Novel methods for detection of bioactive substances and their effects in organisms and in the environmentTatjana Radovanović
, doctoral dissertation
Abstract: Since the concentration of bioactive substances and infectious agents in organisms and in the environment are low highly sensitive techniques such as: chromatography technology coupled with mass spectrometry (GC/MS, LC/MS and LC–MS/MS) and transmission electron microscopy (TEM) are needed for their detection. These techniques are highly sensitive, but time consuming, requiring use of expensive apparatus and large quantities of reagents and organic solvents which are harmful for the environment. Because there is a growing need for analysis of a large number of environmental samples it is necessary to develop new, so called vanguard methods that enable rapid and reliable screening of large numbers of samples in the shortest possible time. Analysis with such “screening” methods are often less accurate or even semi-quantitative, but nevertheless allow reliable identification of nonproblematic samples and in practice they limit the use of demanding classical analytical methods to only a few percent of all the samples.
Therefore, general objectives of the thesis were development of novel methods for sensitive, fast and cost effective detection of pharmaceuticals, viruses and viral particles in waters and biological fluids and for detection of their effects in organisms.
Novel methods were based on the combination of TLS (Thermal Lens Spectrometry), microfluidics and immunological methods such as ELISA. TLS as highly sensitive technique (allowing detection of absorbances of less than 10-6) coupled with microfluidic technology allows detection of very low analyte concentration, shorter time for analysis, higher sample throughput and low consumption of reagents. In such combination microfluidic technology can simplify or speed up antigen-antibody or enzyme-substrate interactions in bioanalytical systems. Decisive advantage of microfluidic systems lies in the fact that small dimensions of such systems, composed of capillaries and micro-reactors with dimensions from about 10 to 100 µm, significantly reduce diffusion time, which is inversely proportional to second power of distance.
However, highly sensitive detection techniques are needed in microfludic systems, because the amounts of analytes in detection volumes are generally small and optical interaction lengths are two to three orders of magnitude shorter than in conventional spectrometric techniques.
By combining microscopic TLS (TLM) with microfluidic technique it is possible to reach very low limits of detection and at the same time shorten ELISA analysis time from 20 h to 20 minutes as was described before in the literature for detection of BNP (brain natriuretic peptide). TLM furthermore allows measurements of extremely small volumes (sub-microliter) as well as fast signal response (milliseconds).
In this Dissertation specific goals were the development of new methods for detection of selected bioactive substances and infectious agents:
-iodinated contrast agents
-NGAL (neutrophil gelatinase associated lipocalin) as a new biomarker of contrast induced nephropathy (CIN)
-antibodies against human papilloma viruses (HPV) viruses and HPV-16 pseudovirions.
For the development of new method for detection of iodinated contrast agents chemical degradation of iodinated contrast agents was investigated as well, as a potential method for their removal from waste water.
For the determination of NGAL, a commercially available ELISA kit was used as the basis for method development. In the initial experiments the final product of the reaction of substrate with enzyme HRP (horse radish peroxidase) was transferred from microtiter plate into a microfluidic system, which served just for the sample transport to TLM detector on microchip. With comparable speed analysis we achieved LOD of 1.4 pg/mL which is 7 times lower in comparison to commercial ELISA test (LOD=10 pg/mL). For further development of the method for detection of NGAL with µFIA-TLM magnetic nanobeads were used as a solid support for primary antibodies of ELISA assay. By applying appropriate magnetic field the antibodies were kept in microfluidic system, which also enabled binding of NGAL, secondary antibodies and reaction of substrate with HRP. Developed method for NGAL detection with LOD of 2.3 pg/mL compares favorably with LOD for commercial ELISA tests (10 pg/mL) in standard microtiter plates and significantly reduces the analysis time. TLM in combination with microchip for NGAL detection reduces the duration of individual incubation steps (from one hour to 5 minutes) and at the same time shortens total analysis time from four hours for commercial ELISA test to 35 minutes allowing higher sample throughput. Analysis of real blood samples was also performed and it has shown good agreement between NGAL concentrations measured by magnetic nanobeads based µFIA-TLM with the concentrations measured by a commercial ELISA test. Such short analysis time of analysis and possible further optimizations are opening new possibilities for application of µFIA-TLM in medical diagnostics and clinical research. By using appropriate antibodies the method for developed NGAL detection could be easily adopted for detection of different pharmaceuticals or pollutants in environmental samples.
