Anthocyanins: Quantification by Tandem Mass Spectrometry.

Mass spectrometry is a high throughput technique widely used for metabolic fingerprinting of plant material. Among the diverse plant metabolites, pigments such as anthocyanins play a determinant role in plant defence mechanisms, protecting them from biotic and abiotic stresses. Anthocyanins are phenolic water-soluble glycosides or acyl-glycosides of anthocyanidins which could be accurately detected and quantified through mass spectrometry. This chapter describes how to extract anthocyanins from higher plant materials and quantify them through a liquid chromatography-mass spectrometry (LC-MS) based method.

Cyanobacteria as Candidates to Support Mars Colonization: Growth and Biofertilization Potential Using Mars Regolith as a Resource.

Cyanobacteria are indicated as organisms that can possibly support Mars colonization, contributing to the production of oxygen and other commodities therein. In this general context, the aim of this work was to evaluate the ability of three species of cyanobacteria (Anabaena cylindrica, Nostoc muscorum, and Arthrospira platensis) and a green microalga (Chlorella vulgaris) to grow using only the resources existing in Mars, i.e., water and Martian regolith stimulant (MGS-1), under an Earth-like atmosphere. A Martian regolith extract was produced and used as a culture medium to grow these species. Their growth was assessed during a period of 25 days, using optical density and fluorometric parameters. After this period, the possible contribution of end-of-life cyanobacteria/microalga as biofertilizing agents was also assessed, using the macrophyte Lemna minor as a vegetable model. Among the three species, N. muscorum showed the best growth performance when compared to the other species, while A. platensis and C. vulgaris were not able to thrive on Mars regolith extract. Therefore, N. muscorum should be the target of future studies not only due to their role in oxygen production but also due to their possible use as a food source, as many members of the Nostoc genus. Cyanobacteria and microalgae (A. platensis and C. vulgaris) showed good abilities as biofertilizing agents, i.e., they stimulated biomass (i.e., dry weight) production at levels comparable to the plants that grew on standard synthetic medium. The highest yield was reached with A. platensis, while the lowest was achieved using the media with N. muscorum. FTIR-ATR (Fourier transform infrared with attenuated total reflectance) spectroscopy showed that the differences between the plants grown on media with or without Martian regolith seem to be related mainly to polysaccharides.

Anthocyanins, Carotenoids and Chlorophylls in Edible Plant Leaves Unveiled by Tandem Mass Spectrometry.

Natural pigments are a quite relevant group of molecules that are widely distributed in nature, possessing a significant role in our daily lives. Besides their colors, natural pigments are currently recognized as having relevant biological properties associated with health benefits, such as anti-tumor, anti-atherogenicity, anti-aging and anti-inflammatory activities, among others. Some of these compounds are easily associated with specific fruits (such as blueberries with anthocyanins, red pitaya with betalain or tomato with lycopene), vegetables (carrots with carotenoids), plant leaves (chlorophylls in green leaves or carotenoids in yellow and red autumn leaves) and even the muscle tissue of vertebrates (such as myoglobin). Despite being less popular as natural pigment sources, edible plant leaves possess a high variety of chlorophylls, as well as a high variety of carotenoids and anthocyanins. The purpose of this review is to critically analyze the whole workflow employed to identify and quantify the most common natural pigments (anthocyanin, carotenoids and chlorophylls) in edible plant leaves using tandem mass spectrometry. Across the literature there, is a lack of consistency in the methods used to extract and analyze these compounds, and this review aims to surpass this issue. Additionally, mass spectrometry has stood out in the context of metabolomics, currently being a widely employed technique in this field. For the three pigments classes, the following steps will be scrutinized: (i) sample pre-preparation, including the solvents and extraction conditions; (ii) details of the chromatographic separation and mass spectrometry experiments (iii) pigment identification and quantification.

Presbylarynx: How Easy Is It To Recognize The Aging Signs? A Prospective Study With Inter- And Intra-Rater Reliability In Laryngoscopic Evaluation.

