Iskanje po repozitoriju

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Opis: In a developed world, water is used in a variety of installations and devices for the improvement of life standard. It is important for these elements to be suitably managed and maintained, otherwise they can present a risk to people's health. Although potable water from a public plumbing system coming via the water supply into an internal plumbing system normally is compliant with the regulations, the quality of water in an internal plumbing system often changes – water gets contaminated. There are several types of microorganisms that can grow in water. They carry a great potential for the growth and proliferation of bacteria from the genus Legionella that cause Legionnaires' disease and Pontiac Fever. Even the fact that Legionella infection has for several years been mentioned in the accident insurance conditions of insurance companies in relation to receiving the insurance fee as compensation for bed day, shows the extent and foremost the seriousness of the disease (5–15% mortality rate). The research part of the master thesis focused mainly on determining the presence of bacteria from the genus Legionella in the water of internal plumbing systems of healthcare facilities. During the entire research period of 8 years, 2,676 samples of cold and hot water were acquired from 23 healthcare facilities. The main purpose of the research was to determine the efficiency or adequacy of applied approaches, i.e. physical disinfection with heat and chemical disinfection with chlorine, in eliminating or reducing the amount of Legionella present in the water of selected healthcare facilities. Another aim was to evaluate the impact of softening potable water with polyphosphates on the proliferation of certain microorganisms in water, and to determine the by-products of chemical disinfection. Upon sampling, an organoleptic examination of water (appearance, odour), electrometric measurements of water temperature, and colorimetric measurements of free chlorine concentrations in water were performed. The data for the analysis of Legionella presence were acquired by water sampling and the isolation of bacteria found in the water. In order to acquire laboratory results of specific physical and chemical parameters, ion chromatography, gas chromatography, spectrophotometry, titration, and inductively coupled plasma – mass detector were used. Although water samples from only two healthcare facilities showed no presence of Legionella, the number of water samples with the presence of Legionella decreased for at least 25% from the beginning to the end of period set. Also, after certain general sanitary and technical measures a noticeable improvement within healthcare facilities after 2008 could be observed. Regarding the initial part of the set period, a more favourable relationship between positive and negative samples, a lower number of samples with highest concentrations of present Legionella, and a higher number of samples with lower concentrations of present Legionella were determined. The research has shown that the existing system of ensuring health-compliant water is fairly efficient; however, the results could be improved additionally by investing more into the preparation and renovation of systems. Due to a limited number of samples, a direct impact of adding polyphosphates to potable water could not be linked to the occurrence of Legionella. None of the samples showed an increased concentration of by-products of potable water disinfection. In its conclusion, the research has shown that the procedures after overheating and after chlorine disinfection result in a similar success. While filtration proved to be most efficient, from the perspective of Legionella infections the use of medical bathtubs can present a high risk to people's health, therefore such devices should be subject to more frequent maintenance and supervision.
Ključne besede: potable water, internal plumbing system, biofilm, health facilities, disinfection, Legionella
Objavljeno v RUNG: 05.09.2016; Ogledov: 5723; Prenosov: 277
Celotno besedilo (1,70 MB)

Opis: The karst ecosystem shows heterogeneity and variability of geology, hydrology, morphology and ecology in space and time. Chemical composition of water in karst plays a crucial role not only in dissolution of karst rocks and deposition but also in ecological processes. The heterogeneous nature of karst aquifers leads to difficulties in predicting groundwater flow and contaminant transport direction and travel times. For its position at the top of karst, epikarst represents the interface between surface and underground. Epikarst ecology, biodiversity and fauna have rarely been systematically studied. The typical absence of enterable passages makes necessary an indirect approach. In this research, epikarst biodiversity in relation to environmental conditions was studied in seven Alpine and Pre-Alpine caves, located at different altitudes, in Slovenia and in Italy: Snežna jama na planini Arto (1556 m a.s.l.), Jama pod Babjim zobom (860 m a.s.l.), Zadlaška jama (300 m a.s.l.) and Pološka jama (500 m a.s.l.), in Slovenian Alps and Pre-alps, and Grotta A del Ponte di Veja (600 m a.s.l.), Covolo della Croce (875 m a.s.l.) and Grotta di Roverè Mille (1005 m a.s.l.) in Lessinian Massif. In these caves, percolating water was monthly monitored for fauna in the period of one year. Temperature, discharge, conductivity and pH were measured at the same time, and water samples for the laboratory analyses of total hardness, concentrations of various ions (calcium, chlorine, nitrate, sulphate and phosphate) and dissolved organic carbon were taken. 66 aquatic and terrestrial, mostly hypogean, species were found. Aquatic fauna was dominant in all caves, with the exception of Covolo della Croce. Caves located at higher elevations harbour less diverse communities. No animals were found when discharge was very low and in correspondence of moonmilk formations. The most abundant and diverse group was Copepoda, with thirteen different species and 776 individuals at different maturity stadia, totally representing 61% of the community (between two and three copepod species per cave in Slovenia and between one and six per cave in Italy). The 62% of copepod species were stygobionts. Some copepod species (e.g. Speocyclops infernus) have wide ecological tolerance to environmental variables, other have more restricted tolerances (e.g. Lessinocamptus insoletus). With regard of copepods, there was a good agreement between the number of observed species and the total number of predicted species, confirming that the sampling method was appropriate and sampling complete. Correlations between the copepod community structure and measured parameters, obtained with Pearson correlation coefficient and principal component analysis (PCA) show different patterns. Presence of copepods was positively correlated with conductivity and dissolved organic carbon in Grotta di Roverè Mille, and with carbonate and nitrate in Pološka jama. In other caves not so high correlations were found between copepod species and measured parameters and between pairs of species. Investigations of the Alpine and Pre-alpine epikarst fauna can help to understand better the ecology of the epikarst fauna and its roles within the large range of different shallow subterranean habitats.
Ključne besede: Epikarst, Alpine and Pre-Alpine caves, percolating water, speleobiology, fauna.
Objavljeno v RUNG: 31.08.2016; Ogledov: 5341; Prenosov: 284
Celotno besedilo (2,00 MB)

