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Emerging infectious diseases that we are witnessing in the modern age are among the leading public health concerns. They most often occur in the form of epidemics or pandemics, and they have not been sufficiently researched. Owing to the current coronavirus disease 2019 (COVID-19) pandemic, the World Health Organization has published various recommendations to prevent the spread of this communicable disease, including a recommendation to wear protective facial masks. Therefore, this study aimed to determine the filtration effectiveness of bacteria, yeasts, and molds on three different commonly and commercially available masks used in children’s educational institutions. In addition, the bacterial content of indoor air bioaerosols was identi-fied. The genera Staphylococcus and Micrococcus were dominant in all samples, whereas bacteria of the genera Bacillus, Acinetobacter, and Corynebacterium were identified at a significantly smaller number. Bacterial, yeast, and mold filtering effectiveness grew from the single-layer cloth mask, which has proven to be the least effective over the surgical mask, to the most effective filtering facepiece type 2 (FFP2) mask. Furthermore, surveys are needed to study the effectiveness of pro-tective measures.

In January 2023, the derogation loophole was closed on “emergency authorisations” for the use of three out of five neonicotinoids in all EU states. In this study, we analysed the sorption/desorption behaviour and kinetic parameters of acetamiprid and thiacloprid, the two neonicotinoids that are still approved for use, either regularly or under emergency authorisations in the EU, and widely used worldwide. Sorption and desorption curves in four soils with different organic matter content were analysed using four kinetic models, namely, Lagergren’s pseudo first-order model, two-site model (TSM), Weber–Morris intraparticle diffusion model and Elovich’s model. Kinetic parameters were correlated to soil physico-chemical characteristics. To determine the mutual influence of soil characteristics and sorption/desorption parameters in the analysed soils, a factor analysis based on principal component analysis (PCA) was performed. Even though the two insecticides are very similar in size and chemical structure, the results showed different sorption/desorption kinetics. The model that best fits the experimental data was TSM. Thiacloprid showed a more rapid sorption compared to acetamiprid, and, in all soils, a higher proportion sorbed at equilibrium. Intra-particle diffusion seemed to be a relevant process in acetamiprid sorption, but not for thiacloprid. Desorption results showed that acetamiprid is more easily and more thoroughly desorbed than thiacloprid, in all soils. The kinetic behaviour differences stem from variations in molecular structure, causing disparate water solubility, lipophilicity, and acid–base properties.

In the pathogenesis of endometriosis, a number of pathological reactions occur. Proteins secreted in the urine are thought to interact with each other and stimulate the pathological processes in endometriosis. Identifying one or more proteins that are specific enough and could serve as biomarkers for endometriosis is both a challenge and a necessity that would facilitate diagnosis. The urine of patients treated in a tertiary university hospital between July 1, 2020 and June 30, 2021 was analyzed. The studied group consists of patients who were treated surgically for endometriosis and in whom the diagnosis was confirmed by pathohistological analysis. The control group consists of patients who were operated for functional ovarian cysts. Urinary proteins were analyzed by chromatography and mass spectrometry (LC-MS/MS). We identified 17 proteins in urine whose concentrations were statistically significantly different in the group with endometriosis (N = 16) compared with the control groups (N = 16). The detected proteins were classified into groups according to their function in invasion, migration and proliferation, proteolysis, immune system, cell adhesion and vascular system. For all mentioned proteins the difference in concentration is statistically significant p < 0.005. Proteins are secreted in the urine of patients with endometriosis that may be involved in the pathogenesis of the disease and are possible biomarkers for endometriosis.

UVOD: Ulcerozni kolitis (UK) je kronična upalna bolest koja zahvaća gastrointestinalni sustav, a čija incidencija je u konstantnom porastu i time predstavlja važan javnozdrvstveni problem. Upalni proces zahvaća sluznicu debelog crijeva, a bolest karakteriziraju razdoblja remisije i pogoršanja. Čimbenici kao što su genetska predispozicija, imunološki sustav, mikroorganizmi, te čimbenici okoliša povezuju se s nastankom UK. Unatoč brojnim istraživanjima etiopatogeneza nije razjašnjena. Istraživanja, kako epidemiološka tako i eksperimentalna, pokazala su povezanost između luteolina, prevencije i razvoja kroničnih bolesti. Pokazano je da luteolin ima široki spektar bioloških učinaka uključujući antioksidacijsko, protuupalno i antitumorsko ali je njegova uloga i mehanizam djelovanja u UK slabo istražena. CILJ: Cilj rada je ispitati utjecaj luteolina na tijek razvoja UK te ispitati ulogu signalnog puta posredovanog izvanstaničnim signalima reguliranih kinaza (ERK) u mehanizam djelovanja luteolina i terapiji UK. MATERIJALI I METODE: Istraživanje je provedeno na eksperimentalnom modelu kolitisa. U istraživanju tijek razvoja kolitisa procijenio se temeljem kliničkih parametara te stupnja oštećenja na razini tkiva debelog crijeva. Izražaj proteina određen je Western blot metodom, a ERK inhibitor primjenjen je kako bi se ispitala uloga ERK signalne molekule u razvoju kolitisa. REZULTATI: Luteolin je ublažio kliničke simptome kolitisa i smanjio intenzitet upalnog procesa u debelom crijevu. Luteolin aktivira ERK protein. Primjena ERK inhibitora pogoršava kliničke simptome kolitisa i smanjuje učinak luteolina. ZAKLJUČAK: Istraživanjem je pokazan protuupalni i antiapoptotski učinak luteolina koji je posredovan ERK signalnim putem.