PURPOSE: To determine if recognition of presbylarynx is easy and straightforward. METHOD: First, a systematic review regarding the structural features of the geriatric larynx through laryngostroboscopy was performed, and its results were presented to the raters in an educational session. Then, video recordings of rigid laryngostroboscopies were randomly selected and presented to a panel of otorhinolaryngologists. Intra- and inter-rater reliability were determined. RESULTS: Based on the main findings of the systematic review, raters were invited to consider the diagnosis of presbylarynx when one or more of the following endoscopic signs were present: atrophic vocal folds (VF), VF bowing, prominence of vocal processes, or spindle-shaped glottal gap. Twenty otorhinolaryngologists from five tertiary hospitals participated in this study (residents and specialists). Among 300 evaluations performed (15 video recordings x 20 raters), the intra-rater agreement was 93.0%. The Cohen´s Kappa for intra-rater reliability was higher than 0.600 for all the raters except for two residents. Cohen´s Kappa was higher among specialists (0.893) than among residents (0.826). The highest Cohen´s Kappa was registered for evaluations performed by specialists with differentiation in laryngology (0.933). Presbylarynx was identified mostly in laryngostroboscopic recordings of subjects older than 65 years. CONCLUSION: we consider the answer to "Presbylarynx: How easy is it to recognize the aging signs" to be "After education on signs of presbylarynx, it is easy." This answer is supported by the high rate of intra- and inter-rater agreement, in a blinded and randomized presentation of laryngostroboscopic recordings, and by the fact that the diagnosis of presbylarynx was mostly attributed to videos regarding subjects aged 65 years and older. It proves that is not difficult to recognize aging signs in the larynx. Thus, in the future, more education regarding presbylarynx and further studies to improve criteria for its diagnosis may improve the management of the aging voice.

Protein discrimination using erythrosin B-based GUMBOS in combination with UV-Vis spectroscopy and chemometrics.

GUMBOS (Group of Uniform Materials Based on Organic Salts) have recently emerged as interesting materials for protein analysis due to their unique features and high tunability. In this regard, four novel erythrosin B (EB)-based GUMBOS were synthesized and their potential to discriminate among proteins with distinct properties (e.g., size, charge, and hydrophobicity) was assessed. These solid-phase materials were prepared using a single-step metathesis reaction between EB and various phosphonium and ammonium cations, namely tetrabutylphosphonium (P4444+), tributylhexadecylphosphonium (P44416+), tetrabutylammonium (N4444+), and benzyldimethylhexadecylammonium (BDHA+). Subsequently, the effect of pH (3.0, 4.5, and 6.0) and reaction time (5, 10, and 15 min) on the discriminatory power of synthesized GUMBOS was evaluated. Absorption spectra resulting from the interaction between EB-based GUMBOS and proteins were analyzed using partial least squares discriminant analysis (PLSDA). Unlike time, the pH value was determined to have influence over GUMBOS discrimination potential. Correct protein assignments varied from 86.5% to 100.0%, and the best discriminatory results were observed for [P4444]2[EB] and [N4444]2[EB] at pH 6.0. Additionally, these two GUMBOS allowed discrimination of protein mixtures containing different ratios of albumin and myoglobin, which appeared as individualized clusters in the PLSDA scores plots. Overall, this study showcases EB-based GUMBOS as simple synthetic targets to provide a label-free, cost-effective, rapid, and successful approach for discrimination of single proteins and their mixtures.

A Review on the Biological Activity of Camellia Species.

Medicinal plants have been used since antiquity to cure illnesses and injuries. In the last few decades, natural compounds extracted from plants have garnered the attention of scientists and the Camellia species are no exception. Several species and cultivars are widespread in Asia, namely in China, Japan, Vietnam and India, being also identified in western countries like Portugal. Tea and oil are the most valuable and appreciated Camellia subproducts extracted from Camellia sinensis and Camellia oleifera, respectively. The economic impact of these species has boosted the search for additional information about the Camellia genus. Many studies can be found in the literature reporting the health benefits of several Camellia species, namely C. sinensis, C. oleifera and Camellia japonica. These species have been highlighted as possessing antimicrobial (antibacterial, antifungal, antiviral) and antitumoral activity and as being a huge source of polyphenols such as the catechins. Particularly, epicatechin (EC), epigallocatechin (EGC), epicatechin-3-gallate (ECG), and specially epigallocatechin-3-gallate (EGCG), the major polyphenols of green tea. This paper presents a detailed review of Camellia species' antioxidant properties and biological activity.

