Relational thinking and the cultural conditionality of human understanding

For Western researchers, the confrontation and understanding of so-called “non-Western” cultural traditions are intricately bound up with linguistic, cultural, historical, and socializational disparity issues. Indeed, the interpretation of various cultures is a process where not only the subject of the interpretation, i.e., the culture, is important, but also interpreter-related factors, e.g., in terms of geography, politics, and economy, play a role. A basic assumption of the present chapter is that Western epistemology embodies only one of the numerous diverse models of human thought and our perception of reality. This chapter presents a specific epistemology, which differs in its characteristics from the norms derived from Euro-American epistemological discourses. The basic paradigms of this model, which I call “relational epistemology,” have been developed in Sinic, especially Chinese, intellectual traditionsit is predicated on the idea of seeing the world as a complex network of interconnections. Such models for interpreting questions of knowledge were thus based on the compatibility of the structures of external reality on the one hand and the structure of the human mind on the other

Application possibilities of antisense oligonucleotides in inducing target gene expression with small activating RNAs

Aktivacija z RNA (angl. RNA activation, RNAa) je mehanizem, pri katerem kratka aktivirajoča RNA (angl. small activating RNA, saRNA) v sodelovanju s proteinom Argonaut 2 (AGO2) in kompleksom RITA, ojača izražanje tarčnih genov preko vezave na njihovo promotorsko regijo. Zaradi zmernega povišanja izražanja genov se zdi obetavna za uporabo v terapiji redkih nevrorazvojnih bolezni, kjer prizadeti geni delujejo v strogo reguliranem območju aktivacije in zato lahko patološki fenotip povzročajo tako ob znižanem kot tudi prekomernem izražanju. V magistrski nalogi smo na podlagi obstoječih saRNA p21 in CEBPA, ki so že v kliničnih poskusih, načrtovali enoverižne protismiselne oligonukleotide (angl. antisense oligonucleotides, ASO) in preverili, če z njimi lahko posnemamo delovanje saRNA. Protismiselni oligunukleotidi so namreč enostavnejši za dostavo v celice in manj imunogeni kot dvoverižne RNA. S protismiselnima oligonukleotidoma ASO p21 in ASO CEBPA smo uspešno aktivirali izražanje tarčnih genov p21 in CEBPA, kar smo preverili s kvantifikacijo mRNA tarčnih genov s qPCR in z detekcijo proteinov s prenosom western. Aktivnost saRNA je odvisna od vezave na protein AGO2, zato smo z metodami imunoprecipitacije in s površinsko plazmonsko resonanco (SPR) preverjali ali se načrtovani protismiselni oligonukleotid specifično veže na AGO2 ter vezavo z SPR tudi potrdili. Nato smo na podlagi obstoječih smernic načrtovali nove saRNA, ki ciljajo razvojno pomembne gene ABCD1, ZC4H2 in SYNGAP1 in CTNNB1, povezane z redkimi, neozdravljivimi monogenskimi boleznimi in spremljali njihovo aktivacijo izražanja s qPCR. Izmed petih novih načrtovanih saRNA smo s tremi kandidati na različnih celičnih linijah dosegli nizko aktivacijo izražanja genov, kar nam bo v prihodnjih raziskavah služilo kot osnova za nadaljnjo optimizacijo zaporedja in načrtovanje protismiselnih oligonukleotidov.RNA activation (RNAa) is a mechanism in which small activating RNA (saRNA) interacts with the Argonaute 2 (AGO2) protein and the RITA complex to enhance the expression of target genes via binding to their promoter region. Due to their moderately enhancing effect on gene expression they hold great promise for application to therapy of rare, neurodevelopmental diseases where the affected genes are tightly regulated to function in a narrow physiological range with both insufficient expression and overexpression leading to a pathogenic phenotype. In this thesis we aimed to design single-stranded antisense oligonucleotides (ASO) based on the existing saRNA p21 and CEBPA in clinical trials in order to overcome delivery issues and immunogenicity associated with double-stranded saRNA. We then monitored their effect on target gene expression to see if they can mimick the effects of saRNA. With antisense oligonucleotides ASO p21 and ASO CEBPA we successfully activated expression of target genes as determined by mRNA quantification with qPCR and protein detection by western blot. As the saRNA activity depends on their interaction with AGO2 protein, we used immunoprecipitation and surface plasmon resonance (SPR) to monitor specific interaction of ASO and AGO2 which we also managed to confirm using SPR. Finally, based on the existing guidelines we designed new saRNA candidates targeting developmentally important genes ABCD1, ZC4H2, SYNGAP1 and CTNNB1, associated with rare, incurable, monogenic diseases and monitor their target gene activation by qPCR. With three of the five designed new saRNA we were able to show low target gene activation which will serve as the basis for further sequence optimisation and eventual ASO design