Opis: With the adjustment of the Slovenian legislation with the European one we have an obligation to increase the reuse and recycling target for municipal solid waste by up to 65% by 2030 and gradual limitation of the landfilling of municipal solid waste to 10% by 2030 (Directive 2008/98 on waste, 2015). This represents a major challenge for municipalities in Slovenia. This study contains a plan for establishing new waste treatment facility in the Municipality of XY. As all new constructions in Slovenia, the presented facility needs to provide all the environmental protective measures predicted by the legislation. Beside the legislation, it is also important to ensure that new waste treatment facilities will have a low or no effect on the environment. Based on the Environmental Protection Act and the Construction Act of Slovenia, measures to ensure long term monitoring of the waste treatment facility as a case study in the Municipality of XY are presented. Before constructing the new facility, the track of “baseline” – a one-year monitoring of the following environmental parameters was crucial: groundwater, air dust deposit, condition of the soil on the area where new facility will operate in the future. The thesis points out the importance of the so-called track “baseline” monitoring of environmental parameters before new waste treatment facility will operate. This will be the basis for the introduction of operating monitoring for the new waste treatment facility in the future. Data for the analysis of presence of pollutants were acquired from groundwater, air dust deposit, soil sampling and containment of chemical pollutants. The following methods of analysis were employed in the acquisition of laboratory results of specific physical and chemical parameters: ion chromatography, gas chromatography, spectrophotometry, titration, inductive coupled plasma – mass detector, measurement with an electrode. It was determined that in the period between 2011 and 2012, the groundwater samples taken from three drilled wells containing pollutants as for example in the P-2 well there were adsorbable organic halogens - AOX, boron and mineral oils found. The results of the dust deposit show increased value of lead and zinc. More concerning are analysis of the soil samples where the quantity of copper at the MM2 measuring point were 530mg/kg, on 20 April 2012 the present critical level was 300 mg/kg. At the same measuring point (MM2) on the same day, the value of zinc was 650 mg/kg and almost reached the critical level of 720 mg/kg. The levels of heavy metals as cadmium, copper, lead and zinc in the soil samples taken at the MM2 and MM3 measuring points were at the warning level. All the detected parameters were evaluated and present a good basis for future operational monitoring of the environment in the area of the new waste treatment facility in the future. This case study has shown that planned steps in the improvement of the waste management in the Municipality of XY are crucial to protect the environment. The analysis and interpretation present an important basis for planning and monitoring a new waste treatment facility which will help to improve our goals and reduce landfilling to 10% and increace recycling by up to 65% by 2030 as predicted by the Directive on waste (Directive 2008/98 on waste, 2015).
Ključne besede: odpadki, krožno gospodarjenje z odpadki, podtalnica, zemljine, prašni delci v zraku, spremljanje stanja okolja.
Objavljeno v RUNG: 20.07.2016; Ogledov: 4587; Prenosov: 309
Celotno besedilo (3,32 MB)