Teste rápido para o HIV durante o pré-natal / Rapid test for HIV during prenatal care / Prueba rápida del VIH durante el prenatal

Objetivo: verificar, na literatura, as características das consultas de pré-natal às gestantes para oferecimento do teste rápido para o Vírus da Imunodeficiência Humana. Método: trata-se de um estudo bibliográfico, descritivo, tipo revisão integrativa da literatura, sem limite temporal, realizado por meio das bases de dados BDENF, LILACS, MEDLINE, CINAHL, Scopus e Web of Science. Incluíram-se artigos originais em português, inglês ou espanhol. Analisaram-se os dados de forma descritiva. Resultados: identificaram-se 1726 publicações, das quais dez compuseram a amostra. Verificaram-se, em geral, as características do estudo e as estratégias para a melhoria das consultas, do aconselhamento e do oferecimento de testes efetivos. Conclusão: conclui-se que a análise dos estudos permitiu a síntese de características, conteúdos, recursos e técnicas de abordagem, além de lacunas e dificuldades tanto dos profissionais quanto das gestantes para a efetivação do aconselhamento para a execução dos testes rápidos para o Vírus da Imunodeficiência Humana em gestantes.(AU)

Objective: to verify, in the literature, the characteristics of prenatal consultations to pregnant women to offer the rapid test for the Human Immunodeficiency Virus. Method: It is a bibliographic, descriptive, integrative literature review type study, without time limit, carried out through the BDENF, LILACS, MEDLINE, CINAHL, Scopus and Web of Science databases. Original articles in Portuguese, English or Spanish are included. The data were analyzed in a descriptive manner. Results: 1726 publications were identified, of which ten composed the sample. In general, the characteristics of the study and the strategies for the improvement of consultations, counseling and offer of effective tests were verified. Conclusion: it is concluded that the analysis of the studies allowed the synthesis of characteristics, contents, resources and techniques of approach, besides gaps and difficulties of both professionals and pregnant women for the realization of counseling for the execution of rapid tests for the Human Immunodeficiency Virus in pregnant women.

Objetivo: verificar, en la literatura, las características de las consultas prenatales a gestantes para ofrecer la prueba rápida del Virus de Inmunodeficiencia Humana. Método: se trata de una revisión bibliográfica, descriptiva, integradora de la literatura, sin límite de tiempo, realizada utilizando las bases de datos BDENF, LILACS, MEDLINE, CINAHL, Scopus y Web of Science. Se incluyeron artículos originales en portugués, inglés o español. Los datos se analizaron de forma descriptiva. Resultados: se identificaron 1726 publicaciones, de las cuales diez componen la muestra. En general, se verificaron las características del estudio y las estrategias para mejorar las consultas, el asesoramiento y la oferta de pruebas efectivas. Conclusión: se concluye que el análisis de los estudios permitió la síntesis de características, contenidos, recursos y técnicas de abordaje, además de vacíos y dificultades tanto de los profesionales como de las gestantes para la efectividad del asesoramiento para la ejecución de las pruebas rápidas para el Virus de la Inmunodeficiencia Humana en mujeres embarazadas.(AU)

Near-Infrared Spectroscopy Applied to the Detection of Multiple Adulterants in Roasted and Ground Arabica Coffee.

Roasted coffee has been the target of increasingly complex adulterations. Sensitive, non-destructive, rapid and multicomponent techniques for their detection are sought after. This work proposes the detection of several common adulterants (corn, barley, soybean, rice, coffee husks and robusta coffee) in roasted ground arabica coffee (from different geographic regions), combining near-infrared (NIR) spectroscopy and chemometrics (Principal Component Analysis-PCA). Adulterated samples were composed of one to six adulterants, ranging from 0.25 to 80% (w/w). The results showed that NIR spectroscopy was able to discriminate pure arabica coffee samples from adulterated ones (for all the concentrations tested), including robusta coffees or coffee husks, and independently of being single or multiple adulterations. The identification of the adulterant in the sample was only feasible for single or double adulterations and in concentrations ≥10%. NIR spectroscopy also showed potential for the geographical discrimination of arabica coffees (South and Central America).

