Selected configuration interaction with truncation energy error and application to the Ne atom

Selected configuration interaction (SCI) for atomic and molecular electronic structure calculations is reformulated in a general framework encompassing all CI methods. The linked cluster expansion is used as an intermediate device to approximate CI coefficients BK of disconnected configurations (those that can be expressed as products of combinations of singly and doubly excited ones) in terms of CI coefficients of lower-excited configurations where each K is a linear combination of configuration-state-functions (CSFs) over all degenerate elements of K. Disconnected configurations up to sextuply excited ones are selected by Brown's energy formula, ΔEK=(E-HKK)BK2/(1-BK2), with BK determined from coefficients of singly and doubly excited configurations. The truncation energy error from disconnected configurations, Δdis, is approximated by the sum of ΔEKS of all discarded Ks. The remaining (connected) configurations are selected by thresholds based on natural orbital concepts. Given a model CI space M, a usual upper bound ES is computed by CI in a selected space S, and EM=E S+ΔEdis+δE, where δE is a residual error which can be calculated by well-defined sensitivity analyses. An SCI calculation on Ne ground state featuring 1077 orbitals is presented. Convergence to within near spectroscopic accuracy (0.5 cm-1) is achieved in a model space M of 1.4× 109 CSFs (1.1 × 1012 determinants) containing up to quadruply excited CSFs. Accurate energy contributions of quintuples and sextuples in a model space of 6.5 × 1012 CSFs are obtained. The impact of SCI on various orbital methods is discussed. Since ΔEdis can readily be calculated for very large basis sets without the need of a CI calculation, it can be used to estimate the orbital basis incompleteness error. A method for precise and efficient evaluation of ES is taken up in a companion paper

Basis set superposition error-counterpoise corrected potential energy surfaces : application to hydrogen peroxide ... X (X=F-, Cl-, Br-, Li+, Na+) complexes

Møller-Plesset (MP2) and Becke-3-Lee-Yang-Parr (B3LYP) calculations have been used to compare the geometrical parameters, hydrogen-bonding properties, vibrational frequencies and relative energies for several X- and X+ hydrogen peroxide complexes. The geometries and interaction energies were corrected for the basis set superposition error (BSSE) in all the complexes (1-5), using the full counterpoise method, yielding small BSSE values for the 6-311 + G(3df,2p) basis set used. The interaction energies calculated ranged from medium to strong hydrogen-bonding systems (1-3) and strong electrostatic interactions (4 and 5). The molecular interactions have been characterized using the atoms in molecules theory (AIM), and by the analysis of the vibrational frequencies. The minima on the BSSE-counterpoise corrected potential-energy surface (PES) have been determined as described by S. Simón, M. Duran, and J. J. Dannenberg, and the results were compared with the uncorrected PES

Automatic generation of active coordinates for quantum dynamics calculations: application to the dynamics of benzene photochemistry

A new practical method to generate a subspace of active coordinates for quantum dynamics calculations is presented. These reduced coordinates are obtained as the normal modes of an analytical quadratic representation of the energy difference between excited and ground states within the complete active space self-consistent field method. At the Franck-Condon point, the largest negative eigenvalues of this Hessian correspond to the photoactive modes: those that reduce the energy difference and lead to the conical intersection; eigenvalues close to 0 correspond to bath modes, while modes with large positive eigenvalues are photoinactive vibrations, which increase the energy difference. The efficacy of quantum dynamics run in the subspace of the photoactive modes is illustrated with the photochemistry of benzene, where theoretical simulations are designed to assist optimal control experiments

Mophological analysis of the human vasculature from medical imaging. Application to cerebral and coranary arteries