U ovom su radu u uvodnom dijelu u osnovnim crtama opisani poli(dimetil-siloksan), zeoliti te teorijska pozadina eksperimentalnih metoda koje su korištene u radu. Složena molekulska dinamika polimernih sustava poput poli(dimetil-siloksana) u kombinaciji sa zeolitima istražena je elektronskom spinskom rezonancijom (ESR), a njihovo termičko ponašanje razlikovnom pretražnom kalorimetrijom (DSC). Utjecaj mehaničke predobrade na kristaliničnost zeolita ispitan je rentgneskom difrakcijom praha (PXRD). Cilj ovog diplomskog rada bio je ispitati utjecaj mehaničke predobrade zeolita na molekulsku dinamiku poli(dimetil-siloksana) (PDMS) u kompozitnim uzorcima. Rezultati pokazuju da prisutnost zeolita usporava lokalnu molekulsku dinamiku u matrici poli(dimetil-siloksana). Pokazano je da mehanička predobrada zeolita u tome igra značajnu ulogu. Učinak zeolita na usporavanje dinamike PDMS lanaca ovisi o molekulskoj masi samog poli(dimetil-siloksana), ali i o mehaničkoj predobradi zeolita te njezinom vremenskom trajanju. Na temelju rezultata ovog istraživanja može se zaključiti da bi zeoliti mogli imati dobru perspektivu kao potencijalna punila za elastomere., In the introduction of this work, the basics of poly(dimethyl-siloxane), zeolites and the theoretical background of the used experimental methods are described. The complex molecular dynamics of polymer systems such as poly(dimethyl-siloxane) in combination with zeolites was investigated by electron spin resonance (ESR), and their thermal behavior by differential scanning calorimetry (DSC). The influence of mechanical pretreatment on the crystallinity of zeolite was examined by powder X-ray diffraction (PXRD). The aim of this work was to examine the influence of mechanical pretreatment of zeolite on the molecular dynamics of poly(dimethyl-siloxane) (PDMS) in composite samples. Results show that the presence of zeolite slows down the local molecular dynamics in the poly(dimethyl-siloxane) matrix. It has been shown that the mechanical pretreatment of zeolite plays a significant role. The effect of zeolite on the restriction of the dynamics of PDMS chains depends on the molecular weight of the poly(dimethyl-siloxane) itself, but also on the mechanical pretreatment of the zeolite and its duration. Based on the results of this research, it can be concluded that zeolites might have a good perspective as potential fillers for elastomers.

Objectives: The purpose of this study was to determine the effects of treating bladder cancer cells with the newly synthesized thieno[2,3-b]pyridine anticancer agents, by focusing on its cytotoxic effects on the investigated human cell line T24. Methods: The T24 bladder cancer cell line was treated with a newly developed thieno[2,3-b]pyridine anticancer compounds: Inhibitor 5 3-amino-N-(3-chloro-2-methylphenyl)-5-oxo-5,6,7,8-tetrahydrothieno[2,3-b]quinoline-2-carboxamide, Inhibitor 6 3-amino-N-(naphthalen-1-yl)-5-oxo-5,6,7,8-tetrahydrothieno[2,3-b]quinoline-2-carboxamide, Inhibitor 8 (E)-3-amino-5-(3-(3-bromophenyl)acryloyl)-N-(3-chloro-2-methylphenyl)-6-methylthieno[2,3-b]pyridine-2-carboxamide and Inhibitor 9 (E)-3-amino-5-(3-(3-bromophenyl)-1-hydroxyallyl)-N-(3-chloro-2-methylphenyl)-6-methylthieno[2,3-b]pyridine-2-carboxamide to determine its cytotoxic effect on T24 cells. The 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to analyze the cellular metabolic activity and determine the cytotoxic effect. Results: Inhibitors 5, 6, 8 and 9 all showed significant cytotoxic effect on the bladder cancer cells in a dose- and time-dependent manner. The most effective was Inhibitor 8 with an IC50 value of 0.4041 μg/mL after 72 hours. Conclusion: Due to their cytotoxic effect on bladder cancer cells, Inhibitors 5, 6, 8 and 9 deserve further attention as a potential treatment for transitional cell cancer., Ciljevi: Svrha ove studije bila je utvrditi učinke tretiranja stanica raka mokraćnog mjehura novosintetiziranim tieno[2,3-b]piridin antitumorskim spojevima, fokusirajući se na njihove citotoksične učinke na ispitivanu humanu staničnu liniju T24. Materijali i metode: T24 stanična linija raka mokraćnog mjehura tretirana je novosintetiziranim tieno[2,3-b]piridin antitumorskim spojevima: Inhibitor 5 - 3-amino-N-(3-kloro-2-metilfenil)-5-okso-5,6,7,8-tetrahidrotieno[2,3-b]kinolin-2-karboksamid, Inhibitor 6 - 3-amino-N-(naftalen-1-il)-5-okso-5,6,7,8-tetrahidrotieno[2,3 -b]kinolin-2-karboksamid, Inhibitor 8 - (E)-3-amino-5-(3-(3-bromofenil)akriloil)-N-(3-klor-2-metilfenil)-6-metiltieno[2, 3-b]piridin-2-karboksamid i Inhibitor 9 - (E)-3-amino-5-(3-(3-bromofenil)-1-hidroksialil)-N-(3-klor-2-metilfenil)-6- metiltieno[2,3-b]piridin-2-karboksamid, kako bi se odredio njihov citotoksični učinak na T24 stanice. Proveden je test 3-(4,5-dimetiltiazolil-2)-2,5-difeniltetrazolij bromida (MTT) kako bi se analizirala stanična metabolička aktivnost i odredio citotoksični učinak. Rezultati: Svi inhibitori 5, 6, 8 i 9 pokazali su značajan citotoksični učinak na stanice raka mokraćnog mjehura i taj učinak je ovisan o dozi i vremenu inkubacije. Najučinkovitiji je bio Inhibitor 8 s IC50 vrijednošću od 0,4041 μg/mL nakon 72 sata. Zaključci: Zbog svog citotoksičnog učinka na stanice raka mokraćnog mjehura, Inhibitori 5, 6, 8 i 9 zaslužuju daljnju pozornost kao potencijalni tretman za rak prijelaznih stanica.