We have also developed a magnetic nanobeads based ELISA assay for detection of anti-HPV-16 L1 antibodies in the sera of HPV-16 infected women. To ensure the selectivity, HPV-16 pseudovirions were used as an antigen for anti-HPV-16 L1 antibodies, which were detected with secondary HRP labeled antibodies. Initially the ELISA assay for antibodies against HPV pseudovirions was performed on a microtiter plate and an LOD of 3.8 ng/mL was achieved by measurement on a microtiter plate reader. When performing a µFIA-TLM measurement of the final ELISA solution the LOD was reduced to 0.9 ng/mL. Similar to the method for NGAL detection based on magnetic nanobeads, these were used as solid support for HPV pseudovirions and after carrying out all the incubation steps of the ELISA test in microfluidic chip the final product of the reaction of substrate with HRP was detected on TLM. With magnetic nanobeads based ELISA assay with µFIA-TLM for measurement of antibodies against PsVs of HPV-16 virus an LOD of 0.6 ng/mL was achieved, which is six times lower in comparison to classic ELISA on microtiter plate. Furthermore, the analysis time was reduced from ten hours to 30 minutes. The novel method was successfully validated by analysis of real sera samples from women who were previously diagnosed for infection with HPV-16 virus.
For determination of iodinated MRI contrast agents we developed a new method based on the measurement of concentration of released iodide which allows indirectly semi-quantitative detection of concentration of iodinated contrast agents. For iodide release from parent molecule of contrast agent we applied a chemical reaction with Cu2+ ions in the presence of H2O2. Released iodide was first oxidized into iodine and then extracted into chloroform. Contrast agents degradation reaction showed 70 % of efficiency for removal of iomeprol, taking into account the 60 % overall efficiency of iodide oxidation and extraction. The extract was injected into microfluidic chip and iodine concentration was determined with TLM. Chloroform as organic solvent with low thermal conductivity and high temperature coefficient of refractive index is a good choice for TLM measurement due to high TLS enhancement factor, which theoretically provides 40 times higher sensitivity of TLM measurements as compared to water and a four time improvement in sensitivity for each milliwatts of excitation power, when compared to spectrophotometry. The developed µFIA-TLM method for indirect determination of contrast agents based on detection of iodine provides around 60 times lower LOD, with low reagent and sample consumption in comparison to spectrophotometry. The LOD of 18 ng/mL for iomeprol achieved with TLM is 16 times lower in comparison to LOD of 294 ng/mL for iomeprol determination with HPLC. In comparison to LOD of 133 ng/mL for iomeprol achieved with detection of released iodide by ion chromatography, µFIA-TLM enables around 7 times lower LOD.
HPLC and HPLC/MS analysis showed that the parent compounds is completely removed after 120 min. of chemical degradation and that different degradation products are formed by cleavage of one or two iodine atoms. By this we have shown that the applied chemical degradation is efficient for removal of iomeprol and could be applied for treatment of waste waters after further optimization and reduction of reaction time.
New analytical methods developed within this work provide limits of detection for the selected compounds which are significantly lower (up to 60 times) in comparison to conventional analytical techniques based on transmission mode measurements. At the same time the new methods allows shorter time of analysis and higher sample throughput for the purpose of fast screening methods. Magnetic nanobeads based µFIA-TLM ELISA assays developed within this work offer several advantages in comparison to commercial ELISA tests on microtiter plates such as: higher surface for antibody binding, lower reagent consumption, and shorter analysis time.
Although the TLS technique didn’t reach appropriate stage of development and applicability for routine chemical analysis, improved methods for detection of NGAL and antibodies against HPV viruses could be applied for clinical studies and development of commercial tests for detection of viruses or other bioactive substances, which are needed for diagnostic purposes in hospitals.
