BOOK OF ABSTRACTS: Cutting Edge – AT THE CROSSROAD OF THREE REALMS Organizator in založnik zbornika: Fakulteta za kemijo in kemijsko tehnologijo, Univerza v Ljubljani Kraj dogodka: Fakulteta za kemijo in kemijsko tehnologijo, Univerza v Ljubljani Uredniki: Jure Jakoš, Klara Klemenčič, Ines Babnik Oblikovanje in prelom: Jure Jakoš, Klara Klemenčič Organizacisjki odbor: Ines Babnik, Jure Jakoš, Klara Klemenčič AT THE CROSSROADS OF THREE REALMS The 2025 Cutting Edge Conference took place on 15 th September 2025 at the Faculty of Chemistry and Chemical Technology, University of Ljubljana. This year’s theme, At the Crossroads of Three Realms, showcased scientific research at the intersection of academia, industry, and public institutions. Through a rich program of lectures spanning chemistry, biochemistry, pharmacy, materials science, and related disciplines, the conference aimed to bring science closer to its participants while supporting them in shaping their current and future career paths. The event welcomed over 100 participants from Slovenia and abroad, representing diverse fields of research. Among them, 71 researchers presented their work within three thematic sessions: Science Behind the Living, Materials of the Future, and Technologies for the Earth and Environment . For the second time in Cutting Edge’s history, a special session titled Young Minds² was organized for secondary school students. A total of 32 young participants had the opportunity to present the outcomes of their school research projects and actively engage in the conference. The program began at 8:30 a.m. with opening remarks by prof. dr. Marko Novinec, the next Dean of the Faculty, and Ines Babnik, President of the Cutting Edge Society. The opening was followed by a series of lectures and poster presentations. The one-day conference concluded with an awards ceremony, where prizes were presented for the best poster and the best oral presentation. After the formal program, participants were invited to an afternoon networking session, offering opportunities to connect with sponsors and fellow researchers. We would like to express our sincere gratitude to the scientific committee members: doc. dr. Jakob Kljun, Uroš Rapuš, Jernej Imperl, Tilen Zorko and CONFERENCE SCHEDULE 8:00 Registration and poster installment Opening ceremony 8:30 Ines Babnik, prof. dr. Marko Novinec LabCore 8:45 Where there’s a will, there’s a way: Start-up in the pharmaceutical and chemical industry dr. Iva Hafner Bratkovič 9:15 Curiosity-driven immunology research inspires novel treatment strategies doc. dr. Luka Ležaić 9:35 From bench to bedside - clinical translation of radiopharmaceuticals in first-in-human trials 9:55 Coffee break Jafral 10:10 The use of bacteriophage technology to address upcoming health & biotechnology challenges dr. Matjaž Mazaj 10:40 Smart materials for a breathable future: Tackling CO₂ indoors dr. Matija Uršič 11:00 What we learn from burning things Lek Pharmaceuticals d.d. 11:20 From vision to reality: Setting up a new state of the art bioinjectables drug product production site 11:35 Coffee break TABLE OF CONTENTS AT THE CROSSROADS OF THREE REALMS 1 Ana Gotvajn 35 Maja Pristavec 2 Ana Šijanec 36 Maja Svete 3 Andrej Bogataj 37 Marcel Tušek 4 Azmat Ullah 38 Mark Loborec 5 Blaž Antonin 39 Martin Jazbec 6 Blaž Kozjek 40 Martina Potočnik 7 Brina Klinar 41 Matic Plut 8 Dane Jemc 42 Matjaž Dlouhy 9 Daša Čebulj 43 Natalija Tomažin 10 Domen Tomc 44 Neja Lesinšek 11 Endis Aletić 45 Nejc Virant 12 Filip Petrovič 46 Nina Krašovec 13 Helena Potočnik 47 Špela Žunec 14 Ines Babnik 48 Tadej Menegatti 15 Iza Rozman 49 Tadej Pirc 16 Jaka Janežič 50 Tal Čarman 17 Jan Hočevar 51 Tia Kralj 18 Jan Kogovšek 52 Tina Kosovel 19 Jan Ocepek 53 Tina Logonder 20 54 Tjaša Likeb Jan Vidergar TABLE OF CONTENTS YOUNG MINDS2 Aljaž Kostevc Redek, Danial Doustmohammadi, 68 Mihajlo Krstić 69 Brina Zver, Klara Grantaša 70 Eva-Arolea Trdan 71 Gregor Čekada, Niko Nolimal 72 Iva Jaklin 73 Jakob Ritlop 74 Jon Milič 75 Katarina Šela, Lara Marzidovšek 76 Lan Dular 77 Lara Širovnik, Petra Ouček 78 Luka Tomažin, Ajk Kalaba, Aljaž Maraž 79 Mija Kapun, Emilija Rojnik 80 Petra Ouček, Lani Habjanič 81 Tisa Lombar, Urban Perko 82 Urban Ocvirk 83 Vid Kodrič, Matic Izak, Anika Gregori Kmecl 84 Vinko Kosten Cold Plasma-activated Water (PAW): Reactive Oxygen and Nitrogen Species (RONS) Among Key Factors Contributing to PAW Antimicrobial Activity Ana Gotvajn*1, Helena Prosen2, Breda Jakovac Strajn1, Štefan Pintarič1 1 Veterinary Faculty, University of Ljubljana, Gerbičeva ulica 60, SI-1000 Ljubljana, Slovenija 2 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenija PAW is a new type of disinfectant that differs from most commonly used chemical disinfectants in that it does not contain any added chemical substances that would form degradation products of themselves after the antimicrobial effect, which could pollute the environment and pose a health risk. PAW is created when water is treated with plasma. Plasma is often defined as the fourth state of matter and is an ionised gas. When plasma interacts with water, chemical reactions take place that Scheme 1: Visual summary of preliminary studies lead to the formation of RONS. (Created in https://BioRender.com) The RONS formed, which are known to contribute to the antimicrobial activity of PAW, can be divided into It is known that the presence of impurities can reduce short-lived and long-lived species. The former, such as the effectiveness of the disinfectants used. Therefore, hydroxyl radicals (•OH), nitric oxide (NO) and future research will focus on the efficacy of the peroxynitrite (ONOO-), usually have a half-life of antimicrobial activity of PAW in the presence of organic nanoseconds to a few seconds and then tend to react and matter, which is in line with chosen standards for testing form more stable molecules. Long-lived species, such as the efficacy of chemical disinfectants. At the same time, hydrogen peroxide (H2O2), ozone (O3), nitrite (NO-2) and we will monitor the pH, ORP and concentration of nitrate (NO-3), have a half-life of minutes to days. selected RONS, thus contributing to a more Physical damage to the vital cellular structures and the comprehensive understanding of the role of key factors in induction of oxidative stress in the bacterial cell are two the antimicrobial activity of PAW. Identifying potentially fundamental principles of the antimicrobial effect of newly formed compounds after disinfection in the RONS. presence of impurities will contribute to evaluating the Lecithin Analysis: Determination of Phospholipids Ana Šijanec,1 Matevž Pompe*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Phospholipids (Figure 1) are amphiphilic molecules in lecithin without the use of reference standards. with different polar head groups and non-polar fatty acid Reversed-phase high-performance liquid tails that differ in length and saturation. As essential chromatography was coupled with high-resolution mass components of cell membranes, they form bilayers in all spectrometry to separate and detect the phospholipid living organisms. Their unique structural properties species. Molecular ions were recorded in full scan mode enable applications such as natural emulsifiers and and analyzed using a custom Python script that generated encapsulating agents in pharmaceutical, food and a list of possible phospholipid structures for each m/z cosmetic formulations, where they facilitate the formation value. The identification was confirmed by matching the of micelles and liposomes to improve the stability, MS/MS fragmentation patterns with the theoretical bioavailability and delivery of active ingredients1. A predictions. common natural source of phospholipids in the industry The proposed method provides a reliable, cost- is lecithin. The performance of lecithin in different effective alternative for phospholipid profiling in formulations is highly dependent on its phospholipid complex lipid matrices and demonstrates that standard- composition. Differences in head group type and fatty free identification is possible using high-resolution acid saturation affect the type of structures lecithin can analytical tools in combination with computational form and its emulsifying behavior. Therefore, support. characterizing the phospholipid profile of lecithin is essential to ensure the stability and functionality of the References: final product2 1. Xie, M. Phospholipids. In Encyclopedia of Food . Chemistry; Melton, L.; Shahidi, F., Varelis, P.; Eds.; Academic Press, 2019, 214–217. 2. Cui, L.; Decker, E.M; Phospholipids in Foods: Prooxidants or Antioxidants? J. Sci. Food Agric. 2015, 96. 3. McHowat, J.; Jones, J. H.; Creer, M. H.; Quantitation of Individual Phospholipid Molecular Species by UV Absorption Measurements. J. Lipid Res. 1996, 37 (11), 2450–2460. Role of Surface Pretreatment in Film Formation and Corrosion Protection of AZ-Series Magnesium Alloys Nina Kovač,1,2 1 1 1 Andrej Bogataj, Ingrid Milošev, Peter Rodič*, 1 Department of Physical and Organic Chemistry, Jožef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia 2 Jožef Stefan International Postgraduate School, Jamova cesta 39, SI-1000 Ljubljana, Slovenia Lightweight and biocompatible, the AZ-series Scheme 1: Experimental workflow for AZ series magnesium alloys, specifically, AZ31, AZ61, and AZ91, magnesium alloys with varying aluminium content have significant potential for biomedical and structural (X = 3, 6, 9 wt.%) and constant zinc content (1 wt.%). applications.1,2 However, their practical use remains The surface morphology and chemical composition limited by high corrosion susceptibility, particularly in were characterised using scanning electron microscopy chloride-rich environments. The corrosion behaviour of coupled with energy-dispersive X-ray spectroscopy magnesium is strongly influenced by (i) its composition (SEM/EDS), both pre- and post-exposure to a corrosive (content of aluminium and zinc) and (ii) the stability of its medium. Potentiodynamic polarization was used to assess surface oxide layer. Therefore, the surface conditions are corrosion behaviour, and it was evaluated in 0.1 M NaCl essential to enhance corrosion resistance.3 This study solution. examines the impact of mechanical surface pretreatment Potentiodynamic measurements revealed a lower and thermal oxidation on the formation of oxide films and corrosion current density and a more positive pitting the corrosion performance of AZ31, AZ61, and AZ91 potential for the thermally oxidised sample, which was alloys. supported by an improved surface morphology after The experimental workflow is outlined in Scheme 1. immersion. These findings highlight the critical role of Each alloy underwent three surface pretreatments: surface condition in passive film formation and (i) ground (e.g., AZ31_G, AZ61_G, AZ91_G), demonstrate that both thermal oxidation and increasing (ii) ground and polished (e.g., AZ31_P, AZ61_P, aluminium content in the alloy series (from AZ31 to AZ91_P), (iii) ground, polished, and thermally oxidised AZ91) contribute to enhanced corrosion resistance in at 200 ̊C (e.g., AZ31_TO, AZ61_TO, AZ91_TO). chloride environments. Future work could focus on investigating how Surface pretreatment different surface pretreatments influence the adhesion of protective surface coatings, enabling improved performance in diverse application environments. Grinding Photodegradation of pharmaceuticals using lignin carbon quantum dots based hybrid photocatalyst under visible light solar simulator Azmat Ullah,1 1 1 1 1 Jan Hočevar, Boštjan Žener , Iskra, Jernej, Urška Lavrenčič Štangar , Jelena Papan Djaniš*,1,2 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Centre of Excellence for Photoconversion, Vinča Institute of Nuclear Sciences, National Institute of the Republic of Serbia, University of Belgrade, 11351 Belgrade, Serbia Pharmaceuticals detected in water environments are posing a serious environmental concern for aquatic life and human beings1. Different methods are used for removal or degradation of pharmaceuticals from water including adsorption, membrane filtration, reverse osmosis and advance oxidation processes or photodegradation2. Among these methods, photodegradation is more promising because it requires only light sources to break down a range of complex Figure 1: a) CQDs/TiO2 photocatalyst and b) pharmaceutical structures 3. The photocatalysts have photodegradation of five pharmaceuticals mixture with major problems with electron hole recombination and CQDs/TiO2 wide band gaps of available efficient photocatalysts. Lignin derived CQDs as a green candidate for References: photocatalysis are extensively explored due to their high 1. Asadzadeh Patehkhor H.; Fattahi M.; Khosravi-Nikou surface area and photoluminescence properties4 M. Synthesis and characterization of ternary chitosan– . These CQDs can be synergistically coupled with the other degradation of tetracycline from pharmaceutical TiO2–ZnO over graphene for photocatalytic photocatalysts to lower their band gap and reduce electron wastewater Sci. Rep. 2021, 11 (1) 1–17. hole recombination like titanium dioxide (TiO2). TiO2 is 2. Jabeen A.; Salem Alsaiari N.; Katubi K. M. S.; Shakir used as a reference photocatalyst due to high efficiency, I.; Alrowaili Z.A.; Al-Buraihi M. S.; Warsi M. F. source due to wide band gap and electron hole photocatalytic materials for the removal of toxic pharmaceutical effluents M. J. Mol. Liq. 2024 , 405 , recombination also affects its efficiency. low cost and toxicity. But TiO Synthesis and characterisation of MXene based-LDH 2 is active in UV light In this study, lignin based CQDs were synthesized 3. Matoh L 125066. Synthesis and characterization of copper(I) complexes with chelating 2- (methylthio)pyrimidines Blaž Antonin 1 1 1 , Uroš Rapuš , Jakob Kljun*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Copper(I) complexes with 2,2’-bipyridine or achieved by using a salt with non-coordinating anion, structurally related N,N-ligands have attracted significant such as PF – 6. Crystal structures of some complexes were attention due to their promising cytotoxicity useful in determined and the geometries of the complexes cancer treatment, as well as interesting optical and synthesized were shown to be deformed tetrahedral. electrochemical properties 1. As copper(I) in most cases exhibits tetrahedral geometry, stoichiometric ratio between metal and bidentate chelating ligand can be either 1:1 or 1:2. In the former case, additional co-ligands must also be present and can provide a way to improve certain properties. One of them is solubility. Nonpolar groups can increase lipophilicity and thus increase bioavailability of compound, for which phosphine ligands are often used2,3. In this work, copper(I) complexes were prepared using 4-heteroaryl-2-(methylthio)pyrimidines, where heteroaryl groups were different heterocycles with nitrogen at position 2’, which provides a coordination mode analogous to 2,2’-bipyridine. Five new complexes from two sets, using six bipyridine-like ligands, were synthesized and Figure 1: Schemes, picture of the product and and an characterized by NMR, IR and UV-VIS spectroscopy, example of a crystal structure of prepared complex in mass spectrometry and CHN elemental analysis. Crystal stoichiometric ratio 1:1 (left) and 1:2 (right). Thermal structures of five of them were determined by X-ray ellipsoids were drawn at 50% probability level. diffraction on single crystal. chloride was reacted with the 1. Constable, E. C.; Housecroft, C. E. The Early Years N , N -ligand and of 2,2′ To prepare the first series of compounds, cuprous References: Intensification of the Carbonization Process Using Microbubbles Blaž Kozjek,1,2 prof. dr. Igor Plazl*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia (igor.plazl@fkkt.uni-lj.si) 2 Belinka Perkemija d.o.o. Zasavska cesta 95, SI-1231 Ljubljana – Črnuče. (ziga.kobal@belinka.si) The intensification of gas–liquid contact processes provides a controllable environment to study gas–liquid plays a central role in the development of sustainable interactions at the microscale and shows significant technologies for carbon capture and utilization (CCU). potential for integration into compact, modular CCU One promising approach is the use of microbubbles to devices. This study lays the groundwork for further enhance the interfacial area for gas–liquid reactions, experiments using CO₂ under similar conditions, where thereby improving mass transfer efficiency and reactor enhanced gas–liquid mass transfer can be quantified using performance. In this study, we investigated the formation the same framework. and stability of air microbubbles in a microchannel as a simplified model for future application in CO₂ absorption and utilization processes.1,3 A Carmin D2 water-powered injector was used to generate a two-phase bulk mixture of air and water in a mixing chamber. Due to the internal design of the injector, air was passively drawn into the flow under a pressure of ~5 bar, forming bubbles with diameters in the range of 30–70 µm. The two-phase mixture was then fed into a polymethyl methacrylate (PMMA) microchannel with dimensions of 2 cm × 0.5 cm × 500 µm, where bubble flow characteristics were monitored under a microscope. A key part of the study was evaluating bubble stability, size distribution, residence time, and the influence of surfactant addition. Tween 20 Figure 1: Visualization of air microbubbles flowing (polyoxyethylene sorbitan monolaurate) was used in through a 0.5 cm-wide microchannel. Conditions: 1% concentrations ranging from 0.1 to 3% w/w to assess its Tween 20, 3000 µL/min total flow rate. Bubbles remain impact on bubble integrity and persistence over time. stable and evenly dispersed, with an average diameter of Insight into the biodiversity of non-biting midges (Chironomidae) in Sečovlje Salterns through DNA barcoding Brina Klinar, 1 1 1 Eva Smrekar, Petra Tavčar Verdev, 1 Marko Dolinar*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Sečovlje salterns have a long history of producing sea The results indicate that the non-biting midges salt. Although the harvests nowadays are typically small infesting petola in the Sečovlje salterns likely belong to and strongly affected by climate and seasonal weather, the genus Chironomus (Table 1). Given that this species salt in Sečovlje is still produced using a traditional 3 is known to inhabit environments with higher salinity, it method that dates back to the 14th 1 century. is plausible that all of the analyzed specimens belong to Petola, a mineralized microbial substrate, plays a Chironomus salinarius. However, the data do not allow vital role in traditional salt production, serving as a us to conclude with complete certainty that all midges in foundation in salt crystallization ponds. Unfortunately, the Sečovlje salterns belong to this species. petola is not immune to pests. Insects such as brine flies (Ephydra) and non-biting midges (Chironomidae) are Table 2: Sequence analysis using BLAST and BOLD degradation of insects morphology, is indicated in the second column. petola and, consequently, a reduction in The results of the BLAST search are presented in the third both the quantity and quality of the harvested salt known to infest databases. The sex of the specimens, determined based on petola with their larvae, leading to 2 . column. The fourth column shows the percentage of In a previous study, the identity of the non-biting identity between the query sequence and the BLAST midges found in the Sečovlje salterns could not be barcode match. The results of the BOLD search are determined beyond the family level 1,2. In this study, an presented in the fifth column. The sixth column shows the effort was made to determine the genus of the specimens percentage of identity between the query sequence and the through DNA barcoding. BOLD barcode match. A total of 13 specimens, comprising insects of both sexes, were selected for analysis. After the isolation of genomic DNA, a specific region of the cytochrome oxidase I gene (COI) was amplified using PCR. This region is widely used as a barcode for species identification in animals. The resulting DNA fragment was inserted into the pJET vector, which was then cloned in Escherichia coli. After the plasmid isolation, the inserts Towards novel delivery agents for boron neutron capture therapy: Dane Jemc,1 Nejc Šercer,1 Filip Angelevski,1 Martin Gazvoda*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia, (martin.gazvoda@fkkt.uni-lj.si) Boron neutron capture therapy (BNCT) is a binary In addition, bioconjugation reagents based on closo-cancer treatment based on the ¹⁰B(n, α)⁷Li nuclear 1,7-dicarbadodecaborane (m-carborane) are being reaction. Non-toxic boron-10 and non-ionizing thermal developed. The reactivity necessary for conjugation with neutron beam have no therapeutic effect individually, but biomolecules can be reached by functionalizing the together they produce high-energy alpha particles and carborane cage with a suitable activating groups. There is lithium ions that travel only slightly less than a cell's potential for further tuning of reagent properties by diameter, thus allowing for highly selective energy introduction of substituents at the non-functionalized deposition and targeted tumor destruction.1 Currently, carbon. Synthesised reagents will be tested for sodium mercapto-undecahydro-closo-dodecaborate conjugation with selected biomolecular targets. (BSH) and L-4-(dihydroxyboryl)phenylalanine (BPA) are the primary boron delivery agents used in clinical BNCT applications. These however suffer from non-ideal tumor uptake and retention.2 Boronated amino acids are interesting, since the tumor uptake is usually high; on the other hand, the boron load is generally low.3 Incorporation of closo-carborane constituents into biomolecules such as peptides and antibodies is an attractive alternative for greater boron delivery; however, problems with lipophilicity and protein aggregation arise Figure 2: Synthetic principle for development of high when boron load is too high.4 boron load biomolecules containing carboranes. Borylation of small molecules can be achieved through halogenation of aromatic systems followed by References: transition-metal-catalyzed or organometallic substitution 1. Valliant, J. F.; Guenther, K. J.; King, A. S.; Morel, P.; reactions. Using this general strategy, we envisioned the Schaffer, P.; Sagbein, O. O.; Stephanson, K. A. The synthesis of a polyborylated phenylalanine derivative as medicinal chemistry of carboranes. Coord. Chem. Esterification of PEG derivate with succinic anhydride Daša Čebulj,1 Matic Rogan,1 Jerneja Kladnik,1 Damijana Urankar,1 Janez Košmrlj*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Succinic anhydride (Scheme 1, 2) is a cyclic oic acid (3) demonstrate that this approach is suitable for anhydride derived from succinic acid. It is industrially esterification of PEG-derived alcohols with succinic produced by catalytic hydrogenation of maleic anhydride. anhydride. The obtained compund shows potential for It plays an important role in organic chemistry as it easily further applications in materials chemistry. reacts with nucelophiles and partcipates in reactions such as hydration, amidation and esterification. Succinic anhydride is of great importance in various industrial processes, serving as a precursor in the production of surfactants, cosmetics, pharmaceuticals, and pigments1,2. Esterification with succinic anhydride is a promising approach in polymer and materials science. For example, Leszczyńska 3 et al. showed that surface modification of cellulose nanocrystals with succinic anhydride pruduces thermally stable succinylated cellulose nanocrystals that may be used as reinforcing agents in polymer composites. Additionally, Cai et al.4 prepared a polyethylene glycol (PEG) prepolymer by reacting PEG with succinic anhydride, followed by coupling with a prepolymer of bi(o-carboxyphenyl)adipate and salicylic acid (SA). The functionalized PEG polymer showed desirable properties Scheme 1: Esterification of PEG-derived alcohol (1) as a drug delivery system, such as high SA loading with succinic anhydride (2). efficiency and slow release rate of SA in gastrointestinal conditions. References: In this work, we explored the esterification of 1. Worberg, A. Succinic Acid and Succinic Anhydride. succinic anhydride ( In Kirk-Othmer Encyclopedia of Chemical 2 ) with a PEG-derived alcohol, tert - butyl(2-(2-hydroxyethoxy)ethyl)carbamate (1), to Technology; John Wiley & Sons, Ltd, 2025; pp 1–20. prepare an ester (3) with potential applications in 2. Succinic Anhydride - an overview | ScienceDirect Improving Repeatability of Gas Diffusion Electrode Measurements for Alkaline Water Electrolysis Domen Tomc 1,2 1,2 1,3 1,3 , Katja Jeraj , Miha Hotko , Nejc Hodnik* 1 Department of Materials Chemistry, National Institute of Chemistry, Hajdrihova ulica 19, SI-1000 Ljubljana, Slovenia 2 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 3 University of Nova Gorica, SI-5000 Nova Gorica, Slovenia The growing concerns over climate change and focusing on its efficiency, stability and repeatability as environmental degradation caused by carbon dioxide 4 can be seen on the figure below. emissions highlight the urgent need to transition toward sustainable energy sources. Among the promising alternatives to fossil fuels, hydrogen stands out due to its high energy density per mass and the fact that its use results in zero emissions. 