Development and validation of a new method for determining trace elements in nail and hair samples

Lasje in nohti so lahko pomembni biološki vzorci za oceno pretekle izpostavljenosti elementom v sledovih, zato se vse pogosteje uporabljajo v kliničnih in epidemioloških raziskavah, v katerih ocenjujejo vpliv okoljskih dejavnikov na zdravje ljudi. Koncentracije elementov v sledovih v Laboratoriju za analitiko encimov in elementov v sledovih, Univerzitetnega kliničnega centra Ljubljana (UKCL), določamo v vzorcih krvi in urina. V okviru magistrske naloge smo razvili in validirali metodo za kvantitativno določanje koncentracij 26 elementov v sledovih v vzorcih las in nohtov, vključno s 7 esencialnimi in 19 neesencialnimi elementi. Na podlagi pregleda literature smo razvili metodo za določanje elementov v sledovih, ki vključuje predanalitske postopke (spiranje, sušenje in kislinski razklop v mikrovalovni pečici) ter analizo tekočih vzorcev z uporabo induktivno sklopljene plazme z masno spektrometrijo (ICP-MS). Metodo smo validirali po smernicah Inštituta za klinične in laboratorijske standarde (CLSI), pri čemer smo določili zmogljivost zaznavanja, natančnost, pravilnost, linearnost in delovno območje metode. Na podlagi validacijskih parametrov smo ugotovili, da metoda izpolnjuje zahteve za predvideno uporabo pri določanju elementov v sledovih v nohtih in laseh. Na vzorcih 140 zdravih oseb obeh spolov smo določili 95 % referenčne intervale za esencialne in neesencialne elemente, ki so bili primerljivi z literaturo, čeprav nekoliko ožji. V nadaljevanju smo ocenili vpliv eksogenih in endogenih dejavnikov ter življenjskih navad in razvad na koncentracije elementov v nohtih na nogi in laseh. Določili smo razlike v koncentracijah V, Mn, Co, Ni, Cu, Zn, As, Se, Sr, Mo, Ag, Cd, Sn med spoloma. S starostjo so se koncentracije večine elementov v vzorcih las in nohtov rahlo nižale. Koncentracije Hg v Obalno-kraški in Goriški regiji so bile povišane, pri ostalih elementih nismo določili pomembnih odstopanj med regijami. Uživanje prehranskih dodatkov je povečalo vsebnost Se v nohtih, uživanje morske hrane pa koncentracije Hg v nohtih in laseh. Kadilci so imeli višje koncentracije Cd in Pb v laseh in nohtih, koncentracije omenjenih elementov pa so korelirale tudi s številom pokajenih cigaret na dan. Lakiranje nohtov ni pokazalo statistično značilnih razlik v koncentraciji elementov, medtem ko je barvanje las vplivalo na koncentracije nekaterih elementov. Mn, Co, Ni, Sr in Ba so bili v umetno barvanih laseh višji, Cu, Zn, As, Se in Mo pa nižji glede na nebarvane. Nazadnje smo primerjali koncentracije elementov v sledovih v nohtih in laseh. Šibko pozitivne korelacije smo določili za elemente Mn, Co, Cu, Se, Al, Ga, As, Sn, Cs, Au V in Pb, močno pozitivno korelacijo pa za element Hg. Nova metoda določanja elementov v sledovih v nohtih in laseh je zanesljiva, vendar so potrebne dodatne usmeritve v standardizacijo in primerljivost rezultatov med različnimi laboratoriji ter opredelitev vpliva barvanja las ter endogenih dejavnikov na večjem številu vzorcev, saj so dosedanji rezultati med študijami nekonsistentni.Hair and nails can serve as important biological samples for assessing past exposure to trace elements, making them increasingly valuable in clinical and epidemiological studies evaluating the effects of environmental factors on human health. In the Laboratory for Enzyme and Trace Element Analytics at the University Medical Centre Ljubljana (UMCL), concentrations of trace elements are typically determined in blood and urine samples. In this master\u27s thesis, we developed and validated a method for the quantitative determination of concentrations of 26 trace elements in hair and nail samples, including 7 essential and 19 non-essential elements. Based on a literature review, we developed a method for trace element determination that incorporates pre-analytical procedures (washing, drying, and acid digestion using a microwave) and liquid sample analysis using inductively coupled plasma mass spectrometry (ICP-MS). The method was validated according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI), assessing detection capability, precision, accuracy, linearity, and working range. Based on the validation parameters, we determined that the method meets the requirements for its intended use in determining trace elements in nails and hair. In samples from 140 healthy individuals of both sexes, we established 95% reference intervals for both essential and non-essential elements, which were comparable to those in the literature, albeit somewhat narrower. We further evaluated the impact of exogenous and endogenous factors on trace element concentrations in toenails and hair. We observed differences in the concentrations of V, Mn, Co, Ni, Cu, Zn, As, Se, Sr, Mo, Ag, Cd, and Sn between genders. With age, the concentrations of most elements in hair and nail samples slightly decreased. Hg concentrations were elevated in the Coastal-Karst and Goriška regions, while no significant regional differences were observed for other elements. Supplement intake increased Se content in nails, and seafood consumption raised Hg concentrations in both nails and hair. Smokers had higher concentrations of Cd and Pb in their hair and nails, with these elements correlating with the number of cigarettes smoked per day. Nail polish did not show statistically significant differences in element concentrations, whereas hair dyeing affected the concentrations of certain elements. Mn, Co, Ni, Sr, and Ba were higher in dyed hair, while Cu, Zn, As, Se, and Mo were lower compared to undyed hair. Finally, we compared trace element concentrations in nails and hair. Weak positive correlations were found for Mn, Co, Cu, Se, Al, Ga, As, Sn, Cs, Au, V, and Pb, while a strong positive correlation was observed for Hg. The newly developed method for determining trace elements in nails and hair is reliable, although further efforts are needed to standardize and harmonize results between different laboratories, as well as to better understand the influence of hair dyeing and endogenous factors based on a larger number of samples, as current study results have been inconsistent