Keywords: ELISA, NGAL, PsVs, kontrastna sredstva, TLM
Published: 02.02.2017; Views: 2410; Downloads: 186
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DEVELOPMENT OF AN ANIMAL MODEL TO STUDY THE ROLE OF THE PHOSPHATASE PTPN22 IN THE PATHOGENESIS OF CHRONIC LYMPHOCYTIC LEUKEMIAÖznur Özlem Ibrahimoğlu
, 2016, doctoral dissertation
Abstract: A polymorphic variant of the phosphatase PTPN22 has recently emerged as a major risk factor for the development of multiple autoimmune diseases. The mechanism how this variant increases the susceptibility for autoimmune diseases is still unclear. A recent study by our lab showed that PTPN22 is markedly overexpressed in chronic lymphocytic leukemia (CLL), which is a common lymphoid malignancy characterized by the clonal outgrowth of autoreactive B lymphocytes. This study also showed that PTPN22 functions primarily as a negative regulator of B cell receptor (BCR) signaling, but can also positively affect the activity of certain downstream signaling pathways.
To understand the role of PTPN22 in the pathogenesis of CLL, we investigated PTPN22 expression in leukemias that develop in the Eµ-TCL1 transgenic mouse model. We observed that PTPN22 is highly overexpressed in these leukemias. We also evaluated expression of PTPN22 in normal murine B cell subsets corresponding to different stages of B cell development and differentiation. We detected high PTPN22 expression in B1 B cells, which are the normal counterparts of CLL B cells, and lower levels in marginal zone B cells, whereas PTPN22 was not expressed or was expressed at extremely low levels in most of the other investigated B cell subsets.We generated knockout mice with targeted disruption of PTPN22. We showed that PTPN22 is not expressed by any B cell subset in PTPN22-/- mice, including B1 B cells. To investigate the impact of PTPN22 deficiency on B1 and marginal zone B cell development and function, we performed immunophenotyping analysis of bone marrow, spleen and peritoneal cavity B cells from a large series of PTPN22-/- and age-matched wild type mice. A small but appreciable reduction in the percentage of marginal zone B cells and an increase in the percentage of B1 B cells in older PTPN22-/- mice was observed. These changes were consistent with the increased levels of natural antibodies typically produced by B1 B cells and lower levels of antibodies typically produced by marginal zone B cells in PTPN22 deficient mice. Altogether, these data suggest that PTPN22 deficiency results in reduced marginal zone and increased B1 B cell immune responses. Importantly, this phenotype is remarkably similar to the phenotype of SHP1-/- mice, although considerably milder. Since SHP1 is the main negative regulator of proximal BCR signaling in B cells, these findings suggest that one important function of PTPN22 could be to provide additional fine-tuning of the intensity of the BCR signal in B1 and marginal zone B cells. In support of this possibility, a modest but consistent increase in anti-IgM-induced calcium mobilization was observed in B1 B cells from PTPN22-deficient compared to wild type mice.To see how PTPN22 deficiency will affect leukemia development and behavior, we crossed PTPN22 knockout mice with E-TCL1 transgenic mice. PTPN22 deficiency accelerated the expansion of the malignant B cells in this model and resulted in earlier leukemia development in comparison to wild type E-TCL1 transgenic mice. These data are consistent with other recent studies from our group showing that a greater capacity to activate downstream signaling pathways upon engagement of the BCR with external ligand is associated with more rapid disease progression in both human CLL and the E-TCL1 model.
In conclusion, we show that PTPN22 is equally overexpressed in normal and leukemic B1 B cells derived from E-TCL1 transgenic mice, suggesting that PTPN22 overexpression is not due to the transforming event, at least in the case of murine CLL. The greater BCR signaling capacity of B1 B cells and the accelerated leukemia development in PTPN22-deficient mice further argues for an important role of BCR signals in determining the aggressiveness of CLL.
Keywords: PTPN22, BCR signalling, B1 B cells, marginal zone B cells, chronic lymphocytic leukemia (CLL), autoimmunity.
Published: 31.01.2017; Views: 1642; Downloads: 156
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Geomorphological characteristics of karst on contact between limestone and dolomite in SloveniaPetra Gostinčar
, 2016, doctoral dissertation
Abstract: Contact karst is a type of karst formed where allogenic waters from the surface influence the karst geomorphic system. Contact karst may be considered in both a strict sense and in a wide sense. In a strict sense, contact karst is the karst phenomena and forms influenced by the contact between a karstifiable rock and a non-karstifiable rock. In a wide sense, contact karst may also be the karst phenomena and forms influenced by the contact between two different karstifiable rocks, for example limestone and dolomite. This thesis focuses on the geomorphological characteristics of contact karst on limestone-dolomite contacts in Slovenia. The purpose of the research was to determine which processes contribute to the development of contact karst on the contact between limestone and dolomite, to define their dynamics, and to identify which surface and underground landforms are developed.