1 Efficient electrocatalysts are required for hydrogen production via electrolysis. Under laboratory conditions, catalytic activity is typically studied using a rotating disk electrode (RDE), which enables controlled environments and straightforward material comparison. However, RDE has a significant limitation, poor mass transport of gaseous reactants to the catalytic site, which restricts the maximum attainable current densities and deviates from the conditions typical of industrial systems.2 As an alternative, gas diffusion electrode (GDE) Figure 1: Repeatability of Electrocatalytic Activity for systems are gaining prominence. These systems enable HER under Identical Conditions the direct diffusion of the gaseous phase through a porous layer to the catalyst, maintaining a high local References: concentration of the reactant and significantly improving 1. Roquea, B. A. C.; Cavalcanti, M. H. C.; Brasileiro, P. mass transport. GDE setups allow for higher current P. F.; Gama, P. H. R. P.; dos Santos, V. A.; Converti, densities, comparable to those in industrial systems, while A.; Benachour, M.; Sarubbo, L. A. Hydrogen- Development of SPE method for bisphenols Endis Aletić*,1 Helena Prosen1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Bisphenols are a group of organic compounds that the coefficients of variations were 1.12% for BPA and play a crucial role in the industrial development of 1.21% for BPS. The average extraction yields were 98.0% polycarbonates and (epoxy) resins as well as many for BPA and 92.4% for BPS (Figure 2). consumer products (e.g. thermal paper).1 On the other hand, they also represent a group of emerging pollutants in (micro)plastics that have several adverse effects on the human body as they are endocrine disruptors. In addition to human health risks, they also pose a challenge when it comes to maintaining healthy aquatic ecosystems. Their effects have been noted to play a role in fertility issues, various developmental and metabolic disorders as well as numerous other health issues.2 Our research was therefore dedicated to the development and optimisation of a solid-phase extraction method for bisphenol A (BPA) and bisphenol S (BPS). The former was the most problematic bisphenol, hence why it had been systematically replaced by BPS over the Figure 1: Determination of breakthrough volumes for past years. Unfortunately, it has since been discovered both compounds that the latter is no less harmful to humans.3 We studied the influence of different extraction parameters in order to achieve the highest possible yields. The influence of six parameters was tested, namely the suitability of the sorbent, choice and volume of elution solvent, varying pH values and ionic strength as well as whether the change in volume of MQ water (during the intermediate wash-out phase) affects the extraction yield. Furthermore, the breakthrough volume for both Genome-scale metabolic modelling of pancreatic ductal adenocarcinoma Filip Petrovič,1 Ana Halužan Vasle,2 Juan Solano,3 Sima Tozandehjani,3 Klementina Fon Tracer,3 Miha Moškon 2 , 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Faculty of Computer and Information Science, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 3 School of Veterinary Medicine, Texas Tech University, 7671 Evans Drive, Amarillo, Texas 79106, United States of America Pancreatic cancer is one of the most lethal malignancies, with a mortality rate nearing 95%. In a significant proportion of cases, tumors arise in the pancreatic ducts, classifying the disease as pancreatic ductal adenocarcinoma (PDAC). Due to the asymptomatic nature of early-stage PDAC and the lack of effective screening methods, diagnosis often occurs at advanced stages, limiting treatment options and contributing to poor patient outcomes1. A deeper understanding of the metabolic alterations associated with pancreatic cancer is crucial for elucidating disease Figure 1: Principle component analysis (PCA) results of progression. In this study, we utilized RNA constructed context-specific GEMs using the determined transcriptomic data from PDAC patients to construct optimal parameters context-specific genome-scale metabolic models (GEMs) using the fast task-driven integrative network inference for tissues (ftINIT) algorithm2. We systematically applied ftINIT to multiple input combinations to identify the optimal parameters for generating biologically relevant metabolic models. Furthermore, we investigated the metabolic impact of melanoma associated antigen A3 (MAGE-A3) overexpression in pancreatic cancer cells. MAGE-A3 is known to contribute to tumorigenesis and is Figure 2: Heatmap showing the most upregulated (blue) 3 associated with poor clinical outcomes ; however, its role and downregulated metabolic subsystems of MAGE-A3 in cancer cell metabolism remains largely unexplored. By positiv transcriptomic samples Selective separation of model compounds from aqueous solution using a microfluidic pervaporation device Helena Potočnik 1 1 , Rok Ambrožič *, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Currently, industries such as the chemical and Additionally, a mathematical model was developed to pharmaceutical sectors are irreplaceable components of better understand and describe the separation process. the modern economy. However, certain production strategies, for example the use of conventional metal catalysts, are gradually becoming obsolete and/or incompatible with the goals of establishing a more sustainable economy due to the depletion of limited natural resources. Therefore, there is an increasing emphasis on adopting more environmentally friendly solutions, for example bioprocesses that utilize enzymes also known as biocatalysts 1. Nevertheless, there is significant potential for improvement of bioprocesses, particularly in the downstream processes (DSPs), considering many production processes currently exhibit inefficiencies in separation and purification steps, which also tend to consume a substantial portion of the overall material and financial resources2a. One of the separation techniques applicable within DSPs for (bio)chemical processes is pervaporation. This membrane-based method enables the separation of multi- component mixtures and operates under relatively mild Figure 1: Molecular-level schematic of pervaporation in conditions, which can lead to reduced energy a microfluidic device consumption compared to other methods such as distillation3 References: . Described phenomena can also be enhanced through P.; Nopens, I.; Dejonghe, W. Investigating 1. Satyawali, Y.; Fernandez del Pozo, D.; Vandezande, the use of microfluidics, a valuable tool for process Pervaporation for In Situ Acetone Removal as Process Atom-economical synthesis and functional diversification of pyridazinone- type heterocycles Ines Babnik, 1 1 1 1 1 Nejc Petek, Jurij Svete, Uroš Grošelj , Bogdan Štefane *, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Pyrazolone-based heterocycles are valuable structural motifs in medicinal chemistry, agrochemicals, and functional materials. Due to their potent biological activity, they have been implemented as antimicrobial, antidiabetic, anticancer, anti-inflammatory, and antifungal 1 agents. Among pyrazolone-based heterocycles, bicyclic pyrazolonepyridazinone systems have emerged as promising enzyme inhibitors, for example as phosphodiesterase 4 (PDE4) inhibitors for the Figure 1: a: pyrazolo[1,2-b]phtalazines, b: pyrazolo[1,2- treatment of inflammatory disorders. 2 b]pyridazine-5,8-diones, c: pyrazolo[1,2- Despite a]pyridazinones. their biological value, some pyrazolopyridazinone derivatives remain challenging to References: synthesize, especially those of the [1,2]-type. While pyrazolo[1,2-b]phtalaiznes3 and pyrazolo[1,2- 1. Dubey, S. Bhosle, P. A. Pyridazinone: An Important 4 a ]pyridazine-5,8-diones are easily accessed via Element of Pharmacophore Possessing Broad multicomponent reactions, pyrazolo[1,2-a ]pyridazinones Spectrum of Activity. Med. Chem. Res. 2015 , 24 (10), are more difficult to prepare. This is due to the 3579–3598. susceptibility of tetrahydropyridazinones to ring 2. Biagini, P., Biancalani, C., Graziano, A., Cesari, N., methodology for the synthesis of pyrazolo[1,2-In this work, we present a novel one-pot Vergelli, C., Crocetti, L., Delcanale, M., Armani, E., Rizzi, A., Puccini, P., Gallo, P. M., Spinabelli, D., contraction under various conditions. Giovannoni, M. P., Cilibrizzi, A., Piaz, V. D., 5 Caruso, P. Functionalized Pyrazoles and a]pyridazinones under mild conditions and with excellent Pyrazolo[3,4-d]Pyridazinones: Synthesis and atom economy. The one-pot procedure enables efficient Evaluation of Their Phosphodiesterase 4 Inhibitory synthesis of desired compounds from easily accessible Activity. Bioorg. Med. Chem. 2010, 18 (10), 3506– starting materials, while bypassing the problems of ring- 3517. From cells to genes: Evaluating the (geno)toxic potential of ferrite-based nanoparticles Iza Rozman*,1,2 1,2 3 3 4 Alja Štern, Álvaro Gallo-Cordova, María del Puerto Morales, Gerardo F. Goya, Bojana Žegura 1,2 , 1 Biotechnical Faculty, University of Ljubljana, Jamnikarjeva ulica 101, SI-1000 Ljubljana, Slovenia 2 National Institute of Biology, Večna pot 121, SI-1000 Ljubljana, Slovenia 3 Instituto de Ciencia de Materiales de Madrid, ICMM, CSIC, Spain 4 Institute of Nanoscience and Materials of Aragón (INMA), CSIC-University of Zaragoza, Zaragoza, Spain Spinel ferrites (SF) are metal oxides with the general consistently downregulated across all samples after 24 formula AB₂O₄, where metal cations A and B occupy hours. tetrahedral and octahedral sites within the crystal These findings contribute to a better understanding of structure. They have attracted significant attention due to the (geno)toxic potential of SF NPs and highlight the need their wide range of applications, including biomedicine, for further investigation into their safety. water treatment, and electronic devices 1. Compared to their bulk counterparts, nanomaterials (particles <100 nm), including SF, exhibit superior properties due to their small size and high surface-to- volume ratio, which enhances their reactivity, thermal stability, mechanical strength, and magnetic behaviour 2. However, data on their safety and potential risks to human health remain limited. In this study, we evaluated the potential cyto- and genotoxic effects of three spinel ferrite nanoparticles (SF NP): γFe2O3, Zn0.7Fe2.3O4, and Mn0.4Fe2.6O4. Toxicity was assessed in vitro using a 3D cell model (spheroids) derived from the HepG2 human liver cancer cell line. Cell viability was measured with the CellTiter-Glo assay, while DNA damage was evaluated using the Comet assay. In addition, reactive oxygen species (ROS) levels were detected with the DCFH-DA fluorescence probe, and Use of HEO as catalysts in Fenton process for treatment of wastewaters Jaka Janežič*,1 Maša Legan,1 Andreja Žgajnar Gotvajn 1 1 University of Ljubljana, Faculty of Chemistry and Chemical Technology, Večna pot 113, SI-1000 Ljubljana, Slovenia High-entropy oxides (HEOs) are a recently discovered group of high-entropy materials (HEMs)1. These one-phase crystalline materials consist of five or more different metal components2. HEOs are structurally complex and can have a variety of useful and unique properties. Due to high tunability, rich magnetic properties, electrical properties and more2, they have attracted significant interest in fields such as energetics, electro-catalysis and catalysis1,2, although their Figure 1: MB solution before (left) and after (right) the capabilities as new materials need to be studied further. Fenton process involving HEO, Fe and H2O2. Therfore, the main idea of the work was to implement HEOs into the Fenton process with the intention of improving it. The Fenton process itself is a type of advanced oxidation process (AOP)3. That means it generates hydroxyl radicals which are used for wastewater treatment. This is achieved by adding peroxide (H 2+ O ) and iron ions (Fe) into the wastewater 2 2 with acidic pH3. The HEO we used for the experiments was synthesised using combustion synthesis and had a composition of (Mg0,2Co0,2Ni0,2Cu0,2Zn0,2)O. As a wastewater substitute we decided to use a 300 mL solution of Methylene blue (MB) with concentration of 10 mg L-1. Firstly, the adsorption properties of HEO were studied. Through various experiments it was concluded, Figure 2: Removal efficiencies for experiments with and that the best results were achieved in a narrow beaker with without added HEO at pH=2. an anchor type mixer at 160 rpm (removal percentage of Phenolic Azobenzenes from Lignin Precursors: Toward Sustainable Photoactive Antimicrobials Jan Hočevar 1 2 1 , Estelle Leonard, Jernej Iskra*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Université de Technologie de Compiègne, ESCOM, TIMR, F-60200 Compiègne, France Lignin is a complex, aromatic biopolymer found in characterised and their antimicrobial activity against plant cell walls, where it provides structural support, various G+ and G- bacterial strains was investigated water resistance and protection against decay. As the (Scheme 1). second most abundant natural polymer after cellulose, it is an important by-product of the pulp and paper industry, but unfortunately most of it is thrown away. 1 Globally, around 50,000,000 tonnes of lignin are produced annually and this is expected to increase to 225,000,000 tonnes by 2030. However, less than 2 % of the total lignin produced is actually utilised. Due to its richness in phenolic structures, lignin offers great potential as a renewable source for the production of valuable molecules, such as azobenzenes. Azobenzenes are organic compounds that have an azo group (-N=N-) linking two benzene rings. Their unique properties are due to a conjugated π-electron system, which significantly influences their optical, electronic and structural behaviour. Of particular interest is their reversible trans–cis photoisomerisation, which makes them useful as photosensitive materials.2 It is because of their unique properties that we consider Scheme 1: The pathway from lignin-based aromatics azobenzenes not only as dyes, but also in everyday and their conversion to azobenzenes and their applications, in particular in molecular electronic and antimicrobial efficacy. photonic devices, medicinal chemistry, photo- pharmacology, drug delivery, etc. Innovative LexKan Selection System for the Discovery of Dimerization Inhibitors Targeting SARS-CoV-2 Main Protease Jan Kogovšek 1 1 , Marko Novinec*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia The SARS-CoV-2 main protease (Mpro, or 3CLpro) is a versatile platform for discovering molecular binders, a cysteine protease responsible for cleaving viral providing a foundation for the development of targeted polyproteins into individual, functional proteins required antiviral strategies. for viral replication. Mpro is active only as a homodimer, making dimerization an essential prerequisite for its enzymatic function. Because its structure and function are highly conserved among coronaviruses, Mpro has emerged as a promising target for antiviral drug development. Traditional inhibition strategies have focused on active site binding, but targeting dimerization offers an alternative approach that may increase specificity and limit resistance1. To address this, we explored the use of affibody Figure 1: (A) Schematic representation of the molecules, small engineered proteins derived from the experimental workflow. A diverse affibody library is Z expressed in E. coli and subjected to selection on medium ‑ domain of Staphylococcal protein A. This domain is naturally involved in binding immunoglobulins, and containing kanamycin using the LexKan system. Selected through protein engineering, it has been repurposed into a clones are expressed, purified by Ni²⁺ affinity versatile scaffold capable of high-affinity binding to a chromatography, and analyzed for the interaction with wide range of target proteins. Affibodies are highly stable, Mpro. (B) Principle of the LexKan selection system. In the non-immunoglobulin binders that are easily produced in absence of a specific affibody, Mpro dimerization enables bacterial systems, making them ideal candidates for LexA-mediated repression of the kanamycin resistance interfering with protein pro gene, preventing bacterial growth. When a specific – protein interactions such as M dimerization 2 pro affibody disrupts M dimerization, repression is . We developed and optimized the LexKan selection relieved, allowing transcription of the resistance gene and system in bacterial survival on kanamycin-containing medium. Escherichia coli to identify affibody binders Characterization of lanthanum doped nickel oxide thin films for oxygen evolution reaction Jan Ocepek*, 1,2 2 1 1 1 Romana Cerc Korošec , Marjan Bele, Nejc Hodnik, Angelja Kjara Surca, 1 National Institute of Chemistry, Hajdrihova 19, SI-1000 Ljubljana, Slovenia 2 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia The Ni(II)/Ni(III) redox couple has long been studied M KOH in a custom-made glass cell with quartz within the framework of the Bode scheme, which maps windows. The potential range employed (0.8 to 1.8 V vs. the electrochemical behavior of nickel hydroxides and RHE) scanned the Ni(II)/Ni(III) redox transformation, as oxyhydroxides through formation of α-Ni(OH)₂/γ- well also the OER region. Measurements were performed NiOOH β-Ni(OH)₂/β-NiOOH phases [1,2]. This redox for pure α-Ni(OH)₂ film and for those with intercalated system gained attention for its reversible charge storage lanthanum (2, 5 and 10 mol.% La) (Graph 1). The properties, making it a key component in Ni-based behavior of powders was also examined using evolved batteries (e.g., Ni-Cd and Ni-MH) and electrochromic gas analysis (EGA) and Raman spectroscopy. (EC) devices. EC devices, composed of two transition oxide films and electrolyte, show distinct color change upon oxidation/reduction [1]. More recently, interest has shifted toward the role of the Ni(II)/Ni(III) transition in electrocatalysis, particularly for the oxygen evolution reaction (OER) in alkaline media [2]. Modern studies explore how structural transformations and phase evolution in the Bode scheme affect catalytic OER activity and stability, also through incorporation of additional elements like Fe, Co, Mn etc. in Ni-based samples. Concurrently, we leaned on the sol-gel synthesis of α-Ni(OH)₂ electrochromic films which showed improved cyclovoltammetric cycling stability after the addition of lanthanum [1]. The films were deposited on FTO glass using the dip coating technique from sols that were synthesized from nickel(II) sulphate heptahydrate. Films Tracking individual degradation mechanisms for Pt-Co nanoparticles Jan Vidergar*,1,2 Ana Rebeka Kamšek*,1 Francisco Ruiz-Zepeda,1 Anja Logar,1,3 Goran Dražić,1 Nejc Hodnik1,3 1 Department of Materials Chemistry, National Institute of Chemistry, Hajdrihova 19, 1000 Ljubljana, Slovenia 2 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, 1000 Ljubljana, Slovenia 3 University of Nova Gorica, Vipavska 13, 5000 Nova Gorica, Slovenia An identical-location STEM (Scanning transmission emerging as the dominant mechanism under the tested electron microscopy) approach was used to track 1 conditions. individual Pt–Co/C nanoparticles before and after electrochemical activation. Nanoparticles in the same field of view were segmented and then paired between the initial and post-activation images using the Hungarian assignment 2 algorithm, yielding a one-to-one correspondence for each particle (Figure 1). Each matched (or unmatched) nanoparticle between images was defined as an event, representing a specific outcome of degradation or stability1. Events were categorized via a decision-tree scheme Figure 1: Matched Pt-Co Nanoparticles Before and (Figure 2) that considered the number of particle masks in After Catalytic Treatment an event (e.g., a single mask if a particle disappeared or appeared, multiple masks if particles merged) as well as changes in particle size (equivalent diameter) and shape (circularity)3. Based on these criteria, each event was assigned to a degradation mechanism category (or marked as no significant change). Degradation modes such as dissolution, detachment, attachment, agglomeration, and Ostwald ripening were thereby distinguished. Particle properties like diameter and circularity were extracted from the IL-STEM images to quantify morphological Mechanical and Biological Evaluation of Alginate-TEMPO Cellulose Hydrogels Jan Vidergar*,1 2 2 2 2 1 Boštjan Vihar , Monika Belak, Ajda Godec, Uroš Maver , Tilen Kopač 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Institute of Biomedical Sciences, Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000 Maribor,Slovenia Hydrogels based on alginate and TEMPO-oxidized tailor mechanical characteristics for specific cell types cellulose have gained increasing attention due to their and applications in regenerative medicine and tissue biocompatibility and tunable mechanical properties1,2. scaffolding. Alginate, a naturally occurring polysaccharide, forms ionically crosslinked networks in the presence of divalent cations such as Ca²⁺, providing a versatile matrix for biomedical applications4. The addition of TEMPO- oxidized cellulose nanofibers (TOCNF) has been shown to influence the structure and stiffness of these materials through hydrogen bonding and electrostatic interactions1,2. This study investigates the mechanical and biological properties of hydrogels composed of alginate and 3% TEMPO-oxidized cellulose nanofibers (TOCNF), combined in various ratios. Hydrogels were prepared by mixing alginate gels of different concentrations with TOCNF gels at a fixed 3% loading, followed by crosslinking with calcium ions (Ca²⁺). Mechanical Figure 1: Tensile strength of 7.5% alginate and 3% properties were evaluated via tensile testing, revealing a TEMPO-oxidized cellulose hydrogels across different nonlinear relationship between the alginate-to-TOCNF mixing ratios. ratio and tensile strength (Figure 1). Optimal mechanical References: performance was observed at intermediate compositions. 