61. Vpliv notranjih in zunanjih okoljskih dejavnikov na nastanek virusov HPVJana Sitar, 2017, undergraduate thesis Abstract: Humani virusi papiloma so eden najpogostejših vzrokov spolno prenosljivih bolezni in nastanka nekaterih vrst raka, kot so rak materničnega vratu, rakava obolenja na spolovilih ter rak glave in vratu. V diplomskem delu smo ugotavljali vpliv notranjih in zunanjih dejavnikov na nastanek in infektivnost virusov HPV-16, določali pa smo tudi strukturne lastnosti virusnega plašča. Kot notranji dejavnik smo uporabili gostiteljski protein SNX17, ki je ključen za okužbo HPV, kot zunanji dejavnik pa nanodelce titanovega dioksida, ki so pogosta sestavina kozmetičnih izdelkov. Pri pridobivanju virusnih delcev HPV-16 v celični liniji 293TT smo ugotovili, da protein SNX17 in nanodelci TiO2 ne vplivajo na količino virusnih delcev, vplivajo pa na njihovo infektivnost. Potrdili smo, da je protein SNX17 ključnega pomena za okužbo HPV, saj so imeli mutirani virusni delci AA2 bistveno zmanjšano stopnjo okužbe. Podobno smo ugotovili pri uporabi nanodelcev TiO2, kjer je bil upad infektivnosti sorazmeren koncentraciji uporabljenih nanodelcev med fazo nastanka virusnih delcev. Pri strukturnih lastnostih virusnega plašča smo analizirali količino in vrsto vključene reporterske DNA, stabilnost plašča in razmerje proteinov L1 : L2 v plašču. Ugotovili smo, da vezava plaščnega proteina L2 s SNX17 in prisotnost nanodelcev TiO2 na te lastnosti ne vplivata. Zaključili smo, da izbrani okoljski dejavniki, protein SNX17 in nanodelci TiO2 ne vplivajo na osnovne značilnosti nastanka virusnih delcev HPV-16, vplivajo pa na njihovo učinkovitost okužbe gostiteljskih celic.
Keywords: HPV, plaščni proteini, okužba, TiO2, SNX17
Published in RUNG: 17.11.2017; Views: 6461; Downloads: 241
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62. VPLIV ATMOSFERSKIH AEROSOLOV NA FIZIOLOŠKI IN VNETNI ODZIV MAKROFAGOVTina Čujec, 2017, master's thesis Abstract: Magistrsko delo obravnava vpliv aerosoliziranih delcev na fiziološki in vnetni odziv makrofagov J774.2. Makrofage smo v prvem delu izpostavili suspenziji filtra z delci črnega ogljika, v drugem delu pa smo postopek izpostavitve ponovili s suspenzijo filtrov, na katere so bili vzorčeni delci iz zunanjega zraka. V obeh primerih smo celice izpostavili različnim koncentracijam suspenzij in analizirali njihov vpliv po 24- urni ali 48- urni inkubaciji. Celično viabilnost smo določili z reagentom PrestoBlue, gensko stabilnost smo ugotavljali z imunofluorescenčno metodo (anti-fosfo-histon H2A.X), oksidativni stres pa smo kvantificirali z barvilom karboksi-H2DCFDA. Vnetni odziv izpostavljenih celic smo opazovali s testom fagocitoze in določanjem citokina TNFα (test ELISA). Rezultati so pokazali, da suspenzija črnega ogljika vpliva na celično živost makrofagov v odvisnosti od koncentracije dodane suspenzije. Dokazali smo povečan nastanek prostih kisikovih radikalov (ROS), znižano gensko stabilnost, ter povečan imunski odziv izpostavljenih celic. Delci črnega ogljika so bistveno povečali obseg fagocitoze in izločanje vnetnega citokina TNFα pri izpostavljenih makrofagih. Pri vseh testih se je odziv večal z višjimi koncentracijami suspenzije črnega ogljika.
Rezultati analiz s suspenzijo realnih vzorcev so pokazali, da je vnetni imunski odziv eden prvih odgovorov makrofagov na izpostavljenost okoljskim aerosolom. Opazili smo tudi večje število poškodb DNA izpostavljenih makrofagov, medtem ko je bila viabilnost teh celic še neprizadeta. Na osnovi rezultatov magistrske naloge smo zaključili, da okoljski aerosoli sprožijo vnetni odziv izpostavljenih makrofagov, negativno pa vplivajo tudi na njihovo gensko stabilnost, viabilnost in nastanek ROS. Predvidevamo, da je izraženost učinkov odvisna tudi od koncentracije in sestave okoljskih aerosolov.