The spatial distribution of contacts between limestone and dolomite in Slovenia was determined in a GIS. Using existing lithological data as a data layer, the extent of carbonate rock cover in Slovenia was calculated. Carbonate rocks cover 47 % of Slovenia’s territory (27 % limestone, 14 % dolomite, and 6 % clastic carbonate or impure carbonate rocks). And, there are 1,353 limestone-dolomite contact lines in the country, totalling a length of 2,625 km.
Study areas were selected based on GIS analysis of the limestone-dolomite contacts. A total of 17 areas in Slovenia were studied in detail. Fieldwork at the study areas consisted of the collection and analysis of rock, sediment, and water samples, allowing each study area to be geomorphologically mapped.
General factors contributing to contact karst development on the lithological contact between limestone and dolomite were determined. The most important factor appears to be the characteristics of the inflow part, formed on the dolomite. Where dolomite functions as a karst rock, the water is dispersedly drained into the karst. In that case, the limestone-dolomite contact does not function as contact karst. Alternatively, where the dolomite functions as fluviokarst, a point recharge, or sinking stream, is formed. In that case, contact karst may be formed. The fluviokarstic character of the dolomite depends on its chemical and mechanical properties. The dolomite bedrock must be positioned at a higher elevation than the neighbouring limestone bedrock. To meet this requirement, dolomite beds, which in Slovenia are generally older than limestone and hence stratigraphically positioned below the limestone beds, need to be positioned above limestone by either folding that leads to inverse stratification, overthrusting, or by displacement along faults. Along faults, the dolomite is more prone to mechanical weathering due to tectonic crushing in addition to its chemical properties. Hence, contact karst is more likely to form at thrust contacts between thrust limestone and dolomite. Limestone-dolomite contact karst develops predominately at higher elevations due to increased precipitation (where allogenic inflow is higher) and greater frost action due to lower temperatures. Intense mechanical weathering of dolomite over limestone directly affects contact karst processes and significantly contributes to the spatial distribution of these types of surfaces. The location of the water table close to the surface is also a leading factor in limestone-dolomite contact karst formation due to enhanced border corrosion.
Landforms typical of contact karst were identified in the study areas during geomorphological analyses. However, they are not as clearly recognizable as those on contact between carbonate and non-carbonate rocks. The reason for this is the fact that allogenic waters from dolomitic catchment areas are by far not as corrosive as those from non-carbonate catchment areas.
Keywords: carbonate rocks, dolomite, limestone, contact karst, allogenic water, karst geomorphology
Published: 19.12.2016; Views: 2940; Downloads: 107
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Ecological niche relations of indigenous and invasive crayfish (Astacoidea) in SloveniaMartina Jaklič
, 2016, doctoral dissertation
Abstract: In this study, realised and fundamental niches were combined between studied ICS (Austropotamobius torrentium (Schrank, 1803), Austropotamobius pallipes (Lereboullet, 1858), Astacus astacus (Linnaeus, 1758)) and NICS (Pacifastacus leniusculus (Dana, 1852), Orconectes limosus (Rafinesque, 1817), Cherax quadricarinatus (von Martens, 1868)). The realised niche was described by population characteristics. P. leniusculus showed similar annual cycle than ICS, but on the other hand it showed higher population density, shorter breeding period, and better individual condition (estimated with SMI) comparing to the ICS. These are its advantages over ICS, which increase its potential for the invasiveness. The most pronounced this threat is to A. astacus, with which it has the most similar/ overlapping realised niche. Similar was observed comparing the ICS and NICS fundamental niches presented by selected physiological measurements (maximal food (Cmax) and oxygen consumption (Rn), concentration of haemocyanin in haemolymph (Hc), electron transport system (ETS) activity and potential growth scope (PGS). As a result of higher plasticity and adaptation to varied environmental conditions, NICS expressed different physiological response to temperature at cellular (ETS activity) and organism (Cmax, Rn, PGS) level. Similar fundamental characteristics (expressed with thermal optimums) were found between P. leniusculus and ICS, while optima for O. limosus and C. quadricarinatus were more narrow and higher (above 22 °C and 30 °C). Modelling of thermal niches showed differences mostly among species on cellular level (in ETS activity). This was a new approach and support for better estimation of success and proliferation of new invaders. Furthermore, C. Quadricarinatus, found for the first time as established population in temperate climate of Europe (in Slovenia in 2009), was used in this study, as a model organism, to show that the combination of fundamental and realised thermal niche, as well as ecophysiological and field research can constitute an important input to the model for estimating the risk associated with C. quadricarinatus. It is rarely possible to observe the development of new invaders in the new environment, therefore monitoring of the C. quadricarinatus through the three growth period (1-lag phase, 2-log phase, 3-stable phase), was unique example in the invasive biology. After extremely increasing the population in 2011 and 2012 (for 7.7 - 10.1 % per month) the population developed and stabilised in March 2012. Based on these high numbers of field and lab data, the TIRI (Thermal Invasive Risk Index) was developed to produce instant results that may be satisfactory for evaluation of different degrees of invasiveness. We found that the oxbow lake Topla had no lethal days (no. of days when water temperature is below or above temperature of death) in comparison with the adjacent rivers where the number of lethal days probably constitutes an important barrier for species spreading. Therefore, we concluded that C. quadricarinatus could potentially reproduce in the adjacent rivers, but survival of the brood is doubtful, especially due to previous results concerning critical min/max temperatures.
Keywords: potočni raki, jelševec, koščak, koščenec, signali rak, trnavec, rdečeškarjevec, sezonska dinamika, termalna toleranca, temeljna ekološka niša, realizirana niša, potencialna invazivnost
Published: 24.11.2016; Views: 2536; Downloads: 243
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Hydrogeological and speleological research of the spring cave Slatinski Izvor and its recharge are (Republic of Macedonia)Biljana Gichevski
, 2016, doctoral dissertation
Abstract: This thesis represents a comprehensive study, giving first extensive information on the hydrogeological and speleological characteristics of the spring cave Slatinski Izvor and its catchment area. The researched spring is located in the Poreče basin of West-Central Macedonia, in the river valley of Slatinska Reka. The wider area around the spring belongs to two tectonic units. The Slatinski Izvor spring is situated on the Pelagonian horst anticlinorium, which is composed of carbonate rocks. The second is the Western Macedonian zone, and it is composed of non-carbonate rocks. The spring represents a significant potential for water supply for the settlement Slatina. Prior to this research only some basic information about the karst aquifer in Precambrian dolomite marbles was available. Therefore, my goal was to ensure better understanding of the functioning of the karst system in the recharge area of the spring, assessment of its vulnerability to various pollution sources, as well as study of karst development of the area.
Because karst aquifers have unique hydrogeological characteristics and specific nature, they also require specifically adapted investigation methods. In order to reach the main goal, speleological investigations, hydrological methods, hydrochemical methods and tracer test were used. A monitoring network was combined with regular samplings for major and trace elements analysis, which enabled considerable advances in understanding the functioning of the karst system. The data from two-year period (December 2011 – November 2013) was compared and analyzed.
The controlling of the karst development in the study area mostly depends on the tectonic and geologic conditions. Incision of Slatinska Reka is the main controlling factor in cave development by lowering the base level of karst terrains. All investigated caves (Gorna Slatinska, Slatinska II, Ovčarska Peštera, Puralo, Slatinski Izvor) have “normal” epigenic karst development. Slatinski Izvor is the youngest cave. The Slatinski Izvor spring serves as a cave entrance for the same cave and, represents the outflow of groundwater from the karst system.
A conceptual model of the karst system of the study area was developed. The Slatinski Izvor spring has typical karst hydrological regime. The karst system is well developed by conduits and rains infiltrated into vadose zone flow rapidly towards the spring. The travel time of low mineralized water within the system corresponds well with the results from an artificial tracer test. Performed artificial tracer test confirmed that the Slatinski Izvor spring is recharged by allogenic stream. The dominant apparent flow velocity of 250 m/h, a single peak of the tracer breakthrough curve and more than 87% of tracer recovered indicate a rapid conduit flow and high vulnerability of the observed drinking water source. Analysis of spatial and temporal variations of physical and chemical parameters show that waters in the study area had different origin and were transmitted along different flow paths. The origin of the water of the Slatinski Izvor spring is from non-carbonate area, but the influence of mixed limestone-dolomite sequence in the carbonate area is significant. The results point out to short residence time of the water in the karst aquifer.