1. Siqueira, P.; Siqueira, É.; de Lima, A. E.; Siqueira, G.; Consistent with previous findings3,5, the inclusion of Pinzón-Garcia, A. D.; Lopes, A. P.; Segura, M. E. C.; TOCNF improved the elastic modulus and reinforced the Isaac, A.; Pereira, F. V.; Botaro, V. R. Three- network structure, particularly at higher alginate content. Dimensional Stable Alginate-Nanocellulose Gels for The influence of copper ions on the binding of N-Acetyl-D-lactosamine to hen egg white lysozyme Jana Bregar*,1 1 1 Timotej Majdi č , Barbara Hribar-Lee, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia, jb00681@student.uni-lj.si Disruption of metal ion homeostasis can lead to modulate the binding of LacNAc by competing for a oxidative stress, which promotes protein misfolding and common binding site. aggregation linked to several pathological conditions 1. The aim of our research was to investigate the influence of copper (Cu 2+) ions on the binding of N-Acetyl-D- lactosamine (LacNAc) to hen egg white lysozyme (HEWL). Fluorescence spectroscopy was used to monitor changes in intrinsic tryptophan fluorescence of HEWL induced by the interaction between LacNAc, Cu2+ ions and HEWL. The effect of copper ions on the secondary structure and the thermostability of HEWL were assessed differential scanning calorimetry (DSC), respectively. Figure 1: Fluorescence emission spectra (λexc = 295 nm) of HEWL in the presence or absence of LacNAc and using circular dichroism (CD) spectroscopy and The location of the binding site of LacNAc and the types 2+ Cu ions. of interactions present were studied using molecular docking. The binding of LacNAc caused an increase in fluorescence intensity and a shift of the emission maximum toward smaller wavelengths, while the addition of Cu2+ ions decreased the fluorescence intensity and did not cause any significant shift of the maximum. CD spectra analysis showed that Cu2+ ions do not cause substantial changes in the protein’s secondary structure Design and development of fluorescent molecular probes for the detection of Alzheimer’s disease biomarkers Jerneja Kladnik,1 1 1 1 1, Matic Rogan, Ross D. Jansen-van Vuuren, Damijana Urankar, Janez Košmrlj* 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia, Janez.kosmrlj@fkkt.uni-lj.si Alzheimer’s disease (AD) is a neurodegenerative further ensures low background fluorescence in aqueous disorder, characterized primarily by cognitive decline and environments, enabling high sensitivity for target dementia. Despite intensive research, there remains a lack detection. of effective therapeutic options and, critically, no widely Looking forward, by integrating rational molecular accessible, non-invasive, and reliable diagnostic methods design this work lays the groundwork for the development for early detection and monitoring of disease progression. of a next-generation “smart molecular probe” for AD The development of such a diagnostic tool would diagnosis – an essential step toward improved patient represent a paradigm shift, enabling identification of at- outcomes and therapeutic development. risk yet asymptomatic individuals and offering the possibility of earlier intervention with existing or future Table 4: Structural motifs of an electron-donating group therapeutics. Furthermore, early and precise diagnosis (EDG), a π-conjugated aromatic linker (HC), and an would significantly accelerate the discovery and clinical electron-withdrawing group (EWG) of prepared validation of new treatment strategies.1 2 fluorescent molecular probes. The Košmrlj research group is addressing this unmet need through the design and synthesis of novel fluorescent molecular probes for the ex vivo detection of AD-related proteins, particularly amyloid β (Aβ) and tau. These probes comprising three essential components: (a) an electron-donating group (EDG), (b) a π-conjugated aromatic linker, and (c) an electron-withdrawing group (EWG) (Table 1). This push–pull configuration facilitates intramolecular charge transfer (ICT), resulting in distinct and tunable photophysical properties. Importantly, the probes should exhibit optical changes – References: To Coordinate New Things Jure Jakoš,1 Elisabeth M. Huf,2 Jakob Kljuna*1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Institute of Inorganic Chemistry, Universität Regensburg, Universitätsstraße 31, DE-93053 Regensburg, Germany For centuries, humans have battled microbial infections, initially employing metal-based treatments such as silver, arsenic, and mercury salts, with varying degree of effectiveness. The discovery of antibiotics by Alexander Fleming in the early 20th century changed the medical paradigm and thus helped save millions of lives. However, the widespread and sometimes inappropriate Figure 1: Structure of two Caerulomycins emergence of antimicrobial resistance References: — a major public 1. Salam, M. A.; Al-Amin, M. Y.; Salam, M. T.; Sigh use of antibiotics and other antimicrobials has driven the health concern identified by the World Health Pawar, J.; Akhter N.; Rabaan, A. A.; Alqumber, M. A. Organization. To counter this, efforts are focusing on A. Antimicrobial Resistance: A Growing Serious smarter use of antibiotics, improving existing drugs, Threat for Global Public Health. Healthcare 2023, 11, searching for new potential terapeutics and exploring 1946. innovative solutions such as metal-based compounds. 2. Funk, A.; Divekar, P. V. Caerulomycin, a New 1 discovered while studying an extract of Production, Isolation, Assay. and Biological Streptomyces Properties. Can. J. Microbiol. 1959 , 5 , 317 – 321. caerules In 1959, a new compound with great potential was Antibiotic from Streptomyces Caeruleus Baldacci, I. . The researchers noticed that something in this 3. Tong, L.; Sun, W.; Wu, S.; Han, Y. Characterization extract inhibited the growth of various filamentous fungi, of Caerulomycin A as a DualTargeting Anticancer yeasts and even some bacteria. After closer inspection of Agent. Eur. J. Pharmacol. 2022, 922, 174914. the exstract and subsiquent isolation and characterization, 4. Singla, A. K.; Krishna Gurram, R.; Chauhan, A.; named this new compound caerulomycin (Cae), later after Caerulomycin A Suppresses Immunity by Inhibiting T Cell Activity. PLOS ONE, 2014 , 9 the discovery of some derivatives they renamed it to they discovered a novel asymmetric 2,2’ Khatri, N.; Vohra, R. M.; Jolly, R. S.; Agrewala, J. N. -bipyridine. They 2 5. Trécourt, F.; Gervais, B.; Mongin, O.; Le Gal, C.; caerulomycin A (CasA) (Figure 1). Further research has Mongin, F.; Quéguiner, G. First Syntheses of shown that these compounds not only have antifungal and Caerulomycin E and Collismycins A and C. A New antibacterial activity, but also exhibit good anticancer, Synthesis of Caerulomycin A. J. Org. Chem. 1998, 63, Understanding Fluorination Effects on Butanol Derivatives Using Molecular Simulations and X-ray Scattering Jure Kovač 1 1 2 1 , Andrej Jamnik, István Szilágyi , Matija Tomšič *, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Department of Physical Chemistry and Materials Science, University of Szeged, Rerrich Béla sq 1., HU-6720 Szeged, Hungary Per- and polyfluoroalkyl substances (PFAS) are a large class of synthetic compounds known for their chemical stability and persistence in the environment. Their widespread use and toxicity have made them a growing concern for environmental science. Among them, fluorinated alcohols can serve as valuable model systems for studying the effects of fluorination on molecular structure and dynamics. In this study, we investigate a series of fluorinated butanol derivatives to determine how variations in fluorine content and substitution patterns affect the organisation of molecules in the liquid phase. We use molecular dynamics (MD) simulations using the OPLS-AA force field in combination with small and wide-angle X-ray scattering (SWAXS) measurements. Theoretical scattering intensities were calculated from the MD trajectories using Figure 1: Experimental and calculated SWAXS the complemented system approach. intensities from MD simulation results for 4,4,4- 1,2 The force field model was validated by trifluorobutan-1-ol (TFB), 4,4,4,3,3-pentafluorobutan-1- comparing the simulated SWAXS curves with ol (PFB), 4,4,4,3,3-pentafluorobutan-2-ol (PFSB), experimental data. A good agreement was found for both 4,4,4,3,2,2-hexafluorobutan-1-ol (HxFB), 4,4,4,3,3,2,2- the peak positions and the overall shape. This confirms heptafluorobutan-1-ol (HFB) and nonafluorotert-butanol that the OPLS-based models reliably capture the most (NFTB). The theoretical data were numerically smeared important structural features of the systems. With to allow direct comparison with the experimental data. the main scattering peak to lower References: q values, indicating that 1. increasing fluorination, we observe a consistent shift of Synthesis of fluorinated homoserine derivatives via Mitsunobu reaction Karin Rot,1 Jure Gregorc,1 Jernej Iskra*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Fluorine is the most electronegative atom with a large reaction in the presence of fluorinated alcohols (primarily inductive effect1. 19F is a ½ spin nucleus, which occurs at perfluoro-tert-butanol). 100 % natural abundance2. Because of its spin, it can be A further aim of this study was to prepare an N- used in magnetic resonance techniques (NMR, MRI, Fmoc-protected building block for subsequent use in MRS)2. 19F has NMR sensitivity that is similar to that of solid-phase peptide synthesis. For this reason, I the 1 1 19 H. Unlike H, F is absent in water and biological exchanged the protective groups of the fluorinated molecules, which makes it a particularly sensitive probe derivative of protected L-homoserine, affording N-Fmoc-because of the absence of background or competing O-(perfluoro-tert-butyl)-L-homoserinate. signals from solvent or from other molecules 3. Most amino acids used as NMR probes are monofluorinated or have a trifluoromethyl group in their structure. In order to improve the signal sensitivity, perfluoro-tert-butyl group can be introduced to natural amino acids. The perfluoro-tert-butyl group introduces nine chemically equivalent fluorine atoms that, because they are not coupled to adjacent protons, result in a sharp singlet. The best approach to install a perfluoro-tert-butyl group on an OH-bearing side chain, is to use Mitsunobu Scheme 2: Removal of the benzyl protecting group and 4 reaction with perfluoro-tert-butanol . exchange of the Boc group for Fmoc. Mitsunobu reaction is a method for the dehydrative coupling of alcohols with acids or pronucleophiles by References: using a combination of an oxidizing azo reagent (DIAD, 1. Marsh, E. N. G.; Suzuki, Y. Using (19)F NMR to DEAD, etc.) and a reducing phosphine reagent (PPh3, n- probe biological interactions of proteins and peptides. Bu 5 P, etc.) under mild conditions. ACS Chem. Bio. 2014, 9 (6), 1242–1250 3 The main focus of this study was to expand and 2. Danielson, M. A.; Falke, J. J. Use of 19F NMR to improve procedures for the synthesis of perfluoro- probe protein structure and conformational changes. tert - synthesis of appropriately protected 195. L-homoserine. 3. Tressler, C. M.; Zondlo, N. J. Perfluoro-tert-butyl Synthesis of protected butyl- Annu. Rev. Biophys. Biomol. Struct. 1996, 25, 163- L -homoserine. First part of the study focused on the PMMA-Siloxane Silica Coating for Corrosion Protection of AA2024-T3 in NaCl solution Nina Kovač, 1,2 1 1 1 1 Ana Kraš, Karolina Mulec, Ingrid Milošev, Peter Rodič*, 1 Jožef Stefan Institute, Department of Physical and Organic Chemistry, Jamova cesta 39, 1000 Ljubljana, Slovenia 2 Jožef Stefan International Postgraduate School, Jamova c. 39, SI-1000 Ljubljana, Slovenia Aluminium alloy (AA)2024-T3 is widely used in the Scheme 1: Stepwise procedure of PMMA siloxane-silica aerospace industry due to its low weight, high mechanical synthesis. The prepared solution was then deposited on strength, and cost-effectiveness. Despite these the AA2024-T3 surface. advantages, its susceptibility to corrosion in chloride-rich The preparation reactions of the coatings were environments, primarily due to the presence of alloying monitored by real-time Fourier transform infrared (FTIR) elements (such as Cu, Fe, and Mn), remains a significant spectroscopy. Following deposition onto AA2024-T3 drawback. Traditionally, corrosion protection has relied substrates, the coatings were analysed using a scanning on chromium(VI)-based compounds; however, their use electron microscope equipped with an energy-dispersive is increasingly restricted worldwide due to their harmful spectrometer (SEM/EDS) to assess their surface impact on human health and the environment. As a safer morphology, topography, elemental composition, and and more sustainable alternative, poly(methyl thickness. Corrosion performance was evaluated in a methacrylate PMMA-siloxane silica sol-gel coatings have 0.1 M NaCl solution using electrochemical impedance gained attention.1,2,3 spectroscopy (EIS). Additionally, the salt spray testing In this study, the corrosion resistance of AA2024-T3 was performed in accordance with the ASTM B117-07A protected with a siloxane-silica sol-gel coating was standard. The findings confirmed the formation of a evaluated. The sol-gel system was synthesised using uniform coating layer, several micrometers thick, tetraethyl orthosilicate (TEOS), methyl methacrylate providing continuous coverage of the alloy surface. These (MMA), 3-(trimethoxysilyl)propyl methacrylate coatings exhibited long-lasting barrier properties against (MAPTMS), and 2,2,2-trifluoroethyl methacrylate corrosion, as indicated by stable EIS responses (FEMA), as schematically presented in Scheme 1. maintained more than 3 months. References: 1. Rodič, P.; Korošec, R.C.; Kapun, B.; Mertelj, A.; Milošev, I. Acrylate-Based Hybrid Sol-Gel Coating for Corrosion Protection of AA7075-T6 in Aircraft Problem-based learning in polymer chemistry using the context of firefighter protective clothing Katja Gubič 1 2 1,3 , * Peter Rodič, Miha Slapničar 1 University of Ljubljana, Faculty of Education, Kardeljeva ploščad 16, 1000 Ljubljana, Slovenia 2 Jožef Stefan Institute, Department of Physical and Organic Chemistry, Jamova 39, 1000 Ljubljana, Slovenia 3 BEC Ljubljana, Cesta v Mestni log 47, SI-1000 Ljubljana, Slovenia Educational psychology shows that authentic, The analysis of textbooks revealed that applied real-life contexts can significantly increase students’ polymer science related to personal protective equipment motivation to learn1. Situational interest triggered by is largely absent from Slovenian secondary education. appealing content can, under the right conditions, develop This work suggests that contextualised PBL can enrich into a lasting individual interest2. Problem-based learning learning by linking theory to safety, materials science and builds on this principle by confronting students with environmental responsibility. complex and realistic problems, fostering deeper In the future, the model could also be extended to understanding, problem-solving skills, collaboration and lower grades and different fields (e.g., technical or non- intrinsic motivation3. technical oriented schools). In secondary school, scientific concepts such as polymers, wettability, hydrophobicity and surface tension are often presented in abstract terms, making it difficult for students to recognize their practical significance. This study presents a problem-orientated learning module that embeds these topics in the authentic example of firefighters' protective clothing. These multi-layer garments, with an outer layer typically made of high- performance aramid fibres such as Nomex® and Kevlar, provide fire resistance and ballistic protection. To enhance protective properties, the surface is treated with Scheme 1: Model representing interdependence of three fluorinated finishes that provide water repellency, fire levels of chemistry concepts5. resistance or antimicrobial performance. Such surface treatments can create superhydrophobic properties by References: lowering the surface free energy. As a result, water 1. Schraw, G.; Flowerday, T.; Lehman, S. Increasing Postsynthetic Ligand Exchange Effects on CO2 Performance Klara Klemenčič,1,2 Nataša Zabukovec Logar,1,2 Matjaž Mazaj,1 1 National Institute of Chemistry, Hajdrihova 19, 1000 Ljubljana, Slovenia 2 School of Science, University of Nova Gorica, Vipavska 13, 5000 Nova Gorica, Slovenia Metal–Organic Frameworks (MOFs) are a versatile class of porous crystalline materials characterized by unique chemical and physical properties, making them suitable for a broad range of applications. In recent years, MOFs have gained considerable attention as promising candidates for selective CO 1 capture and separation. One 2 of the most compelling features of MOFs lies in their tunable synthesis. Their highly ordered structures, large surface areas, and adjustable porosity allow for precise tailoring to enhance CO 2 affinity. A widely adopted 2 the framework by introducing additional functional Figure 1: Illustration of the fine-tuning approach strategy to improve CO2 uptake involves functionalizing groups into the organic linkers. For example, the applied to the NICS-24 structure. incorporation of amino groups has been shown to significantly increase CO 3 References: adsorption capacity . 2 1. Yang, S. Q.; Hu, T. L.; Chen, B. Microporous metal- This enhancement has been demonstrated in a series organic framework materials for efficient capture and of functionalized triazolate-based MOFs, including separation of greenhouse gases. Sci. China Chem. CALF-20, CALF-15, and NICS-24 4. The introduction of 2023, 66, 2181–2203. capture performance compared to CALF-15, with both Serre, C. Synthetic Aspects and Characterization materials outperforming CALF-20 under low CO 2 Needs in MOF Chemistry an extra amine group in NICS-24 resulted in superior CO2 2. Achenbach, B.; Yurdusen, A.; Stock, N.; Maurin, G.; concentration conditions. – from Discovery to Applications. Adv. Mater. 2025 , 2411359. To further enhance the performance of NICS-24, we 3. Lee, G.; Ahmed, I.; Jhung, S. H. CO2 adsorption using propose functionalizing the framework with linkers functionalized metal–organic frameworks under low exhibiting greater hydrophobicity (Figure 1). This pressure: Contribution of functional groups, excluding modification aims to systematically tune the pore amines, to adsorption. Chem. Eng. J. 2024, 481, Investigating Protein Interactors of the TDP-43 Pathology-Linked Protein Klara Razboršek,1 Jerneja Nimac,2 Boris Rogelj*,1,2 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Department of Biotechnology, Jozef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia, (boris.rogelj@ijs.si) Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease primarily affecting motor neurons. Initial symptoms typically present as muscle weakness, often starting in distal limbs and progressing with disease advancement. The average survival time post-onset is approximately three years, with respiratory failure being the most common cause of death. Around 50 % of patients also exhibit non-motor symptoms, with 10–15% diagnosed with frontotemporal dementia (FTD), and an additional 35–45 % showing mild cognitive and/or behavioral changes1. To date, more than 20 genes have been associated with ALS. The most frequent genetic causes include hexanucleotide repeat expansions in C9orf72 and mutations in SOD1, TARDBP, FUS, and TBK1. A key neuropathological hallmark of ALS is the accumulation of cytoplasmic aggregates of the TDP-43 protein, which are also found in patients with FTD and other neurodegenerative diseases1,2. TDP-43 is an essential DNA/RNA-binding protein from the heterogeneous nuclear ribonucleoprotein (hnRNP) family, involved in RNA metabolism, mRNA transport, stress granule formation, and central nervous system development. Mislocalization of TDP-43 to the cytoplasm is closely linked to both gain- and loss-of-function pathogenic peroxidase for labelling protein interactors of TDP-43. Scheme 1: Overview of the experiments using ascorbate mechanisms2,3. Synthesis and characterization of niobium-doped titania photocatalysts Klara Urankar,1 Andrijana Sever Škapin,2,3 Urška Lavrenčič Štangar*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, 1000 Ljubljana, Slovenia 2 Slovenian National Building and Civil Engineering Institute, Dimičeva ulica 12, 1000 Ljubljana, Slovenia 3 Faculty of Polymer Technology, Ozare 19, 2380 Slovenj Gradec, Slovenia Water pollution is a global problem that affects ecosystems, the economy and human health. The most common pollutants include microplastics, persistent organic compounds, heavy metals and other compounds that enter water systems through domestic and industrial wastewater. These compounds are difficult to degrade in the environment, and some of them are not even degradable by conventional water treatment methods. One of the methods for degradation of persistent micropollutants is heterogeneous photocatalysis, in which irradiation and suitable catalysts (usually metal oxides) are used to convert poorly degradable compounds into smaller and less harmful compounds1. Titanium(IV) oxide (TiO2) is a commonly used catalyst due to its good photocatalytic properties. Its modifications, such as Figure 1: Nitrogen adsorption-desorption isotherms of doping with different metal ions, can enhance its the analyzed catalysts 2 efficiency . The characterization involved nitrogen physisorption Catalyst degradation [%] analysis, X-ray powder diffraction, UV-Vis diffuse TiO 2 33 reflection spectroscopy, Fourier transform infrared Ti 0.95 Nb 0.05 O 2 34 spectroscopy and scanning electron microscopy. In order Ti 0.90 Nb 0.10 O 2 42 to determine the catalytic activity, the degradation Ti 0.85 Nb 0.15 O 2 42 reaction of the water pollutant phenytoin, a representative modified with various amounts of niobium (Nb) dopants reaction 3 using the sol-gel method and then to characterize them . Phenytoin The purpose of this work was to synthesize TiO2 Table 5: Phenytoin degradation (%) after 3 hours of Comparison of GATK and Deep Learning-Based DeepVariant in diagnostic Next-generation Sequencing (NGS) Variant Calling Klemen Klopčič 1 2 *, Aleš Maver, 1 Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana, Slovenia, (kk80756@student.uni-lj.si) 2 Clinical Institute of Genomic Medicine, University Medical Centre Ljubljana, Šlajmerjeva 4, SI-1000 Ljubljana, Slovenia Analysis of next-generation sequencing (NGS) potentially reduced interpretation workload. Notably, requires comprehensive bioinformatic analysis to derive DeepVariant did not identify any new pathogenic variants meaningful results. Genome analysis toolkit (GATK) 1 has in our cohort, indicating comparable clinical sensitivity. long been the standard for variant calling, but deep learning tools like DeepVariant 2 are increasingly used for their strong performance. Since each tool utilises different algorithms, their outputs can differ significantly. While previous studies have compared GATK and DeepVariant on trio exome sequencing (ES)3 4 and whole genome sequencing (GS) datasets, the critical challenge remains the accurate identification of pathogenic variants that enable precise genetic diagnoses in the clinical setting. We compared GATK and DeepVariant on 782 exomes and 69 genomes from patients without previously identified pathogenic variants. In addition to comparing Figure 1: Comparison of discovered variants (GATK = variant count, we searched for clinically relevant variants Genome analysis toolkit, DV = DeepVariant) that might explain the patients’ phenotypes. identified a similar total number of variants in the GS 1. McKenna, A.; Hanna, M.; Banks, E.; Sivachenko, A.; Cibulskis, K.; Kernytsky, A.; Garimella, K.; As shown in Figure 1, GATK and DeepVariant References: GATK identifying a higher proportion of unique calls The Genome Analysis Toolkit: A MapReduce Framework for Analyzing next-Generation DNA than DeepVariant (8.19 % and 3.79 % of algorithm- Sequencing Data. Genome Res. 2010 , 20 (9), 1297 – specific variants respectively). All observed differences 1303. each tool also detected a distinct subset of variants, with Altshuler, D.; Gabriel, S.; Daly, M.; DePristo, M. A. datasets, with a large overlap between the two. However, Comparison of mechanical properties of alkali-activated materials made of different alternative alkali activators Kaja Zupančič 1 1 2 , Liam Manevski, Majda Pavlin*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Slovenian National Building and Civil Engineering Institute, Dimičeva ulica 12, SI-1000 Ljubljana, Slovenia The construction industry is recognized as one of the sodium and potassium hydroxide solutions. The resulting most significant contributors to global greenhouse gas AAAs were used to prepare various AAM mixtures, using emissions, and within this sector, the cement industry metallurgical slag and metakaolin as precursors. These alone is responsible for approximately 8% of global materials were then tested for early compressive strength carbon dioxide (CO₂) emissions. The production process after three days. Additionally, FTIR (Fourier-transform of ordinary Portland cement is highly carbon-intensive: infrared) spectroscopy was used to analyze the structural for every tone of cement produced, approximately 0.7 to features of the hardened AAMs. For comparison, 1 tone of CO₂ is emitted into the atmosphere. In response reference samples were also prepared using commercial to growing concerns about climate change and activators (Geosil and Silvez) and standard sodium and environmental degradation, researchers and engineers potassium hydroxide solutions. have been actively seeking more sustainable alternatives. One such promising alternative is the use of alkali- activated materials (AAMs)1, which are emerging as potential low-carbon binders to replace traditional cement. AAMs require an alkali solution (activator) to initiate the chemical reaction that leads to hardening and strength development. In commercial applications, this activator is often a solution of potassium or sodium silicate. However, Figure 1: Alkali activated materials prepared of the production of these commercial alkali activators is different alternative activators. also associated with high carbon emissions. For example, producing just one kilogram of sodium silicate can References: generate approximately 1.5 kilograms of CO₂2 1. Provis, J.; Deventer, J. Alkali Activated Materials: , which AAMs. To address this issue, researchers have begun 2. Turner, L. K.; Collins, F. G. Carbon Dioxide Equivalent (CO2-e) Emissions: A Comparison exploring the use of alternative alkaline activators undermines some of the environmental benefits of using State-of-the-Art Report, RILEM TC 224-AAM; 2014. H/D Exchange Enabled by Polystyrene-Supported Iridium Catalyst Luka Jedlovčnik*,1 Ross Jansen-van Vuuren,1 Damijana Urankar,1 Janez Košmrlj1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia The synthesis of deuterium-labeled organic resulting heterogeneous catalyst was employed in the compounds typically follows two principal strategies. deuteration of a range of substrates previously One involves indirect methods, such as total synthesis investigated using the corresponding homogeneous using deuterated substrates or reagents. The other counterpart. We will discuss the comparative employs direct techniques, including hydrogen/deuterium performance of the homogeneous and heterogeneous (H/D) exchange or deutero-defunctionalization within the systems, including recyclability and overall potential for target molecule. Among these, direct H/D exchange is practical applications. often more efficient and cost-effective, particularly when introducing deuterium into complex molecules like pharmaceutical compounds.1,2 In both direct and indirect methods, catalysts based on precious metals are commonly employed, with iridium Figure 1: General scheme for H/D exchange with being among the most prevalent due to its high activity in hydrogen isotope exchange reactions. However, 2 immobilised catalysts. homogeneous iridium catalysts pose challenges in recovery and reuse, as they are difficult to separate from 1. Kopf, S.; Bourriquen, F.; Li, W.; Neumann, H.; Junge, References: reaction mixtures and can deactivate through aggregation. K.; Beller, M. Recent Developments for the Immobilizing homogeneous catalysts onto solid Deuterium and Tritium Labeling of Organic supports (e.g., polymers or silica) offers a sustainable Molecules. Chem. Rev. 2022, 122 (6), 6634–6718. simplifying purification. Deuterated Drugs and Biomarkers in the COVID-19 3 This strategy tackles challenges Pandemic. ACS Omega 2022 , 7 (46), 41840 – 41858. like catalyst recovery, aggregation, and deactivation. approach to catalysis by enhancing recyclability and 2. Jedlovčnik, L.; Košmrlj, J.; Massey, T. E.; Derdau, V. 3. Jedlovčnik, L.; Höfferle. J.; Pashaj, F.; Kladnik, J.; Immobilized catalysts are easily separated, often by Košmrlj, J.; Derdau, V.; Jansen-van Vuuren, R. D. filtration, and reused with minimal activity loss, reducing Sustainable synthetic routes to deuterium-labelled precious metal use and aligning with green chemistry organic compounds using immobilized and recyclable principles. It also limits leaching and product (bio)catalysts. GSC 2025, 6 (1), 1–35. combining the performance of homogeneous catalysts Homogeneous Catalysts for Fine Chemicals Production, Springer Netherlands, Dordrecht, 2010. 4 contamination, crucial in pharmaceutical synthesis, while 4. Barbaro, P.; Liguori, F. eds., Heterogenized Alkali-Silica Reaction in Concrete –Focus on Silicate Aggregates Used in Slovenia Maja Pristavec ,1* 1 2 2 1 , Primož Oprčkal , Radmila Milačič Ščančar , Janez Ščančar , Ana Mladenovič 1 Slovenian National Building and Civil Engineering Institute, Dimičeva ulica 12, 1000 Ljubljana, Slovenia 2 Department of Environmental Sciences, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana Concrete, with over 4.1 billion tonnes produced in neglected in Slovenia, recent degradation of high- 2023, is an essential construction material for modern performance concretes has highlighted its relevance and infrastructure including transport, energy, and urban the urgent need for systematic assessment. In development, and there is no material that can replace it collaboration with Alpacem Cement we will aim to in the near future. Concrete is a mixture of binder (pure develop innovative concrete mixes using silicate cement or cement with supplementary cementitious aggregates and appropriate cement-SCM combinations materials-SCMs) aggregate, water, and chemical that will effectively mitigate ASR without compromising admixtures.1 It is usually reinforced to compensate for its its durability and long-term performance. low tensile strength and ductility. Among the various mechanisms affecting concrete durability, the alkali-silica reaction (ASR) is recognized as one of the most critical due to its expansive nature and long-term progression. ASR is a chemical reaction between reactive silica in aggregates and alkalis (Na⁺, K⁺) in concrete pore solution, forming crystalline or amorphous gel products that expand and cause micro- and later macrocracks in concrete. 1,2 The later act as physical catalysts for further degradation processes. Reactivity of silica depend on the silica type – it is – high in amorphous forms (e.g. opal, volcanic glass), moderate in cryptocrystalline or microcrystalline quartz, and lower in deformed quartz found in metamorphic rocks. In Slovenian aggregate, microcrystalline and deformed quartz are prevalent. Geology in Slovenia is predominantly carbonate-based (limestone, dolomite), but in the northeast (Pohorje, Mura Determination of Liquid Products from Electrochemical CO2 Reduction using GC-MS Maja Svete, 1 1,2 1 1,2,3 Blaž Tomc* , Dušan Strmčnik , Nejc Hodnik 1 Laboratory for Electrocatalysis, Department of Materials Chemistry, National Institute of Chemistry, 1000 Ljubljana, Slovenia 2 University of Nova Gorica, 5000 Nova Gorica, Slovenia 3 Institute of Metals and Technology, 1000 Ljubljana, Slovenia CO2 is a major greenhouse gas, whose rising concentration in the atmosphere is the primary driver of today’s climate change. 1 Reducing CO emissions and 2 producing valuable chemicals are key goals for sustainable development.2 Electrochemical CO2 reduction (CO2RR) offers a promising route to reduce CO2 concentrations while producing value-added products such as ethanol. Accurate quantification of liquid products formed during this process is essential.1 Typically techniques for this determination are nuclear magnetic resonance (NMR), high performance Figure 1: Chromatogram of liquid products from liquid chromatography (HPLC), and gas chromatography 1 CO2RR using GC-MS coupled with mass spectrometry (GC-MS). NMR is widely used due to its high sensitivity and ability to detect References: unknown products, however, it has limitations when it 1. Nitopi, S.; Bertheussen, E.; Scott, S. B.; Liu, X.; comes to detecting products at very low concentrations. Engstfeld, A. K.; Horch, S.; Seger, B.; Stephens, I. E. HPLC, on the other hand, has limited sensitivity for L.; Chan, K.; Hahn, C.; Nørskov, J. K.; Jaramillo, T. volatile aldehydes and alcohols, which are the primary F.; Chorkendorff, I. Progress and Perspectives of products.1,3 Electrochemical CO2 Reduction on Copper in In this work, we explore the capabilities of headspace Aqueous Electrolyte. Chem. Rev. 2019, 119 (12), GC-MS, a well-established method that is less commonly 7610–7672. used in CO2RR, for the detection of volatile products with 2. Tomc, B.; Bele, M.; Azeezulla Nazrulla, M.; Šket, P.; lower detection limits.3 We found that alcohols, Finšgar, M.; Surca, A. K.; Kamšek, A. R.; Šala, M.; Use of specific pyrazole derivatives for characterization of L-threonine dehydrogenase Marcel Tušek,1 1 Marko Novinec*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Pyrazole-based compounds are gaining prominence Figure 1: Effect of compound I1 concentration on in pharmaceutical research due to their notable reaction rates. The blue curve is 100 µM of L-Thr, antimicrobial effects. The synthesis of new pyrazole orange curve is 250 µM of L-Thr and the green curve is derivatives offers promising ways for the development of 500 µM of L-Thr novel antibiotics1. An example of such a compound is 4- (2-aminoethyl)-1-(pyridine-2-yl)-1H-pyrazole-5-ol (I1), which was found to inhibit the growth of Escherichia coli 1,0 and the activity of L-threonine dehydrogenase (TDH). ]/s This enzyme is crucial for amino acid metabolism and UF biofilm production. As TDH is absent in human cells and R 0,5 [ lacks a human homologue, inhibitors like I1 present a v valuable starting point for future drug discovery2. This study focused on evaluating four types of pyrazole derivatives for their ability to inhibit TDH and 0,0 0 50 100 influence biofilm formation. Enzyme activity was c(I2) [ μ M] monitored through a fluorometric method, based on the Figure 2 + : Effect of compound I2 concentration on conversion of NAD to NADH, which allows for real time reaction rates. The blue curve is 100 µM of L 3-Thr, activity tracking . Among the tested molecules, I1 proved orange curve is 250 µM of L-Thr and the green curve is to be the most effective inhibitor. A second compound, 500 µM of L-Thr I2, showed comparable inhibition but could not be concentrations References: extensively analysed due to limited solubility at higher 4. 1. Naim, M. J.; Alam, O.; Nawaz, F.; Alam, M. J.; Alam, Further experiments demonstrated that all tested P. Current Status of Pyrazole and Its Biological derivatives reduced biofilm formation. However, only I1 activities. J. Pharm. Bioallied. Sci. 2016, 8, 2–17. and I2 were shown to exert a direct effect on TDH 2. Mikulič Vernik, N.: Karakterizacija interakcije med activity. These results support a potential association Disulfide-based conformational locks for studies of alpha-actinin-1 actin cross-linking mechanism Mark Loborec,1 1 1 Jošt Hočevar, Miha Pavšič*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia The actin cytoskeleton plays an important role in cell verification. The protein was then expressed in E. coli and structure, adhesion and mobility1. Consequently, actin is purified via nickel-affinity, ion-exchange and size- one of the earliest evolved proteins, dating back to the exclusion chromatography. SDS-PAGE of the mutants common ancestor of life. It is also one of the most under reducing vs. non-reducing conditions was then used abundant proteins in many cells, accounting for up to to check for the formation of cross-chain disulfide bonds. 10 % of protein mass in most cells2. Formation of such bonds was confirmed in one of the Alpha-actinin is an actin-binding protein of the mutants, while actin-bundling activity tests are ongoing. spectrin family, and likely also its originator. It consists of three major domains: an actin-binding domain (ABD), a calcium-binding calmodulin-like domain consisting of 4 two EF-hand motifs (CaM-like domain) and a rod domain which connects the two3. It dimerizes in an anti-parallel fashion, with the CaM-like domain of one monomer regulating the ABD of the other. There are four human isoforms of alpha-actinin, two of which are calcium- sensitive (isoforms 1 and 4) and can be found in most 4 cells, while the other two are calcium-insensitive (isoforms 2 and 3) and can be found only in muscle cells, Figure 1: A model of a special "half-dimer form" of localized to the Z-disc of the sarcomeres human actinin 1 in a calcium-bound state. The ABD and 3 . As there is currently no accurate model of the SR 1 and 2 domains would belong to one chain in a conformational changes responsible for loss of actin- typical dimer, while the EF and SR 3 and 4 domains bundling activity upon calcium binding in non-muscle would belong to the other chain. isoforms of alpha actinin, we have set to identify mutations which would conformationally lock parts of the References: structure, and measure its actin-bundling activity in the 1. Foley, K. S.; Young, P. W. The Non-Muscle Frequency up-converted white to green emission tuning in 1-D Photonic crystal (MC) structures for LED applications Martin Jazbec,1,2 1 1 1 Prasenjit P. Sukul*, Lu í s F. Santos, Rui M. Almeida, 1 Centro de Química Estrutural, Institute of Molecular Sciences and Departamento de Engenharia Química, Instituto Superior Tecnico, Universidade de Lisboa, Av. Rovisco Pais, PT-1049-001, Lisboa, Portugal 2 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia This study investigates one-dimensional photonic crystal (1D-PC) structures fabricated on silicon substrates, specifically focusing on a Fabry-Pérot microcavity (MC) designed for enhanced light emission at selected wavelengths within the white to green spectral range. The microcavity was constructed using alternating layers of titania and lanthanide (Ln)-doped aluminosilicate glass, synthesized via sol-gel (SG) processing. The aluminosilicate sols were tri-doped with Tm³⁺/Er³⁺/Yb³⁺ in a 0.5/0.5/5.0 mol% rat 1 io, serving as the low-refractive-index (n=1.48) component, while titania Figure 1: SEM image of Fabry-Pérot microcavity (MC). sol provided the high-refractive-index counterpart. The It shows five Bragg mirror pairs on each side of the MC structure consists of five Bragg mirror pairs on each defect layer side of a defect layer (composed of a double layer of Ln- doped 2 aluminosilicate). Cross-sectional SEM images of the FP MC is shown in the Figure 1 and Figure 2 is shows the UV-Vis Reflectance data of the MC sample designed with defect layer targeting 525 nm for 0o angle. In alignment with green chemistry principles, we further explored low-temperature processing methods by introducing zinc oxide (ZnO)3 as an alternative high- refractive-index thin-film material. The influence of heat treatment temperature on ZnO’s crystalline structure was systematically examined. In summary, this work Modeling-guided process development of a capillary force driven microfluidic distillation device Martina Potočnik 1 1 , Rok Ambrožič *, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Distillation is one of the oldest and most frequently MBD approach. Model optimization is economically used separation processes, based on the different boiling more efficient and faster than empirical testing of points of components in miscible liquid mixtures. Due to 3 manufactured prototypes. its simplicity, high efficiency for various mixtures and the Within this research, microfluidic distillation device possibility of continuous separation, this method is shown in Figure 1 was developed, and a corresponding widely applied in numerous industrial sectors, such as the mathematical model was created to describe the process. pharmaceutical, petroleum, food and chemical industries. This was followed by numerical model solving and Despite its many advantages, distillation also has certain optimization of the distillation process to improve limitations. It is an energy-intensive process, which separation efficiency, reduce energy consumption, and conflicts with sustainable development guidelines. The enable system scale-up. equipment required to ensure adequate productivity is large, meaning the process also demands significant operational space. Additionally, distillation systems are stationary, reagent consumption is relatively high even in pilot laboratories, and as a result, distillation is a relatively expensive process 1. It is possible to improve the distillation process with miniaturization, which is a promising approach for process intensification. Micro-structured systems are characterized by large specific surface areas (high surface-to-volume ratio), that enhances heat and mass transfer within the system, enabling faster separation, easier temperature regulation and, most importantly, greater energy efficiency. Firstly, due to smaller reaction volumes, the consumption of chemicals and energy is reduced, along with waste production and environmental Synthesis and testing of molecularly imprinted polymers for electrochemical determination of neonicotinoids Matic Plut, 1 1 1 1 Maksimiljan Dekleva, Gregor Marolt, Helena Prosen 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Neonicotinoids are a class of insecticides that have been in use since the end of the 20. century. Their use has increased rapidly, particularly in Africa and Asia due to their high efficiency and target specificity. However, widespread application has led to major imbalances in ecosystems, especially in insect and pollinator populations. In Europe their use has already been restricted and further restrictions are expected for the US. 1–3 Thiacloprid and acetamiprid are important representatives of this group of pesticides, but their poor electroactivity due to the absence of a NO 3 group makes 2 direct electrochemical detection methods inapplicable for their determination. In our work, several molecularly imprinted polymers (MIPs) for thiacloprid and acetamiprid have been synthesised and further evaluated Figure 1: Cyclic voltammograms recorded by working for electrochemical detection. The specific binding electrodes before and after accumulation in 0.1 M been screened and quantified by calculating the modified electrode without MIP, E5 – modified imprinting factor (IF). The polymer with the highest IF electrode with MIP). efficiencies of the polymers for the target analytes have acetamiprid solution (E1 – untreated electrode, E4 – for each analyte was applied to the working electrode of the screen-printed electrode via drop-casting. References: Analysis of cyclic voltammograms of 1. Jeschke, P.; Nauen, R.; Schindler, M.; Elbert, A. hexacyanoferrate redox probe at the i) untreated Overview of the Status and Global Strategy for electrode, ii) modified electrode without the molecularly Neonicotinoids. J. Agric. Food Chem. 2011, 59 (7), Analysis of Air Pollutants in Ljubljana Bežigrad and Murska Sobota during Summer 2025 Tai Lupscha,1 2 Matjaž Dlouhy* , 1 Srednja poklicna in tehniška šola Murska Sobota, Šolsko naselje 12, SI-9000 Murska Sobota, Slovenia 2 Gimnazija Ledina, Resljeva cesta 12, SI-1000 Ljubljana, Slovenia Environmental chemistry examines the chemical composition, transformations, and fate of substances in the natural environment, with atmospheric chemistry forming a critical branch that investigates pollutant generation, transport, and removal. Air quality research is central to this field, as atmospheric pollutants undergo complex chemical reactions affecting both human health and climate. Between July 27 and August 9, 2025, we conducted a comparative study of atmospheric pollutants at two Slovenian weather stations: Ljubljana Bežigrad (urban) and Murska Sobota (less urban/rural). Concentrations of PM₁₀, PM₂.₅, NO₂, and O₃ were measured alongside meteorological parameters such as temperature, wind, and precipitation. Ljubljana exhibited higher mean concentrations of PM₁₀, PM₂.