In this project, we have investigated new ways of modelling and analysis of human vasculature from Medical images. The research was divided in two main areas: cerebral vasculature analysis and coronary arteries modeling. Regarding cerebral vasculature analysis, we have studed cerebral aneurysms, internal carotid and the Circle of Willis (CoW). Aneurysms are abnormal vessel enlargements that can rupture causing important cerebral damages or death. The understanding of this pathology, together with its virtual treatment, and image diagnosis and prognosis, includes identification and detailed measurement of the aneurysms. In this context, we have proposed two automatic aneurysm isolation method, to separate the abnormal part of the vessel from the healthy part, to homogenize and speed-up the processing pipeline usually employed to study this pathology, [Cardenes2011TMI, arrabide2011MedPhys]. The results obtained from both methods have been also compared and validatied in [Cardenes2012MBEC]. A second important task here the analysis of the internal carotid [Bogunovic2011Media] and the automatic labelling of the CoW, Bogunovic2011MICCAI, Bogunovic2012TMI]. The second area of research covers the study of coronary arteries, specially coronary bifurcations because there is where the formation of atherosclerotic plaque is more common, and where the intervention is more challenging. Therefore, we proposed a novel modelling method from Computed Tomography Angiography (CTA) images, combined with Conventional Coronary Angiography (CCA), to obtain realistic vascular models of coronary bifurcations, presented in [Cardenes2011MICCAI], and fully validated including phantom experiments in [Cardene2013MedPhys]. The realistic models obtained from this method are being used to simulate stenting procedures, and to investigate the hemodynamic variables in coronary bifurcations in the works submitted in [Morlachi2012, Chiastra2012]. Additionally, another preliminary work has been done to reconstruct the coronary tree from rotational angiography, and published in [Cardenes2012ISBI].

The Fry method applied to an augen orthogneiss: Problems and results

The application of the Fry method to measure strain in deformed porphyritic granites is discussed. This method requires that the distribution of markers has to satisfy at least two conditions. It has to be homogeneous and isotropic. Statistics on point distribution with the help of a Morishita diagram can easily test homogeneity. Isotropy can be checked with a cumulative histogram of angles between points. Application of these tests to undeformed (Mte Capanne granite, Elba) and to deformed (Randa orthogneiss, Alps of Switzerland) porphyritic granite reveals that their K-feldspars phenocrysts both satisfy these conditions and can be used as strain markers with the Fry method. Other problems are also examined. One is the possible distribution of deformation on discrete shear-bands. Providing several tests are met, we conclude that the Fry method can be used to estimate strain in deformed porphyritic granites. (c) 2006 Elsevier Ltd. All rights reserved.

Development of a Method to Determine Pavement Damage Due to Detours, Project 02-105, Apirl 2005

The research presented in this report provides the basis for the development of a new procedure to be used by the Iowa DOT and cities and counties in the state to deal with detours. Even though the project initially focused on investigating new tools to determine condition and compensation, the focus was shifted to traffic and the gas tax method to set the basis for the new procedure. It was concluded that the condition-based approach, even though accurate and consistent condition evaluations can be achieved, is not feasible or cost effective because of the current practices of data collection (two-year cycle) and also the logistics of the procedure (before and after determination). The gas tax method provides for a simple, easy to implement, and consistent approach to dealing with compensation for use of detours. It removes the subjectivity out of the current procedures and provides for a more realistic (traffic based) approach to the compensation determination.

Body fluid and tissue analysis using filter paper sampling support prior to LC-MS/MS: application to fatal overdose with colchicine

