Interphase cytogenetics using fluorescence in situ hybridization: an overview of its application to diffuse and solid tissue

Interphase cytogenetics, utilizing fluorescence in situ hybridization (FISH) techniques, has been successfully applied to diffuse and solid tissue specimens. Most studies have been performed on isolated cells, such as blood or bone marrow cells; a few have been performed on cells from body fluids, such as amniotic fluid, urine, sperm, and sputum. Mechanically or chemically disaggregated cells from solid tissues have also been used as single cell suspensions for FISH. Additionally, intact organized tissue samples represented by touch preparations or thin tissue sections have been used, especially in cancer studies. Advantages and pitfalls of application of FISH methodology to each type of specimen and some significant biological findings achieved are illustrated in this overview.

Connexin hemichannels and cell-cell channels: comparison of properties

Connexin46 (Cx46) forms functional hemichannels in the absence of contact by an apposed hemichannel and we have used these hemichannels to study gating and permeation at the single channel level with high time resolution. Using both cell-attached and -excised patch configurations, we find that single Cx46 hemichannels exhibit some properties expected of half of a gap junction channel, as well as novel properties. Cx46 hemichannels have a large unitary conductance (~300 pS) and a relatively large pore as inferred from permeability to TEA. Both monovalent cations and anions can permeate, but cations are substantially more permeable. The open channel conductance shows marked inward rectification in symmetric salts. We find that the conductance and permeability properties of Cx46 cell-cell channels can be explained by the series addition of two hemichannels. These data suggest that the pore structures of unapposed hemichannels and cell-cell channels are conserved. Also like cell-cell channels, unapposed Cx46 hemichannels are closed by elevated levels of H+ or Ca2+ ions on the cytoplasmic face. Closure occurs in excised patches indicating that the actions of these agents do not require a soluble cytoplasmic factor. Fast (<0.5 ms) application of H+ to either side of the open hemichannel causes an immediate small reduction in unitary conductance followed by complete closure with latencies that are dependent on H+ concentration and side of application; sensitivity is much greater to H+ on the cytoplasmic side. Closure by cytoplasmic H+ does not require that the hemichannel be open. Thus, H+ ions readily permeate Cx46 hemichannels, but at high enough concentration close them by acting at a cytoplasmic site(s) that causes a conformational change resulting in complete closure. Extracellular H+ may permeate to act on the cytoplasmic site or act on a lower affinity extracellular site. Thus, the unapposed hemichannel is a valuable tool in addressing fundamental questions concerning the operation of gap junction channels that are difficult to answer by existing methods. The ability of Cx46, and perhaps other connexins, to form functional unapposed hemichannels that are opened by moderate depolarization may represent an unexplored role of connexins as mediators of transport across the plasma membrane.

Clinical features of panic patients sensitive to hyperventilation or breath-holding methods for inducing panic attacks

Our aim was to compare the clinical features of panic disorder (PD) patients sensitive to hyperventilation or breath-holding methods of inducing panic attacks. Eighty-five PD patients were submitted to both a hyperventilation challenge test and a breath-holding test. They were asked to hyperventilate (30 breaths/min) for 4 min and a week later to hold their breath for as long as possible, four times with a 2-min interval. Anxiety scales were applied before and after the tests. We selected the patients who responded with a panic attack to just one of the tests, i.e., those who had a panic attack after hyperventilating (HPA, N = 24, 16 females, 8 males, mean age ± SD = 38.5 ± 12.7 years) and those who had a panic attack after breath holding (BHPA, N = 20, 11 females, 9 males, mean age ± SD = 42.1 ± 10.6 years). Both groups had similar (chi² = 1.28, d.f. = 1, P = 0.672) respiratory symptoms (fear of dying, chest/pain disconfort, shortness of breath, paresthesias, and feelings of choking) during a panic attack. The criteria of Briggs et al. [British Journal of Psychiatry, 1993; 163: 201-209] for respiratory PD subtype were fulfilled by 18 (75.0%) HPA patients and by 14 (70.0%) BHPA patients. The HPA group had a later onset of the disease compared to BHPA patients (37.9 ± 11.0 vs 21.3 ± 12.9 years old, Mann-Whitney, P < 0.001), and had a higher family prevalence of PD (70.8 vs 25.0%, chi² = 19.65, d.f. = 1, P = 0.041). Our data suggest that these two groups - HPA and BHPA patients - may be specific subtypes of PD.