₅, and NO₂, attributable to more intense anthropogenic emissions from traffic and heating systems. In contrast, Murska Sobota consistently displayed higher ozone (O₃) concentrations, illustrating the “ozone paradox” where urban areas with elevated nitrogen oxide (NO) emissions promote O₃ depletion through the titration reaction NO + O₃ → NO₂ + O₂.1 Rural areas, with fewer NO sources, have less ozone destruction, allowing photochemically generated ozone to persist. During the study period, anticyclonic conditions Optimization of horseradish peroxidase immobilization into cross-linked enzyme aggregates using microfluidics Natalija Tomažin 1 1 1 , Marko Božinović, Polona Žnidaršič-Plazl*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Slovenia, polona.znidarsic@fkkt.uni-lj.si Efficient, carrier-free immobilization is pivotal for activity of 99 % was achieved at a retention time of 0.34 building robust biocatalysts that can be reused and min with acetone and glutaraldehyde concentrations of 90 operated continuously. Cross-linked enzyme aggregates % (v/v) and 1 mM, respectively. The resulting CLEA- (CLEAs) are attractive because they combine high HRPs exhibited an average particle radius of 150 nm, volumetric activity with simple, low-cost preparation via determined using dynamic light scattering. precipitation followed by intra-aggregate crosslinking, The optimized CLEA-HRP preparation provides a while typically enhancing stability and enabling catalyst robust, reusable biocatalyst suitable for future continuous recycling1. However, conventional batch-scale oxidation reactions. These findings demonstrate the precipitation and crosslinking that requests centrifugation potential of microfluidics-assisted CLEA generation for for CLEAs separation from the reaction mixture often creating high-activity immobilized enzymes tailored for suffer from poorly controlled mixing and agglomeration, advanced biocatalytic applications. leading to highly polydisperse particle populations, variable recovered activity, and mass-transfer limitations that hamper reproducibility and scale-up. Microfluidic platforms mitigate these issues by imposing well-defined residence times and rapid micromixing, which narrow particle size distributions, improve transport, and seamlessly interface with continuous-flow operation2. Leveraging these advantages, we aimed to optimize the immobilization of horseradish peroxidase (HRP) into CLEAs within a microfluidic platform, generating highly Figure 1: Set-up for a continuous CLEA-HRP uniform, high-activity, and reusable nanoparticles generation in a microfluidic system. suitable for cost-effective continuous biocatalytic processes. Immobilization in this setup not only enhances References: operational stability and catalyst reuse but also enables 1. Sheldon, R. A.; van Pelt, S. Enzyme Immobilisation Synthesis, characterization and interaction between lignin nanoparticles and nanoplastics Neja Lesinšek 1 1 1 1 1,2 1 *, Azmat Ullah, Jan Hočevar, Boštjan Žener, Jelena Papan Djaniš, Jernej Iskra, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Centre of Excellence for Photoconversion, Vinča Institute of Nuclear Sciences, National Institute of the Republic of Serbia, University of Belgrade, RS-11351 Belgrade, Serbia Plastic pollution is a global problem and has a major nanoparticles. Representative SEM images of lignin impact on the environment. While plastic materials nanoparticles and nanoplastics are shown in Figure 1. improve everyday life and are widely used across Interactions will be studied using zeta potential industries due to their resilience and durability, these measurements and DLS at various pH values. This study properties also contribute to their persistence in the is a starting point for future research on the interaction environment for centuries. Humans are exposed to micro- between lignin nanoparticles and nanoplastics. A better and nanoplastics through tap and bottled water, personal understanding of these interactions could help reduce care products and contaminated marine organisms that environmental pollution. have ingested plastic particles present in aquatic environments. Marine-based sources of plastic pollution include recreational cruises, oil extraction, product transportation and fishing activities1. Nanoplastics in water can interact with naturally occurring polyphenolic . compounds, such as lignin. In this study, we focused on Figure 1: SEM: Lignin nanoparticles from organosolv the synthesis and characterization of lignin nanoparticles lignin (left), nanoplastic particles (right) (LNPs) and nanoplastics (NPs) to establish a model system for studying their potential interactions. Figure 1: Representative SEM images of : a) Lignin Lignin was isolated from milled straw biomass using nanoparticles from organosolv lignin, b) nanoplastic an organosolv extraction method. Biomass was mixed particles with ethyl acetate and hydrochloric acid and heated at acetone and distilled water to precipitate lignin, which Chem. 2019, 114, 196–201. 2. Pourbaba, R.; Abdulkhani, A.; Rashidi, A.; Ashori, A. was then filtered and dried. Lignin nanoparticles (LNPs) Lignin nanoparticles as a highly efficient adsorbent were prepared via a hydrotropic synthesis method for the removal of methylene blue from aqueous 2 reported in . media. Sci. Rep. 2024 , 14 , 9039. Nanoplastics (NPs) were synthesized by dissolving 3. filtered, and the solvent was removed using a rotary 1. Almeida, M.; Oliveira, M. The why and how of micro(nano)plastic research. TrAC, Trends Anal. evaporator. The resulting residue was treated with 130°C for 3 hours. After cooling, the mixture was References: Heptanuclear zinc(II) complex with 3-cyanopyridine Nejc Virant,1 Nina Podjed Rihtaršič,1 Barbara Modec*1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Zinc(II) plays a crucial biological role, acting as an essential cofactor in over 300 enzymes, enabling proper functioning of protein synthesis, nucleic acid metabolism, and cellular signaling etc. Additionally, zinc(II) contributes to the structural stability of proteins, which is vital for precise regulation of gene expression and the maintenance of functionality in various biological systems1. 3-Cyanopyridine (3-PyCN) is a solid, colorless organic compound with the molecular formula C6H4N2. The molecule consists of a pyridine ring with a cyano group (–CN) attached at the third position. Figure 1: ORTEP drawing of the heptanuclear complex molecule [Zn7O2(CH3COO)10(3-PyCN)2], as found in 1. Analogous heptanuclear zinc(II) complexes with acetate and nitrogen donor ligands such as 1-methylimidazole or pyrazine were prepared before2,3. Herein, we will present the crystal structures of both Scheme 1: Structural formula of 3-cyanopyridine 1 compounds, along with the infrared and H NMR spectra, (3-PyCN). as well as the TG-DSC and elemental analysis of compound 1. 3-Cyanopyridine can bind to transition metal ions in a monodentate manner through the ring nitrogen or, less commonly, also through the cyano group in a bridging References: manner. Surprisingly, the coordination chemistry of 1. Parkin G. Synthetic analogues relevant to the structure zinc(II) with 3-cyanopyridine is relatively poorly and function of zinc enzymes. Chem. Rev. 2004, 104, Cyclic anhydrides as multi-purpose compounds Nina Krašovec,1 Matic Rogan,1 Jerneja Kladnik,1 Damijana Urankar,1 Janez Košmrlj*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia, (janez.kosmrlj@fkkt.uni-lj.si) Cyclic anhydrides are commonly 5- or 6-member esterification reaction with a derivative of diethylene ring molecules, conventionally derived from glycol to yield an elongated carboxyl derivative (Figure corresponding dicarboxylic acids by an intramolecular 1). For a successful reaction procedure an excess of dehydration reaction. Normally, the use of toxic triethylamine was added, which served as a base as well dehydration reagents and harsh conditions is required, as a solvent. This procedure resulted in a successful though recently new procedures have been developed synthesis of the desired compound (2,2-dimethyl-4,15- which employ Lewis acids as catalysts1 or allow dioxo-3,8,11,14-tetraoxa-5-azaoctadecan-18-oic acid, (1) dicarboxylic acids to be directly transformed into cyclic with little side products, which were easily removed by anhydrides via electrolysis2. dissolving the reaction mixture in dichloromethane and Cyclic anhydrides act as electrophiles and are able to washing it with 1 M HCl solution and brine. Synthesized react with nucleophilic compounds, such as amines and molecule exhibits potential use as a linking element and alcohols to form amides and esters, respectively. Due to in nanoparticle surface functionalization. facile activation of one carbonyl group in anhydride bond, reaction can proceed under mild conditions and without the presence of coupling reagents, which makes cyclic anhydrides valuable tools in organic and bioconjugate chemistry1. They are commonly used for polymer functionalization, hydrogel formation and in some cases Figure 1: Synthesized compound 2,2-dimethyl-4,15-polymer synthesis. Protein modifications with cyclic dioxo-3,8,11,14-tetraoxa-5-azaoctadecan-18-oic acid (1). anhydrides lead to increased protein solubility and can reverse the total charge of the biomolecule. Their pH- References: selectively target low pH tissues. In addition, cyclic Anhydrides as Powerful Tools for Bioconjugation and Smart Delivery. Bioconjugate Chem. 2021 , 32 , 482 – anhydrides are often used as cross-linkers between sensitivity is exploited in drug delivery systems to 1. Spanedda, M. V.; Bourel-Bonnet, L. Cyclic various molecules . 2. Schneider, J.; Häring, A. P.; Waldvogel, S. R. 1 496. Development of extraction method for perfluoroalkyl pollutants in water Špela Žunec,1 Helena Prosen*,1 Ida Kraševec1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Per- and polyfluoroalkyl substances (PFAS) are compounds with strong carbon–fluorine (C-F) bonds that 1,2 cause their exceptional stability and resistance to ery 1 degradation. Therefore, they are widely used in industrial cov 0,8 HLB re applications. However, their persistence and 0,6 ed WAX bioaccumulation have raised significant concerns due to 0,4 aliz their adverse effects on human health and the 0,2 MAX rm o environment.¹ Determination of PFAS typically involves n 0 extraction and preconcentration, followed by analysis PFBA PFPeA PFHxA PFHpA PFOA using sensitive techniques such as liquid PFAS with different chain length chromatography–tandem mass spectrometry (LC- Figure 1: Comparison of normalized recoveries for MS/MS).² One of the most common extraction techniques different SPE cartridges is solid-phase extraction (SPE), which was also used in this work for the extraction of five PFAS (C4-C8) from water samples. The method was optimized by evaluating 1. Ng, C.; Cousins, I. T.; DeWitt, J. C.; References: Glüge, J.; sorbent type (Fig. 1), elution solvent, and extraction Goldenman, G.; Herzke, D.; Lohmann, R.; Miller, M.; conditions (pH and salt concentration). The final method Patton, S.; Scheringer, M.; Trier, X.; Wang, Z. consisted of sample pH and salt content adjusted to 2-3 Addressing Urgent Questions for PFAS in the 21st were used with 5% NH 12765. 3 in methanol as the elution 2. Liu, Y.; Bao, J.; Hu, X.-M.; Lu, G.-L.;Yu, and 3.5 %, respectively. Cartridges with WAX sorbent Century. Environ. Sci. Technol. 2021, 55, 2755– solvent. The breakthrough volume exceeded 500 mL and W.-J.; extraction recoveries were above 95% for all analytes. A the analysis of PFOA and PFOS in the salty matrices Meng, Z.-H. Optimization of extraction methods for seawater sample was analysed, revealing the presence of during the wastewater treatment. Anal. Bioanal. PFAS at concentrations below the limit of quantification, Chem. 2020, 412, 1234–1245. approximately 5-6 ng/L. Characterization of ATP-dependent amide bond synthetase for selective amidation Tadej Menegatti*,1 1 2 2 1 , Žiga Gerdina , András Kotschy , Gábor Tasnádi, Polona Žnidaršič Plazl , 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Servier Research Institute of Medicinal Chemistry, Zahony str. 7, HU-1031 Budapest, Hungary Amide bond formation is a central transformation in clear route toward intensified and scalable biocatalytic pharmaceutical synthesis, representing up to 16 % of all processes4. reactions in medicinal chemistry. Conventional approaches typically rely on stoichiometric activation of carboxylic acids with coupling reagents, which often results in poor atom economy and the generation of hazardous by-products1. In contrast, enzymatic methods operating in aqueous media without toxic reagents offer a greener and more sustainable alternative2. Among these, ATP-dependent amide bond synthetases (ABS) have emerged as promising catalysts, capable of performing both acid adenylation and subsequent amidation within a single active site, requiring only a small excess of the amine partner1,3. Here, we describe process development for the ATP- dependent coupling of trans-cinnamic acid with 1,2- phenylenedimethanamine catalyzed by an ABS from Streptomyces TR1341 in the presence of inorganic Figure 1: Reaction scheme of ABS-catalyzed amide pyrophosphatase (PPase). The reaction proceeds through synthesis from trans-cinnamic acid and 1,2-an acyl – AMP intermediate, which undergoes phenylenedimethanamine. nucleophilic attack by the amine to form the amide product. References: Higher temperatures increased activity but also diamide 1 Tang, Q.; Petchey, M.; Rowlinson, B.; Burden, T. J.; formation; 30 °C was chosen to balance both. The enzyme Fairlamb, I. J. S.; Grogan, G. Broad Spectrum was most active at pH 7.50–7.82. At 30 % (v/v) co- Enantioselective Amide Bond Synthetase from Optimisation of extrusion based on a rheological analysis Tadej Pirc*,1 Rok Ambrožič,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia This abstract summarizes research investigating the process changes while maintaining equivalent rheological optimisation of the extrusion process for paraffin-based behavior to the current product standard. The optimised rodent bait using rheological analysis as a tool for formulation not only matches the mechanical and systematic process improvement. The study is grounded functional requirements but also results in reduced in industrial practice and aims to address specific issues cost and greater process robustness. Examples of cost in the manufacturing process 1 — namely, mechanical stress reduction with fixed tan(δ) are shown in Table 1. on the extruder, mixture inhomogeneity, and unnecessarily high production costs—while preserving product functionality and mechanical integrity1. Rheological measurements were carried out using a plate–plate geometry on a rotational rheometer under controlled strain amplitude. The bait, being a viscoelastic solid, required careful sample preparation to ensure consistent results. A customized steel mold and hydraulic press were used to produce test specimens, and an optimal compaction pressure was determined through statistical evaluation of measurement repeatability. Key rheological parameters such as the elastic modulus (G′), viscous modulus (G″), and tan(δ) were used to quantify the material's response under small-strain oscillatory conditions, with tan(δ) serving as the principal metric for distinguishing the viscoelastic properties of various polymers1. A Plackett Figure 1: Comparison of regression model predictions – Burman design of experiments (DoE) and experimental results for rheological parameters was employed to screen nine variables: seven formulation 1 components, extrusion temperature, and shear rate2,3. Table 6: Relative deviations of predicted rheological Through statistical analysis including multiple linear regression and ANOVA, the study identified the most 1 parameters from experimental values for selected Selective Extraction of Magnesium from Slovenian Dolomite Tal Čarman*,1 Mitja Kolar,1 Vilma Ducman,2 Gorazd Žibret,3 Lea Žibret,2 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia, (tal.charman@gmail.com) 2 Slovenian National Building and Civil Engineering Institute, Dimičeva ulica 12, SI-1000 Ljubljana, Slovenia 3 Geological Survey of Slovenia, Dimičeva ulica 14, SI-1000 Ljubljana, Slovenia Magnesium (Mg) is a critical metal with applications Mg leaching with 7.5 mL and 105 % equivalent of H2SO4 in the automotive, aerospace, electronics, chemical and followed by selective Mg precipitation with 20 mL of pharmaceutical industries. 1 It is currently the third most NaOH. It achieved an overall Mg yield of 98.3 % and a highly produced metal globally2 and demand is likely to molar ratio of 155 : 1. Selective leaching with 30 mL and increase as it is one of the most promising metals for the 105 % equivalent of H2C2O4 was more selective with development of more sustainable batteries.3 Sources of Mg : Ca = 457 : 1, but Mg yield was only 62,5 %. Mg range from seawater to many minerals, of which Overall, the developed procedures are highly dolomite (CaMg(CO3)2) is among the most suitable. efficient and selective for Mg. Moreover, only readily Around 90 % of the world Mg supply is produced in available and inexpensive reagents are needed and the China from dolomite, using the energy intensive and energy expenditure is minimal. polluting Pidgeon process.2,4 The aim of our work was to develop a hydrometallurgical procedure for the selective extraction of Mg from Slovenian dolomite. The emphasis was on high efficiency and selectivity for Mg over Ca, while other impurities were only monitored. The dolomite sample, identified as dolomite rock, was collected from a Slovenian quarry and prepared by grinding to a fine powder (performed by the Geological Survey of Slovenia). The elemental composition of the sample was then determined using microwave digestion and ICP-OES analysis. The sample was close to ideal Figure 1: Mg yield and molar ratio Mg/Ca for leaching dolomite with metal impurities of less than 0.15 % by with tartaric (T) and oxalic (O) acid. Leaching with weight and the molar ratio Mg : Ca of 0.93 : 1. sulfuric (VI) (S) acid was followed by precipitation with Electrochemical Monitoring of Polystyrene Nanoplastics–Fungal Biomass Interactions for Advanced Sensing Applications Tia Kralj*,1 2,3 1 Andrej Gregori, Gregor Marolt, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana, Slovenia 3 MycoMedica Ltd, Podkoren 72, SI-4280 Kranjska Gora, Slovenia Nanoplastics (NPs), ultrafine synthetic polymer TV biomass was introduced into 5.0 mL of 0.8 µg/L particles, are emerging as a concerning class of pollutants PSNPs suspensions, no decrease in ipa was observed, due to their high mobility, persistence, and ability to confirming a nearly-complete adsorption and thus penetrate biological membranes and accumulate within removal of PSNPs from solution. This process was aquatic ecosystems1. Their nanoscale dimensions observed in real time by the sensor without any (< 1 µm) and intricate interactions in water make their interferences of TV biomass particles, as confirmed by detection challenging, often requiring complex analytical additional control experiments using CV. This study procedures and sophisticated instrumentation techniques. presents a portable electrochemical sensor for real-time Their small size facilitates uptake by organisms and detection of PSNPs and their adsorption onto TV integration into food webs, raising concerns over biomass, offering a sustainable approach for on-site ecological and human health impacts2. In this work, we monitoring and mitigation of NPs pollution in aquatic present a novel, integrated approach to i) remove environments. polystyrene nanoplastics (PSNPs) from water using fungal biomass and ii) electrochemically monitor PSNPs for the evaluation of their adsorption onto the biomass surface. The mycosorption on a medicinal fungus Trametes versicolor (TV) particles was systematically evaluated under various operational parameters. Its effectiveness as a sustainable and efficient biosorbent was monitored and confirmed with the electrochemical sensor, based on a gold screen-printed electrode (Au- SPE), modified with a mesoporous silica thin film via electro-assisted deposition and subsequently 3,4 Figure 1: PSNPs detection via pore blocking on Establishing Enzyme Assays for the Detection of Tyrosinase and Hyaluronidase Inhibitors in Marine Algae Tina Kosovel,1 1 1 1 Eva Smrekar, Petra Tavčar Verdev, Marko Dolinar*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia We introduced to our laboratory and adapted two simple, robust, and rapid enzyme assays to determine whether algae from Sečoveljske soline contain substances that inhibit enzymes linked to skin aging. The assays need to be sensitive to low concentrations of inhibitors while remaining resistant to interference from other algal components. Tyrosinase catalyzes two steps in melanin production, including a rate-limiting one, making it a prime target for reducing hyperpigmentation. Hyaluronidase breaks down hyaluronic acid, and its Figure 1: Inhibition curve obtained by the optimized Inhibiting these two enzymes may help prevent both skin concentration of 60 U/ml. Data represent the mean ± SD from three independent experiments. conditions increased activity with age contributes to wrinkles. tyrosinase assay with kojic acid at an enzyme 1. For the tyrosinase inhibitory assay, we adapted a method based on the spectrophotometric detection of the reaction product2. In this assay, tyrosinase oxidizes dopamine to dopamine quinone, which exhibits a maximum absorbance at 475 nm. We modified the original assay by replacing the substrate L-tyrosine with dopamine, which is converted to the target product in a single step. Additionally, we optimized the concentrations of both enzyme and substrate to maximize the differential absorbance value. The assay was validated using kojic acid, a well-known tyrosinase inhibitor. Figure 2: Inhibition curve obtained by the hyaluronidase For the hyaluronidase assay, we adapted the method assay with tannic acid at an enzyme concentration of Investigating FHL2 and β-catenin Interaction by Cross-Linking Mass Spectrometry Tina Logonder1 1 , Aljaž Gaber * 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Epithelial Cell Adhesion Molecule (EpCAM) plays a crucial role in epithelial morphogenesis and carcinogenesis. It is one of the most utilized carcinoma biomarkers as its overexpression is linked to poor prognosis in various cancers. The intracellular domain of EpCAM (EpIC: aa 289–314) interacts with FHL2 and β- catenin, key proteins in the canonical Wnt signaling pathway. This complex induces the expression of oncogenes (Figure 1), making it a promising target for therapies1. Despite its functional relevance, structural Figure 1: Schematic representation of intracellular information of the complex with FHL2, β-catenin and signaling with EpIC EpIC remains limited. We aim to elucidate the structures using an integrative structural biology approach by employing Cross-Linking Mass Spectrometry (XL-MS) with BS3 and Small-Angle X-ray Scattering (SAXS). FHL2 (32.2 kDa) is composed of four and a half LIM domains, each containing 2 zinc fingers. β-catenin (85.4 kDa) consists of intrinsically disordered termini and the core region with 12 armadillo repeats (ARMs: aa 138- 664). The interface between FHL2 and β-catenin has not been precisely defined. It was investigated by yeast two- hybrid assay and coimmunoprecipitation. Results from two separate studies only partly overlap. The first one Figure 2: Cross-links between FHL2 and β-catenin showed that the N-terminus and ARM1 within β-catenin detected by XL-MS analysis. are crucial for binding. The second study determined that Analysis of adhesive joint using beech wood Tjaša Likeb*,1 Martin Capuder,2 Tina Skalar,1 Andreja Pondelak,2 Ermelinda Maçoas,3 Amelia Almeida4 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia (tjasa.likeb@gmail.com) 2 Slovenian National Building and Civil Engineering Institute, Dimičeva ulica 12, 1000 Ljubljana, Slovenia 3 CQE, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais 1, 1049-001 Lisbon, Portugal 4 CeFEMA, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais 1, 1049-001 Lisbon Portugal Wood is a porous, anisotropic material distinguished presence of growth rings, application time and pressure, by a complex array of natural anatomical features. In and the intrinsic wood structure. Moreover, after three softwoods, the main structural elements are longitudinal months of natural aging under the given conditions, no tracheids, while in hardwoods, they primarily consist of significant changes were observed in the physical, vessel elements and longitudinal fibers. The lumens of chemical, or mechanical properties of the joints. This these cells are generally large enough to permit the suggests that extended aging studies are necessary to gain infiltration of liquid adhesives, a process crucial for deeper insights into the long-term behavior of these forming strong and durable bonds. Despite this, adhesive bonded systems. joints are often regarded as the weakest components in wooden assemblies, as they are essential for transferring loads across the bonding interface. This highlights the importance of thoroughly investigating adhesive performance and behavior in such systems.1 In this study, beech wood