to ALD progression. Both oxidative stress and mitochondrial dysfunction compromise essential cellular functions and energy generation and hence are important pathogenic mechanisms of ALD. An important process mediating the mitochondrial disruption induced by alcohol intake is the trafficking of cholesterol to mitochondria, mediated by acid sphingomyelinase-induced endoplasmic reticulum stress, which contributes to increased cholesterol synthesis and StARD1upregulation. Mitochondrial cholesterol accumulation not only sensitizes to oxidative stress but it can contribute to the metabolic reprogramming in ALD, manifested by activation of the hypoxia inducible transcription factor 1 and stimulation of glycolysis and lactate secretion. Thus, a better understanding of the mechanisms underlying alcohol-mediated mitochondrial impairment and oxidative stress may lead to the identification of novel treatments for ALD. The present review briefly summarizes current knowledge on the cellular and molecular mechanisms contributing to alcohol-induced mitochondrial dysfunction and cholesterol accumulation and provides insights for potential therapeutic targets in ALD., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

This paper is a comprehensive review grouping the information on the extraction, characterization, and quantitation of olive and olive oil proteins and providing a practical guide about these proteins. Most characterized olive proteins are located in the fruit, mainly in the seed, where different oleosins and storage proteins have been found. Unlike the seed, the olive pulp contains a lower protein content having been described a polypeptide of 4.6 kDa and a thaumain-like protein. Other important proteins studied in olive fruits have been enzymes which could play important roles in olives characteristics. Part of these proteins is transferred from the fruit to the oil during the manufacturing process of olive oil. In fact, the same polypeptide of 4.6 kDa found in the pulp has been described in the olive oil and, additionally, the presence of other proteins and enzymes have also been described. Protein profiles have recently been proposed as an interesting strategy for the varietal classification of olive fruits and oils. Nevertheless, there is still a lot of knowledge without being explored requiring new studies focused on the determination and characterization of these proteins.

In recent years, scopolamine has become a drug of common use for recreational and predatory purposes and several ways of administration have been devised. A method for the rapid analysis of suspicious samples was developed, using a portable capillary electrophoresis with contactless conductivity detection. The method allows the separation of scopolamine from atropine which has a similar structure and is present along with scopolamine in some samples. The method was demonstrated to be useful for the fast analysis of several types of evidential items which have recently been reported to have been abused with fatal consequences or employed for criminal purposes. An infusion of Datura stramonium L., in which scopolamine and atropine naturally coexist, was analyzed for being frequently consumed for recreational purposes. A spiked moisturizing cream and six spiked alcoholic beverages were also analyzed. In spite of the complexity of the specimens, the sample pre-treatment methods developed were simple and fast., (© 2013.)

A comparative study on the retentiveness of two plastic bags (DUO and Royal Pack) has been carried out by gas chromatography with mass spectrometry detection. Two types of dynamites were packed in both plastic bags. The bags were placed into glass jars and headspace analyses were performed over 11 weeks to detect whether the volatile constituents of the dynamites were released from the bags. DUO plastic bags showed much better retentiveness than Royal Pack plastic bags. Ethylene glycol dinitrate (EGDN) was quickly detected in the headspace of the glass jars containing Royal Pack plastic bags after 1 week of storage. On the contrary, only a weak signal of EGDN, which was not detectable in the total ion chromatogram, was detected after 11 weeks of storage. Moreover, DUO plastic bags have shown less background signals than the Royal Pack bags, being the former bags much more suitable for the storage of dynamites., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