Remote sensing for three-dimensional modelling of hydromorphology

Successful management of rivers requires an understanding of the fluvial processes that govern them. This, in turn cannot be achieved without a means of quantifying their geomorphology and hydrology and the spatio-temporal interactions between them, that is, their hydromorphology. For a long time, it has been laborious and time-consuming to measure river topography, especially in the submerged part of the channel. The measurement of the flow field has been challenging as well, and hence, such measurements have long been sparse in natural environments. Technological advancements in the field of remote sensing in the recent years have opened up new possibilities for capturing synoptic information on river environments. This thesis presents new developments in fluvial remote sensing of both topography and water flow. A set of close-range remote sensing methods is employed to eventually construct a high-resolution unified empirical hydromorphological model, that is, river channel and floodplain topography and three-dimensional areal flow field. Empirical as well as hydraulic theory-based optical remote sensing methods are tested and evaluated using normal colour aerial photographs and sonar calibration and reference measurements on a rocky-bed sub-Arctic river. The empirical optical bathymetry model is developed further by the introduction of a deep-water radiance parameter estimation algorithm that extends the field of application of the model to shallow streams. The effect of this parameter on the model is also assessed in a study of a sandy-bed sub-Arctic river using close-range high-resolution aerial photography, presenting one of the first examples of fluvial bathymetry modelling from unmanned aerial vehicles (UAV). Further close-range remote sensing methods are added to complete the topography integrating the river bed with the floodplain to create a seamless high-resolution topography. Boat- cart- and backpack-based mobile laser scanning (MLS) are used to measure the topography of the dry part of the channel at a high resolution and accuracy. Multitemporal MLS is evaluated along with UAV-based photogrammetry against terrestrial laser scanning reference data and merged with UAV-based bathymetry to create a two-year series of seamless digital terrain models. These allow the evaluation of the methodology for conducting high-resolution change analysis of the entire channel. The remote sensing based model of hydromorphology is completed by a new methodology for mapping the flow field in 3D. An acoustic Doppler current profiler (ADCP) is deployed on a remote-controlled boat with a survey-grade global navigation satellite system (GNSS) receiver, allowing the positioning of the areally sampled 3D flow vectors in 3D space as a point cloud and its interpolation into a 3D matrix allows a quantitative volumetric flow analysis. Multitemporal areal 3D flow field data show the evolution of the flow field during a snow-melt flood event. The combination of the underwater and dry topography with the flow field yields a compete model of river hydromorphology at the reach scale.

New methods for controlling twin configurations and characterizing twin boundaries in 5M Ni-Mn-Ga for the development of applications