In the era of escalating antimicrobial resistance, the need for antibacterial drugs with novel or improved modes of action (MOAs) is a health concern of utmost importance. Adding or improving the chelating abilities of existing drugs or finding new, nature-inspired chelating agents seems to be one of the major ways to ensure progress. This review article provides insight into the modes of action of antibacterial agents, class by class, through the perspective of chelation. We covered a wide scope of antibacterials, from a century-old quintessential chelating agent nitroxoline, currently unearthed due to its newly discovered anticancer and antibiofilm activities, over the commonly used antibacterial classes, to new cephalosporin cefiderocol and a potential future class of tetramates. We show the impressive spectrum of roles that chelation plays in antibacterial MOAs. This, by itself, demonstrates the importance of understanding the fundamental chemistry behind such complex processes.

The development of recombinant insulin protein as a drug was a major step in the history of medicine. Today, the use of recombinant biological drugs is growing, which includes recombinant antibodies, a third of which is produced in Escherichia coli. Although these prokaryotic cells do not possess all the mechanisms for posttranslational modifications of proteins, E. coli has an oxidative environment for proteins in its periplasmic space where proteins can be directed for proper structuring. An important form of recombinant antibody, characterized by a very low molecular weight with preserved target molecule recognition properties, is the single-chain variable antibody fragment (scFv). The aim of this work is to investigate which factors result in higher yield and purity during scFv production in E. coli. Three methods of protein isolation from periplasm were used. Plasmid pOPE101 was used to transform E. coli pREP-DE3 strain with the scFv protein gene. The identity of the pOPE101-scFv expression vector was verified by restriction analysis, and high protein expression was achieved by overnight induction at 18°C. The scFv expression was confirmed by SDS-PAGE technique and Western blot analysis. We have shown that protein isolation from the periplasmic space is improved by vigorous stirring after cell resuspension and that double osmotic shock is the best technique for releasing soluble scFVs. It was observed that a large amount of scFv still remains in insoluble form. The obtained scFvs were also successfully isolated using the immobilized metal affinity chromatography (IMAC) and binding of the His tag, without loss during chromatography. To remove residual contaminants, a subsequent purification step based on size exclusion is recommended. Razvoj rekombinantnog inzulinskog proteina kao lijeka bio je veliki korak u povijesti medicine. Danas je sve veća upotreba rekombinantnih bioloških lijekova, što uključuje rekombinantna protutijela, od kojih se trećina proizvodi u Escherichia coli. Iako ove prokariotske stanice ne posjeduju sve mehanizme za posttranslacijske modifikacije proteina, E. coli ima oksidativno okruženje za proteine u svom periplazmatskom prostoru gdje se proteini mogu usmjeriti na pravilno strukturiranje. Važan oblik rekombinantnog protutijela, kojeg karakterizira vrlo niska molekularna težina sa očuvanim svojstvima prepoznavanja ciljne molekule, je jednolančani varijabilni fragment protutijela (scFv). Cilj ovog rada je istražiti koji čimbenici rezultiraju većim prinosom i čistoćom tijekom proizvodnje scFv u E. coli. Korištene su tri metode izolacije proteina iz periplazme. Plazmid pOPE101 korišten je za transformaciju E. coli pREP-DE3 soja s scFv proteinskim genom. Identitet ekspresijskog vektora pOPE101-scFv verificiran je restrikcijskom analizom, a visoka ekspresija proteina postignuta je indukcijom preko noći na 18°C. Ekspresija scFv potvrđena je SDS-PAGE tehnikom i Western blot analizom. Pokazali smo da se izolacija proteina iz periplazmatskog prostora poboljšava snažnim miješanjem nakon resuspenzije stanica i da je dvostruki osmotski šok najbolja tehnika za otpuštanje topljivih scFV-a. Uočeno je da velika količina scFv još uvijek ostaje u netopivom obliku. Dobiveni scFvs također su uspješno izolirani korištenjem imobilizirane metalne afinitetne kromatografije (IMAC) i vezanja His oznake, bez gubitka tijekom kromatografije. Za uklanjanje zaostalih kontaminanata preporučuje se naknadni korak pročišćavanja koji se temelji na isključenju veličine.