Development of immunoproteasome inhibitors and modulators of immune signaling pathways with anti-tumor and anti-inflammatory activity

Imunski sistem ima ključno, a kompleksno vlogo pri zaščiti organizma. V določenih okoliščinah lahko postane prekomerno aktiven ali premalo odziven, kar vodi v razvoj kroničnih vnetij, avtoimunskih bolezni in rakavih obolenj. Z zaviranjem pretiranega imunskega odziva lahko zdravimo avtoimunske bolezni, medtem ko lahko z imunomodulatorji, ki dodatno aktivirajo imunski sistem, okrepimo odziv proti okužbam ali rakavim celicam. V našem delu smo se osredotočili na razvoj novih imunomodulatornih učinkovin, s katerimi smo ciljali različne farmakološke tarče vpletene v patogenezo omenjenih obolenj: TLR7 in 8, TAK1 ter imunoproteasom. Načrtovanja in optimizacije novih učinkovin smo se lotili z različnimi pristopi. Sintetizirali smo serijo spojin z izoksazolo[5,4-d]pirimidinskim skeletom, kjer so najbolje delujoči analogi izkazali selektivno agonistično delovanje na TLR7 v nizko mikromolarnih koncentracijah ter povečali sproščanje vnetnih citokinov. Z načrtovanjem na osnovi fragmentov smo pripravili obsežno knjižnico 2-cianobenzotiazolov, benzimidazolov ter 1-metilbenzimidazolov z različnimi elektron privlačnimi ter elektron donorskimi skupinami vezanimi na obroč. Določili smo reaktivnost in stabilnost sintetiziranih derivatov ter razvili analoge, ki bodo koristni za prihodnje aplikacije v biokonjugaciji in kemijski biologiji. Pripravljeni analogi so izkazali selektivno zaviralno aktivnost do podenote ß5i imunoproteasoma z vrednostmi IC50 v nizkem mikromolarnem območju. Prav tako so izkazali sposobnost znižanja sproščanja citokinov ter selektivno citotoksično delovanje na celice multiplega mieloma. Kot prvi smo načrtovali, sintetizirali ter biološko ovrednotili himerne razgrajevalce (PROTAC) TAK1, ki so povzročili razgradnjo TAK1 v s TNF-? stimulirani celični liniji raka dojke ter monocitni levkemični celični liniji. Spojini, ki sta izzvali najvišjo razgradnjo TAK1, sta izkazovali signifikanten vpliv na zmanjšanje živosti omenjenih celičnih linij stimuliranih s TNF-?, ter izzvali celično apoptozo. Selektivno delovanje molekul PROTAC le pod vnetnimi pogoji predstavlja velik potencial za nadaljnji razvoj selektivnih terapij za različna obolenja povezana z neustreznim imunskim odzivom. Na vseh treh področjih smo dosegli zadane cilje in razvili nove selektivne modulatorje z aktivnostmi v nizko mikromolarnih ter nanomolarnih koncentracijah.The immune system plays an important but complex role in protecting the organism against patogens. Under certain circumstances, it can become overactive or underactive, which can lead to the development of chronic inflammation, autoimmune diseases, allergies, and cancer. Autoimmune diseases can be treated by suppressing an excessive immune response, while immunomodulators, which additionally activate the immune system, can strengthen the response against infections or cancer cells. In our work, we have focused on the development of new immunomodulatory agents for different pharmacological targets involved in the development of autoimmune diseases and cancers: TLR7 and 8, TAK1 and the immunoproteasome. We synthesized a series of compounds with an isoxazolo[5,4-d]pyrimidine scaffold, with the most potent analogs showing selective agonist activity on TLR7 at low micromolar concentrations and increasing the secretion of inflammatory cytokines. Using fragment-based design, we prepared a large library