Keywords: Makrofagi, črni ogljik, okoljski aerosoli, vnetni odziv, genotoksičnost.
Published in RUNG: 13.10.2017; Views: 5826; Downloads: 286
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64. Vimentin Modulates Infectious Internalization of Human Papillomavirus 16 PseudovirionsGeorgia Schäfer, Lisa M. Graham, Dirk M. Lang, Melissa J. Blumenthal, Martina Bergant Marušič, Arieh A. Katz, 2017, original scientific article Keywords: HPV16, vimentin, pseudovirions, infectious internalization
Published in RUNG: 17.08.2017; Views: 4308; Downloads: 248
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66. Characterizing the spatio-temporal role of sorting nexin 17 in human papillomavirus traffickingMartina Bergant Marušič, Špela Peternel, David Pim, Justyna Broniarczyk, Lawrence Banks, 2017, original scientific article Keywords: HPV, L2, intracellular trafficking, sorting nexin 17, capsid disassembly
Published in RUNG: 23.05.2017; Views: 4370; Downloads: 0
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67. The VPS4 component of the ESCRT machinery plays an essential role in HPV infectious entry and capsid disassemblyJustyna Broniarczyk, David Pim, Paola Massimi, Martina Bergant Marušič, Anna Gozdzicka-Jozefiak, Colin Crump, Lawrence Banks, 2017, original scientific article Abstract: Human Papillomavirus (HPV) infection involves multiple steps, from cell attachment, through endocytic trafficking towards the trans-Golgi network, and, ultimately, the entry into the nucleus during mitosis. An essential viral protein in infectious entry is the minor capsid protein L2, which engages different components of the endocytic sorting machinery during this process. The ESCRT machinery is one such component that seems to play an important role in the early stages of infection. Here we have analysed the role of specific ESCRT components in HPV infection, and we find an essential role for VPS4. Loss of VPS4 blocks infection with multiple PV types, suggesting an evolutionarily conserved critical step in infectious entry. Intriguingly, both L1 and L2 can interact with VPS4, and appear to be in complex with VPS4 during the early stages of virus infection. By using cell lines stably expressing a dominant-negative mutant form of VPS4, we also show that loss of VPS4 ATPase activity results in a marked delay in capsid uncoating, resulting in a defect in the endocytic transport of incoming PsVs. These results demonstrate that the ESCRT machinery, and in particular VPS4, plays a critical role in the early stages of PV infection.
Keywords: HPV, ESCRT machinery, infection
Published in RUNG: 08.05.2017; Views: 4472; Downloads: 271
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68. 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 in RUNG: 24.11.2016; Views: 5473; Downloads: 346
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69. Diclofenac removal by simulated solar assisted photocatalysis using TiO2-based zeolite catalyst; mechanisms, pathways and environmental aspectsSubhan Salaeh, Daria Juretic Perisic, Martina Biosic, Hrvoje Kušić, Sandra Babich, Urška Lavrenčič Štangar, Dionysios D. Dionysiou, Ana Lončarić Božić, 2016, original scientific article Abstract: The study explores the potential of immobilized TiO2-based zeolite composite photocatalyst (TiO2-FeZ)
made of commercial AEROXIDE TiO2 P25 and iron-exchanged zeolite of ZSM5 type (FeZ), for solar assisted
treatment of diclofenac (DCF), pharmaceutical included in the ‘‘watch list” during last prioritization in
water legislation by EU.
In this study the efficiency of applied photocatalytic treatment, solar/TiO2-FeZ/H2O2, of DCF water
solution was evaluated on basis of DCF removal and conversion kinetics, as well as the changes of
common parameters for assessing water quality. Hence, the changes in the removal and mineralization
of overall organic content, biodegradability, toxicity to Vibrio fischeri, dechlorination of DCF and
its formed by-products, were monitored during the treatment. The obtained data were correlated with
the evolution of DCF by-products, identified and monitored during the treatment by HPLC/MSMS
analysis. In order to estimate the influence of water matrix, all experiments were performed in the
presence of chloride or sulphate as counter ions. The obtained data revealed that degradation
mechanism of DCF by applied treatment process using immobilized TiO2-FeZ includes the adsorption
onto photocatalyst surface and consequent degradation. The contribution of homogeneous Fenton
reaction due to leached iron ions was found to be negligible. The adsorption and degradation pathway of DCF were influenced by the type of counter ions, which was reflected in the observed changes of
water quality parameters.
Keywords: Solar photocatalysis, TiO2-FeZ catalyst, Diclofenac, Degradation pathway, Biodegradability, Toxicity
Published in RUNG: 21.07.2016; Views: 5499; Downloads: 0
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