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L’entérotoxine B staphylococcique (SEB) est une toxine entérique hautement résistante à la chaleur et est responsable de plus de 50 % des cas d’intoxication d’origine alimentaire par une entérotoxine. L’objectif principal de ce projet de maîtrise est de développer et valider une méthode basée sur des nouvelles stratégies analytiques permettant la détection et la quantification de SEB dans les matrices alimentaires. Une carte de peptides tryptiques a été produite et 3 peptides tryptiques spécifiques ont été sélectionnés pour servir de peptides témoins à partir des 9 fragments protéolytiques identifiés (couverture de 35 % de la séquence). L’anhydride acétique et la forme deutérée furent utilisés afin de synthétiser des peptides standards marqués avec un isotope léger et lourd. La combinaison de mélanges des deux isotopes à des concentrations molaires différentes fut utilisée afin d’établir la linéarité et les résultats ont démontré que les mesures faites par dilution isotopique combinée au CL-SM/SM respectaient les critères généralement reconnus d’épreuves biologiques avec des valeurs de pente près de 1, des valeurs de R2 supérieure à 0,98 et des coefficients de variation (CV%) inférieurs à 8 %. La précision et l’exactitude de la méthode ont été évaluées à l’aide d’échantillons d’homogénat de viande de poulet dans lesquels SEB a été introduite. SEB a été enrichie à 0,2, 1 et 2 pmol/g. Les résultats analytiques révèlent que la méthode procure une plage d’exactitude de 84,9 à 91,1 %. Dans l’ensemble, les résultats présentés dans ce mémoire démontrent que les méthodes protéomiques peuvent être utilisées efficacement pour détecter et quantifier SEB dans les matrices alimentaires. Mots clés : spectrométrie de masse; marquage isotopique; protéomique quantitative; entérotoxines / Staphylococcal enterotoxin B is a highly heat-resistant enteric toxin and it is responsible for over 50% of enterotoxin food poisoning. It represents a particular challenge during food processing since, even if the bacteria have been destroyed, the biological activity of the toxin remains unchanged. The objective of this study was to develop and validate a new method based on a novel proteomic strategy to detect and quantify SEB in food matrices. Tryptic peptide map was generated and 3 specific tryptic peptides were selected and used as surrogate peptides from 9 identified proteolytic fragments (sequence coverage of 35%). Peptides were label with light and heavy form of acetic anhydride to create an isobaric tag that will allow quantification. The linearity was tested using mixtures of different molar ratios and the results showed that measurements by LC-MS/MS were within generally accepted criteria for bioassays with slope values near to 1, values of R2 above 0.98 and less than 8% coefficient of variation (%CV). The precision and accuracy of the method were assessed using chicken meat homogenate samples spiked with SEB at 0.2, 1 and 2 pmol/g. The results indicated that the method can provide accuracy within 84.9 – 91.1% range. Overall, the results presented in this thesis show that proteomics-based methods can be effectively used to detect, confirm and quantify SEB in food matrices. Keywords: mass spectrometry; stable isotope labeling; quantitative proteomics; enterotoxins