Uvod: Ulcerozni kolitis upalna je bolest koja zahvaća debelo crijevo te uzrokuje niz patoloških promjena na sluznici debelog crijeva. Promjene na sluznici jedan su od rizičnih čimbenika koji utiru put ka karcinomu debelog crijeva. Bolest je najčešće kroničnog tijeka s izmjenom razdoblja težih manifestacija i mirnog stanja. Neki od glavnih simptoma su krvavi proljevi, mučnina, glavobolja, malaksalost i gubitak tjelesne mase. Trenutačno dostupni tretmani ulceroznog kolitisa su nezadovoljavajući, dok se prirodni bioaktivni spojevi poput flavonoida smatraju terapijskim agensima u razvoju. Luteolin je jedan od spojeva iz velike skupine flavonoida, a ističe se po protutumorskom, protuupalnom, antioksidativnom i antiapoptotskom učinku u mnogim bolestima. Cilj: na pokusnom modelu ulceroznog kolitisa ispitati učinak luteolina na razvoj i intenzitet kolitisa te proces apoptoze. Materijal i metode: Model ulceroznog kolitisa izazvan je u C57BL/6 miševa primjenom 3 % otopine natrijevog dekstran sulfata (DSS) tijekom sedam dana u vodi za piće. Kako bi se ispitalo terapijsko djelovanje luteolin se primijenio u dvije doze od trećeg do sedmog dana pokusnog perioda, per os. Tijekom pokusnog perioda odnosno tijekom razvoja kolitisa pratile su se kliničke promjene i stupanj oštećenja tkiva debeloga crijeva. Izražaj specifičnih proteina određen je Western blot metodom i imunohistokemijski. Rezultati: Luteolin je ublažio simptome DSS-induciranog kolitisa kod miševa, zacijelio oštećenje tkiva debelog crijeva te smanjio upalu. Luteolin je značajno smanjio izražaj nuklearnog čimbenik-kapaB podjedinice p65 i proapoptotskih proteina. Zaključak: Rezultati potvrđuju prethodno pokazane učinke luteolina te sugeriraju da se luteolin može koristiti kao novi terapeutski kandidat u liječenju ulceroznog kolitisa ili kao dodatak postojećoj terapiji., Introduction: Ulcerative colitis is an inflammatory disease that affects the colon and causes a number of pathological changes in the colon mucosa. Mucosal changes are one of the risk factors in the development of colon cancer. The disease has a chronic course and it is characterized by periods of severe manifestations and a remission state. Some of the main symptoms are bloody diarrhea, nausea, headache, and weight loss. Currently used treatments of ulcerative colitis are unsatisfactory, while natural bioactive compounds like flavonoides are considered to be emerging therapeutic agents. Luteolin is a natural compound with beneficial effects on a variety of diseases, however, its effect on ulcerative colitis has been poorly studied. Luteolin is one of the members of a large group of flavonoids, and is distinguished by its antitumor, anti-inflammatory, antioxidant, and antiapoptotic effect in many diseases. Aim: using an experimental model of ulcerative colitis investigate the effect of luteolin on the development and intensity of ulcerative colitis and the process of apoptosis. Material and methods: The ulcerative colitis was induced by the administration of 3% dextran sulfate sodium salt (DSS) in drinking water for seven days. To investigate the effect of luteolin on the development of colitis two doses of luteolin are administrated between day 3 and day 7 of the experiment period, orally. During the development of colitis, clinical changes were monitored as well as the degree of damage of the colon tissue by histopathological analysis. The expression of the investigated proteins was determined by the Western blot method and immunohistochemically. Results: Luteolin attenuated symptoms of DSS-induced colitis in mice, ameliorated colon tissue damage, and reduced inflammation. Luteolin significantly reduced the expression of nuclear factor-kappa of subunit p65 and proapoptotic proteins. Conclusions: The results confirmed the anti-inflammatory and anti-apoptotic effects of luteolin. This suggested that luteolin can be used as a novel therapeutic candidate in the treatment of ulcerative colitis or could be used as a supplement to existing therapy.