Opis: Studies on rotavirus biology have always suffered for the lack of a reverse genetics system applicable to all genome segments and independent from the use of helper virus. One of the main reasons proposed to explain the difficulties encountered is the poor expression of the transfected cDNAs. In this work we investigated the role of 5’UTRs in rotavirus cDNAs expression in un-infected cells. We showed that in the 5’UTR of several (but not all) genome segments of rotavirus is present a 5’-terminal inhibitory motif (IM = 5’-GGY(U/A)UY-3’) that, independently from the ORF or the 3’UTR, downregulates both transcription and translation of rotavirus cDNAs when expressed exploiting T7 polymerase-encoding recombinant vaccinia virus. We mapped two mutations (insertion of a G upstream 5’UTR and the U to A mutation of the fifth nucleotide of IM) that are capable of making the inhibitory motif non-functional restoring a satisfying yield of protein synthesis. After the analysis of all genome segment 5’UTR of SA11 strain, we created two distinct sets of mutated rotavirus genome segments containing non-functional IM. We planned to employ these sets in the development of a plasmid-based reverse genetics system that exploit vaccinia virus-encoded T7 polymerase. In an attempt to produce in vivo biotinylated Tripled-Layered Particles (TLPs), we created a recombinant VP4 fused to the Biotin Acceptor Peptide (BAP) that, when co-expressed with the biotin ligase BirA, is efficiently in vivo biotinylated. We exploited recombinant vaccinia virus system to achieve high levels of VP4-BAP and the enzyme BirA in cells. Upon infection with rotavirus, VP4-BAP would be incorporated in the newly forming viral particles. In addition, we constructed a full-length genome segment 4 cDNA encoding the recombinant VP4-BAP to be used with a VP4 temperature sensitive rotavirus in order to generate a recombinant rotavirus encoding VP4-BAP. Thus, the results obtained allowed us to design two distinct possible ways of generating recombinant rotavirus. The first was a genetic strategy for the incorporation of an exogenous genomic segment, with the characteristic of being helper-virus free and applicable to all genomic segments. This was the consequence of the description of an inhibitory motif present in many viral mRNAs and the identification of two mutations that abrogate its inhibitory activity allowing expression of viral proteins. The second focused on the expression of a recombinant viral structural protein expressed in virus-infected cells for the consequent incorporation into newly made virions.
Ključne besede: rotavirus, UTR, reverse genetics, T7 polymerase, in vivo biotinylation
Objavljeno v RUNG: 08.06.2016; Ogledov: 4797; Prenosov: 284
Celotno besedilo (2,87 MB)

Opis: TDP-43 inclusions are important histopathological features of various neurodegenerative disorders, including Amyotrophic Lateral Sclerosis. However, the relation of these inclusions with the pathogenesis of the disease is still unclear. Various hypotheses have been proposed. For instance, it was suggested that the inclusions are (1) primary toxic species, (2) part of the normal cellular protective response to toxic intermediates and (3) responsible for the nuclear depletion of TDP-43. Understanding the relationship between TDP-43 aggregation and neurodegeneration is crucial for the eventual management of the disease. TDP-43 is a protein that has a marked tendency to unfold and become insoluble. In particular, its C-terminal end has a so-called “prion-like domain”, a sequence rich in Glutamine (Q) and Asparagine (N) that is involved in both the interactions with other proteins and the self-aggregation process. A cellular model of aggregation has been previously developed by our group, using the TDP-43 Q/N rich amino acid sequence repeated 12 times (12xQ/N) fused to EGFP reporter. The EGFP-12xQ/N cellular inclusions are capable of sequestering wild type TDP-43 both in non-neuronal and neuronal cells. In this study we went further with ALS modeling, creating a Drosophila model with EGFP-12xQ/N-induced aggregates. We show here that Drosophila melanogaster TDP-43 ortholog (TBPH) overexpression in Drosophila eye using GMR-Gal4 driver, is neurotoxic and causes necrosis and loss of function of the eye. More important, the neurotoxicity of TBPH can be abolished by its incorporation to the insoluble aggregates induced by EGFP-12xQ/N. This data indicates that aggregation is not toxic per se and instead has a protective role, modulating the functional TBPH available in the tissue. Notwithstanding the fact that aggregation is protective in presence of an excess of TBPH, we wanted to further understand the role of the aggregates in an environment where just the endogenous TBPH is present. For this purpose, we induced EGFP-12xQ/N transgene constitutively in CNS using ELAV-Gal4 driver. The flies were born and went through the larval stage without differing from control flies in any significant feature of their 8 phenotype. However, during aging the locomotion ability and survival rate of EGFP-12xQ/N flies were impaired. Interestingly, the climbing deficit was correlated with a physiological reduction in the endogenous TBPH levels. Thus, the aggregation, when coupled with low TDP-43 levels generates phenotypic consequences in our Drosophila model, probably due to a TDP- 43 loss of function. In sum, these data suggest that although the aggregates may be a result of neuroprotection in a context where TBPH is in excess, at a certain stage they become responsible for the pathology, likely due to the TBPH loss of function. If we consider that TDP-43/TBPH inclusions act as a sink for the newly formed soluble TDP-43/TBPH, the modulation of these inclusions could be used as a potential therapeutic approach, as this would restore the normal levels of TDP-43/TBPH and its function. Consequently, in the last part of the study we were interested to understand if the clearance of TDP- 43/TBPH aggregates could be an effective strategy to treat ALS, by recovering TDP-43/TBPH function. For this purpose, using the previously established cell-based TDP-43 aggregation models we analyzed aggregate clearance after treatment with several FDA approved drugs. Three of these drugs were found to significantly reduce aggregation through the proteasome pathway. Furthermore, one of the drugs (nortriptyline) was shown to rescue EGFP-12xQ/N dependent locomotor dysfunction in the Drosophila model. Altogether these data indicate that the clearance of TDP-43 aggregates may be a novel therapeutic strategy for ALS treatment.
Ključne besede: TDP-43/TBPH, aggregation, Drosophila melanogaster, ALS
Objavljeno v RUNG: 03.03.2016; Ogledov: 5413; Prenosov: 85
Celotno besedilo (53,65 MB)