The anthropogenic impact in the study area is insignificant which is reflected in the good water quality. In order to preserve it, land surface zoning was performed in terms of groundwater and spring water vulnerability to pollution according to the hydrogeological research.
A comprehensive research with a combined use of various research tools presents an innovative approach and a new contribution to the Macedonian karstological science. Applied methods proved to be successful for to study of the karst system. Finally, the results of the study have clear applicative significance in terms of drinking water management.
Keywords: karst hydrology, cave, speleological investigation, hydrochemistry, tracer test, spring Slatinski Izvor
Published: 14.10.2016; Views: 15459; Downloads: 76
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Transformacija antičnih motivov v slovenski književnosti devetnajstega in dvajsetega stoletjaUrša Prša
, doctoral dissertation
Abstract: Antična mitologija že stoletja navdihuje evropski prostor ter se v njem na različne načine transformira in interpretira v različnih vrstah umetnosti. Antični motivi so odmevali v slovenski književnosti od srednjega veka pa vse do sodobnosti. Slovenska literatura jih je v različnih fazah svojega razvoja sprejemala ali zavračala ter iz njih oblikovala svoje motivno-tematske zasnove. Posebna pozornost te doktorske disertacije je posvečena tipom literarne recepcije, ki se je udejanjila v slovenskem slovstvu romantike in v dvajsetem stoletju, ko so antični liki in motivi postali simboli in prispodobe moderne subjektivnosti. Po krajšem pregledu tuje in domače znanosti o mitologiji s poudarkom na teoretski obravnavi mita v literaturi sledi opazovanje recepcije antične motivike v slovenskem in evropskem literarnem prostoru od antike naprej. Posebna pozornost bo namenjena motivom Medeje, Odiseja in Orfeja v izbranih delih slovenske književnosti 19. in 20. stoletja, saj je vsak lik s svojo univerzalnostjo in enkratnostjo pomembno vplival na slovensko literaturo. Ob pomoči medbesedilne metode so analizirani namen avtorjeve uporabe izbranega antičnega motiva, njegova vloga, interpretacija in morebitna transformacija ali aktualizacija.
Keywords: antični motivi, transformacija, slovenska književnost, Medeja, Odisej, Orfej
Published: 11.10.2016; Views: 2746; Downloads: 262
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THE AUTOBIOGRAPHICAL EXPERIENCE OF MIGRATION IN THE LITERARY WORKS OF WOMEN WRITERS OF THE SLOVENIAN LITERARY POLYSYSTEMMegi Rožič
, 2016, doctoral dissertation
Abstract: People have travelled and migrated since the early periods of history and the phenomenon of migration, defined as “the movement of large numbers of people, birds or animals from one place to another” (Oxford Advanced Dictionary, 7th ed.) has a history of hundreds of years (Pourjafari, Vahidpour, 2014) and is by no means new. But the way of travelling, migrating and mobility in contemporary reality does have many specific aspects and recently it has been studied in new ways, with new concerns. The experience of migration in the contemporary world is a fundamental characteristic of human societies. “It is a system in which the circulation of people, sources and information follows multiple paths. The energy and barriers that alter the course or deflect the contemporary patterns of movement have both obvious and hidden features. While nothing is utterly random, the consequences of change are often far from predictable” (Papastergiadis, 2000: 1). This unpredictability and multidirectionality has led to changes in approaches to the study of migration in recent decades. In the last few decades the study of migration has increasingly been accompanied by a tendency to study it on the individual level − at the level of personal life stories (Milharčič Hladnik, 2007). These are also markedly expressed in the medium of literature.