Darbo tikslas: Įvertinti KTU Cheminės technologijos fakulteto studentų žinias ir požiūrį apie genetiškai modifikuotą maistą, jo ženklinimą. Darbo uždaviniai: 1. Nustatyti KTU Cheminės technologijos fakulteto studentų informavimo šaltinius apie genetiškai modifikuotą maistą ir jo ženklinimą. 2. Ištirti KTU Cheminės technologijos fakulteto studentų žinias ir požiūrį apie genetiškai modifikuotą maistą ir jo ženklinimą. 3. Parengti rekomendacijas siekiant pagerinti genetiškai modifikuoto maisto ir jo ženklinimo informuotumą. Tyrimo metodika: Tyrimo objektas: Žinios ir požiūris apie genetiškai modifikuotą maistą ir jo ženklinimą. Tyrimo metodai: Mokslinės literatūros analizė, teisinių dokumentų analizė, anketinė apklausa. Tyrimo metu buvo išplatinta 120, atsako dažnis - 80,83 proc. Statistinės analizės metodai: Apklausos rezultatai buvo analizuojami naudojant Microsoft office Excel 2007 ir kompiuterinį SPSS 13.0 versijos statistinį duomenų analizės paketą. Statistinis duomenų reikšmingumas buvo tikrinamas pagal chi kvadrato kriterijų ir laisvės laipsnių skaičių. Požymius analizuojant kito rodiklio atžvilgiu skirtumai laikyti statistiškai reikšmingais, kai reikšmingumo lygmuo p<0,05. Rezultatai: Dauguma respondentų dažniausiai gauna informaciją apie genetiškai modifikuotą maistą ir jo ženklinimą internete (15,5 proc. moterų ir 11,3 proc. vyrų), 30,9 proc. moterų ir 35,1 proc. vyrų žino šiek tiek, tačiau norėtų būti daugiau informuojamas,-a. Didžioji dalis apklaustųjų mano, jog... [toliau žr. visą tekstą] / Aim of study: to evaluate student‘s knowledge and attitude to genetically modified food and its labeling in KTU faculty of chemical technology. Objectives: 1. To survey the repositories of genetically modified food and his labeling. 2. To survey the knowledge and attitude to genetically modified food and its labeling. 3. To prepare recomendations to increase privity of genetically modified food and its labeling. Methods: Object of the research - knowledge and attitude to genetically modified food and its labeling. The questionnaire was compiled after making an analysis of the latest Lithuanian and foreign scientific literature and legislation. There were totally 120 students of KTU faculty of chemical tecnology interviewed using the anonymous questionnaire (response rate 80,83 percentage). Data analysis was performed using the statistical data analysis package SPSS (version 13.00) (the chi-squared test, df) and Microsoft office Excel 2007. Differences between the indices were considered statistically significant for p-values less than 0.05. Results: The most of respondents get information about genetically modified food and its labeling commonly in internet (15.5 percentage women and 11.3 percentage men), 30.9 percentage women and 35.1 men know a little bit, but they want to get more information. In addition to this, respondents think, that genetically modified food is harmful (33.7 percentage women and 32.3 percentage men) because of lack information about its safety (33.6... [to full text]

Žalieji pirkimai – tai tokie pirkimai, kurie integruoja aplinkosauginius reikalavimus į viešojo pirkimo sąlygas, siekdami sumažinti neigiamą poveikį aplinkai. Žalieji pirkimai yra nauja sfera viešojo administravimo studijose. Nors koncepcija teoriškai yra pagrįsta ir analizuota, tačiau praktiniai jos įgyvendinimo aspektai literatūroje nėra plačiai analizuoti, kadangi Lietuvoje žaliųjų pirkimų tiesioginis reglamentavimas pradėtas 2007 m. antroje pusėje. Darbo tikslas yra išanalizuoti žaliųjų pirkimų politikos įgyvendinimą teisinio reglamentavimo, instituciniu, finansiniu aspektu, išskiriant įgyvendinimą įtvirtinančius instrumentus. Darbas sudarytas iš penkių dalių, kurios pagal analizės objektą suskirstytos į smulkesnius skyrelius. Pirmajame skyriuje aptariamas viešosios politikos ciklas, kurio vienas iš etapų yra įgyvendinimas, aprašomi galimi įgyvendinimo mechanizmai ir instrumentai, kuriais remiasi žaliųjų pirkimų politikos įgyvendinimas. Antrajame skyriuje analizuojama ES reguliacinė politika, kurią pasitelkiama įgyvendinant žaliuosius pirkimus, bei Komisijos vaidmuo įgyvendinimo etape; teisinė bazė – ES direktyvos, jų perkėlimas į LR įstatymus, svarbiausios ES ir Lietuvos strategijos, programos, skirtos žaliųjų pirkimų įgyvendinimui. Trečiajame skyriuje nagrinėjamos Lietuvos institucijos, dalyvaujančios įgyvendinimo procese ir nevyriausybinių organizacijų veikla. Kadangi žaliųjų pirkimų įgyvendinimas yra lėšų reikalaujanti sritis, apžvelgiami įgyvendinimo finansiniai... [toliau žr. visą tekstą] / Green procurement is a way of public procurement, which integrates environmental considerations in public procurement process, seeking to reduce negative influence on environment. The relevancy of this paper is determined by novelty of green procurement policy in public administration studies. Even the concept is theoretically grounded but practical aspects of implementation are not strongly analysed, because reglamentation in Lithuania was started in second part of 2007. The subject of this paper focuses on analysis of green procurement policy reglamentation, institutional, financial aspects, and exclusion instruments of implementation. The paper is composed of the five main parts which are divided into smaller sections depending on the analyzing subject. The system of public policy cycle, which includes implementation stage and instruments of implementation, which are used in green procurement policy are analysed in the first part. EU regulation policy and Commission role in implementation of green procurements are analysed in the second part. This part of paper reveals policy reglamentation in EU directives and transfer of it to Lithuanian legislation. The most important are EU and Lithuanians strategies, programs, which implement green procurements. Lithuanian institutions and NGO are analysed in the third part of paper. This part focuses on financial instruments, because green procuremement is financal resources required policy... [to full text]