Because of the various matrices available for forensic investigations, the development of versatile analytical approaches allowing the simultaneous determination of drugs is challenging. The aim of this work was to assess a liquid chromatography-tandem mass spectrometry (LC-MS/MS) platform allowing the rapid quantification of colchicine in body fluids and tissues collected in the context of a fatal overdose. For this purpose, filter paper was used as a sampling support and was associated with an automated 96-well plate extraction performed by the LC autosampler itself. The developed method features a 7-min total run time including automated filter paper extraction (2 min) and chromatographic separation (5 min). The sample preparation was reduced to a minimum regardless of the matrix analyzed. This platform was fully validated for dried blood spots (DBS) in the toxic concentration range of colchicine. The DBS calibration curve was applied successfully to quantification in all other matrices (body fluids and tissues) except for bile, where an excessive matrix effect was found. The distribution of colchicine for a fatal overdose case was reported as follows: peripheral blood, 29 ng/ml; urine, 94 ng/ml; vitreous humour and cerebrospinal fluid, < 5 ng/ml; pericardial fluid, 14 ng/ml; brain, < 5 pg/mg; heart, 121 pg/mg; kidney, 245 pg/mg; and liver, 143 pg/mg. Although filter paper is usually employed for DBS, we report here the extension of this alternative sampling support to the analysis of other body fluids and tissues. The developed platform represents a rapid and versatile approach for drug determination in multiple forensic media.

Nursing Activities Score: an updated guideline for its application in the Intensive Care Unit

ABSTRACT Objective To describe nursing workload in Intensive Care Units (ICU) in different countries according to the scores obtained with Nursing Activities Score (NAS) and to verify the agreement among countries on the NAS guideline interpretation. Method This cross-sectional study considered 1-day measure of NAS (November 2012) obtained from 758 patients in 19 ICUs of seven countries (Norway, the Netherlands, Spain, Poland, Egypt, Greece and Brazil). The Delphi technique was used in expertise meetings and consensus. Results The NAS score was 72.8% in average, ranging from 44.5% (Spain) to 101.8% (Norway). The mean NAS score from Poland, Greece and Egypt was 83.0%, 64.6% and 57.1%, respectively. The NAS score was similar in Brazil (54.0%) and in the Netherlands (51.0%). There were doubts in the understanding of five out 23 items of the NAS (21.7%) which were discussed until researchers’ consensus. Conclusion NAS score were different in the seven countries. Future studies must verify if the fine standardization of the guideline can have a impact on differences in the NAS results.

A subsampling approach to estimating the distribution of diversing statistics with application to assessing financial market risks

In this paper we propose a subsampling estimator for the distribution ofstatistics diverging at either known rates when the underlying timeseries in strictly stationary abd strong mixing. Based on our results weprovide a detailed discussion how to estimate extreme order statisticswith dependent data and present two applications to assessing financialmarket risk. Our method performs well in estimating Value at Risk andprovides a superior alternative to Hill's estimator in operationalizingSafety First portofolio selection.

A pseudo-spectral method for the simulation of poro-elastic seismic wave propagation in 2D polar coordinates using domain decomposition

We present a novel numerical approach for the comprehensive, flexible, and accurate simulation of poro-elastic wave propagation in 2D polar coordinates. An important application of this method and its extensions will be the modeling of complex seismic wave phenomena in fluid-filled boreholes, which represents a major, and as of yet largely unresolved, computational problem in exploration geophysics. In view of this, we consider a numerical mesh, which can be arbitrarily heterogeneous, consisting of two or more concentric rings representing the fluid in the center and the surrounding porous medium. The spatial discretization is based on a Chebyshev expansion in the radial direction and a Fourier expansion in the azimuthal direction and a Runge-Kutta integration scheme for the time evolution. A domain decomposition method is used to match the fluid-solid boundary conditions based on the method of characteristics. This multi-domain approach allows for significant reductions of the number of grid points in the azimuthal direction for the inner grid domain and thus for corresponding increases of the time step and enhancements of computational efficiency. The viability and accuracy of the proposed method has been rigorously tested and verified through comparisons with analytical solutions as well as with the results obtained with a corresponding, previously published, and independently bench-marked solution for 2D Cartesian coordinates. Finally, the proposed numerical solution also satisfies the reciprocity theorem, which indicates that the inherent singularity associated with the origin of the polar coordinate system is adequately handled.