A Front Line on Klebsiella pneumoniae Capsular Polysaccharide Knowledge: Fourier Transform Infrared Spectroscopy as an Accurate and Fast Typing Tool.

Genomics-based population analysis of multidrug-resistant (MDR) Klebsiella pneumoniae motivated a renewed interest on the capsule as an evolutionary and virulence marker of clinically relevant strains. Whole-genome sequencing (WGS)-based approaches have provided great insights into the genetic variability of the capsular locus, but genotypic-biochemical capsular (K)-type correlations are lacking, hindering the establishment of a reliable framework for K-type characterization and typing. To fill this gap, we combined molecular, comparative genomics, and multivariate data analysis tools with biochemical data on the capsular locus to support the usefulness of Fourier transform infrared (FT-IR) spectroscopy as a reliable K typing tool. To validate our approach, we used a representative collection of well-defined MDR K. pneumoniae lineages involved in local or nationwide epidemics in multiple countries. With this, we demonstrate a high accuracy and resolution of our FT-IR-based spectroscopy approach for K-type discrimination that is even higher than that provided by WGS. Moreover, the specific associations established between certain K types and specific K. pneumoniae lineages with high clinical relevance, together with the accuracy, simplicity, short time to result, and inexpensive features of the method, support the value of the developed FT-IR-based approach for an easy, fast, and cost-effective strain typing. This fulfills a still unmet need for tools to support real-time monitoring and control of K. pneumoniae infections. In addition, the genotypic-biochemical correlations established provide insights on sugar composition/structure of newly defined K. pneumoniae capsular types.IMPORTANCE Klebsiella pneumoniae is nowadays recognized as one of the most defiant human pathogens, whose infections are increasingly more challenging to treat and control. Whole-genome sequencing (WGS) has been key for clarifying the population structure of K. pneumoniae, and it is still instrumental to provide insights into potential pathogenicity and evolutionary markers, such as the capsular locus. However, this information and WGS are still far from being accessible and translated into routine clinical microbiology laboratories as quick and cost-efficient strain diagnostic tools. Here, we propose a biochemical fingerprinting approach based on Fourier transform infrared spectroscopy (FT-IR) and multivariate data analysis tools for K. pneumoniae capsular typing that, because of its high resolution, speed, and low cost, can be an asset to provide enough information to support real-time epidemiology and infection control decisions. Besides, it provides a simple framework for phenotypic/biochemical validation of K. pneumoniae capsular diversity.

Author Correction: Escherichia coli and Salmonella Enteritidis dual-species biofilms: interspecies interactions and antibiofilm efficacy of phages.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Escherichia coli and Salmonella Enteritidis dual-species biofilms: interspecies interactions and antibiofilm efficacy of phages.

Escherichia coli and Salmonella Enteritidis are foodborne pathogens forming challenging biofilms that contribute to their virulence, antimicrobial resistance, and survival on surfaces. Interspecies interactions occur between species in mixed biofilms promoting different outcomes to each species. Here we describe the interactions between E. coli and S. Enteritidis strains, and their control using specific phages. Single-species biofilms presented more cells compared to dual-species biofilms. The spatial organization of strains, observed by confocal microscopy, revealed similar arrangements in both single- and dual-species biofilms. The EPS matrix composition, assessed by Fourier-transform infrared spectroscopy, disclosed that the spectra extracted from the different dual-species biofilms can either be a combination of both species EPS matrix components or that the EPS matrix of one species predominates. Phages damaged more the single-species biofilms than the mixed biofilms, showing also that the killing efficacy was greatly dependent on the phage growth characteristics, bacterial growth parameters, and bacterial spatial distribution in biofilms. This combination of methodologies provides new knowledge of species-species and phage-host interactions in biofilms of these two major foodborne pathogens.