with two grain orientations (radial and tangential) was bonded using polyurethane adhesive (PUR). Specimens were prepared in accordance with the EN 302-12 standard and subjected to natural weathering for three months in Lisbon, Portugal. The research aimed to evaluate both the depth and distribution of adhesive penetration in plates with different fiber and growth ring configurations. Additionally, the study examined the effects of aging by Where does Pb in the blood of children from the upper Meža Valley (Slovenia) come from? Tjaša Žerdoner 1,2 1 1 1 1 , Darja Mazej, Marta Jagodic Hudobivnik, Janja Snoj Tratnik, Neža Palir, Ingrid Falnoga,1 1,2 1,2 Milena Horvat, Tea Zuliani*, 1 Jožef Stefan Institute, Department of Environmental Sciences, Jamova cesta 39, SI-1000 Ljubljana, Slovenia 2 Jožef Stefan International Postgraduate School, Jamova cesta 39, SI-1000 Ljubljana, Slovenia Lead (Pb) is a naturally occurring, non-essential, and Pb isotope analysis of samples from the upper Meža toxic element. Contamination with it is both a persistent Valley revealed that local samples (e.g., soil, river global environmental and public health issue. An area in sediment, ores, mine waste) have a distinct Pb isotope Slovenia that has been significantly affected by Pb composition that differs from that of anthropogenic pollution is the upper Meža Valley. There lies the largest samples (e.g., Pb-battery components manufactured in the Pb and Zn ore deposit in Slovenia with a long history of area). Similar to the Pb isotope composition of PM10 mining and ore processing. During processing, a large samples, which showed a mix of local and anthropogenic amount of mine waste with high concentrations of Pb and sources, Pb isotope ratios in children's blood also showed other potentially toxic elements (PTEs) was produced and mixed values (Figure 1), suggesting that inhalation of deposited in abandoned mine shafts and near small contaminated air is a significant exposure pathway streams along the upper Meža Valley. As a result, the area alongside soil and dust ingestion. was excessively polluted with Pb, Zn, and other PTEs1,2. In addition to mining, the industry for the production and recycling of Pb-batteries has been present since 1956. As Pb is transmitted to the body through the inhalation and ingestion of contaminated dust particles, food, and water, surrounding inhabitants have had high levels of Pb in their blood. Especially at risk were young children, so the first testing for Pb concentrations in blood was carried out from 1974 to 1976, and the median Pb concentration in the blood of preschool children was over 400 µg/L3. In 1990, the concentrations decreased to (41– 284) µg/L. In 2001–2002, more than one-third of tested children still had elevated blood Pb concentrations Figure 1: Pb isotope ratios in the blood of children from Cancer-Associated Fibroblast Overgrowth Disrupts Lung Cancer Spheroid Formation: Characterization by Flow Cytometry Tonja Oman Sušnik 1,2 1,2 2 2,3 , Anamarija Habič, Barbara Breznik*, Metka Novak, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 National Institute of Biology, Večna pot 121, SI-1000 Ljubljana, Slovenia 3 Jožef Stefan International Postgraduate School, Jamova cesta 39, SI-1000 Ljubljana, Slovenia Non-small cell lung cancer (NSCLC) accounts for The outcome of the study highlights the problem of approximately 85 % of all lung cancer cases. It is CAFs overgrowth and the importance of their exclusion characterized by high aggressiveness, late detection and from primary NSCLC cell cultures. difficult treatment, which often results in poor prognosis 1. Our aim was to optimize the preparation of viable spheroids of repeatable sizes by centrifugation and to characterize cell cultures by analyzing the expression of cell markers. Three-dimensional (3D) tumor models, such as spheroids, are important for NSCLC studies, since they better mimic tumor microenvironment in patients than Scheme 1: Average diameter of LC004 and LC005 adherent cell lines2. We established spheroids from spheroids formed from 10,000 cells over a 3-day period. different numbers of cells from cell cultures LC004 and LC005, obtained from tumor biopsies of patients with NSCLC. After optimizing the protocol, we established spheroids from 10,000 cells and then continued their cultivation in ClinoStar bioreactor. We monitored the growth of 12 spheroids from each culture using light microscopy and analyzed spheroid diameter and surface area during cultivation. The results showed that the Scheme 2: Percentage of cells positive for tumor overall growth of spheroids was negligible. We noticed reduction in spheroid size (scheme 1) and significant cell napsin A, the epithelial marker EpCAM, and the markers cytokeratin 5 (CK5), cytokeratin 7 (CK7), p63, migration into the medium. Despite those results, fibroblast marker αSMA determined by flow cytometry. spheroids were of repeatable sizes. After the cultivation The effects of biodegradable and conventional microplastics on aquatic organisms Urša Košak 1 1 1,2 , Barbara Klun, Gabriela Kalčikova 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Faculty of Mechanical Engineering, Brno University of Technology, Brno, Czech Republic Microplastics are widespread pollutants that can oxygen and nutrients were also measured. These negatively affect aquatic ecosystems by accumulating in remained mostly stable across all groups, with only slight the organisms, disrupting physiological functions and variations observed in the P3HB treatment. acting as carriers for toxic substances. While The results indicate that the presence of microplastics biodegradable plastics are often marketed as can have effects on the aquatic ecosystems, even if they environmentally friendly alternatives, their degradation in are biodegradable. Future studies should consider long aquatic environments is often incomplete and slow, term studies and critical evaluation of the biodegradable allowing them to persist long enough to cause biological materials in aquatic environments. effects. Our study examined the impact of three differently persistent types of microplastics polyethylene terephthalate (PET, non-biodegradable), polylactic acid (PLA, low biodegradable) and poly(3-hydroxybutyrate) (P3HB, fast biodegradable), on two freshwater organisms Daphnia magna and Lemna minor. To mimic environmentally more relevant ecosystem, both organisms were combined in one microcosm. Over a 21- day period, the organisms were exposed to a concentration of 10 mg/L of microplastics with particle size of less than 80µm for all three types. The most visible effects were found at the presence of P3HB, where the reproduction of Daphnia magna was noticeably increased and accelerated. P3HB also had statistically significantly more fronds, longer root length and lower chlorophyll a Figure 1: Daphnia magna under the microscope (SEM) content in Lemna minor than control samples without with PLA microplastics visible in the gut (100x). microplastics after 21-day period. Analysis under Contaminants of Emerging Concern in the Ljubljanica River Basin: A Study of the Presence of Pesticides Veronika Plut,1,2 1,2 2,3,4 5 ,2 Jan Hočevar , Ester Heath, Jurij Trontelj, Vesna Mislej* 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Slovenian Chemical Society, Section for the Environment, Hajdrihova 19, SI-1000 Ljubljana, Slovenia 3 International Postgraduate School Jožef Stefan, Jamova cesta 39, SI-1000 Ljubljana, Slovenia 4 Jožef Stefan institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia 5 Faculty of Pharmacy, Aškerčeva cesta 7, SI-1000 Ljubljana, Slovenia A key element of water management in Slovenia is Table 7: Comparison of parameters MEC and LOQ with the National River Basin Management Plan, which chronic (C) and acute (A) PNEC in 2009–2023 (Note: provides a systematic assessment of the status of water *AA; **MAC). bodies. Accordingly, surface waters are evaluated in P L P MEC µg µg µg L NE ONE O RQ terms of their chemical and ecological status 1 . L L L Q Q C C-1-1-1 /2 In this study, we analysed publicly available 1 1 3 ≥ 2 surveillance monitoring data (2009–2023) on pesticides C C Tebuconazole 0.010* 1.2 0.01 √ 0.008 tested in the Ljubljanica River, at sampling points Azoxystrobin ˂0.002 * 0.676 C 0.01 √ 0.0015 C downstream of treated municipal wastewater discharges Propiconazole 0.046** 0.00085 A 0.002 X 54.1 A within the Ljubljana catchment, but upstream of the C C AMPA 0.13* 3.0 0.1 √ 0.043 confluence with the Sava River 1 . The dataset includes Metalaxyl ˂0.001 * 6.25 C 0.01 √ 0.0001 C 1,994 chemical measurements of pesticides, which C C Metazachlor 0.011* 0.0022 0.008 X 5.0 represent 63.8 % of all measurements carried out for 165 Flufenacet 0.048** 4.0 A 0.01 √ 0.012 A different groups of pollutants. The number of pesticides tested in the given period was 75. They are grouped as follows: i) year of their The results contribute to a more comprehensive measurement, ii) chemical properties and application, and understanding of the chemical status of the Ljubljanica iii) EU-authorised or banned River and support the development of more effective 1 . Of these, 97.5 % of the detected pesticides were below the limit of quantification methods for monitoring and assessing water pollution. (LOQ). Parameters such as measured environmental References: concentration (MEC) as the highest maximum annual concentration (MAC, acute impact) and the highest Moste-Podgrad, 1. Waters, Archive data - water quality, Ljubljanica, Pharmaceutical Contaminants in the Ljubljanica River Basin: Insights from National Surveillance Monitoring Jan Hočevar, 1,2 1,2 2,3,4 5 2 Veronika Plut, Ester Heath, Jurij Trontelj, Vesna Mislej*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Slovenian Chemical Society, Section for the Environment, Hajdrihova 19, SI-1000 Ljubljana, Slovenia 3 International Postgraduate School Jožef Stefan, Jamova cesta 39, SI-1000 Ljubljana, Slovenia 4 Jožef Stefan institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia 5 Faculty of Pharmacy, Aškerčeva cesta 7, SI-1000 Ljubljana, Slovenia Pharmaceutical active compounds (PhACs) are 2,3,4 literature) indicate that diclofenac, clarithromycin, among the major environmental pollutants in surface azitromycin, venlafaxine, and O-desmethylvenlafaxine waters because they are designed to affect living things present a high risk to the aquatic environment. directly. The results provide a clearer understanding of the In this study, we analysed parameters tested in the chemical status of the Ljubljanica River, particularly Slovenian national surveillance monitoring used to detect since several PhACs covered here will be regularly new pollutants in surface waters. The study looked at analysed under the new EU Urban Wastewater Treatment publicly available data for the period between 2015 and Directive, requiring quaternary treatment with at least 80 2023, focusing on sampling points downstream of treated % removal efficiency of the selected PhACs from municipal wastewater discharges (TMWD) in the wastewater. Ljubljana catchment, before the confluence of the Ljubljanica and Sava Rivers1. The dataset includes 148 chemical measurements of 58 substances, categorised by chemical properties and applications. The tested PhACs represented 35.2 % of all tested pollutants, while the proportion of their results was only 4.73 %, which indicates the technical complexity and cost of the measurement methods used1. The PhACs originating from various sources are collecting in TMWDs (Figure 1). In particular, we reviewed and compared parameters Figure 1: Sources of Pharmaceuticals in Waters. such as limit of quantification (LOQ), predicted no effect concentration (PNEC) and the risk quotient (RQ) . 1. 1,2,3,4 References: Determining the relevant PNEC for a specific PhAC Podgrad, http://arso.si/vode/podatki/, accessed: 2023 Waters, Archive data - water quality, Ljubljanica, Moste- Valorization Pathways for Sodium Sulfate: Addressing One of the Abundant By-products in Battery Industry Vinz Ymannuelle C. Tatad,1,2,3 1,2,3 1,2,3 Cédric Tassel, Jacob Olchowka*, 1 University of Bordeaux, CNRS, Bordeaux INP, ICMCB, UMR 5026, F-33600 Pessac, France 2 Réseau sur le Stockage Electrochimique de l’Energie (RS2E), CNRS FR 3459, Hub de l’Energie, Rue Baudelocque, 80039 Amiens Cedex, France 3 ALISTORE-European Research Institute, CNRS FR 3104, Hub de l’Energie, Rue Baudelocque, 80039 Amiens Cedex, France The rapid expansion of the lithium-ion battery (LIB) Together, these strategies aim to shift Na2SO4 from a industry has resulted in a significant increase in sodium costly waste disposal problem to an opportunity for sulfate (Na2SO4) production, making it one of the most resource recovery and circular economy integration, abundant inorganic by-products across the entire battery creating both economic value and environmental benefits value chain. Na2SO4 waste production occurs at multiple for the rapidly growing battery industry. stages of the LIB life cycle. The first major source is lithium refining, where hard-rock spodumene ores are processed into battery-grade LiOH or Li2CO3. The second is the production of precursor cathode-active materials (pCAM), such as Ni1−x−yMnxCoy(OH)2 or Ni1−x−yMnxCoyCO3 for use in NMC-type batteries. A third significant source is LIB recycling, particularly during the hydrometallurgical process. Over the full cycle of an NMC battery, approximately 2.5-3.5 kg of Na2SO4 is generated per kilogram of NMC cathode material. Projections indicate that by 2030, the LIB sector alone could produce as much as 8.8×103 metric tons of Na2SO 1 4 annually. Managing this growing waste stream through conventional methods (discharge through rivers) will be both costly2 2 and environmentally Figure 1 : Schematic overview of NaSO4 generation unsustainable, with potential threats to aquatic across the LiB (NMC type) lifecycle and proposed ecosystems and plant life3. solutions to address them. Design and Synthesis of Functionalized Carbazoles for Optoelectronic Applications Vitan Šlamberger 1 1 2 1 , Griša Grigorij Prinčič, Marko J ošt, Jernej Iskra*, 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Faculty of Electrical Engineering, University of Ljubljana, Tržaška cesta 25, SI-1000 Ljubljana, Slovenia Carbazole-based molecules have emerged as experimental approach enables the targeted optimization versatile and high-performing materials for interface of carbazole-based interfacial layers for improved engineering in optoelectronic devices, particularly as performance in PSCs, paving the way for more stable and hole-transporting self-assembled monolayers (SAMs) in efficient next-generation optoelectronic technologies. p–i–n perovskite solar cells (PSCs) (Figure 1). The widespread use of these molecules can be attributed to the unique combination of favorable properties, including favorable optoelectronic characteristics, high thermal and morphological stability as well as synthetic tunability that allows precise modification of both electronic and structural features. 1,2 The overall performance, efficiency, and long-term stability of PSCs are strongly influenced by the structural and electronic features of the SAM layer. Therefore, precise control over molecular orientation, electronic structure, and anchoring group design is essential for Figure 1: Schematic representation of a p-i-n perovskite oxide (ITO) integrity and stability while also ensuring molecules are flipped by 90 ° adopting a vertical optimal alignment between the SAM’s highest occupied orientation resulting in an increased dipole moment achieving efficient hole transport, maintaining indium tin solar cell and SAM interface tuning. Carbazole molecular orbital (HOMO) level and the electrode work function (WF).2,3 References: While many previous studies have focused on 1. Konidena, R. K.; Thomas, K. R. J.; Park, J. W. horizontal carbazole architectures,3 less is known about Recent Advances in the Design of Multi-Substituted the impact of orientation and dipole alignment at the Carbazoles for Optoelectronics: Synthesis and Understanding macronutrients through the three levels of chemical representation Zala Cvitanič 1 2 1,3 , Matjaž Dlouhy, Miha Slapničar* 1 University of Ljubljana, Faculty of Education, Kardeljeva ploščad 16, 1000 Ljubljana, Slovenia 2 Ledina Upper Secondary School, Resljeva cesta 12, 1000 Ljubljana, Slovenia 3 BEC Ljubljana, Cesta v Mestni log 47, 1000 Ljubljana, Slovenia Learning chemical concepts is based on the 14 years (M = 14.63, SD = .49) from a primary school in observation of natural phenomena. For the correct Ljubljana in the school year 2023/2024. construction of mental models of chemical concepts, it is Knowledge of the content of macronutrients and essential that both teaching and learning involve the possible misconceptions were measured using a paper- simultaneous integration of the three levels of and-pencil knowledge test. Students’ individual interest representation (Figure 1).¹ in learning this content was assessed using a purposive questionnaire, while motivation was explored through structured interviews. We interviewed the top 10% of students (high achievement) and the bottom 10% (low achievement) based on their performance on the knowledge test. The results show that half of the students (51%) Figure 1 achieved at least 50% of the total score in the knowledge : Johnstone’s model, the triangle of the triple nature of the chemical concept 1 test. A t-test revealed no statistically significant difference between the genders in the number of points achieved in It is known that students today tend to use learning the test (t(79) = −.87, p > .05). Almost half of the students strategies that allow them to make quick decisions with (48%) had misconceptions in at least one question. The minimal cognitive effort. However, these strategies can analysis of variance (ANOVA) showed no statistically oversimplify the learning process and lead to significant effect of individual interest on the students' misconceptions or incomplete understanding of, for test results, F(2, 79) = .42, p = .66, η² = .01. example, the relationship between the submicroscopic The surveys showed that students who scored well structure of a substance and its macroscopic properties. on the knowledge test generally had a stronger external Research shows that students who have developed orientation to learning chemistry. Regardless of their test misconceptions often justify their reasoning with a scores, all students indicated that experiments were the Expression and purification of recombinant ORF1p from LINE1 Zala Kramar*,1 Vera Župunski,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia The ORF1 protein (ORF1p) is an essential component of the LINE-1 retrotransposon, acting as a high-affinity, non-sequence-specific RNA-binding protein with nucleic acid chaperone activity. It contains an intrinsically disordered N-terminal region (NTR), a coiled coil domain (CC) responsible for trimerization, a central RNA recognition motif (RRM), and a C-terminal domain (CTD), which together mediate nucleic acid binding1. Our aim was to express and purify recombinant ORF1p fused to a maltose-binding protein (MBP) tag to assess construct stability and obtain enough protein for further studies. We constructed pMCSG7-MBP-ORF1p and pMCSG7-ORF1p-MBP vectors by PCR and in vivo assembly2. These constructs were expressed in E. coli BL21[DE3], and the resulting MBP-ORF1p and ORF1p- MBP fusion proteins were purified by immobilized metal ion affinity chromatography (IMAC), exploiting the His₆- tag, or MBP affinity chromatography targeting MBP. Figure 1: Structure of LINE1 ORF1p homotrimer (PDB After induction, both constructs showed bands near ID: 6FIA, 2YKP; AlphaFold DB ID: AF-Q9UN81-F1). the expected size (~83 kDa), but the C-terminal fusion (ORF1p-MBP) also displayed a lower band at ~66 kDa, suggesting possible proteolysis. We therefore focused on the MBP-ORF1p construct. To evaluate purification efficiency, we compared isolation of MBP-ORF1p using HisTrap and MBPTrap columns. In both cases, a substantial amount of the target Photoreaction of functionalized poly(β-benzyl-L-aspartate) with tetrazole and maleimide groups Zala Trbežnik 1 2 1 , Petra Utroša*, Jernej Iskra 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Department of Polymer Chemistry and Technology, National Institute of Chemistry, Hajdrihova ulica 19, SI-1000 Ljubljana, Slovenia Polypeptides are polymers with amide bonds (– correspond to the expected pyrazoline product. CONH–) between α-amino acid repeating units. Synthetic Presumably, NITEC photoreaction took place and polypeptides can be efficiently, cost-effectively, and on a pyrazoline initially formed but then rearomatized into large scale prepared from 2 N-carboxyanhydride pyrazole. This explains observed fluorescence, but the monomers via ring-opening polymerization.1 Various NMR spectra had no signals for pyrazole; since the functional groups can be implemented into the pyrazole structure contains double bonds in positions polypeptide structure to allow further post- without hydrogen atoms, there are no corresponding polymerization reactions. Especially interesting are protons for NMR detection. photochemical reactions as we can control when and Our work demonstrates successful poly(β-benzyl-L-where new bonds are formed. In this work, we explore a aspartate) functionalization with tetrazole and maleimide light-induced nitrile imine mediated tetrazole-ene groups, and shows promising results for NITEC cycloaddition (NITEC). In this 1,3-dipolar cycloaddition, photoreaction between the functionalized polypeptide photoexcited tetrazoles release nitrogen to generate chains. reactive nitrile imines in situ, which then react with various alkenes or alkynes to form pyrazoline-based covalent bonds.2 The goal of our work was to functionalize polypeptide chains with a tetrazole and a maleimide group, and demonstrate linking of the two polypeptides together via NITEC reaction. To obtain a polymer with functional groups suitable for NITEC, we first transformed our starting material, β- benzyl-L-aspartate, into β-benzyl-L-aspartate N- carboxyanhydridemonomer. The 1H nuclear magnetic resonance (NMR) spectroscopic data of the synthesized Coordination of deprotonated furosemide to zinc(II) ion Zarja Uranjek,1 Barbara Modec,1 Nina Podjed Rihtaršič*,1 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Furosemide (IUPAC name: 4-chloro-2-(furan-2- ray diffraction analysis. The results will be presented to ylmethylamino)-5-sulfamoylbenzoic acid) is a loop shed light on the possible composition of the unidentified diuretic used to treat hypertension and edema, caused by compound. cardiac, renal and hepatic failure. It is a weak acid with a pK 1 value of 3.8. In addition to the carboxyl group, a furosemide molecule contains a chlorine-substituted phenyl ring, a furan ring, a sulphonamide group and an amine group (Figure 1). These functional groups enable the coordination of furosemide to metal centers. Surprisingly, only two crystal structures have been reported in the Cambridge Structural Database 2 featuring coordination compounds with furosemide anion. The latter is coordinated either in a monodentate or bidentate chelating manner via carboxylate oxygens. Figure 4: ORTEP drawing of [Zn(fur)2(NH3)2] References: 1. Granero, G. E.; Longhi, M. R.; Mora, M. J.; Junginger, Figure 3 H. E.; Midha, K. K.; Shah, V. P.; Stavchansky, S.; : Structural formula of furosemide. Dressman, J. B.; Barends, D. M. Biowaiver remains relatively unexplored with this pharmaceutically Zinc(II), a biologically important