Traditional methods for studying the magnetic shape memory (MSM) alloys Ni-Mn-Ga include subjecting the entire sample to a uniform magnetic field or completely actuating the sample mechanically. These methods have produced significant results in characterizing the MSM effect, the properties of Ni-Mn-Ga and have pioneered the development of applications from this material. Twin boundaries and their configuration within a Ni-Mn-Ga sample are a key component in the magnetic shape memory effect. Applications that are developed require an understanding of twin boundary characteristics and, more importantly, the ability to predictably control them. Twins have such a critical role that the twinning stress of a Ni-Mn-Ga crystal is the defining characteristic that indicates its quality and significant research has been conducted to minimize this property. This dissertation reports a decrease in the twinning stress, predictably controlling the twin configuration and characterizing the dynamics of twin boundaries. A reduction of the twinning stress is demonstrated by the discovery of Type II twins within Ni-Mn-Ga which have as little as 10% of the twinning stress of traditional Type I twins. Furthermore, new methods of actuating a Ni-Mn-Ga element using localized unidirectional or bidirectional magnetic fields were developed that can predictably control the twin configuration in a localized area of a Ni-Mn-Ga element. This method of controlling the local twin configuration was used in the characterization of twin boundary dynamics. Using a localized magnetic pulse, the velocity and acceleration of a single twin boundary were measured to be 82.5 m/s and 2.9 × 107 m/s2, and the time needed for the twin boundary to nucleate and begin moving was less than 2.8 μs. Using a bidirectional magnetic field from a diametrically magnetized cylindrical magnet, a highly reproducible and controllable local twin configuration was created in a Ni-Mn-Ga element which is the fundamental pumping mechanism in the MSM micropump that has been co-invented and extensively characterized by the author.

Nichesourcing The Uralic Languages For The Benefit Of Linguistic Research And Lingual Societies