Improved estimation of the covariance matrix of stock returns with an application to portofolio selection

This paper proposes to estimate the covariance matrix of stock returnsby an optimally weighted average of two existing estimators: the samplecovariance matrix and single-index covariance matrix. This method isgenerally known as shrinkage, and it is standard in decision theory andin empirical Bayesian statistics. Our shrinkage estimator can be seenas a way to account for extra-market covariance without having to specifyan arbitrary multi-factor structure. For NYSE and AMEX stock returns from1972 to 1995, it can be used to select portfolios with significantly lowerout-of-sample variance than a set of existing estimators, includingmulti-factor models.

Flexible multivariate GARCH modeling with an application to international stock markets

The goal of this paper is to estimate time-varying covariance matrices.Since the covariance matrix of financial returns is known to changethrough time and is an essential ingredient in risk measurement, portfolioselection, and tests of asset pricing models, this is a very importantproblem in practice. Our model of choice is the Diagonal-Vech version ofthe Multivariate GARCH(1,1) model. The problem is that the estimation ofthe general Diagonal-Vech model model is numerically infeasible indimensions higher than 5. The common approach is to estimate more restrictive models which are tractable but may not conform to the data. Our contributionis to propose an alternative estimation method that is numerically feasible,produces positive semi-definite conditional covariance matrices, and doesnot impose unrealistic a priori restrictions. We provide an empiricalapplication in the context of international stock markets, comparing thenew estimator to a number of existing ones.

Méthodologie intégrée de microgravimétrie en milieu urbain : test d'application au génie civil sur le métro M2 de Lausanne