Lewis acid, monographs for immediate release solid oral dosage forms: furosemide. J. Pharm. Sci. 2010 , 99 , 2544 – 2556. active ligand. Therefore, this study investigates zinc(II) 2. Groom, C. R.; Bruno, I. J.; Lightfoot, M. P.; Ward, S. coordination chemistry with deprotonated furosemide. C. The Cambridge Structural Database. Acta Immobilization of glucose oxidase into cross-linked enzyme aggregates using microfluidic system Žiga Gerdina 1 1 1 *, Borut Šketa , Polona Žnidaršič Plazl , 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia Biocatalysis offers a sustainable and efficient alternative to conventional chemical synthesis, enabling reactions with high selectivity and yield under mild, environmentally friendly conditions1. However, the industrial application of enzymes is often limited by their poor operational stability, sensitivity to process conditions, and high cost2. Immobilization techniques, such as the formation of cross-linked enzyme aggregates (CLEAs), have been developed to overcome these limitations by improving enzyme robustness, simplifying product separation, and allowing enzyme reuse3. When combined with microfluidic systems, CLEA synthesis benefits from precise control over reaction parameters, Figure 5: CLEA particles synthesized under optimal rapid mixing, and scalability for continuous-flow conditions; observed by SEM at 25000× magnification. operation4. CLEAs from glucose oxidase (GOx) 1. Bai, J.; Huang, C.; Liu, Y.; Zheng, X.; Liu, J.; Zhou, — a model enzyme L.; Liu, J.; Jiang, Y. Integrating Biocatalysis with with broad applicability in biosensors, antimicrobial In this study, a microfluidics-based generation of References: 5 Continuous Flow: Current Status, Challenges, and systems, and biofuel cells —was optimized. Using the Future Perspectives. J. Adv. Res. 2025. Box–Behnken experimental design, the influence of 2. Žnidaršič-Plazl, P. Biocatalytic Process enzyme, precipitant, and cross-linker concentrations on Intensification via Efficient Biocatalyst investigated. At the optimized conditions, monodisperse Integration. Curr. Opin. Green Sustain. Chem. 2021, 32 (100546), 100546. CLEA-GOx particles with an average hydrodynamic the properties of the resulting particles was systematically Immobilization, Miniaturization, and Process 3. Sheldon, R. A.; van Pelt, S. Enzyme Immobilisation radius of 113 ± 1.37 nm were produced and an in Biocatalysis: Why, What and How. Chem. Soc. Rev. immobilization yield of 100% was reached. The 2013, 42 (15), 6223–6235. N-Substituted 2-phenylimidazoles as substrates in the catalytic C–H bond functionalization Žiga Močnik 1 1 1 1 1 , Uroš Grošelj, Jurij Svete, Bogdan Štefane, Franc Požgan * 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, 1000 Ljubljana, Slovenia Catalytic C–H activation has emerged as a products (with mono:di ratios ranging from groundbreaking strategy for the formation of new carbon- approximately 2:1 to 3.7:1), the benzyl group carbon (C–C) bonds, particularly in the synthesis of predominantly directed the reaction towards the biaryls, which are crucial building blocks in formation of the monosubstituted product. Furthermore, pharmaceuticals, agrochemicals, and materials science. for the substrate bearing a (2-pyridyl)methyl group, we This approach offers significant advantages over classical observed an intriguing additional C–H activation on the cross-coupling reactions, such as enhanced atom and step imidazole ring itself, however most of products formed in economy, and reduced waste generation. While palladium trace amounts. All resulting products were catalysts have traditionally been at the forefront of C–H comprehensively characterized using 1H NMR, 13C NMR, activation, cheaper and air-stable ruthenium catalysts IR spectroscopy, and high-resolution mass spectrometry have become an attractive alternative, enabling access to (HRMS). otherwise challenging reaction pathways. However, the widespread application of C–H arylation still faces challenges, including issues with regioselectivity, the frequent requirement for specific directing groups, and often elevated reaction temperatures that are unsuitable for thermally labile substrates.1,2,3 Scheme 1: Two-step reaction scheme The aim of our research was to investigate the influence of various N-substituents of 2-phenylimidazole Table 8: Ratios of mono- and diaryl products on the outcome of catalytic ortho C–H arylation. We R Mono:Di synthesized a series of N-functionalized 2-phenyl-1H- Me 2:1 imidazoles (including N-methyl, N-ethyl, N-butyl, N- Et 3.7:1 benzyl, and N-pyridylmethyl derivatives) starting from Bu 2.8:1 commercially available 2-phenylimidazole and Bn Mono appropriate organic halides, following established pyridylmethyl / literature procedures. These products were then subjected NAC supplement development for the reduction of the health gap Product development and business plan Aljaž Kostevc Redek 1 1 1 1 *, Danial Doustmohammadi, Diana Doustmohammadi, Mihajlo Krstić , Andraž Velušček,1 1 Mentor: Alenka Mozer* 1 Gimnazija Vič, Tržaška cesta 72, 1000 Ljubljana, Slovenia In Slovenia, only half of the low-income population follow instructions, and diverse sales and marketing is in good health, compared to only a quarter in Lithuania. channels that address the price-sensitive market. The 70–80% of people with high incomes report good health. business model targets value-driven consumers through Significant health gap can also be observed between retail partnerships with discount stores, online sales via different levels of education. To help reduce the health our own website featuring AI chatbot support, and gap of the disadvantaged population, we (1) developed a strategic collaborations with local pharmacies and health product mix to address common health issues and boost centers. Competitive pricing significantly differentiates the immune system, and (2) prepared a business idea, us. Our sales strategy initially targets Slovenia and the targeting those people (OECD, 2025 and Eurostat, 2025). EU. We expect to reach target market in 3 years. Primary goal was to develop a scientifically backed, price-competitive, synergetic antioxidative mix—a Table 1: Developed products and supportive services product designed for daily use that boosts the immune Product system and neutralizes free radicals through a carefully RespiraNAC NAC Protect selected combination of antioxidants. Secondly, we have 200 mg N-acetilcistein 400 mg N-acetilcistein (NAC)*, Koencim Q10, created a synergetic mucolytic mix that focuses on (NAC)*, Vitamin B3, Vitamin C, Vitamin E, thinning mucus and supporting respiratory health, making Vitamin A, Vitamin B7 Vitamin B2, Selenometionin breathing easier during respiratory discomfort. Lastly, we Support for coughing up Strengthens the natural have established a comprehensive customer support thick mucus. Contributes immune response. Works in provide scientifically grounded information and promote breathing environments. Restores levels of the body's antioxidant - transparen system to easier both water and fatty , including a website, chatbot, and live support, to t communication about the products’ effects glutathione and potential side effects in a user-friendly manner. Last, Supportive services we prepared a comprehensive business plan, studying the https://wellforall.netlify.app customer needs, market analysis, and preparing a Web page With Customer support (AI production and financial plan for a start-up company. chatbot and expert support) and Web- shop Methodology. We reviewed scientific literature on *serving size is 2 capsules NAC and other ingredients, analyzed health and market Microplastics found in food from cutting boards Brina Zver*,1 Klara Grantaša*,1 Mentors: dr. Marija Meznarič,1 dr. Manca Kivač Viršek,2 1 Gimnazija Franca Miklošiča Ljutomer, Prešernova ulica 34, 9240 Ljutomer 2 National Institute of Biology, Marine Biology Station Piran, Fornače 41, 6330 Piran Microplastics are all plastic particles smaller than 5 mm. Based on their shape, they are divided into six basic groups: fragments, films, pellets, granules, filaments and foams. The most common sources of it are plastic containers, cutlery, straws, tea bags, plastic food packaging and canned coatings. Traces of microplastics are also obtained to our food through plastic cutting boards. According to a study by the Chemical Association, an individual is on average exposed to up to 79.4 million microplastic particles per year. Microplastic particles can be introduced into our Figure1: Average number of microplastic particles body in different ways: oral (most commonly), created depending on food hardness and cutting board respiratory and topical. On average (oral and respiratory wear milk, tea bags and seafood due to plastic pollution of the formed particles of microplastics and their standard deviation ocean. They bring physical (e.g. damage to the body when PP+AA PP+AA PP+PE PP+PE PP N plastic particles rub against the tissue), chemically (e.g. microplastic particles annually. It is present in water, Table 9: Minimum, maximum and average size of intake) an individual ingests between 74,000 and 121,000 PP W N W N W consequences of chemicals contained in plastics) and Min. size 67,2 70,9 66,2 74,4 47,4 95,1 biologically (e.g. transmission of pathogens) dangers to Max. 1766,4 1636,4 1119,9 2166,7 728,6 3050,8 our body. size We found differences in the excretion of the Average 567,1 582,3 515,5 531,3 288,2 774,4 amount and size of microplastic particles according to the size Standard wear and chemical composition of the plastic cutting 532,2 434,6 268,0 488,6 200,4 682,4 deviation boards. We compared polypropylene, a mixture of polypropylene and polyethylene, and a mixture of References: Galvanic Coupling of Aluminium Alloy and Copper for Electric Vehicle Applications Peter Rodič 1 2 3 1 3 *, Eva-Arolea Trdan, Andraž Logar , Ingrid Milošev, Damjan Klobčar , 1 Jožef Stefan Institute, Department of Physical and Organic Chemistry, Jamova c. 39, SI-1000 Ljubljana, Slovenia 2 Gimnazija Ledina, Resljeva c. 12, SI-1000 Ljubljana, Slovenia 3 University of Ljubljana, Faculty of Mechanical Engineering, Laboratory for Welding, Aškerčeva cesta 6, SI-1000 Ljubljana, Slovenia The increasing integration of aluminium and copper polymeric sealants, or mechanical design modifications to components in electric vehicles (EVs) and battery interrupt electrical continuity at dissimilar-metal systems presents significant corrosion challenges arising junctions. Among these, anodizing is particularly from galvanic coupling effects.¹ When two dissimilar promising, forming a dense, adherent oxide layer that metals are in direct electrical contact within a conductive reduces both general corrosion and the galvanic potential electrolyte, the less noble metal (aluminium) acts as the difference. anode and corrodes preferentially, whereas the more noble metal (copper) functions as the cathode and remains largely unaffected.² Such interactions are of particular concern in EV battery modules, busbars, cooling systems, and structural assemblies, where weight reduction, electrical conductivity, and thermal management are 2 cm critical to performance.¹ While the use of aluminium offers substantial mass savings, its electrochemical Figure 1: An example of galvanically coupled AA6082 activity in the presence of copper poses a durability risk, and Cu-ETP mounted in 3D printed holder during testing particularly in chloride-containing environments. in the salt spray chamber (after 192 hours). This study investigates galvanic corrosion between AA6068 and Cu-ETP, both widely applied in high- This research is also a restudy for developing durable performance EV assemblies. Cylindrical specimens with protective technologies to extend the operational lifespan diameters of 6.8 mm, 9.4 mm, and 11.4 mm were of EV energy and power systems. Future work will precision-machined and mounted in a custom 3D-printed include systematic evaluation of multiple surface polymer holder (Figure 1), ensuring reproducible, flush treatments under prolonged salt spray and cyclic metallic contact at the coupling interface. The coupled corrosion exposures, combined with electrochemical Compartment-based pharmacokinetic model of phytocannabinoids in chronic users Niko Nolimal,1 Gregor Čekada,1 Patricija Zupan,3 5 Mentors: Miha Slapničar , 1,2 Nejc Umek, 4 Janez Mavri* 1 Biotechnical Education Centre Ljubljana, Cesta v Mestni log 47, 1000 Ljubljana, Slovenija 2 Faculty of Education, University of Ljubljana, Kardeljeva ploščad 16, 1000 Ljubljana, Slovenija 3 Gymnasium Kranj, Koroška cesta 13, 4000 Kranj, Slovenija 4 Faculty of Medicine, Institute of Anatomy, University of Ljubljana, Korytkova 2, 1000 Ljubljana, Slovenija 5 National Institute of Chemistry, Hajdrihova ulica 19, 1000 Ljubljana, Slovenija In our society, the use of marijuana is increasing psychoactive and is excreted directly via the urine. For among the young population. To investigate the long- this reason, it is the only metabolite that we have included term effects of marijuana on chronic users, we performed in our model. compartment-based pharmacokinetic modelling of the There is virtually no gradual accumulation of THC THC molecule and its metabolites THC−OH and and its metabolites THC−OH and THC−COOH in users THC−COOH. who smoke marijuana infrequently. In chronic marijuana The THC molecule occurs naturally in various plants users, however, the amount of THC ingested is greater of the Cannabis genus, such as Cannabis indica, Cannabis than the number of cannabinoids metabolised and sativa and Cannabis ruderalis. The highest concentration excreted. This effect can lead to false positive results in of THC is found in Cannabis sativa. tests for marijuana use conducted by law enforcement and Pharmacokinetics includes ingestion, metabolism medical personnel. and excretion. When marijuana is smoked, the smoke is absorbed through the alveoli directly into the bloodstream, which means that smoked THC bypasses first-pass metabolism (it does not pass through the liver). It then passes directly into adipocyte tissues such as the brain and fat1. In the brain, it affects the endocannabinoid nervous system. The endocannabinoid nervous system consists of neurotransmitters, receptors and various enzymes that synthesise and break down the neurotransmitters. A Optimization and study of milk protein coloration, as a sustainable alternative to traditional plastic materials Iva Jaklin,1 1 1 Anže Škrget , Mentor: mag. Ni na Žuman 1 Gimnazija Franca Miklošiča Ljutomer, Prešernova ulica 34, 9240 Ljutomer, Slovenia Write abstract text here. Use Times New Roman; font squares showed that the products made from protein milk size 10; justified; line spacing 1,15 (Normal style). Total had the best ratings in terms of strength and temperature abstract length is limited to one page. Please delete the resistance. We also formed beads from some of the instructions page before submission. Cite1 2a references in samples, which we tested as dog toys – this allowed us to the following2 3 fashion. explore additional practical applications of the Plastic waste is an increasingly serious biopolymer. Overall, a reduction in the amount of environmental problem, so the development of precipitated casein was observed in all types of milk, biodegradable materials is necessary to reduce pollution likely due to protein degradation by proteases, and the consumption of fossil resources. In the study, we denaturation of casein and/or formation of insoluble examined the possibility of preparing a biopolymer from aggregates, and changes in the colloidal structure of milk denatured casein, a milk protein, as a sustainable caused by microbiological activity. alternative to synthetic plastic. We determined how the We can conclude that protein milk has the greatest type of milk, amount and concentration of vinegar, and potential for producing bioplastics based on casein. milk temperature affect the utilization of casein and the Incorporating surplus or waste milk into the process properties of the resulting solid substance. further enhances the environmental and economic Casein is a natural phosphoprotein that forms 3 justification for using this material. colloidal micelles in milk that are sensitive to pH changes1. At a pH reduction to the isoelectric point (around 4.6), the micelles destabilize and precipitate. The precipitation process was carried out using alcohol vinegar, and the resulting precipitate was dried and formed into blocks. We examined the mechanical properties of the products, including strength, odor, heat resistance, and practicality. We used different types of milk: whole, low-fat, protein (with higher protein content), soy milk, and Figure 1: Condition of squares made from different Water electrolysis in connection with fuel cells Maj Matjašec,1 Jakob Ritlop,1 Mentor: Nina Žuman*,1 1 Gimnazija Franca Miklošiča Ljutomer, Prešernova ulica 34, SI-9240 Ljutomer, Slovenia Fossil fuels and their excessive use remain one of the principle of converting hydrogen into electrical energy, key problems of modern society, contributing to although the output was limited due to the simplicity of environmental pollution, resource depletion, and climate the setup. change. As an alternative, hydrogen offers a cleaner and Overall, our project highlighted the potential of more sustainable energy solution, especially when used in hydrogen as a sustainable and clean energy source for the fuel cells, which produce only water and heat during future, though further research is needed to improve operation. This greatly reduces waste compared to efficiency, reduce costs, and enable large-scale conventional energy sources and helps lower harmful applications. emissions. In our research project, we investigated the production of hydrogen through the electrolysis of water, where an electric current splits water into hydrogen and oxygen gases. We examined how different factors affect gas production, including the type of electrolyte (sodium hydrogen carbonate – NaHCO₃ vs. potassium iodide – KI), the applied voltage and current, and the duration of the process. Our experiments showed that potassium iodide (KI) is a more effective electrolyte than sodium hydrogen carbonate, resulting in greater gas production. We also confirmed that higher voltage and current increase the Scheme 1: Volume of products amount of gas produced, and that gas formation is proportional to time, following a consistent linear trend. References: (Proton Exchange Membrane) fuel cell using materials Elsevier Academic Press: Burlington, 2005 2. As a practical application, we built a simple PEM 1. Barbir, F. PEM Fuel Cells: Theory and Practice; available in the school laboratory. The fuel cell Smrdu, A. Snov in spremembe 2; Založništvo Jutro: Ljubljana, 2007; pp 100 – 101. successfully generated 0.14 V, demonstrating the basic Antimibiofilm and antibiofilm activity of natural substances against Escherichia Coli bacteria Jon Milič 1 1 2 , Mentors: dr. Marija Meznarič, prof. bio. in kem. , izr. prof. dr. Martina Oder, dipl. san. inž. 1 Gimnazija Franca Miklošiča, Prešernova ulica 34, SI-9240 Ljutomer, Slovenia 2 Zdravstvena fakulteta Univerze v Ljubljani, Zdravstvena pot 5, SI-1000 Ljubljana, Slovenia Nowadays, people are increasingly exposed to influenced by environmental factors, as we detected microorganisms that have various effects on our health. differences between individual ethereal charcoals, Due to the increasing use of antibiotics, cleaning agents depending on the year the plants were harvested. and other synthetic substances, pathogenic microorganisms are becoming resistant. This makes it difficult to remove them, which in turn can lead to more and more bacterial diseases. For this reason, we decided to study alternatives to modern cleaners in the research. Essential oils were used in the research. We determined the effectiveness of essential oils of lavender, ginger and spruce tops in removing Escherichia coli bacteria and the ability of lavender essential oil in removing biofilms. We used 100% concentration of all extracts, and for lavender Figure 1: Escherichia coli essential oils we also used oils diluted to 50% and 25%. effectiveness of these oils obtained from plants in 2022, 1. Nazzaro, F., Fratianni, F., De Martino, L., Coppola, R., De Feo, V., Effects of essential oils on In the case of lavender essential oil, we also checked the References: 2023 and 2024. We first prepared an overnight culture of pathogenic bacteria. Pharmaceuticals 2013, 6, 1451– E. coli bacteria on a nutritious microbiological medium, 1474 which we later used in the experimental work. Discs 2. Nour, A. H., Modather, R. H., Yuns, R. M., Elnour, soaked with essential oils were applied to the cultures and A. A. M., Ismail, N. A., Characterization of bioactive the antimicrobial effectiveness of each substance was compounds in patchouli oil using microwave-assisted part of the experiments, we transferred the bacteria to Prod. 2024, 208, 117901 3. Oder, M., Piletić, K., Fink, R., Marijanović, Z., microtiter plates, in which we grew a biofilm, which was determined based on the inhibition zones. In the second and traditional hydrodistillation methods. Ind. Crops Krištof, R., Bićanić, L., Tomić Linšak, D., Gobin, I. treated with essential oils in the next steps. After exposing A synergistic anti-bacterial and anti-adhesion activity Electrochemical determination of bisphenol S in thermal paper Lara Marzidovšek,1 Katarina Šela,1 Mentors: Damijana Gregorič*,1 doc. Kristina Žagar Soderžnik*,2 1 Srednja šola Slovenska Bistrica, Ulica dr. Jožeta Pučnika 21, 2310 Slovenska Bistrica 2 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, 1000 Ljubljana, Slovenia Endocrine disruptors are substances that are foreign people (OEP) such as shopkeepers and restaurant to the body but, due to their structural similarity to natural workers, the value is greater than 0.00002 ng/day. In the hormones, can mimic, enhance, or block their function. EU, the Tolerable Daily Intake (TDI) for BPS has not yet When their harmful effects are proven, they are referred been determined. If the TDI limit were set the same as for to as endocrine-disrupting chemicals. Among them are BPA (0.2ng/kg body weight/day), occupationally bisphenols, a group of organic compounds that are widely exposed people could exceed this limit just from contact used in industry, particularly in the production of plastics with thermal paper, bearing in mind that this does not and epoxy resins. Due to their properties – such as include other possible sources of exposure. strength, impact resistance, and transparency – bisphenols are essential for the manufacture of everyday products, such as plastic bottles, packaging, and children's toys. Bisphenols are also used as an organic acid in thermal paper, which is used to print invoices, labels, declarations, ATM receipts, airline tickets and parking tickets. In this form, they can be particularly problematic, as they can be Scheme 1: The process of preparing real samples more easily absorbed into food, our bodies and the environment as monomers. Table 10: Estimated TOTAL daily intake of BPS due to The purpose of the present research work was to contact with thermal paper study the presence of bisphenol S (BPS) in thermal paper, EDI EDI(OEP) All EDI EDI (OEP) which is often encountered in shops, restaurants, [ng/day/kg [ng/day/kg samples [ng/day] [ng/day] drugstores and other everyday environments. The aim Overall bw] bw] was to determine whether BPS is present in most samples 0,0000003 0,000022 0,004 0,31 average and to check the absence of bisphenol A, which is banned in the European Union. Using cyclic voltammetry, we analysed 26 thermal References: paper samples and prepared BPS standard solutions of 1. Bisfenoli – ECHA https://echa.europa.eu/sl/hot- Water never lies: Wastewater-Based Epidemiology as a Tool to Understand Psychoactive Substance Use Among Youth Lan Dular, 1 1 2 3 1 Aljaž Maraž, Nejc Murn Blažić, Mentors: dr. Ana Kroflič*, mag. Nika Cebin*, Stana Kovač Hace*,2 2 dr. Uroš Ocepek*, 1 Gimnazija Ledina, Resljeva cesta 12, 1000 Ljubljana 2 Srednja tehniška in poklicna šola Trbovlje, Šuštarjeva kolonija 7a, 1420 Trbovlje, 3 Kemijski inštitut Ljubljana, Hajdrihova ulica 19, 1000 Ljubljana Understanding patterns of psychoactive substance (PAS) use among youth remains a public health and environmental challenge. In this interdisciplinary study, we employed wastewater-based epidemiology (WBE) to analyze wastewater samples from treatment plants in Ljubljana and Trbovlje, Slovenia, aiming to detect the presence of PAS and correlate them with survey-based data collected from high school students. Solid phase extraction was used to concentrate the wastewater samples before applying rapid immunoassay urine tests. We also conducted an anonymous online survey (N = 532) and analyzed the results using cluster analysis and AI-based data mining. Our findings confirm the presence of cocaine, THC, and morphine in wastewater samples, particularly after Figure 1: Solid-phase extraction cartridges (Oasis sample concentration. Initial unprocessed samples MCX) used for sample concentration from wastewater yielded false negatives due to low analyte levels, samples collected in Ljubljana and Trbovlje. significant peer influence and accessibility of PAS in References: 1. Castiglioni, S.; Zuccato, E.; Crisci, E.; Chiabrando, confirming our hypothesis. The survey revealed urban areas. Data mining identified four distinct user C.; Fanelli, R.; Bagnati, R. Environ. Sci. Technol. groups differing in experience, attitudes, and risk 2006, 40 (12), 357 – 363. perception. 