Antioxidant Activity of Blueberry (Vaccinium spp.) Cultivar Leaves: Differences Across the Vegetative Stage and the Application of Near Infrared Spectroscopy.

Blueberries production has increased in the last few years boosted somehow by the World Health Organization (WHO) recommendations for a healthier nutrition and their recognized potential to treat several diseases. The production increase lead to high amounts of discarded leaves that could be very valuable. In this context, the antioxidant activity of Vaccinium spp. leaves, by means of the total phenolic (TPC) and flavonoid (TFC) content and total antioxidant capacity (TAC) was determined. Adult leaves of twenty-seven Vaccinium cultivars collected in three geographic regions and three seasons of the year were included. The antioxidant activity was additionally estimated with near infrared (NIR) spectroscopy and data transferability across the regions and seasons was evaluated. The TPC, TFC and TAC ranged from 39.6-272.8 mg gallic acid, 41.2-269.1 mg catechin and 22.6-124.8 mM Trolox per g of dry leaf, respectively. Globally through the seasons, the higher values of the three parameters were obtained in December. Regarding the geographic region, region A provided the cultivars with the higher antioxidant content. Titan was the cultivar with higher TPC and TAC and Misty the one with the higher TFC. NIR spectroscopy combined with the partial least squares analysis was able to successfully predict the antioxidant activity with coefficients of determination and range error ratios ranging from 0.84-0.99 and 11.2-26.8. Despite some identified limitations on data transferability, NIR spectroscopy proved to be a reliable, low cost and quick method to predict the antioxidant activity of the considered cultivar leaves.

Differentiation of Taxonomically Closely Related Species of the Genus Acinetobacter Using Raman Spectroscopy and Chemometrics.

In recent years, several efforts have been made to develop quick and low cost bacterial identification methods. Genotypic methods, despite their accuracy, are laborious and time consuming, leaving spectroscopic methods as a potential alternative. Mass and infrared spectroscopy are among the most reconnoitered techniques for this purpose, with Raman having been practically unexplored. Some species of the bacterial genus Acinetobacter are recognized as etiological agents of nosocomial infections associated with high rates of mortality and morbidity, which makes their accurate identification important. The goal of this study was to assess the ability of Raman spectroscopy to discriminate between 16 Acinetobacter species belonging to two phylogroups containing taxonomically closely related species, that is, the Acinetobacter baumannii-Acinetobacter calcoaceticus complex (six species) and haemolytic clade (10 species). Bacterial spectra were acquired without the need for any sample pre-treatment and were further analyzed with multivariate data analysis, namely partial least squares discriminant analysis (PLSDA). Species discrimination was achieved through a series of sequential PLSDA models, with the percentage of correct species assignments ranging from 72.1% to 98.7%. The obtained results suggest that Raman spectroscopy is a promising alternative for identification of Acinetobacter species.

Antioxidant capacity of Camellia japonica cultivars assessed by near- and mid-infrared spectroscopy.

MAIN CONCLUSION: Camellia japonica antioxidant capacity highly differs among its cultivars and could be successfully predicted by near- and mid-infrared spectroscopy. Camellia japonica is a Theaceae family species which are mainly used as an ornamental plant due to its colourful flowers presenting over than 32,000 recognized cultivars. However, this species have been somehow neglected due to the popular tea source, Camellia sinensis. In this study, the antioxidant profile (total phenolic and flavonoid content and total antioxidant capacity-TPC, TFC and TAC) of 31 C. japonica cultivars leaves was determined and further assessed by near- and mid-infrared spectroscopy. The leaves' antioxidant profile was revealed to be highly dependent on the cultivars analysed being in some cases distinct even for different trees of the same cultivar. Near- and mid-infrared spectroscopy proved to be suitable techniques to predict the total phenolic and flavonoid content as well as the total antioxidant capacity. The best results were obtained with near-infrared spectroscopy whose root mean square error of the prediction set samples was of 5.7 mg of gallic acid/g dry leaf; 3.5 mg catechin/g dry leaf and 3.3 mM Trolox/g dry leaf for TPC, TFC and TAC (with coefficients of the determinations equal to or higher than 0.93). Moreover, the range error ratios were higher than 15 meaning that the developed partial least-squares models are very good for calibration and quantification determinations according to the guidelines for near-infrared models development and maintenance. In this work, the antioxidant profile of several C. japonica cultivars leaves was determined for the first time, being that a rapid and low cost spectroscopic-based method was also proposed for its determination.