The methods for perpetrating forgery and alteration of documents are becoming increasingly more sophisticated. Forensic examinations of questioned documents routinely involve physical and chemical analysis of inks. Raman spectroscopy is a very attractive technique for ink analysis because it combines chemical selectivity with ease and fast analysis and it does not require sample preparation nor leads to destruction of the evidence. However, some limitations of this technique include low sensitivity and the overwhelming phenomenon of fluorescence, which can be solved by resonance Raman spectroscopy and surface-enhanced Raman spectroscopy. This article aims to demonstrate the great potential of the Raman-based techniques by providing an overview of their application to forensic examinations of ink evidence from pens and printers. Moreover, it is also addressed the chemistry of ink-paper interactions and the problematic of intersecting lines., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Gunshot residues (GSR) are valuable evidence which provide the forensic analyst with useful information about a crime scene when proper analytical methods are used. Nowadays, the method of choice for analyzing GSR is scanning electron microscopy coupled with energy dispersive X-ray analysis (SEM/EDX). However, SEM/EDX presents limitations when the GSR identification of "non-toxic" ammunition types is performed. To overcome this drawback, Raman spectroscopy has been recently proposed as a complementary technique to SEM/EDX. However, for the time being, it can only be used in a limited number of casework (e.g. examining the macroscopic GSR produced at short distance over victim's clothes) and further research to know when this technique could support SEM-EDX results is required. In the present work, the memory effect of the weapon, which plays an important role to link the GSR found and the ammunition fired, is studied. Twenty shots were fired at close distance (~30 cm) at paper targets using the same weapon with two different types of ammunition. The first, third, ninth, and twentieth shots were fired with the first ammunition and the shots among them using the second ammunition. The macroscopic GSR produced by the first ammunition were analyzed using Raman spectroscopy. First, the spectra obtained were visually differentiated by taking into account the band at about 1342 cm(-1). This first approach shown that in the first shot were no GSR particles from the second ammunition, but 1.5-7.5% of analyzed particles corresponded to the second ammunition in the third, ninth, and twentieth shots. Additionally, the same differentiation was then performed by discriminant analysis using the spectral range from 1800 to 800 cm(-1). Although using this second approach only one GSR was identified as the second ammunition, was remarkable that after the shots with different ammunition the GSR obtained shows greater variability. The results obtained suggest that the memory effect of the weapon has not a significant influence when the organic analysis of macroscopic GSR on targets by Raman spectroscopy is performed., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Alcohol-induced liver disease (ALD) is a major health concern of alcohol abuse and a leading cause of liver-related morbidity and mortality. The pathogenesis of ALD is multifactorial and still ill characterized. One of the hallmarks of ALD common for both patients and experimental models is the alteration in the architecture and function of mitochondria. Due to their primordial role in energy production, metabolism and cell fate decisions, these changes in mitochondria caused by alcohol are considered an important contributory factor in ALD. A better understanding of the mechanisms underlying alcohol-mediated mitochondrial alterations may shed light on ALD pathogenesis and provide novel avenues for treatment. The purpose of the current review is to briefly update the latest developments in ALD research regarding morphological and functional mitochondrial regulation including mitochondrial dynamics and biogenesis, mitochondrial protein acetylation and evidence for an endoplasmic reticulum stress-mitochondrial cholesterol link of potential relevance for ALD.

This work is focused on a novel procedure to discriminate nitrocellulose-based samples with non-explosive and explosive properties. The nitrocellulose study has been scarcely approached in the literature due to its special polymeric properties such as its high molar mass and complex chemical and structural characteristics. These properties require the nitrocellulose analysis to be performed by using a few organic solvents and in consequence, they limit the number of adequate analytical techniques for its study. In terms of identification of pre-blast explosives, mass spectrometry is one of the most preferred technique because it allows to obtain structural information. However, it has never been used to analyze polymeric nitrocellulose. In this study, the differentiation of non-explosive and explosive samples through nitrocellulose fingerprints obtained by capillary electrophoresis was investigated. A batch of 30 different smokeless gunpowders and 23 different everyday products were pulverized, derivatized with a fluorescent agent and analyzed by capillary electrophoresis with laser-induced fluorescence detection. Since this methodology is specific to d-glucopyranose derivatives (cellulosic and related compounds), and paper samples could be easily found in explosion scenes, 11 different paper samples were also included in the study as potential interference samples. In order to discriminate among samples, multivariate analysis (principal component analysis and soft independent modeling of class analogy) was applied to the obtained electrophoretic profiles. To the best of our knowledge, this represents the first study that achieve a successful discrimination between non-explosive and explosive nitrocellulose-based samples, as well as potential cellulose interference samples, and posterior classification of unknown samples into their corresponding groups using CE-LIF and chemometric tools., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In this work, a new purpose-made portable CE instrument with a contactless conductivity detector was used for the determination of degradation products of nitrogen mustards in different water samples. The capillary was coated with poly(1-vinylpyrrolidone-co-2-dimethylaminoethyl methacrylate) to avoid analyte-wall interactions. The coating procedure was studied to obtain the best repeatability of the migration time of the analytes. Four different coating procedures were compared; flushing the capillary with the copolymer at 100 psi for 2 min at 60°C provided the best RSD values (<4%). The analytical method was also optimized. The use of 20 mM of MES adjusted to pH 6.0 with His as running buffer allowed a good baseline separation of the three analytes in different water samples without matrix interferences. The method permitted the detection of the three degradation products down to 5 μM., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