RESUME Dès le printemps 2004, la construction d'une 2ème ligne de métro est entreprise dans la ville de Lausanne en Suisse. En reliant Ouchy, au bord du lac Léman (alt. 373 m) à Epalinges (alt. 711 m), le nouveau métro "M2" traversera dès 2008 l'agglomération lausannoise du Sud au Nord sur une distance de 6 km. Depuis l'avant-projet, en 1999, une grande quantité de données géologiques a été récolté et de nombreux forages exécutés sur le site. Ceci nous a donné une occasion unique d'entreprendre une étude de microgravimétrique urbaine de détail. Le mode de creusement du tunnel dépend fortement des matériaux à excaver et il est classiquement du domaine du géologue, avec ses connaissances de la géologie régionale et de la stratigraphie des forages, de fournir à l'ingénieur un modèle géologique. Ce modèle indiquera dans ce cas l'épaisseur des terrains meubles qui recouvrent le soubassement rocheux. La représentativité spatiale d'une information très localisée, comme celle d'un forage, est d'autant plus compliquée que le détail recherché est petit. C'est à ce moment là que la prospection géophysique, plus spécialement gravimétrique, peut apporter des informations complémentaires déterminantes pour régionaliser les données ponctuelles des forages. La microgravimétrie en milieu urbain implique de corriger avec soin les perturbations gravifiques sur la mesure de la pesanteur dues aux effets de la topographie, des bâtiments et des caves afin d'isoler l'effet gravifique dû exclusivement à l'épaisseur du remplissage des terrains meubles. Tenant compte de l'intensité des corrections topographiques en milieu urbain, nous avons donné une grande importance aux sous-sols, leurs effets gravifiques pouvant atteindre l'ordre du dixième de mGal. Nous avons donc intégré ces corrections celle de topographie et traité les effets des bâtiments de manière indépendante. Nous avons inclus dans le modèle numérique de terrain (MNT) la chaussée et les sous-sols afin de construire un modèle numérique de terrain urbain. Nous utiliserons un nouvel acronyme « MNTU »pour décrire ce modèle. Nous proposons d'établir des cartes de corrections topographiques préalables, basées sur les données à disposition fournies par le cadastre en faisant des hypothèses sur la profondeur des sous-sols et la hauteur des bâtiments. Les deux zones de test choisies sont caractéristiques des différents types d'urbanisation présente à Lausanne et se révèlent par conséquent très intéressantes pour élaborer une méthodologie globale de la microgravimétrie urbaine. Le but était d'évaluer l'épaisseur du remplissage morainique sur un fond rocheux molassique se situant à une profondeur variable de quelques mètres à une trentaine de mètres et d'en établir une coupe dans l'axe du futur tracé du métro. Les résultats des modélisations se sont révélés très convaincants en détectant des zones qui diffèrent sensiblement du modèle géologique d'avant projet. Nous avons également démontré que l'application de cette méthode géophysique, non destructive, est à même de limiter le nombre de sondages mécaniques lors de l'avant-projet et du projet définitif, ce qui peut limiter à la fois les coûts et le dérangement engendré par ces travaux de surface. L'adaptabilité de la technique gravimétrique permet d'intervenir dans toutes les différentes phases d'un projet de génie civil comme celui de la construction d'un métro en souterrain. KURZFASSUNG Seit dem Frühling 2004 ist in der Stadt Lausanne (Schweiz) die neue U-Bahn "M2" in Konstruktion. Diese soll auf 6 km Länge die Lausanner Agglomeration von Süd nach Nord durchqueren. Die dem Projekt zu Grunde liegende technische Planung sieht vor, daß die Bahnlinie hauptsächlich in der Molasse angesiedelt sein wird. Seit dem Vorentwurf (1999) ist eine große Anzahl geologischer Angaben gesammelt worden. Daraus ergab sich die einmalige Gelegenheit, die Informationen aus den damit verbundenen zahlreichen Bohrungen zu einer detaillierten mikrogravimetrischen Studie der Stadt Lausanne zu erweitern und zu vervollständigen. Das Ziel bestand darin, die Mächtigkeit der die Molasseüberdeckenden Moräneablagerung abzuschätzen, um eine entsprechendes geologisches Profile entlang der künftigen Bahnlinie zu erstellen. Weiterhin sollte gezeigt werden, daß die Anwendung dieser nicht-invasiven geophysikalischen Methode es ermöglicht, die Anzahl der benötigten Bohrungen sowohl in der Pilotphase wie auch im endgültigen Projekt zu reduzieren, was zu wesentlichen finanziellen Einsparungen in der Ausführung des Werkes beitragen würde. Die beiden in dieser Studie bearbeiteten Testzonen befinden sich im Nordteil und im Stadtzentrum von Lausanne und sind durch eine unterschiedliche Urbanisierung charakterisiert. Das anstehende Gestein liegt in verschiedenen Tiefen: von einigen Metern bis zu etwa dreißig Metern. Diese Zonen weisen alle Schwierigkeiten einer urbanen Bebauung mit hoher Verkehrsdichte auf und waren daher massgebend bei der Ausarbeitung einer globalen mikrogravimetrischen Methodologie für die Stadt Lausanne. Die so entwickelte Technik ermöglicht, die störenden Auswirkungen der Topographie, der Gebäude, der Keller und der Öffentlichen Infrastrukturen sorgfältig zu korrigieren, um so die ausschließlich auf die Mächtigkeit des Lockergesteins zurückzuführenden Effekte zu isolieren. In Bezug auf die Intensität der Auswirkungen der topographischen Korrekturen im Stadtgebiet wurde den Untergeschossen eine besonders grosse Bedeutung zugemessen da die entsprechenden Schwerkrafteffekte eine Grösse von rund einem Zehntel mGal erreichen können. Wir schlagen deshalb vor, vorläufige Karten der topographischen Korrekturen zu erstellen. Diese Korrekturen basieren auf den uns vom Katasterplan gelieferten Daten und einigen Hypothesen bezüglich der Tiefe der Untergeschosse und der Höhe der Gebäude. Die Verfügbarkeit einer derartigen Karte vor der eigentlichen gravimetrischen Messkampagne würde uns erlauben, die Position der Meßstationen besser zu wählen. Wir sahen zudem, daß ein entsprechenden a priori Filter benutzt werden kann, wenn die Form und die Intensität der Anomalie offensichtlich dem entsprechenden Gebäude zugeordnet werden können. Diese Strategie muß jedoch mit Vorsicht angewandt werden, denn falls