The emerging technologies have recently challenged the libraries to reconsider their role as a mere mediator between the collections, researchers, and wider audiences (Sula, 2013), and libraries, especially the nationwide institutions like national libraries, haven’t always managed to face the challenge (Nygren et al., 2014). In the Digitization Project of Kindred Languages, the National Library of Finland has become a node that connects the partners to interplay and work for shared goals and objectives. In this paper, I will be drawing a picture of the crowdsourcing methods that have been established during the project to support both linguistic research and lingual diversity. The National Library of Finland has been executing the Digitization Project of Kindred Languages since 2012. The project seeks to digitize and publish approximately 1,200 monograph titles and more than 100 newspapers titles in various, and in some cases endangered Uralic languages. Once the digitization has been completed in 2015, the Fenno-Ugrica online collection will consist of 110,000 monograph pages and around 90,000 newspaper pages to which all users will have open access regardless of their place of residence. The majority of the digitized literature was originally published in the 1920s and 1930s in the Soviet Union, and it was the genesis and consolidation period of literary languages. This was the era when many Uralic languages were converted into media of popular education, enlightenment, and dissemination of information pertinent to the developing political agenda of the Soviet state. The ‘deluge’ of popular literature in the 1920s to 1930s suddenly challenged the lexical orthographic norms of the limited ecclesiastical publications from the 1880s onward. Newspapers were now written in orthographies and in word forms that the locals would understand. Textbooks were written to address the separate needs of both adults and children. New concepts were introduced in the language. This was the beginning of a renaissance and period of enlightenment (Rueter, 2013). The linguistically oriented population can also find writings to their delight, especially lexical items specific to a given publication, and orthographically documented specifics of phonetics. The project is financially supported by the Kone Foundation in Helsinki and is part of the Foundation’s Language Programme. One of the key objectives of the Kone Foundation Language Programme is to support a culture of openness and interaction in linguistic research, but also to promote citizen science as a tool for the participation of the language community in research. In addition to sharing this aspiration, our objective within the Language Programme is to make sure that old and new corpora in Uralic languages are made available for the open and interactive use of the academic community as well as the language societies. Wordlists are available in 17 languages, but without tokenization, lemmatization, and so on. This approach was verified with the scholars, and we consider the wordlists as raw data for linguists. Our data is used for creating the morphological analyzers and online dictionaries at the Helsinki and Tromsø Universities, for instance. In order to reach the targets, we will produce not only the digitized materials but also their development tools for supporting linguistic research and citizen science. The Digitization Project of Kindred Languages is thus linked with the research of language technology. The mission is to improve the usage and usability of digitized content. During the project, we have advanced methods that will refine the raw data for further use, especially in the linguistic research. How does the library meet the objectives, which appears to be beyond its traditional playground? The written materials from this period are a gold mine, so how could we retrieve these hidden treasures of languages out of the stack that contains more than 200,000 pages of literature in various Uralic languages? The problem is that the machined-encoded text (OCR) contains often too many mistakes to be used as such in research. The mistakes in OCRed texts must be corrected. For enhancing the OCRed texts, the National Library of Finland developed an open-source code OCR editor that enabled the editing of machine-encoded text for the benefit of linguistic research. This tool was necessary to implement, since these rare and peripheral prints did often include already perished characters, which are sadly neglected by the modern OCR software developers, but belong to the historical context of kindred languages and thus are an essential part of the linguistic heritage (van Hemel, 2014). Our crowdsourcing tool application is essentially an editor of Alto XML format. It consists of a back-end for managing users, permissions, and files, communicating through a REST API with a front-end interface—that is, the actual editor for correcting the OCRed text. The enhanced XML files can be retrieved from the Fenno-Ugrica collection for further purposes. Could the crowd do this work to support the academic research? The challenge in crowdsourcing lies in its nature. The targets in the traditional crowdsourcing have often been split into several microtasks that do not require any special skills from the anonymous people, a faceless crowd. This way of crowdsourcing may produce quantitative results, but from the research’s point of view, there is a danger that the needs of linguists are not necessarily met. Also, the remarkable downside is the lack of shared goal or the social affinity. There is no reward in the traditional methods of crowdsourcing (de Boer et al., 2012). Also, there has been criticism that digital humanities makes the humanities too data-driven and oriented towards quantitative methods, losing the values of critical qualitative methods (Fish, 2012). And on top of that, the downsides of the traditional crowdsourcing become more imminent when you leave the Anglophone world. Our potential crowd is geographically scattered in Russia. This crowd is linguistically heterogeneous, speaking 17 different languages. In many cases languages are close to extinction or longing for language revitalization, and the native speakers do not always have Internet access, so an open call for crowdsourcing would not have produced appeasing results for linguists. Thus, one has to identify carefully the potential niches to complete the needed tasks. When using the help of a crowd in a project that is aiming to support both linguistic research and survival of endangered languages, the approach has to be a different one. In nichesourcing, the tasks are distributed amongst a small crowd of citizen scientists (communities). Although communities provide smaller pools to draw resources, their specific richness in skill is suited for complex tasks with high-quality product expectations found in nichesourcing. Communities have a purpose and identity, and their regular interaction engenders social trust and reputation. These communities can correspond to research more precisely (de Boer et al., 2012). Instead of repetitive and rather trivial tasks, we are trying to utilize the knowledge and skills of citizen scientists to provide qualitative results. In nichesourcing, we hand in such assignments that would precisely fill the gaps in linguistic research. A typical task would be editing and collecting the words in such fields of vocabularies where the researchers do require more information. For instance, there is lack of Hill Mari words and terminology in anatomy. We have digitized the books in medicine, and we could try to track the words related to human organs by assigning the citizen scientists to edit and collect words with the OCR editor. From the nichesourcing’s perspective, it is essential that altruism play a central role when the language communities are involved. In nichesourcing, our goal is to reach a certain level of interplay, where the language communities would benefit from the results. For instance, the corrected words in Ingrian will be added to an online dictionary, which is made freely available for the public, so the society can benefit, too. This objective of interplay can be understood as an aspiration to support the endangered languages and the maintenance of lingual diversity, but also as a servant of ‘two masters’: research and society.