With the objective of investigating the influence of structural modifications of the polyketide chain of the bengamides upon their antitumoral activities, we targeted the preparation of bengamide E analogues with modification of the stereochemistry at C-2 and at C-3, the substituent at the C-2 position, and the presence of oxirane rings. For the synthesis of these analogues, a new synthetic method for asymmetric epoxidation, developed in our laboratories, was employed utilizing the chiral sulfonium salts 22 and 23. In order to access 2-epi-bengamide E from these epoxy amides, a synthetic methodology, developed by Miyashita, allowed an oxirane-ring-opening reaction with a double inversion of the configuration. Alternatively, an aldol reaction provided access to the same analogue in a shorter and more efficient manner. Finally, biological evaluation of all of these bengamide E analogues demonstrated that the polyketide chain is essential for the antitumor activity of these natural products, not being amenable to structural or configurational modifications.

The differentiation of blue ballpoint pen inks written on documents through an LA-ICP-MS methodology is proposed. Small common office paper portions containing ink strokes from 21 blue pens of known origin were cut and measured without any sample preparation. In a first step, Mg, Ca and Sr were proposed as internal standards (ISs) and used in order to normalize elemental intensities and subtract background signals from the paper. Then, specific criteria were designed and employed to identify target elements (Li, V, Mn, Co, Ni, Cu, Zn, Zr, Sn, W and Pb) which resulted independent of the IS chosen in a 98% of the cases and allowed a qualitative clustering of the samples. In a second step, an elemental-related ratio (ink ratio) based on the targets previously identified was used to obtain mass independent intensities and perform pairwise comparisons by means of multivariate statistical analyses (MANOVA, Tukey's HSD and T2 Hotelling). This treatment improved the discrimination power (DP) and provided objective results, achieving a complete differentiation among different brands and a partial differentiation within pen inks from the same brands. The designed data treatment, together with the use of multivariate statistical tools, represents an easy and useful tool for differentiating among blue ballpoint pen inks, with hardly sample destruction and without the need for methodological calibrations, being its use potentially advantageous from a forensic-practice standpoint. To test the procedure, it was applied to analyze real handwritten questioned contracts, previously studied by the Department of Forensic Document Exams of the Criminalistics Service of Civil Guard (Spain). The results showed that all questioned ink entries were clustered in the same group, being those different from the remaining ink on the document., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Background: Mitochondria are the powerhouse of mammalian cells and the main source of reactive oxygen species (ROS) associated with oxygen consumption. In addition, they also play a strategic role in controlling the fate of cells through regulation of death pathways. Mitochondrial ROS production fulfills a signaling role through regulation of redox pathways, but also contributes to mitochondrial damage in a number of pathological states., Scope of Review: Mitochondria are exposed to the constant generation of oxidant species, and yet the organelle remains functional due to the existence of an armamentarium of antioxidant defense systems aimed to repair oxidative damage, of which mitochondrial glutathione (mGSH) is of particular relevance. Thus, the aim of the review is to cover the regulation of mGSH and its role in disease., Major Conclusions: Cumulating evidence over recent years has demonstrated the essential role for mGSH in mitochondrial physiology and disease. Despite its high concentration in the mitochondrial matrix, mitochondria lack the enzymes to synthesize GSH de novo, so that mGSH originates from cytosolic GSH via transport through specific mitochondrial carriers, which exhibit sensitivity to membrane dynamics. Depletion of mGSH sensitizes cells to stimuli leading to oxidative stress such as TNF, hypoxia or amyloid β-peptide, thereby contributing to disease pathogenesis., General Significance: Understanding the regulation of mGSH may provide novel insights to disease pathogenesis and toxicity and the opportunity to design therapeutic targets of intervention in cell death susceptibility and disease. This article is part of a Special Issue entitled Cellular functions of glutathione., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Based on our previously described synthetic strategy for bengamide E, a natural product of marine origin with antitumor activity, a small library of analogues modified at the terminal olefinic position was generated with the objective of investigating the effect of structural modifications on antitumor properties. Biological evaluation of these analogues, consisting of IC50 determinations against various tumor cell lines, revealed important aspects with respect to the structural requirements of this olefinic position for activity. Interestingly, the analogue possessing a cyclopentyl group displayed greater potency than the parent bengamide E, representing a key finding upon which to base further investigations into the design of new analogues with promising biological activities., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Apart from powerful explosives, dynamites are complex samples with an intricate analysis. These mixtures of compounds of diverse chemical nature present a challenge to the analyst, and as a result, several analytical techniques need to be applied currently for their analysis. Taking into account that presently there are almost no methods for dynamite analysis in the literature, it is crucial to develop analytical methods that could be applied for the analysis of these samples. This study introduces the use of Raman spectroscopy to analyze dynamites. Two different dynamites made up of ethylene glycol dinitrate and ammonium nitrate, among other minor components, were analyzed by Raman spectroscopy. First, confocal Raman spectroscopy allowed the identification of different components easily distinguished by eye (ammonium nitrate, ethylene glycol dinitrate, and sawdust). Then, Raman mapping was used to show the distribution of the main components throughout the dynamite mass. Finally, several minor components were identified after flocculation (nitrocellulose) or precipitation (sawdust, CaCO3, and flour). The results obtained demonstrate the huge potential of this technique for the analysis of such a complex and tricky sample.