weitere Anomalien dazukommen, können bedeutende Verschiebungen durch Übèrlagerungen der Schwerewirkung verschiedener Strukturen entstehen. Die Ergebnisse der Modellierung haben sich als sehr überzeugend erwiesen, da sie im Voraus unbekannte sensible Zonen korrekt identifiziert haben. Die Anwendbarkeit der in dieser Arbeit entwickelten gravimetrischen Technik ermöglicht es, während allen Phasen eines Grossbauprojekts, wie zum Beispiel bei der Konstruktion einer unterirdischen U-Bahn, einzugreifen. ABSTRACT Since Spring of 2004 a new metro line has been under construction in the city of Lausanne in Switzerland. The new line, the M2, will be 6 km long and will traverse the city from south to north. The civil engineering project determined that the line would be located primarily in the Molasse. Since the preparatory project in 1999, a great quantity of geological data has been collected, and the many drillings made on the site have proved to be a unique opportunity to undertake a study of urban microgravimetry. The goal was to evaluate the thickness of the morainic filling over the molassic bedrock, and to establish a section along the axis of the future line. It then had to be shown that the application of this nondestructive geophysical method could reduce the number of mechanical surveys required both for a preparatory and a definitive project, which would lead to real savings in the realization of a civil engineering project. The two test zones chosen, one in the northern part of the city and one in the city centre, are characterised by various types of urbanisation. Bedrock is at a depth varying from a few metres to about thirty metres. These zones well exemplify the various difficulties encountered in an urban environment and are therefore very interesting for the development of an overall methodology of urban microgravimetry. Microgravimetry in an urban environment requires careful corrections for gravific disturbances due to the effects of topography, buildings, cellars, and the infrastructure of distribution networks, in order to isolate the gravific effect due exclusively to the thickness of loose soil filling. Bearing in mind the intensity of the topographic corrections in an urban environment, we gave particular importance to basements. Their gravific effects can reach the order of one tenth of one meal, and can influence above all the precision of the Bouguer anomaly. We propose to establish preliminary topographic correction charts based on data provided to us by the land register, by making assumptions on the depths of basements and the heights of buildings. Availability of this chart previous to a gravimetry campaign would enable us to choose optimum measuring sites. We have also seen that an a priori filter can be used when the form and the intensity of the anomaly correspond visually to the corresponding building. This strategy must be used with caution because if other anomalies are to be associated, important shifts can be generated by the superposition of the effects of different structures. The results of the model have proved to be very convincing in detecting previously unknown sensitive zones. The adaptability of the gravimetry technique allows for application in all phases of a civil engineering project such as the construction of an underground metro line. RIASSUNTO Dalla primavera 2004 una nuova linea metropolitana é in costruzione nella città di Losanna in Svizzera. La nuova metropolitana "M2" traverserà per la lunghezza di 6 km il centro urbano di Losanna da sud a nord. II progetto d'ingegneria civile prevedeva un tracciato situato essenzialmente nel fondo roccioso arenaceo terziario (molassa). Dalla redazione del progetto preliminare, avvenuta nel 1999, una grande quantità di dati geologici sono stati raccolti e sono stati eseguiti numerosi sondaggi. Questo sì é presentato come un'occasione unica per mettere a punto uno studio microgravimetrico in ambiente urbano con lo scopo di valutare lo spessore dei terreni sciolti di origine glaciale che ricoprono il fondo roccioso di molassa e di mettere in evidenza come l'applicazione di questo metodo geofisico non distruttivo possa limitare il numero di sondaggi meccanici nella fase di progetto preliminare ed esecutivo con conseguente reale risparmio economico nella realizzazione di una tale opera. Le due zone di test sono situate una nella zona nord e la seconda nel centro storico di Losanna e sono caratterizzate da stili architettonici differenti. II fondo roccioso é situato ad una profondità variabile da qualche metro ad una trentina. Queste due zone sembrano ben rappresentare tutte le difficoltà di un ambiente urbano e ben si prestano per elaborare una metodologia globale per la microgravimetria in ambiente urbano. L'applicazione di questa tecnica nell'ambiente suddetto implica la correzione attenta delle perturbazioni sulla misura dell'accelerazione gravitazionale, causate dalla topografia, gli edifici, le cantine e le infrastrutture dei sottoservizi, per ben isolare il segnale esclusivamente causato dallo spessore dei terreni sciolti. Tenuto conto, dell'intensità delle correzioni topografiche, abbiamo dato grande importanza alle cantine, poiché il loro effetto sulle misure può raggiungere il decimo di mGal. Proponiamo quindi di redigere una carta delle correzioni topografiche preliminare all'acquisizione, facendo delle ipotesi sulla profondità delle cantine e sull'altezza degli edifici, sulla base delle planimetrie catastali. L'analisi di questa carta permetterà di scegliere le posizioni più adatte per le stazioni gravimetriche. Abbiamo anche osservato che un filtro a priori, qualora la forma e l'intensità dell'anomalia fosse facilmente riconducibile in maniera visuale ad un edificio, possa essere efficace. Tuttavia questa strategia deve essere utilizzata con precauzione, poiché può introdurre uno scarto, qualora più anomalie, dovute a differenti strutture, si sovrappongano. I risultati delle modellizzazioni si sono rivelati convincenti, evidenziando zone sensibili non conosciute preventivamente. L'adattabilità della tecnica gravimetrica ha mostrato di poter intervenire in differenti fasi di un progetto di ingegneria civile, quale è quella di un'opera in sotterraneo.