U suvremeno doba poljoprivreda se suočava s velikim problemom primjene prekomjernih količina kemijskih sredstava (insekticida) u suzbijanju raznih štetnika i njihovih razvojnih oblika, često puta bez ikakve kontrole i provođenja redovitih monitoringa njihovih ostataka u okolišu. U skupinu najčešće primjenjivanih insekticida prvenstveno zbog visoke učinkovitosti, selektivnosti i dugotrajne zaštite nasada ubraja se neonikotinoidni insekticid imidakloprid. U Hrvatskoj imidakloprid je učinkovito sredstvo u zaštiti šećerne repe, ali svoju primjenu pronašao je i u maslinicima Kvarnera i Istre u zaštiti od bolesti i štetnika na kulturama maslina, pogotovo u suzbijanju maslinove muhe (Bactrocera oleae). Iako je aktivan pri niskim dozama, njegova višekratna primjena tijekom sezone rasta i nekontrolirana uporaba predstavljaju javno-zdravstveni problem zbog ulaska u ciklus kruženja tvari u okolišu s toksičnim potencijalom prema vodnim resursima, tlu, zraku i živim bićima. Proučavana je postojanost imidakloprida u tlima maslinika s područja Istre i otoka Krka u ovisnosti o koncentraciji insekticida i fizikalno-kemijskim karakteristikama tla te kinetika razgradnje uz primjenu matematičkih modela. Rezultati ukazuju da je brža razgradnja imidakloprida u tlu Istra nego u tlu Krk, uzrokovana višim kationskim izmjenjivačkim kapacitetom i višem udjelu organskog ugljika i gline. Pri višoj dozi imidakloprida (5 mg/kg) Model kinetike prvoga reda najbolje opisuje razgradnju u tlu Krk dok se u tlu Istra uočava bifazna razgradnja. Bifazno ponašanje razgradnje je uočeno kod oba tla pri nižoj dozi imidakloprida (0,5 mg/kg). Vrijeme poluživota od 91 dan ubraja imidakloprid u umjereno postojane pesticide. Pretpostavka je da se u tlu Istra razgradnja imidakloprida odvijala uz abiotičku kiselu hidrolizu te da u ovom blago kiselom tlu, veliki udio gline uzrokuje vezu između brzina sorpcije i razgradnje i na taj način olakšava i ubrzava reakciju hidrolize. U tlu Krk prisutnost mikrobne populacije vjerojatno je utjecala na postojanost imidakloprida, a istodobno nije postignuta poveznica s procesima sorpcije. Rezultati ukazuju da se matematičko modeliranje može koristiti kao vrlo prikladan alat za predviđanje postojanosti imidakloprida kojim se omogućuje praćenje ostataka imidakloprida u onečišćenom tlu. Nadalje, primjena imidakloprida treba biti strogo kontrolirana, posebno u tlima s niskim sadržajem organske tvari gdje je rizik od onečišćenja tla i podzemnih voda visok zbog dulje postojanosti i mogućnosti površinskog ispiranja i procjeđivanja kroz tlo., Nowadays, agriculture industry faces with the great problem of applying excessive chemicals (insecticides) amounts in the controling of various pests and their developmental forms, often without any control and regular monitoring of their residues in the environment. One the most commonly used insecticides, primarily due to its high efficiency, selectivity and long-term plantation protection is the neonicotinoid insecticide imidacloprid. In Croatia, imidacloprid is most used as an an effective mean in the protection of sugar beet, but its application has been recorded in Kvarner and Istria olive orchards for the protection of olive tree against diseases and pests such as olive fruit fly (Bactrocera oleae). Although imidacloprid is active at low doses, its repeated application during the growing season and uncontrolled use, poses it as a public health problem since it was entered to the matter cycle in the environment with toxic effect for aquatic resources, soil, air and living beings. The imidacloprid persistence in the soils from Istria and the Krk island olive orchards as a function of the insecticide concentration and physicochemical soil characteristics was studied and analyzed by mathematical modelling. The results of the study indicate that the degradation of imidacloprid was more rapid in the Istria soil than in the Krk soil what is caused by higher cation exchange capacity and higher organic carbon and clay content in Istria soil. At a higher concentration used (5 mg/kg), the first-order kinetics model gives the best description of imidacloprid degradation in the Krk soil, while the biphasic degradation is observed in the Istria soil. Biphasic degradation was observed in both soils at a lower imidacloprid concentration (0.5 mg/ kg). Whit half-life of 91 days in analyzed soils, imidacloprid belongs to the group of moderately persistent pesticides. It can assumed that in the Istria soil, imidacloprid degradation took place by abiotic acidic hydrolysis where high clay amount stimulated imidacloprid sorption and accelerated the hydrolysis reaction. In the Krk soil, the presence of microbial populations probably affected the imidacloprid persistence, and no connection between degradation and sorption processes was achieved. The results of the study indicate that mathematical modeling can be used as a suitable tool for prediction of imidacloprid persistence, which allows monitoring of imidacloprid residues in contaminated soils. Furthermore, the use of imidacloprid should be carried out under strictly controlled conditions, especially in soils with low organic matter content where the potential of soil and groundwater contamination by imidacloprid is high due to its longer persistence and surface leaching and moving through the soil profile.