The work presented in this thesis describes methodologies developed for integration and accurate interpretation of barcoded DNA, to empower large-scale-omics analysis. The objectives mainly aim at enabling multiplexed proteomic measurements in high-throughput format through DNA barcoding and massive parallel sequencing. The thesis is based on four scientific papers that focus on three main criteria; (i) to prepare reagents for large-scale affinity-proteomics, (ii) to present technical advances in barcoding systems for parallel protein detection, and (iii) address challenges in complex sequencing data analysis. In the first part, bio-conjugation of antibodies is assessed at significantly downscaled reagent quantities. This allows for selection of affinity binders without restrictions to accessibility in large amounts and purity from amine-containing buffers or stabilizer materials (Paper I). This is followed by DNA barcoding of antibodies using minimal reagent quantities. The procedure additionally enables efficient purification of barcoded antibodies from free remaining DNA residues to improve sensitivity and accuracy of the subsequent measurements (Paper II). By utilizing a solid-phase approach on magnetic beads, a high-throughput set-up is ready to be facilitated by automation. Subsequently, the applicability of prepared bio-conjugates for parallel protein detection is demonstrated in different types of standard immunoassays (Papers I and II). As the second part, the method immuno-sequencing (I-Seq) is presented for DNAmediated protein detection using barcoded antibodies. I-Seq achieved the detection of clinically relevant proteins in human blood plasma by parallel DNA readout (Paper II). The methodology is further developed to track antibody-antigen interaction events on suspension bead arrays, while being encapsulated in barcoded emulsion droplets (Paper III). The method, denoted compartmentalized immuno-sequencing (cI-Seq), is potent to perform specific detections with paired antibodies and can provide information on details of joint recognition events. Recent progress in technical developments of DNA sequencing has increased the interest in large-scale studies to analyze higher number of samples in parallel. The third part of this thesis focuses on addressing challenges of large-scale sequencing analysis. Decoding of a huge DNA-barcoded data is presented, aiming at phase-defined sequence investigation of canine MHC loci in over 3000 samples (Paper IV). The analysis revealed new single nucleotide variations and a notable number of novel haplotypes for the 2nd exon of DLA DRB1. Taken together, this thesis demonstrates emerging applications of barcoded sequencing in protein and DNA detection. Improvements through the barcoding systems for assay parallelization, de-convolution of antigen-antibody interactions, sequence variant analysis, as well as large-scale data interpretation would aid biomedical studies to achieve a deeper understanding of biological processes. The future perspectives of the developed methodologies may therefore stem for advancing large-scale omics investigations, particularly in the promising field of DNA-mediated proteomics, for highly multiplex studies of numerous samples at a notably improved molecular resolution. / <p>QC 20150203</p>