of 2-cyanobenzothiazoles, benzimidazoles and 1-methylbenzimidazoles with different electron-withdrawing and electron-donating substituents. We determined the reactivity and stability of synthesized derivatives and developed analogs that will be useful for future applications in bioconjugation and chemical biology. The prepared analogs showed selective inhibitory activity against the β5i-subunit of the immunoproteasome with IC50 values in the low micromolar range. They also showed a reduction in the release of cytokines and selective cytotoxicity on multiple myeloma cells. We were the first to develop, synthesize, and biologically study chimeric TAK1 degraders (TAK1 PROTACs), which induced TAK1 degradation in a TNF-α-stimulated breast cancer cell line and a monocytic leukemia cell line. The compounds that induced the strongest degradation of TAK1 showed a significant effect on reducing the viability of the aforementioned TNF-α-stimulated cell lines and caused cell apoptosis. The selective effect of PROTACs only under inflammatory conditions represents great potential for the further development of selective therapies for various conditions related to the improper immune response. To conclude, we achieved our goals in all three areas, and developed new selective modulators with activities at low micromolar and nanomolar concentrations

Informatization of container terminals in SZ - Tovorni promet

Diplomsko delo naj obsega glavne funkcionalnosti podane v specifikaciji podjetja SŽ-Tovorni promet za izdelavo spletne aplikacije za informatizacijo kontejnerskih terminalov. Aplikacija naj vključuje avtentikacijo uporabnika, urejanje pravic in vlog uporabnikov, upravljanje s podatki, možnost izvoza in uvoza podatkov, 3D vizualizacijo terminalskega prostora, ter avtomatsko sortiranje kontejnerjev. Poskrbite tudi za intuitivno uporabniško izkušnjo in izgled aplikacije.The thesis should cover the main functionalities given in the specification of the company SŽ-Tovorni promet for the development of a web application for the informatisation of container terminals. The application should include user authentication, editing of user rights and roles, data management, data export and import, 3D visualisation of the terminal space, and automatic container sorting. Make sure that the user experience is intuitive and pay attention to the design of the application

Correlating 0nubetabeta0nubetabeta decays and flavor observables in leptoquark models

In this paper, we investigate minimal scalar leptoquark models that dynamically generate neutrino Majorana masses at the one-loop level and examine their implications for low-energy processes. We show that these models can produce viable neutrino masses, consistent with neutrino oscillation and cosmological data. By using leptoquark couplings f ixed by neutrino data, we predict additional contributions to neutrinoless double-beta decays ($0nubetabeta$), which are chirality enhanced and compete with the standard contributions from the Majorana masses. Our analysis demonstrates that these effects are sizable for leptoquark masses as large as $mathcal{O}$(300 TeV), potentially increasing or decreasing the $0nubetabeta$ half-life, and creating an ambiguity between the normal and inverted mass ordering scenarios. Furthermore, we explore the correlation between $0nubetabeta$ and flavor observables, such as kaon decays and $mu to e$ conversion in nuclei, emphasizing that the latter is complementary to $0nubetabeta$ decays