INTRODUCTION: Diabetes mellitus is a metabolic disorder with multiple etiology, characterised by chronic hyperglycemia with disturbances of metabolism of carbohydrates, fats and proteins. It results from defects in insulin secretion, action or both. Dipeptidyl-peptidase IV/CD26 (DPP IV/CD26) molecule is known to be involved in an array of physiological and pathological processes, in the regulation of glycaemia as well. A variety of complications caused by diabetes can cause hospitalizations as infections and gangrene. DPP IV/CD26 inhibitors such as alogliptin, linagliptin and saxagliptin can be used to treat diabetes mellitus. The aim of this reseaarch was to investigate the processes of wound healing in conditions of CD26 deficiency in experimental hyperglycaemia. MATERIALS AND METHODS: In this experiment two strains of mice were used, wild type and CD26- deficient mice. Experimental diabetes was induced using a solution of streptozotocine in citrate buffer (50mg/kg, i.p.) during five days. Serm samples as well as pancreas, cutaneous (control) tissue and wound tissue were analyzed. Pathohistological, immunohistochemical and immunobiochemical analyses were performed on wound samples and control skin. Serum samples were analysed for DPP IV/CD26 activity and concentration of target angiogenic factors. RESULTS: Results of this study revealed that DPP IV/CD26 has an important role in the regulation of blood glucose concentration. Inactivation of DPP IV/CD26 improves the state associated with hyperglycemia. The process of cutaneous wound healing is improved in conditions of CD26 deficiency. CONCLUSION: The inhibition of DPP IV/CD26 has positive effects on the wound healing process in hyperglycemia. DPP IV/CD26 inhibition can be suggested as a therapeutic option for treatment of diabetes mellitus and its complications like delayed wound healing.

Cilj istraživanja: Cilj je istraživanja bio ispitati potencijalno citotoksično djelovanje fenolnih spojeva izoliranih iz pupa masline sorti Leccino i Frantoio na dvije stanične linije humanih karcinoma dojke: MDA-MB-231 i MCF-7. Broj karcinomskih stanica nakon izlaganja uzorcima pretpostavljeno se trebao smanjiti u odnosu na kontrolnu skupinu. Materijali i metode: Ispitivanje citotoksičnosti fenolnih spojeva na dvjema staničnim linijama karcinoma dojke rađeno je MTT testom nakon 4, 24, 48 i 72 sata inkubacije. Usporedbom apsorbancije nastalog formazana kod stanica tretiranih spojevima dobivenih iz uzoraka masline te apsorbancije kontrolne skupine, odnosno netretiranih stanica, dobiven je postotak preživljenja karcinomskih stanica. Rezultati: Rezultati izlaganja stanica karcinoma dojke (stanične linije MDA-MB-231 i MCF- 7) ekstraktima masline E3 (sorta Leccino) i E4 (sorta Frantoio) prikazani su grafički u odnosu postotka metabolički aktivnih stanica i koncentracije spoja. Najznačajniji učinak na staničnu liniju MDA-MB-231 postignut je nakon inkubacije od 48 sati pri koncentraciji od 200 μg/mL za obje sorte, a na staničnu liniju MCF-7 nakon inkubacije od 72 sata pri koncentraciji od 100 μg/mL za sortu E3 i koncentraciji od 200 μg/mL za sortu E4. Zaključak: Ekstrakti pupa masline pokazuju određen citotoksični utjecaj na stanične linije humanih karcinoma dojke MDA-MB-231 i MCF-7, koji ovisi o vremenu izlaganja i koncentraciji, te u određenoj mjeri smanjuju preživljenje stanica karcinoma. Učinak ekstrakata nije uvijek razmjeran povećanju koncentracije. Ekstraktima sorti maslina korištenih u našem istraživanju nije dostignuta IC50 vrijednost., Objective: The aim of the research was to examine the potential cytotoxic effects of phenolic compounds isolated from Leccino and Frantoio olive buds on two cell lines of human breast cancer: MDA-MB-231 and MCF-7. The number of carcinoma cells was assumed to be reduced after exposure to phenolic compounds in comparison with the control group. Materials and methods: Cytotoxic effects of phenolic compounds on breast cancer cell lines were determinged with MTT assay, after the periods of 4, 24, 48 and 72 hours of incubation. The comparison between the absorbance of created formazan in cells treated with samples obtained from the olive buds and the absorbance of the control group (cells untreated with olive bud samples) established the survival rates of carcinoma cells. Results: The results of breast cancer cells (cell lines MDA-MB-231 and MCF-7) exposure to olive extracts E3 (Leccino sort) and E4 (Frantoio sort) are shown graphically as relation of the percentage of metabolically active cells to the concentration of the compound. The most significant effect on the MDA-MB-231 cell line was achieved after incubation of 48 hours at a concentration of 200 μg/mL for both sorts, and on the MCF-7 cell line after incubation of 72 hours at a concentration of 100 μg/mL for the E3 sort and concentration of 200 μg/mL for the E4 sort. Conclusion: Olive bud extracts show a certain cytotoxic effect on human breast cancer cell lines MDA-MB-231 and MCF-7, which depends on exposure time and concentration, and to a certain extent reduce the survival of said cancer cells. The effect of extracts is not always proportional to the increase in concentration. The extracts of the cultivars used in our research did not reach the IC50 value.