The gunpowder age is information of great importance that could help to establish safety regulations related to the propellants use and manipulation. In this work, a forced aging treatment (65°C for 120 days) was applied to four gunpowders stabilized with diphenylamine (DPA). The evolution of DPA and derivatives (N-nitroso-DPA, 2-nitro-DPA, 4-nitro-DPA, and 4-4'-dinitro-DPA) concentration during the days was leaded by High Performance Liquid Chromatography (HPLC). The variation with time of the peak areas of these compounds was used to construct different statistical models that could predict the gunpowders age. These models were validated using nitrocellulose-based gunpowders of known manufacture date. Models that best predicted the gunpowder age provided prediction errors lower than 6, 4, and 2 years for single-base gunpowders with dinitrotoluene (≥ 10%(m/m)), single-base gunpowders and double-base gunpowders, respectively., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background: Treatment with parenteral iron causes oxidative stress, inflammation and endothelial dysfunction. Ferric carboxymaltose (FCM) is a new preparation of non-dextran iron which, due to its pharmacokinetics and stability, may induce less toxicity than other iron molecules. The aim of this study was to analyse the effect of FCM on inflammation and adhesion molecules in chronic kidney disease (CKD)., Methods: Forty-seven patients with predialysis CKD and iron-deficiency anaemia received a single dose of FCM (15 mg/kg, maximum dose 1 gram). At baseline and after 60 minutes (acute effect) and after 3 weeks and 3 months (sub-acute effect), we determined inflammatory markers: C-reactive protein (CRP), interleukin-6 (IL-6) and endothelial dysfunction: intercellular adhesion molecule (ICAM) and vascular adhesion molecule (VCAM)., Results: Treatment with FCM was associated with a significant increase in haemoglobin levels: 10 (0.7) vs. 11.4 (1.3)g/dl, p<.0001. CRP, IL-6, ICAM and VCAM levels did not correlate with baseline haemoglobin or ferritin levels and there was no relationship between changes in these markers and those of haemoglobin after administration of FCM. No significant, acute or sub-acute changes occurred in any of the inflammatory or endothelial markers studied. Statin therapy was associated with lower VCAM concentrations., Conclusions: Treatment with high doses of FCM in patients with predialysis CKD has no proinflammatory effect and does not alter levels of adhesion molecules ICAM and VCAM in this population.