Arterial spin labeling is a completely non-invasive method for blood-flow measurement techniques. Alzheimer's disease pathology includes microvascular abnormalities in addition to practically all risk factors having a vascular component that reduces cerebral perfusion. Hemodynamic parameters of cerebral blood flow and arterial transit time were estimated via single-compartment modeling of pseudo continuous arterial spin labeling data and neurocognitive test scores (Alzheimer's disease assessment scale and mini-mental state examination) were compared between a group of healthy (N=20) and early Alzheimer's disease (N=25) subjects before and six months after the Alzheimer's subjects began treatment with cholinesterase inhibitors. The early Alzheimer's group showed improved CBF after 6 months' treatment in every Alzheimer's-prone region except the medial and lateral temporal lobes. No difference in arterial transit time was found between groups, indicating that the pathophysiological process causing hypoperfusion in Alzheimer's disease may differ from vascular dementia.

Arterial spin labeling is a completely non-invasive method for blood-flow measurement techniques. Alzheimer's disease pathology includes microvascular abnormalities in addition to practically all risk factors having a vascular component that reduces cerebral perfusion. Hemodynamic parameters of cerebral blood flow and arterial transit time were estimated via single-compartment modeling of pseudo continuous arterial spin labeling data and neurocognitive test scores (Alzheimer's disease assessment scale and mini-mental state examination) were compared between a group of healthy (N=20) and early Alzheimer's disease (N=25) subjects before and six months after the Alzheimer's subjects began treatment with cholinesterase inhibitors. The early Alzheimer's group showed improved CBF after 6 months' treatment in every Alzheimer's-prone region except the medial and lateral temporal lobes. No difference in arterial transit time was found between groups, indicating that the pathophysiological process causing hypoperfusion in Alzheimer's disease may differ from vascular dementia.

Automated analysis of temporal data is a task of utmost importance for intelligent machines. For example, ubiquitous computing systems need to understand the intention of humans from the stream of sensory information, and health-care monitoring systems can assist patients and doctors by providing automatically annotated daily health reports. We present a set of extensions of switching linear dynamic systems (SLDSs) which provide the ability to capture the higher-order temporal structures within data and to produce more accurate results for the tasks such as labeling and estimation of global variations within data. The presented models are formulated within a dynamic Bayesian network formulation along with the inference and learning methods thereof. First, segmental SLDSs (S-SLDSs) produce superior labeling results by capturing the descriptive duration patterns within each LDS segment. The encoded duration models describe data more descriptively and allow us to avoid the severe problem of over-segmented labels, which leads to superior accuracy. Second, parametric SLDSs (P-SLDSs) allows us to encode the temporal data with global variations. In particular, we have identified two types of global systematic variations : temporal and spatial variations. The P-SLDS model assumes that there is an underlying canonical model which is globally transformed in time and space by the two associated global parameters respectively. Third, we present hierarchical SLDSs (H-SLDSs), a generalization of standard SLDSs with hierarchic Markov chains. H-SLDSs are able to encode temporal data which exhibits hierarchic structure where the underlying low-level temporal patterns repeatedly appear among different higher-level contexts. The developed SLDS extensions have been applied to two real-world problems. The first problem is to automatically decode the dance messages of honey bee dances where the goal is to correctly segment the dance sequences into different regimes and parse the messages about the location of food sources embedded in the data. The second problem is to analyze wearable exercise data where we aim to provide an automatically generated exercise record at multiple temporal and semantic resolutions. It is demonstrated that the H-SLDS model with multiple layers can be learned from data, and can be successfully applied to interpret the exercise data at multiple granularities.

Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 5 uppsatser.