Effects of peeling methods on the quality of cubiu fruits

Cubiu (Solanum sessiliflorum Dunal) is an Amazonian Basin native fruit. Its importance comes from its high contents of pectin. Currently, processing technologies are necessary for the substitution of the traditional system (small crops and small-scale processing) for a larger scale system and thus increase the use of biodiversity and promote the implementation of Local Productive Arrangements of agribusiness in the Amazon. This research aims to evaluate the methods of peeling cubiu. Ripe fruits were divided into lots (150 each) and subjected to the following treatments: immersion in 2.5% NaOH boiling solution for 5 minutes, exposure to water vapor, and immersion in water at 96 ºC for 5, 10, 15 and 20 minutes. The peel released during heat treatment and immediately removed under running tap water. In the control treatment, the fruits were manually peeled (unheated) with a stainless steel knife. The treatments were evaluated for completeness and ease of peeling, tissue integrity, texture, and peroxidase activity. The immersion in 2.5% NaOH boiling solution (5 minutes) stood out as the best treatment since it inhibited the enzymatic browning and intensified the natural yellow color of the cubiu fruit and easily and fully peeled the whole fruit more rapidly without damaging its tissues. This treatment was chosen as the most advantageous because it can promote simultaneous peeling and bleaching. Therefore, it is recommended for cubiu industrial processing.

Comparison of capillary electrophoresis and high performance liquid chromatography methods for caffeine determination in decaffeinated coffee

Decaffeinated coffee accounts for 10 percent of coffee sales in the world; it is preferred by consumers that do not wish or are sensitive to caffeine effects. This article presents an analytical comparison of capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) methods for residual caffeine quantification in decaffeinated coffee in terms of validation parameters, costs, analysis time, composition and treatment of the residues generated, and caffeine quantification in 20 commercial samples. Both methods showed suitable validation parameters. Caffeine content did not differ statistically in the two different methods of analysis. The main advantage of the high performance liquid chromatography (HPLC) method was the 42-fold lower detection limit. Nevertheless, the capillary electrophoresis (CE) detection limit was 115-fold lower than the allowable limit by the Brazilian law. The capillary electrophoresis (CE) analyses were 30% faster, the reagent costs were 76.5-fold, and the volume of the residues generated was 33-fold lower. Therefore, the capillary electrophoresis (CE) method proved to be a valuable analytical tool for this type of analysis.

The Competitive Analysis of Selected Biorefineries in Finland

Biorefineries is a perspective field of study that covers many opportunities of a successful business unit with respect to sustainability. The thesis focuses on the following key objective: identification of a competitive biorefineries production process in small and medium segments of the chemical and forest industries in Finland. The scope of the research relates to the selected biorefineries operations in Finland and the use of hemicellulose, as a raw material. The identification of the types of biorefineries and the important technical and process characteristics opens the advantage in the company’s competitive analysis. The study concentrates on the practical approach to the scientific methods of the market and companies research with the help of Quality Function Deployment and House of Quality tool. The thesis’s findings provide mindset version of the expert’s House of Quality application, identification of crucial biorefineries technical and design characteristics’ correlation and their effect on the competitive behavior of a company. The theoretical background helps to build the picture of the problematic issues within the field and provides scientific possible solutions. The analysis of the biorefineries’ market and companies operations bring the practical-oriented aptitude of the research. The results of the research can be used for the following investigations in a field and may be applied as a company’s management analytic and strategic application.

Marketing a Mobile Application in a Highly Competitive Global Marketplace

The purpose of this research was to study the marketing of mobile applications. The main objective was to find out what are the most efficient ways of marketing to increase the sales for a mobile application within a highly competitive marketplace. The marketplaces, app stores, are studied from the perspective of size, ease of entry, competition and customers and their purchasing process. The study also includes research on what are some of the main marketing methods used in mobile app marketing in general. The study consists of two parts, theoretical and empirical research. Theoretical research was done by studying past scientific research on the chosen subjects. As the subject is very new, the research was also extended to other publications from the field of mobile technology. The empirical part was done through interviews and empirical experiments with a case-company, which were used to answer the main objective of this study. These experiments showed that the chosen methods of mobile app marketing, app store optimization, localization and selected social media marketing activities, created the most sales when used together. Positive results were seen also when the activities were conducted by themselves, but together they were able to push the case company to their all time best results. However the key to succeeding and hitting high positions in the app store rankings would most likely require creating a solid marketing strategy, trying out other marketing activities alongside the ones used here, without forgetting to stay on top of mobile technology trends.