The present dissertation presents literary oeuvres of seven women writers, who thematise the autobiographical experience of migration, it offers an individually, woman-centred experience and view of migration. In selected oeuvres personal views are expressed on that experience, different strategies of coping with life in new realities and regarding relationships in these environments. The thematisation of the experience of migration in selected oeuvres is also connected with a problematisation of other concepts: the concepts of belonging, borders, nation-state, culture and language. It is also vitally connected with the personal identity construction of the lyrical subjects and literary characters and also because of the autobiographical character of selected literary works of the literary artists themselves. The experience of migration in selected literary works leads to unique identity formations, which in themselves combine elements of different cultural backgrounds and traditions. In their literary works these selected women writers also shape their relationship toward time and space dimensions, tradition and interpersonal relationships through the experience of migration.
The experience of migration, the relocation of the subject and a change of the geographical area in selected oeuvres, does not only represent a change of the geographical position: it also allows a deviation from other rigid and seemingly fixed and unvarying patterns and virtual realities that accompany human life. Migration can also present an alienation effect from traditionalisms and determinants that define human lives. In the literary oeuvres of these selected women writers, migration is only in part tied to the traditional concepts related to migrants, with the loss of roots and rupture with the place of origin. In the selected literary oeuvres, migrants are rarely considered to be uprooted and unable to find their anchor or confidently start a new chapter in their lives in a new environment. Migration is mostly connected with the possibility of expanding the horizon of insights and perspectives of looking at life, with the acceptance of its complexities, ambiguities and incompleteness. Mostly, the experience of migration is seen as a new, creative option, which opens and examines the wide range of other issues and dilemmas. The condition of uprooted loss has traditionally negative connotations, but in the selected oeuvres vagueness and fluidity allow a real insight into the real, complex nature of life and human existence.
Keywords: The experience of migration, women writers, Maruša Krese, Ifigenija Zagoričnik Simonović, Brina Švigelj-Mérat − Brina Svit, Gabriela Babnik, Stanislava Chrobáková Repar, Erica Johnson Debeljak, Lidija Dimkovska, literary polysystem, nomadic entity, locational feminism, fluid identity, transnationalism, transculturalism
Published: 11.10.2016; Views: 3385; Downloads: 347
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General framework for the conservation of historical rural landscape. Case study of Qadisha Valley in Lebanon.Bachir Zarif Keyrouz
, 2016, doctoral dissertation
Abstract: The international community has reacted to the challenges faced by historic cities and cultural landscapes. In 2011, UNESCO adopted the international recommendation on the Historic Urban Landscape in order to draw the attention of governments and local communities to the need to improve the management of historic urban areas. The aim of the present dissertation is to expand the UNESCO definition to Historic Rural Landscapes, and to explore how the proposed methodology could be applied to the management of rural areas. The dissertation takes the Qadisha Valley in Lebanon as an example, where some elements have already been recognised for their outstanding universal value. It is considered however that these historic-cultural elements are part of the larger context of the Qadisha Valley, which has its specificity and its qualities that merit being recognised. It is also considered that the management of the elements already recognised by UNESCO can only be successfully safeguarded if the management is understood in the context of the entire valley. Consequently, the thesis is articulated taking into account the following parameters:
1. UNESCO has already set the restoration plan to solve the problems of Qadisha Valley. Based on this plan, the question can be raised whether or not the rural planning in Qadisha is well managed at the present?
2. In case Qadisha Zone is not well conserved, what improvements could be brought about and implemented in order to safeguard the cultural heritage and historical rural landscape, and what should be the guiding policies of restoration?
3. Once a restoration plan is implemented in Qadisha Zone, what management plan should be brought up and implemented to safeguard its cultural heritage and historical rural landscape?
4. How can conservation of cultural heritage in Qadisha Zone be done culturally and in a socially acceptable way?
5. What sectors of the nation are to be involved in the Management plan? How can the MP preserve and restore Qadisha Zone?
The aim of the dissertation has been to examine the possibilities of involvement of the local community and relevant authorities in a more structured management of their properties within the context of the entire valley and its exceptional historic and landscape qualities. It is hoped that the study can generate more interest in the management of the rural context within the increasingly globalising society, and draw attention to the qualities of historic rural landscapes as the setting for an improved quality of life.
Keywords: Historic Rural Landscape, Qadisha World Heritage, Heritage Community, Sustainable Tourism, Strategic Urban Management, Outstanding Universal Values & Conservation Process.