Kvasci su pripadnici carstva gljiva (Fungi) kao i plijesni i mesnate gljive. Eukariotski su organizmi koji se često nazivaju jednostaničnim gljivama, a razmnožavaju se pupanjem. Saccharomyces cerevisiae neinfektivni je mikrob čiji je eukariotski genom bio prvi sekvencioniran. Svojim fenotipskim svojstvima poput učinkovitosti parenja ili sporulacije, doprinio je napretku u istraživanjima. Često korišteni modelni organizam upravo je soj Saccharomyces cerevisiae W303 koji je usko povezan s prvim potpuno sekvencioniranim sojem kvasca S288C te se stoga naziva njegovim potomkom. Mitohondriji, kao organeli većine eukariotskih organizama razlikuju se između različitih vrsta eukariota, po veličini, ali i sadržaju genoma. Cilj ovog rada je analizirati proteom mitohondrija kvasca Saccharomyces cerevisae W303. Stanice kvasca uzgajane su na kvaščevoj podlozi do ulaska u ranu stacionarnu fazu. Iz takvih su stanica izolirani mitohondriji, a iz njih proteini. Ekstrahirani proteini mitohondrija su nakon pripreme analizirani LC/MS metodom. Nakon identifikacije, dobiveni su proteini sistematizirani i utvrđena im je uloga i važnost u funkcioniranju stanice kvasca., Yeasts are members of the fungi kingdom (Fungi), as well as molds and fleshy mushrooms. They are eukaryotic organisms often called unicellular fungi, and they reproduce by budding. Saccharomyces cerevisiae is a non-infectious microbe whose eukaryotic genome was the first to be sequenced. With their phenotypic properties such as mating efficiency or sporulation, they have contributed to scientific research progress. A frequently used model organism is the strain Saccharomyces cerevisiae W303, which is closely related to the first fully sequenced yeast strain S288C and is therefore called its descendant. Mitochondria, as organelles of most eukaryotic organisms, differ between different types of eukaryotes in terms of size and genome content. The aim of this work was to analyze the mitochondrial proteome of the yeast Saccharomyces cerevisae W303. Yeast cells were grown on yeast medium until they reached the early stationary phase. Mitochondria and proteins were isolated from such cells. After preparation, extracted mitochondrial proteins were analyzed by the LC/MS method. After identification, the obtained proteins were systematized and their role and importance in the functioning of the yeast cell was determined.

Ulcerative colitis (UC) is a multifactorial condition characterized by a destructive immune response that failed to be attenuated by common regulatory mechanisms which reduce inflammation and promote mucosa healing. The inhibition of CD26, a multifunctional glycoprotein that controls the immune response via its dipeptidyl peptidase (DP) 4 enzyme activity, was proven to have beneficial effects in various autoimmune inflammatory diseases. The polarization of macrophages into either pro-inflammatory M1 or anti-inflammatory M2 subclass is a key intersection that mediates the immune-inflammatory process in UC. Hence, we hypothesized that the deficiency of CD26 affects that process in the dextran sulfate sodium (DSS)-induced model of UC. We found that mRNA expression of M2 markers arginase 1 and Fizz were increased, while the expression of M1 marker inducible NO synthase was downregulated in CD26−/− mice. Decreased STAT1 mRNA, as well as upregulated pSTAT6 and pSTAT3, additionally support the demonstrated activation of M2 macrophages under CD26 deficiency. Finally, we investigated DP8 and DP9, proteins with DP4-like activity, and found that CD26 deficiency is not a key factor for the noted upregulation of their expression in UC. In conclusion, we demonstrate that CD26 deficiency regulates macrophage polarization toward the anti-inflammatory M2 phenotype, which is driven by STAT6/STAT3 signaling pathways. This process is additionally enhanced by the reduction of M1 differentiation via the suppression of proinflammatory STAT1. Therefore, further studies should investigate the clinical potential of CD26 inhibitors in the treatment of UC.

Cilj istraživanja: Cilj je istraživanja usporedba upalnog odgovora kod različito liječenih pacijenata oboljelih od ulceroznog kolitisa. Materijali i metode: Uzorci krvi svih sudionika uzeti su za analizu. Uzorak je tretiran Fc-receptor blokirajućim reagensom te je uzorak inkubiran 20 minuta bez izvora svjetlosti pri sobnoj temperaturi uz dodatak anti-humanog CD14s PerCP-Cy5.5-konjugiranog protutijela i protutijela konjugiranog fikoeritrinom, reaktivnog na humani CD16 te dodatak mišjih antitijela reaktivnim na humani CD44, koji je konjugiran s fluorescein izotiocijanatom. Nakon lize stanica, stanice su analizirane pomoću protočne citometrije. Podatci su obrađeni korištenjem FlowLogic Software-a. Rezultati: Pacijenti oboljeli od ulceroznog kolitisa liječeni biološkom terapijom imali su značajno smanjenje medijana intenziteta fluorescencije CD44 biljega na CD14+CD16++ subpopulaciji monocita. Medijan intenziteta fluorescencije CD44 biljega na CD44+CD14+ limfocitima gotovo je dvostruko veći kod pacijenata liječenih biološkom terapijom u usporedbi s pacijentima liječenih nebiološkom terapijom. Postotci CD44+CD14+ limfocita nisu se pokazali različitima između navedenih skupina. Zaključak: Opisana je različita ekspresija CD44 biljega na neklasičnim monocitima pacijenata, s obzirom na terapiju koju pacijent prima. Osim neklasičnih monocita, zahvaćeni su i CD44+CD14+ limfociti – uočena je različita ekspresija CD44 biljega ovisno o terapiji., The aim of research: The aim of this research was comparison of inflammatory response between differently treated patients with ulcerative colitis. Materials and methods: Blood samples of all participants were taken for analysis. The sample was treated with Fc receptor blocking solution, then incubated for 20 minutes without a light source at room temperature with the addition of anti-human CD14s PerCP-Cy5.5-conjugated antibodies and phycoerythrin-conjugated antibodies reactive to human CD16 and with murine antibodies reactive to human CD44, conjugated with fluorescein isothiocyanate. After cell lysis, cells were analyzed by flow cytometry. Data were processed using FlowLogic Software. The results: Ulcerative colitis patients treated with biological therapy had a significant decrease in the median fluorescence intensity of the CD44 marker on the CD14+CD16++ subpopulation of monocytes. The median fluorescence intensity of the CD44 marker on CD44+CD14+ lymphocytes is almost twice as high in patients treated with biological therapy compared to patients treated with non-biological therapy. The percentages of CD44+CD14+ lymphocytes did not differ between the mentioned groups. Conclusions: Different expression of the CD44 marker on non-classical monocytes of patients was described, with regard to the therapy the patient received. In addition to non-classical monocytes, CD44+CD14+ lymphocytes are also affected – different expression of the CD44 marker was observed depending on the therapy.