Minimum requirements for application of ink dating methods based on solvent analysis in casework

Several ink dating methods based on solvents analysis using gas chromatography/mass spectrometry (GC/MS) were proposed in the last decades. These methods follow the drying of solvents from ballpoint pen inks on paper and seem very promising. However, several questions arose over the last few years among questioned documents examiners regarding the transparency and reproducibility of the proposed techniques. These questions should be carefully studied for accurate and ethical application of this methodology in casework. Inspired by a real investigation involving ink dating, the present paper discusses this particular issue throughout four main topics: aging processes, dating methods, validation procedures and data interpretation. This work presents a wide picture of the ink dating field, warns about potential shortcomings and also proposes some solutions to avoid reporting errors in court.

Application of real-time PCR for total airborne bacterial assessment : comparison with epifluorescence microscopy and culture-dependent methods

Traditional culture-dependent methods to quantify and identify airborne microorganisms are limited by factors such as short-duration sampling times and inability to count nonculturableor non-viable bacteria. Consequently, the quantitative assessment of bioaerosols is often underestimated. Use of the real-time quantitative polymerase chain reaction (Q-PCR) to quantify bacteria in environmental samples presents an alternative method, which should overcome this problem. The aim of this study was to evaluate the performance of a real-time Q-PCR assay as a simple and reliable way to quantify the airborne bacterial load within poultry houses and sewage treatment plants, in comparison with epifluorescencemicroscopy and culture-dependent methods. The estimates of bacterial load that we obtained from real-time PCR and epifluorescence methods, are comparable, however, our analysis of sewage treatment plants indicate these methods give values 270-290 fold greater than those obtained by the ''impaction on nutrient agar'' method. The culture-dependent method of air impaction on nutrient agar was also inadequate in poultry houses, as was the impinger-culture method, which gave a bacterial load estimate 32-fold lower than obtained by Q-PCR. Real-time quantitative PCR thus proves to be a reliable, discerning, and simple method that could be used to estimate airborne bacterial load in a broad variety of other environments expected to carry high numbers of airborne bacteria. [Authors]