Optimal Recycling Combination of Ash in South-East Finland

The present world energy production is heavily relying on the combustion of solid fuels like coals, peat, biomass, municipal solid waste, whereas the share of renewable fuels is anticipated to increase in the future to mitigate climate change. In Finland, peat and wood are widely used for energy production. In any case, the combustion of solid fuels results in generation of several types of thermal conversion residues, such as bottom ash, fly ash, and boiler slag. The predominant residue type is determined by the incineration technology applied, while its composition is primarily relevant to the composition of fuels combusted. An extensive research has been conducted on technical suitability of ash for multiple recycling methods. Most of attention was drawn to the recycling of the coal combustion residues, as coal is the primary solid fuel consumed globally. The recycling methods of coal residues include utilization in a cement industry, in concrete manufacturing, and mine backfilling, to name few. Biomass combustion residues were also studied to some extent with forest fertilization, road construction, and road stabilization being the predominant utilization options. Lastly, residues form municipal solid waste incineration attracted more attention recently following the growing number of waste incineration plants globally. The recycling methods of waste incineration residues are the most limited due to its hazardous nature and varying composition, and include, among others, landfill construction, road construction, mine backfilling. In the study, environmental and economic aspects of multiple recycling options of thermal conversion residues generated within a case-study area were studied. The case-study area was South-East Finland. The environmental analysis was performed using an internationally recognized methodology — life cycle assessment. Economic assessment was conducted applying a widely used methodology — cost-benefit analysis. Finally, the results of the analyses were combined to enable easier comparison of the recycling methods. The recycling methods included the use of ash in forest fertilization, road construction, road stabilization, and landfill construction. Ash landfilling was set as a baseline scenario. Quantitative data about the amounts of ash generated and its composition was obtained from companies, their environmental reports, technical reports and other previously published literature. Overall, the amount of ash in the case-study area was 101 700 t. However, the data about 58 400 t of fly ash and 35 100 t of bottom ash and boiler slag were included in the study due to lack of data about leaching of heavy metals in some cases. The recycling methods were modelled according to the scientific studies published previously. Overall, the results of the study indicated that ash utilization for fertilization and neutralization of 17 600 ha of forest was the most economically beneficial method, which resulted in the net present value increase by 58% compared to ash landfilling. Regarding the environmental impact, the use of ash in the construction of 11 km of roads was the most attractive method with decreased environmental impact of 13% compared to ash landfilling. The least preferred method was the use of ash for landfill construction since it only enabled 11% increase of net present value, while inducing additional 1% of negative impact on the environment. Therefore, a following recycling route was proposed in the study. Where possible and legally acceptable, recycle fly and bottom ash for forest fertilization, which has strictest requirements out of all studied methods. If the quality of fly ash is not suitable for forest fertilization, then it should be utilized, first, in paved road construction, second, in road stabilization. Bottom ash not suitable for forest fertilization, as well as boiler slag, should be used in landfill construction. Landfilling should only be practiced when recycling by either of the methods is not possible due to legal requirements or there is not enough demand on the market. Current demand on ash and possible changes in the future were assessed in the study. Currently, the area of forest fertilized in the case-study are is only 451 ha, whereas about 17 600 ha of forest could be fertilized with ash generated in the region. Provided that the average forest fertilizing values in Finland are higher and the area treated with fellings is about 40 000 ha, the amount of ash utilized in forest fertilization could be increased. Regarding road construction, no new projects launched by the Center of Economic Development, Transport and the Environment in the case-study area were identified. A potential application can be found in the construction of private roads. However, no centralized data about such projects is available. The use of ash in stabilization of forest roads is not expected to increased in the future with a current downwards trend in the length of forest roads built. Finally, the use of ash in landfill construction is not a promising option due to the reducing number of landfills in operation in Finland.