Published: 07.10.2016; Views: 2952; Downloads: 48
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Building the shared dimension of the rural landscape: tools, principles and methodsCaterina Groli
, 2016, doctoral dissertation
Abstract: This research investigates the rural landscape of the lagoon of Venice and its surrounding territory focusing on the forms of exploitation of its natural resources, the transformations that occurred throughout the course of history and the consequent formation of its primary economical sector.
New economic sectors, new agricultural policies and other spatial and sectorial drivers transformed the primary sector itself which changed and adapted its productive schemes to the main economic patterns, heavily affecting the former rural asset of the landscape of the lagoon by changing its demographic, economic activities and land use practices.
The study carries out a systematic and detailed analysis of all economic, cultural and social features that have interacted and impacted this specific landscape, relying mainly on the comparison of historical and current land-use maps through three different historical spans: 1823-1840 (pre-industrialization phase), 1933-1966 (industrializing process, considered as a determining factor of the transformation of this landscape) and 2013 (present post-industrialized condition). The maps, combined with relative data, are compared with the aid of the Geographical Information System (GIS) computer software. As a result, a set of new maps has been obtained, which point out the historical landscape permanencies, defined through in situ surveys on the territory. A multitude of other archival documents from direct and indirect sources have been examined in order to build up a sound and detailed socio-economic context and the overall historical background.
Resources, goods and services provided by the rural landscape and its development are an important part of regional and EU policy instruments and also an essential factor of identity and growth for local communities. The consumption of resources, their availability and maintenance over time and accessibility through property regulations are all factors that shape the common and public dimension of the rural landscape, which represents another study challenge of this work.
This study proposes the establishment of the rural park of the lagoon of Venice, through the implementation of the multi-functional role of agriculture, for the fulfilment of two fundamental conditions of the peri-urban rurality: the need of safeguarding the landscape and its significance, by preserving the authenticity and integrity of the rural characters and the dynamism of the primary sector, which is strictly related to the survival of the rural landscape.
Keywords: Rural landscape of the lagoon of Venice, UNESCO Management Plan 2012–2018, European Landscape Convention, Common Agricultural Policy (CAP), Italian Constitution, Cultural Heritage and Landscape Code, authenticity and integrity of the rural heritage, intangible heritage, landscape evaluation, land-use cartography, Geographical Information System (GIS), fish-farming, multifunctional agriculture, new rural community, common pool resources, public goods, club goods, property rights, rural park
Published: 04.10.2016; Views: 2350; Downloads: 71
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SYMBIOSIS ECOLOGY OF SELECTED SCYPHOZOALucija Raspor Dall'Olio
, 2016, doctoral dissertation
Abstract: Scyphozoa with symbionts have an advantage in oligotrophic environments due to the additional source of nutrients provided by their symbiontic algae, just as corals have benefits from their symbionts. The literature, however, has thus far devoted far less attention to the association between scyphozoan hosts and Symbiodinium sp. than it does to corals. This thesis investigated the identity of symbionts from scyphozoan medusae (Cotylorhiza tuberculata, Phyllorhiza punctata and Cassiopea xamachana) using a phylogenetic approach. Two scyphozoan species, P. punctata and C. tuberculata, were sampled over the Mediterranean Sea, while Cassiopea xamachana was sampled in the Atlantic Ocean. Symbionts were identified from live medusae and their identity and phylogenetic relationships were determined by analysing two nuclear markers, ITS2 and 28S rDNA, from symbionts. Symbiodinium sp. sequences belong to clades A, B, and C based on markers ITS2 and 28S rDNA. Moreover, individual medusae host only one type Symbiodinium (A, B or C). Host species from the Mediterranean Sea hosted Symbiodinium from clade A and B (C. tuberculata) or only from clade A (Phyllorhiza punctata), while the host from the Atlantic Ocean (C. xamachana) hosted Symbiodinium from clade B or C. The phylogeography of C. tuberculata medusae was analysed using mtCO1. All the haplotypes sampled over the Mediterranean Sea were clustered together without any sign of phylogeographic structuring.
Keywords: Scyphozoa, Cotylorhiza tuberculata, Symbiodinium sp., symbiosis, Mediterranean Sea, phylogeography, CO1, 28S rDNA, ITS regions
Published: 03.10.2016; Views: 2923; Downloads: 276
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