A review on the application of vibrational spectroscopy to the chemistry of nuts.

Nuts are highly appreciated due to their nutritional relevance and flavour, being the source of many desirable and healthy compounds as polyunsaturated fatty acids and antioxidants. Their characterization became the target of many studies in the last years through conventional analytical techniques as chromatographic ones. Due to the limitations associated to these techniques, as time, cost and environmental concerns, spectroscopic techniques have been increasingly pointed as reliable alternatives. Either applied to raw materials quality control or to more complex process, as industrial in-line monitoring, spectroscopic techniques, namely vibrational spectroscopy, are gathering strong acceptance. This paper presents a review on the application of vibrational spectroscopy, infrared and Raman, to nuts characterization. Estimates of several qualitative and quantitative parameters, origin authentication and/or adulteration in almonds, peanuts, pine nuts, hazelnuts, walnuts, Brazil nuts, cashews, chestnuts and pistachios will be covered. Advantages and limitations of these techniques and future trends will also be discussed.

Discrimination of non-typhoid Salmonella serogroups and serotypes by Fourier Transform Infrared Spectroscopy: A comprehensive analysis.

Simpler, quick and low-cost methods for routine Salmonella enterica typing are required for epidemiologic surveillance of this important zoonotic pathogen. In this study, using a comprehensive isolate collection, we investigated the potential of Fourier transform infrared spectroscopy (FTIRS) to discriminate the most clinically-relevant serogroups and serotypes of non-typhoid Salmonella. Moreover, the role of O-units composition on the FTIRS Salmonella discrimination was also explored. S. enterica isolates (n = 325; 2002-2015; different sources and countries), of 57 serotypes and 15 serogroups [including the most frequent ones, B-n = 122; C-n = 108; D-n = 43 and E-n = 33)] were analysed by FTIRS. Infrared spectra were analysed by Partial Least Square Discriminant Analysis (PLSDA) and/or Principal Component Analysis (PCA). The polysaccharides region provided the spectral sharpest differences being used in the subsequent Salmonella typing. Serogroups (B, C, D and E) discrimination was achieved with high accuracy (99.6% of correct assignments; PLSDA model). Differences in the O-unit structures composition of those serogroups are likely justifying the discrimination achieved. Other serogroups (G, H, K, L, M, N, O, T, U, Y, Z) were correctly predicted as not belonging to serogroups B, C, D nor E, except for 3 isolates of serogroups H (S. Sundsvall, n = 1) and K (S. Cerro, n = 2). In fact, O-unit structure of serogroup H and K shows some similarity with sub-serogroup C1 with the remaining serogroups presenting marked differences in this cellular component. The sub-serogroups discrimination was successfully achieved for C1, C2 and C3 (using PCA), and for E1-E2-E3 and E4 (by PLSDA). Appropriate serotype discrimination was obtained for most of S. Rissen from the remaining C1 serotypes (91.5%-PLSDA), and S. Enteritidis (D1) from the remaining D1/D2 serotypes (93.4%-PLSDA). The lack of available O-unit composition for particular serotypes prevents the elucidation of the role of this cellular component on the discrimination at serotype level obtained. FTIRS was able to discriminate relevant serogroups (B, C, D and E), sub-serogroups (C1, C2 and C3; E1-E2-E3 and E4) and particular important serotypes (S. Enteritidis, S. Rissen and S. Senftenberg). Further studies on O-antigen composition would clarify the fundaments of discrimination obtained by FTIRS.

Citrus species and hybrids depicted by near- and mid-infrared spectroscopy.