Peanut is recognized as a potent food allergen producing one of the most frequent food allergies. This fact has originated the publication of an elevated number of scientific reports dealing with peanut allergens and, especially, the prevalence of peanut allergy. For this reason, the information available on peanut allergens is increasing and the debate about peanut allergy is always renewed. This article reviews the information currently available on peanut allergens and on the techniques used for their chemical characterization. Moreover, a general overview on the current biotechnological approaches used to reduce or eliminate peanut allergens is also provided.

A new type of chiral sulfonium salts that are characterized by a bicyclic system has been designed and synthesized from α-amino acids. Their corresponding ylides, which were prepared by basic treatment of the sulfonium salts, reacted smoothly with a broad array of simple and chiral aldehydes to provide trans-epoxy amides in reasonable to very good yields and excellent stereoselectivities (>98%). The obtained epoxy amides were found to be useful as synthetic building blocks. Thus, they were reduced into their corresponding epoxy alcohols and subjected to oxirane-ring-opening reactions with different types of nucleophiles., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Oat (Avena sativa L.) is an important crop produced in various regions of Europe and North America. Oat lipids are a heterogeneous mixture of acyl lipids and unsaponifiable components. The neutral lipids are mainly triacylglycerols and account for 50-60% of total oat lipids. Oat oil is also rich in polar lipids, that is, phospholipids and glycolipids. Characterization of oat polar lipids has largely been performed by thin-layer chromatography (TLC), but the composition of phospholipid classes has been poorly studied. The aim of our work was the determination of different phospholipids in Romanian oat samples. For that purpose, one commercial sample (Comun) and four pure varieties (Jeremy, Lovrin 1, Lovrin 27-T, and Mures) were used. High-performance liquid chromatography combined with an evaporative light scattering detector results allowed us to establish that phosphatidylethanolamine was the most representative phospholipid in all of the oat samples. In addition, high-performance liquid chromatography combined with electrospray ionization mass spectrometry analysis showed that C16:0, C16:1, C18:0, C18:1, C18:2, C18:3, C20:0, and C20:1 were the fatty acids bound to the glycerol backbone. Using first-preparative TLC and later gas chromatography, it was demonstrated that linoleic acid (C18:2) was the main fatty acid of the phospholipid fraction in all of the samples.