Study of heat capacity measurement methods for small samples

Methods of measuring specific heats of small samples were studied. Three automated methods were explored, two of which have shown promising results. The adiabatic continuous heating method, has provided smooth well behaved data but further work is presently underway to improve on the results obtained so far . The decay method has been success fully implemented demonstrating reasonable agreement with accepted data for a copper test sample.

Bayesian Analysis for a Theory of Random Consumer Demand: The Case of Indivisible Goods

McCausland (2004a) describes a new theory of random consumer demand. Theoretically consistent random demand can be represented by a \"regular\" \"L-utility\" function on the consumption set X. The present paper is about Bayesian inference for regular L-utility functions. We express prior and posterior uncertainty in terms of distributions over the indefinite-dimensional parameter set of a flexible functional form. We propose a class of proper priors on the parameter set. The priors are flexible, in the sense that they put positive probability in the neighborhood of any L-utility function that is regular on a large subset bar(X) of X; and regular, in the sense that they assign zero probability to the set of L-utility functions that are irregular on bar(X). We propose methods of Bayesian inference for an environment with indivisible goods, leaving the more difficult case of indefinitely divisible goods for another paper. We analyse individual choice data from a consumer experiment described in Harbaugh et al. (2001).

On the Manipulability of Proportional Representation

This paper presents a new model of voter behaviour under methods of proportional representation (PR). We abstract away from rounding, and assume that a party securing k percent of the vote wins exactly k percent of the available seats. Under this assumption PR is not manipulable by any voter aiming at maximisation of the number of seats in the parliament of her most preferred party. However in this paper we assume that voters are concerned, first and foremost, with the distribution of power in the post-election parliament. We show that, irrespective of which positional scoring rule is adopted, there will always exist circumstances where a voter would have an incentive to vote insincerely. We demonstrate that a voter’s attitude toward uncertainty can influence her incentives to make an insincere vote. Finally, we show that the introduction of a threshold - a rule that a party must secure at least a certain percentage of the vote in order to reach parliament - creates new opportunities for strategic voting. We use the model to explain voter behaviour at the most recent New Zealand general election.

Investigation of femtosecond laser technology for the fabrication of drug nanocrystals in suspension

La technique du laser femtoseconde (fs) a été précédemment utilisée pour la production de nanoparticules d'or dans un environnement aqueux biologiquement compatible. Au cours de ce travail de maîtrise, cette méthode a été investiguée en vue d'une application pour la fabrication de nanocristaux de médicament en utilisant le paclitaxel comme modèle. Deux procédés distincts de cette technologie à savoir l'ablation et la fragmentation ont été étudiés. L'influence de la puissance du laser, de point de focalisation, et de la durée du traitement sur la distribution de taille des particules obtenues ainsi que leur intégrité chimique a été évaluée. Les paramètres ont ainsi été optimisés pour la fabrication des nanoparticules. L’évaluation morphologique et chimique a été réalisée par microscopie électronique et spectroscopie infrarouge respectivement. L'état cristallin des nanoparticules de paclitaxel a été caractérisé par calorimétrie differentielle et diffraction des rayons X. L'optimisation du procédé de production de nanoparticules par laser fs a permis d'obtenir des nanocristaux de taille moyenne (400 nm, polydispersité ≤ 0,3). Cependant une dégradation non négligeable a été observée. La cristallinité du médicament a été maintenue durant la procédure de réduction de taille, mais le paclitaxel anhydre a été transformé en une forme hydratée. Les résultats de cette étude suggèrent que le laser fs peut générer des nanocristaux de principe actif. Cependant cette technique peut se révéler problématique pour des médicaments sensibles à la dégradation. Grâce à sa facilité d'utilisation et la possibilité de travailler avec des quantités restreintes de produit, le laser fs pourrait représenter une alternative valable pour la production de nanoparticules de médicaments peu solubles lors des phases initiales de développement préclinique. Mots-clés: paclitaxel, nanocristaux, laser femtoseconde, ablation, fragmentation