BACKGROUND: Citrus trees are among the most cultivated plants in the world, with a high economic impact. The wide sexual compatibility among relatives gave rise to a large number of hybrids that are difficult to discriminate. This work sought to explore the ability of infrared spectroscopy to discriminate among Citrus species and/or hybrids and to contribute to the elucidation of its relatedness. RESULTS: Adult leaves of 18 distinct Citrus plants were included in this work. Near- and mid-infrared (NIR and FTIR) spectra were acquired from leaves after harvesting and a drying period of 1 month. Spectra were modelled by principal component analysis and partial least squares discriminant analysis. Both techniques revealed a high discrimination potential (78.5-95.9%), being the best results achieved with NIR spectroscopy and air-dried leaves (95.9%). CONCLUSION: Infrared spectroscopy was able to successfully discriminate several Citrus species and/or hybrids. Our results contributed also to enhance insights regarding the studied Citrus species and/or hybrids. Despite the benefit of including additional samples, the results herein obtained clearly pointed infrared spectroscopy as a reliable technique for Citrus species and/or hybrid discrimination. © 2018 Society of Chemical Industry.

An Overview of the Evolution of Infrared Spectroscopy Applied to Bacterial Typing.

The sustained emergence of new declared bacterial species makes typing a continuous challenge for microbiologists. Molecular biology techniques have a very significant role in the context of bacterial typing, but they are often very laborious, time consuming, and eventually fail when dealing with very closely related species. Spectroscopic-based techniques appear in some situations as a viable alternative to molecular methods with advantages in terms of analysis time and cost. Infrared and mass spectrometry are among the most exploited techniques in this context: particularly, infrared spectroscopy emerged as a very promising method with multiple reported successful applications. This article presents a systematic review on infrared spectroscopy applications for bacterial typing, highlighting fundamental aspects of infrared spectroscopy, a detailed literature review (covering different taxonomic levels and bacterial species), advantages, and limitations of the technique over molecular biology methods and a comparison with other competing spectroscopic techniques such as MALDI-TOF MS, Raman, and intrinsic fluorescence. Infrared spectroscopy possesses a high potential for bacterial typing at distinct taxonomic levels and worthy of further developments and systematization. The development of databases appears fundamental toward the establishment of infrared spectroscopy as a viable method for bacterial typing.

Experimental and Computational Thermochemical Study of Maleic Anhydride and Vinylene Carbonate.

The standard molar enthalpies of formation of maleic anhydride and vinylene carbonate in gaseous phase, at T = 298.15 K, were derived from the standard molar enthalpies of formation of the compounds in condensed phase combined with the phase transition enthalpies. The standard molar enthalpies of formation in condensed phase were obtained from the enthalpies of combustion measured using static bomb combustion calorimetry and mini-bomb combustion calorimetry for vinylene carbonate and maleic anhydride, respectively. Phase transition enthalpies were obtained by Calvet microcalorimetry. High level quantum calculations were performed at the composite G3 level of theory in order to estimate the standard molar enthalpies of formation of both compounds in gaseous phase. Good agreement was obtained between experimental and computational results. In addition, analysis of the factors affecting the relative stability of both systems has been carried out in the framework of the ab initio valence bond (VB) theory in order to clarify the aromaticity/antiaromaticity issues involving these molecular systems.

Rapid detection of high-risk Enterococcus faecium clones by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

We aimed to explore the potential of matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for early identification of dominant Enterococcus faecium (Efm) clones involved in human infections. Well-characterized Efm isolates (n=77), analyzed by pulsed-field gel electrophoresis and multilocus sequence typing(eBURST and BAPS [Bayesian analysis of population structure] algorithms), and belonging to different hospital (n=53) and community (n=24) phylogenomic groups, were tested. Mass spectra (Bruker) were analyzed by visual inspection and different chemometric tools. Discrimination between groups comprising isolates commonly found in hospitals (BAPS 2.1a, 3.3a1, 3.3a2) and community (BAPS 2.1b and 3.2) was achieved with >99% accuracy, while identification of sequence types belonging to different BAPS subgroups was associated with >95% correct predictions. Our work is a proof of concept with regard to the suitability of MALDI-TOF MS in the identification of high-risk Efm clones. Further studies including strains from a wider variety of clones and sources will strengthen the potential of the workflow here described.