The industrial application of nitrocellulose depends on its nitrogen content. When nitrocellulose presents high nitrogen content is used in the manufacture of explosives whereas nitrocellulose with low nitrogen content is used to make a wide range of daily and non-explosive products (e.g. cigarettes, paints, lacquers). This fact makes really important to develop a method for the determination and discrimination of nitrocellulose samples. This work reports, for the first time, the qualitative determination of nitrocellulose previously derivatized with 8-aminopyrene-1,3,6-trisulfonic acid (APTS) by capillary electrophoresis (CE-LIF) with laser-induced fluorescence detection (CE-LIF). APTS-labeled nitrocellulose was determined in lowly and highly nitrated nitrocellulose samples present in collodions and smokeless gunpowders, respectively, after their pulverization in liquid nitrogen. The method described enables the visual discrimination of different nitrocelluloses on the basis of the different electrophoretic profiles obtained, and provides a useful tool to determine nitrocellulose. Additionally, the use of field-amplified sample injection (FASI) enabled enhanced sample detection, which made it possible to determine nitrocellulose contained in ∼15 μg of gunpowder., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Olive proteins are not well known and there are still a lot of unknown information requiring further studies focused on the determination and characterization of these proteins. Despite the widely use of gel electrophoresis, this is the first time that capillary gel electrophoresis (CGE) is applied to separate proteins extracted from olive fruits. Seven common peaks were identified in the twenty olive varieties studied in this work. According to their migration times, these seven peaks could correspond to molecules with molecular masses of 11.0±0.4, 13.9±0.5, 16.3±0.8, 22.1±0.6, 30±1, 48±1, and 53±2 kDa. All of the determined molecular masses could be attributed to proteins and four of them have been previously observed by SDS-PAGE. The electrophoretic profiles were also evaluated for their capability to differentiate olive varieties according to their presumed geographical origin. Results demonstrated that this method could successfully classify the studied olive varieties by its combination with multivariate chemometrics tools., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The ability to unequivocally identify a gunshot residue (GSR) when a firearm is discharged is a very important and crucial part of crime scene investigation. To date, the great majority of the analyses have focused on the inorganic components of GSR, but the introduction of "lead-free" or "nontoxic" ammunitions makes it difficult to prevent false negatives. This study introduces a fast methodology for the organic analysis of GSR using Raman spectroscopy. Six different types of ammunition were fired at short distances into cloth targets, and the Raman spectra produced by the GSR were measured and compared with the spectra from the unfired gunpowder ammunition. The GSR spectrum shows high similarity to the spectrum of the unfired ammunition, allowing the GSR to be traced to the ammunition used. Additionally, other substances that might be found on the victim's, shooter's, or suspect's clothes and might be confused with GSR, such as sand, dried blood, or black ink from a common ballpoint pen, were analyzed to test the screening capability of the Raman technique. The results obtained evidenced that Raman spectroscopy is a useful screening tool when fast analysis is desired and that little sample preparation is required for the analysis of GSR evidence.

After the train bombing in Madrid (Spain) on 11 March 2004, methenamine was detected in some of the specimens of Goma-2 ECO dynamite submitted to the forensic laboratories when analyzed by gas chromatography coupled with mass spectrometry detection (GC-MS). Methenamine is synthesized from formaldehyde and ammonia through a condensation reaction. However, neither methenamine nor any of these compounds were used to manufacture Goma-2 ECO dynamite. Four different experiments were designed in order to explain the presence of methenamine detected in the dynamite samples analyzed. In the first one, GC-MS was used to analyze the individual components of Goma-2 ECO provided by the manufacturer and the components mixed in a raw paste. Methenamine was detected in the manufacturer's ammonium nitrate and in the raw paste. The other experiments were designed to find the precursors sources for methenamine generation in Goma-2 ECO. Results revealed that these sources could be ammonium nitrate for ammonia and sawdust for formaldehyde. Under heating conditions, dynamite could produce these precursors, which could condense in the injection port of the GC-MS system and generate methenamine. However, methenamine was not always detected in these dynamites. This was explained by the existence of two opposite effects: (a) dynamite stability makes difficult that ammonium nitrate releases ammonia and (b) there is a gradual loss of formaldehyde in sawdust along the time. Both effects can prevent the formation of an amount of methenamine large enough to be detected., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Fourier Transform Infrared (FTIR) and Raman spectroscopic techniques were used to perform a comparative study of the spectral profiles of single-base, double-base and triple-base smokeless gunpowders. Preliminary results based on visual comparison of the spectra point out that spectra obtained by both vibrational techniques were useful for a rapid identification of gunpowders containing dinitrotoluene as one of the major components and triple-base gunpowders. Additionally, the Raman spectra of gunpowders with diphenylamine in its primary composition showed a characteristic band, assigned to 2-nitro-diphenylamine, allowing the identification of this type of gunpowders. Further differentiation among the spectra of different types of smokeless gunpowders obtained by both vibrational spectroscopic techniques was investigated by discriminant analysis. Different analyses were applied to spectral data considering the different composition of gunpowders. The presence or absence of different compounds (such as dinitrotoluene, diphenylamine or dibutyl phthalate) or the type of gunpowder according to the number of active components (single-base or double-base gunpowder) has been taken into account. FTIR and Raman spectroscopy in combination with discriminant analyses were successful tools of forensic interest for the classification of gunpowders and the possible identification of unknown samples of gunpowders., (Copyright © 2011 Elsevier B.V. All rights reserved.)