2. Castiglioni, S.; Bijlsma, L.; Covaci, A.; Emke, E.; This study highlights the complementary nature of Hernández, F. Sci. Total Environ. 2011, 409, 3564– Analysis of awareness of “zero waste” and Maraton treh src Petra Ouček1, Lara Širovnik*1, Mentor: Mateja Godec1 1 Gimnazija Franca Miklošiča Ljutomer, Prešernova ulica 34, SI-9240 Ljutomer, Slovenia Recycling practices have been known for thousands very supportive of the marathon's "zero waste" operation of years. With the development of industry, we have and encouraged more events like this. become a community that exceeds 8 million tons of waste annually in Slovenia alone.1 Despite the increasing amount of waste, attitudes towards it have begun to change – we have created a concept that tries to preserve resources through production, consumption, and reuse, without burning or otherwise endangering the health of the ecosystem and people. We started using the term “zero waste” for it. 2 Most of the movement in Slovenia is promoted by the organization Ekologi brez meja, which aims to guide individuals and organizations in preventing waste generation through awareness and networking. And one such event where they help is the Maraton treh src.3 Scheme 1: Percentage of participants that knew that Maraton treh src is a sports event organized by Maraton treh src was and is a “zero waste” event Športno društvo Tri srca since 1981. At the same time, it is also the first running event in Slovenia with a "zero We also conducted two interviews. One was with waste" vision and the "zero waste" event title it received Marko Pintarič, the main organizer of the marathon, are responsible for waste management, proper sorting, waste” needs various adjustments and more financial 4 and educating visitors on proper waste sorting. resources, since environmentally friendly materials are in 2022. At the event itself, members of “zelena straža” where he told us that organizing such a big event as “zero The first research method was the observation of the often more expensive. The second interview was with “zelena straža” on how visitors handle waste. We noticed Katja Sreš, a representative of the association Ekologi that a large number of visitors would have thrown away brez meja. She expressed the importance of “zelena their trash incorrectly if it weren’t for the intervention of straža” helpers at these events are of great meaning and MR-Pingvini vs. Pasosaurs: A Mixed Reality Game for Raising Awareness about Psychoactive Substances Luka Tomažin, 1 1 2 1 1 Ajk Kalaba, Aljaž Maraž , Mentors: d r. Uroš Ocepek *, Stana Kovač Hace, mag. Nika Cebin,2 1 Srednja tehniška in poklicna šola Trbovlje, Šuštarjeva kolonija 7a, 1420 Trbovlje, Slovenia, 2 Gimnazija Ledina, Resljeva cesta 12, 1001 Ljubljana, Slovenia Rapid technological advancements, particularly in With this study, we demonstrated that young people artificial intelligence (AI) and mixed reality (MR), offer can effectively engage their peers in understanding and new opportunities to create engaging educational tools. embracing social values. As Peter Drucker famously said, Our project aimed to develop an innovative card game "In times of turbulent change, the issue is not the enhanced by mixed reality to raise awareness among turbulence, but rather acting with yesterday's logic." adolescents about the risks of psychoactive substances (PAS). Each card in the game represents a creature: penguins, symbolizing resistance strategies against PAS, and "pasosaurs", dinosaurs alike monsters, representing specific psychoactive substances—designed using generative AI to reflect the substance’s characteristics and effects visually. Considering the increasing popularity of tobacco and nicotine products among youth, we conducted a survey involving 532 students aged 14–19 from STPŠ Trbovlje and Gimnazija Ledina. Our results indicated that curiosity and peer influence are primary reasons behind adolescents' experimentation with new nicotine products Figure 1: Students playing the MR-enhanced card game, such as vapes and nicotine pouches, surpassing traditional experiencing interactive learning through mixed reality cigarette usage. technology. The developed game integrates digital MR elements via AR.js and p5.js libraries, enhancing interactivity and References: engagement. Through gameplay, participants confront 1. Jannah, N. L.; Wasis, W.; Wiryanto, W.; Suryanti, S.; realistic scenarios regarding substance effects, fostering Widodo, W.; Murni, A. W.; Farokhah, L.; Penehafo, Microbiological analysis of concrete surfaces at Jože Plečnik Gymnasium Ljubljana Mija Kapun,1 1 1 2 2 Emilija Rojnik, Mentors: mag. Darja Silan*, doc. dr. Polona Zalar, doc. dr. Martina Turk, 1 High school Jože Plečnik Ljubljana, Šubičeva 1, SI-1000 Ljubljana, Slovenia, SI-1000 Ljubljana, Slovenia 2 Biotechnical Faculty, University of Ljubljana, Department of Biology, Večna pot 111, SI-1000 Ljubljana, Slovenia In this study, we investigated the presence and potential impact of micro-organisms on concrete, focusing on their growth, metabolic properties and ability to dissolve and form minerals. We sampled different locations of the concrete fence on the terrace of the Jože Plečnik Gymnasium and also took concrete samples at selected sample points. When observing the growth of micro-organisms, we found that higher microbial diversity and abundance occur on wetter surfaces. By analysing the concrete samples under a scanning electron Picture 2: Overview of microbial growth on R2A, HA, microscope (SEM), we detected biological debris and MEA + Ch, DRBC and BHI media from all sampling calcium carbonate (CaCO3) deposition, with the highest sites after 144 hours of incubation. deposition detected on the sample with the higher moisture content, which also had the highest growth and biodiversity, indicating that the CaCO3 is formed due to microorganism activity. Further analysis showed that isolated micro- organisms can have both positive and negative effects on concrete. Some isolates form pigments that can cause undesirable aesthetic changes to concrete surfaces, while others are capable of dissolving CaCO3, which can lead Picture 3: Crystal formation by bacterial isolate B-2. to material degradation. We have also identified micro- organisms that exhibit positive properties for concrete, such as the ability to biomineralise, which allows the References: concrete to self-harden. All fungi and two selected 1. Agarwal, H.; Bajpai, S.; Mishra, A.; Kohli, I.; Varma, bacterial isolates are urease-positive, which means that A.; Fouillaud, M.; Dufossé, L.; Joshi, N. C. Bacterial they form minerals by the process of ureolysis. The results pigments and their multifaceted roles in contemporary of our research also suggest ways to protect concrete biotechnology and pharmacological applications. Synthesis of silicon dioxide nanoparticles with incorporated fluorescent dyes Petra Ouček 1 1 1 2,3 *, Lani Habjanič , Mentors: Mateja Godec, Sebastjan Nemec 1 Gimnazija Franca Miklošiča Ljutomer, Prešernova ulica 34, SI-9240 Ljutomer, Slovenia 2 Materials Research Laboratory, University of Nova Gorica, Vipavska cesta 11c, 5270 Ajdovščina, Slovenia 3 Department for materials synthesis, Jožef Stefan Institute, Jamova cesta 39, 1000 Ljubljana, Slovenia Nanotechnology is an interdisciplinary and fast individual silane-functionalized fluorescent dyes.3 developing field focused on the research and creation of Furthermore, the NP were colloidally stabilized by materials, usually nanoparticles (NP), with distinctive attaching polyethylene (PEG) molecules on their surface. properties in the nanometer scale. The high surface-to- The NP were characterized with several analytical volume ratio of such materials enables innovations methods. Their dimensions were determined by electron beyond the possibilities of conventional materials of microscopy (Figure A and B) and their hydrodynamic larger size.1 diameter was measured by dynamic light scattering. We Silicon dioxide, referred to as silica, is a material found that the fluorescein-embedded NP had the largest widely present in nature and everyday life – as silica gel diameter (147 nm) and rhodamine-embedded ones had packets, used to adsorb moisture, and as quartz sand in the smallest (118 nm). As expected, the attachment of construction and glassmaking. Moreover, silica has low PEG to the NP surface increased the hydrodynamic toxicity and is biocompatible, which enables its use in diameter (by ~10 nm). Moreover, the surface charge of pharmaceutical, cosmetic and food industries. Silica the NP was determined with zeta potential measurements. nanoparticles are widely researched for applications in The rhodamine-embedded NP had zeta potential of -45 biomedical, analytical, optical, and environmental fields.2 mV, which was 14 mV lower than fluoresceine or Eu- complex embedded NP (zeta around -31 mV). By attaching PEG-molecules to the surface, the zeta potential value did not change significantly, since PEG is uncharged. Lastly, the NP suspensions were exposed to UV light at 254 and 366 nm, where we qualitatively confirmed the successful incorporation of dyes by observing the distinctive emission of colored light from dye-embedded silica NP samples (Figure C and D). In our study we successfully synthesized silica NP Dimerization of T7 lysozyme using antiparallel coiled coils from protein Sso10a Tisa Lombar,2 2 ,1 1 2 Urban Perko, Mentors: Marko Novinec,* Ana Obaha, Alenka Mozer 1 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia 2 Gimnazija Vič, Tržaška c. 72, SI-1000 Ljubljana, Slovenia The objective of the study was to modify the tertiary exhibits broad-spectrum bacteriolytic activity against structure of an enzyme in order to achieve greater stability both Gram-positive and Gram-negative bacteria, making and increased activity. More than 35% of the proteins in it a promising candidate in the development of novel the cell are found in their oligomeric form, which has antimicrobial strategies. Given the alarming rise in been shown to increase thermostability1,2. We multidrug-resistant pathogens, particularly among Gram- investigated whether we could achieve an increase in 6 negative species, T7 lysozyme offers significant stability and catalytic activity by dimerization of T7 therapeutic potential. When used in combination with lysozyme, an enzyme naturally found in bacteriophage existing antibiotics or treatments, it can markedly enhance T7, whose role is to break down peptidoglycans in the cell efficacy. Its unique lytic mechanism makes the wall, causing cell lysis 3. development of bacterial resistance highly unlikely, After numerous attempts, we were able to create a positioning T7 lysozyme as a promising breakthrough in self-assembling dimeric enzyme by fusing T7 lysozyme the fight against antibiotic-resistant infections. and the coiled-coil structure of protein Sso10a4. Coiled 1 coils are naturally occurring protein structures composed of two or more α 0,8-helices connected through hydrophobic and electrostatic interactions5. The antiparallel coiled coil 0,6 s found in protein Sso10a is recognized as one of the most ab 0,4 stable two-stranded coiled coils known. By fusing it with T7 lysozyme, we obtain a dimer with two active sites, 0,2 increasing the chance of a successful collision with the 0 substrate. Using collision theory, we can therefore predict 0 20 40 60 80 E. coli WT + Zn2+ time (min) an increase in catalytic efficiency. mod + Zn2+ mod Figure 2: The graph shows the activity of wild-type and mutant lysozyme with added Zn2+ ions and the mutant A method of producing sodium metal on a laboratory scale Urban Ocvirk,1 Mentor: Slavko Pečar*,1 1 Gimnazija in srednja šola Rudolfa Maistra, Novi trg 41a, 1241 Kamnik, Slovenia This research article focuses on the production of tube. For the apparatus, a cylindrical design was chosen to sodium metal through high-temperature distillation from fit the type of furnace used. The setup consisted of a common and inexpensive reagents. Sodium is a very useful reaction vessel with a cover plate screwed on top. A element that can be used on its own to dry certain organic graphite gasket was placed between the reaction vessel and solvents or to produce special sodium compounds such as the cover plate to prevent the metal parts from fusing sodium hydride and sodium amide. The synthesis begins together while also ensuring a gas-tight seal. The cover with choosing the right reducing agents. The cheapest, plate featured two pipes: an argon feed pipe and a most widely available, and most effective are aluminum condenser pipe. At the end of the condenser pipe, a separate and carbon, but for the reaction to proceed properly, both connected pipe served as an off-gas outlet to prevent reducing agents must be used in a specific ratio. pressure buildup. Beside the off-gas pipe, a receiving flask Unfortunately, it is not practical to use only one of the collected the distilled sodium. During the reaction, the reducing agents without the other. My experiments have heating was increased in stages. Initially, the temperature shown that aluminum behaves too aggressively at higher was raised to 400 °C to fully pyrolyze any remaining wood temperatures, producing a runaway thermite reaction that, residues in the charcoal. It was then gradually increased to despite yielding a decent amount of sodium, is very 1150 °C, the furnace’s maximum limit. The first drops of dangerous and places excessive demands on the apparatus. sodium appeared at slightly below 1000 °C after 23 Carbon, on the other hand, is too mild. The reaction with minutes. For the remainder of the reaction, the temperature carbon alone can be useful and has historically been used was maintained at 1150 °C to maximize sodium recovery. industrially to produce sodium, but it requires much higher Throughout the process, insulation covered the top of the temperatures to run smoothly, which would necessitate a reaction vessel, and temperature-monitoring equipment different apparatus and heating system1. The solution to was used to track the reaction conditions. This apparatus is this problem is to use both reducing agents simultaneously not limited to producing sodium alone. In a similar—or in a 4:1 carbon-to-aluminum ratio. This achieves a reaction even identical—setup, other alkali metals (except lithium) that has a manageable activation energy but does not turn could be prepared. Besides alkali metals, other elements into a runaway thermite reaction. When both reducing and compounds such as mercury and phosphorus could agents are used together, aluminum provides the energy for also be produced with this apparatus. Mercury could be the reaction, while carbon moderates it, creating an ideal obtained by decomposing mercury sulfide in an oxygen balance. The reaction also does not take very long to atmosphere (argon would be replaced with oxygen), while Effect of CnGL lectin from the Clouded Funnel Mushroom on the growth of Escherichia coli and Listeria innocua Vid Kodrič ,1 1 1 2 2 ,* Matic Izak, Anika Gregori Kmecl, Mentors: Jerica Sabotič, Nika Zaveršek 1 Gimnazija Vič Grammar School, Tržaška cesta 72, SI-1000 Ljubljana, Slovenia, (vidkodric9@gmail.com) 2 Department of Biotechnology, Jožef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia Due to the overuse of antibiotics in the past, we are would need to be investigated further. On the contrary, the now facing one of the greatest threats to public health – lectin had a growth-promoting effect on E. coli. However, antibiotic resistance in microorganisms. Therefore, it is bacterial growth was lower at a higher lectin crucial to find new antibacterial substances, and relatively concentration than at a lower concentration, suggesting unexplored fungi are an important source of such that further studies on the effects of higher CnGL lectin compounds. In our study, we focussed on lectins which concentrations on bacteria would be worthwhile. C. are proteins that can reversibly bind to sugars. Various nebularis proved to be an interesting source of substances lectins with insecticidal activity and effects on cancer that influence bacterial growth. cells have already been isolated from the Clouded Funnel Mushroom ( 1 Clitocybe nebularis ). One example is lactose-binding lectin CNL, which serves as a defence protein, as its nematotoxic and cytotoxic properties have been proven2,3. In our study, we isolated a lectin that binds to glucose (CnGL) from C. nebularis using affinity chromatography. The purity of the isolated lectin was verified by gel electrophoresis, which confirmed our assumption that the glucose-binding lectin would be present in our sample. We also found sepharose-binding lectin (CnSepLs), which was removed from our sample by further affinity chromatography. In the second part of the study, we investigated how Figure 1: Growth curves of Listeria innocua: control the isolated lectin affects the growth of the bacterial sample (blue), sample with added antibiotic ampicillin species Listeria innocua and Escherichia coli. We also (green) and sample with added CnGL lectin (red). investigated the effect of the C. nebularis extract on the bacteria and compared this effect with that of the References: Sustainable Synthesis of Turbostratic Graphene from Waste Carbon Fiber Composites via Flash Joule Heating Vinko Kosten, 1 Mentors: Nika Cebin*,¹ Boštjan Genorio² 1 Gimnazija Ledina, Resljeva cesta 12, 1000 Ljubljana, Slovenia 2 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot 113, SI-1000 Ljubljana, Slovenia The growing environmental burden of carbon fiber for future industrial applications in green energy, composites, widely used in industries due to their catalysis, and environmental protection. excellent mechanical properties, highlights an urgent need for sustainable recycling solutions. This research explores the transformation of waste carbon fiber composites into turbostratic graphene using the Flash Joule Heating (FJH) method—a promising, cost- effective, and energy-efficient process. By exposing crushed carbon fiber composites to high-intensity electric discharges, the material rapidly heats to over 3000 K in milliseconds, resulting in the exfoliation of amorphous carbon into high-quality turbostratic graphene. The study aimed to optimize the FJH process to achieve maximal conversion efficiency and minimal energy loss. Systematic experimentation was conducted to fine-tune critical parameters, including preheating Figure 1: SEM image of optimized turbostratic intensity, sample resistance, voltage, and pulse duration. graphene showing layered and wrinkled morphology Optimal conditions were identified as a 200 ms electrical (magnification 20,000×). pulse at 150–175 V on preheated (10 A, 3×2 s) 40 mg samples with 120 Ω initial resistance. Characterization References: via Scanning Electron Microscopy (SEM) confirmed the 1. Luong, D. X.; Bets, K.V.; Algozeeb, W. A.; Stanford, formation of turbostratic graphene with characteristic M. G.; Kittrell, C.; Chen, W.; Salvatierra, R. V.; Ren, layered morphology and significantly increased surface M.; McHugh, E. A.; Advincula, P. A.; Wang, Z.; energy utilization from the capacitor bank, validating the The optimized process demonstrated near-complete Yakobson, B.I.; Tour, J. M. Gram–scale bottom–up flash graphene synthesis. Nature 2020 , 577 , 647 – 651. area. Bhatt, M.; Guo, H.; Mancevski, V.; Shahsavari, R.; pH-Sensitive Fluorescent Probes for Detecting Urinary Tract Infections Živa Anderluh*,1 Jošt Dolinar,1 Mentors: assoc. prof. Janez Mravljak, M. Pharm., Ph.D.2, Alenka Mozer*,1 1 Vič Grammar School, Tržaška cesta 72, SI-1000 Ljubljana, Slovenia, (ziva.anderluh@gmail.com) 2 Faculty of Pharmacy, University of Ljubljana, Aškerčeva cesta 7, SI-1000 Ljubljana, Slovenia Urinary tract infections are among the most common and even after washing, a portion of the compound bacterial infections, with uropathogenic Escherichia coli remained bound to them. being the most frequent causative agent.3 Pathogens bind to glycoproteins on the surface of uroepithelial cells with No their lectin FimH. This leads to the ability to oppose wash urinary flow, and thus prolong the infection time. FimH is a mannose-binding adhesin located at the tip of type 1 pili.3 Current treatment approach is the usage of antibiotics. This method leads to development of strains resistant against most of the currently available 1 × 3 antimicrobials. New treatment strategies are already wash being developed, among which antiadhesive therapy represents an attractive alternative. New diagnostic therapies are also emerging.1,4 Within this research project, mannose-based FimH antagonists were synthesized. D-mannose`s relatively Figure 1: Bacterial imaging of two final compounds, weak affinity for FimH can be improved by incorporating showing their presence in the sample before and after various aglycones to the D- rinsing with buffer. The two final compounds are mannopyranose via α - glycosidic bond. Therefore, fluorescent probes presented in the upper pictures. were attached to D-mannose. All the fluorescent dyes are References: (rhodamine B, rhodamine 6G and 5-carboxyfluorescein) pH-sensitive due to changing the shape of the molecule in Technologies for the Diagnosis of Urinary Tract 1. Harris, M.; Fasolino, T. New and Emerging a different environment.2 Compound synthesized within Infections. J. Lab. Med. 2022, 46 (1), 3–15. the research project have an aglycon attached via N- 2. Janc, T. Chemical Glycosylation of Rhodamine B and glycosidic bond, which is known to be more stable in 6G with D-Mannose and D-Glucose; Master’s Thesis,