Glioblastoma multiforme (GBM) is the most common and fatal primary brain tumor, and is highly resistant to conventional radiotherapy and chemotherapy. Therefore, the development of multidrug resistance and tumor recurrence are frequent. Given the poor survival with the current treatments, new therapeutic strategies are urgently needed. Radiotherapy (RT) is a common cancer treatment modality for GBM. However, there is still a need to improve RT efficiency, while reducing the severe side effects. Radiosensitizers can enhance the killing effect on tumor cells with less side effects on healthy tissues. Herein, we present our pioneering study on the highly stable and amphiphilic metallacarboranes, ferrabis(dicarbollides) ([o-FESAN]- and [8,8'-I2-o-FESAN]-), as potential radiosensitizers for GBM radiotherapy. We propose radiation methodologies that utilize secondary radiation emissions from iodine and iron, using ferrabis(dicarbollides) as iodine/iron donors, aiming to achieve a greater therapeutic effect than that of a conventional radiotherapy. As a proof-of-concept, we show that using 2D and 3D models of U87 cells, the cellular viability and survival were reduced using this treatment approach. We also tested for the first time the proton boron fusion reaction (PBFR) with ferrabis(dicarbollides), taking advantage of their high boron (11B) content. The results from the cellular damage response obtained suggest that proton boron fusion radiation therapy, when combined with boron-rich compounds, is a promising modality to fight against resistant tumors. Although these results are encouraging, more developments are needed to further explore ferrabis(dicarbollides) as radiosensitizers towards a positive impact on the therapeutic strategies for GBM., The authors received support from the Spanish Ministerio de Economía y Competitividad (PID2019-106832RB-100), the Generalitat de Catalunya (2017SGR1720), FCT - Fundação para a Ciência e a Tecnologia, in the scope of the project UID/Multi/04349/2019 and the projects LISBOA-01-0247-FEDER-045904 and UTAP-EXPL/FMT/0020/2021 of Centro de Ciências e Tecnologias Nucleares/IST, PTDC/BTM-TEC/29256/2017, UIDP/04565/2020 of iBB/IST, UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences – UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy – i4HB. The Metrology Laboratory of Ionizing Radiation team of Centro Tecnológico e Nuclear, Instituto Superior Técnico (CTN/IST) is acknowledged for their support in the irradiation setups. Miquel Nuez-Martínez is enrolled in the PhD program of the UAB. MQM and VMA acknowledge financial support by the Spanish Government MCIN/AEI/10.13039/501100011033 (project 2019-108434GB-I00 to VMA and project IJC2018-035283-I to MQM), and Universitat Jaume I (project UJI-B2018-53 to V. M. A. and project UJI-A2020-21 to MQM). SV thanks Croatian Science Foundation (project IP-2018-01-3168). Catarina I.G. Pinto is enrolled in the PhD scholarship 689 DFA/BD/07119/2020., With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000917-S).

Kvasac Saccharomyces cerevisiae godinama se koristi kao modelni organizam u cijelom nizu istraživanja. Tako se koristi i za proučavanje molekularnih mehanizama oksidativnog stresa te za proučavanje učinaka metalnih iona na metaboličku aktivnost stanica. Danas postoje raznolike tehnologije i materijali koji se koriste u dentalnoj medicini. Glavne karakteristike „dobrih“ dentalnih materijala su biokompatibilnost i fizikalne kvalitete (mehanička i termička otpornost). Dentalne legure u sebi najčešće sadrže nikal, titanij, krom, molibden, kobalt i sl. Poznato je da je nikal najalergeniji metal, da je krom često povezivan s oksidativnim stresom, no i ostali metali koji se nalaze u dentalnim legurama mogu biti potencijalno toksični. Legure "rematitan" i "remanium" često se koriste u izradi dentalnih lukova zbog svoje superelastičnosti, biokompatibilnosti i lakog rukovanja prilikom postavljanja ortodontskog aparata. U ovom istraživanju proučavano je kako će stanice kvasca reagirati na ione metala koji se otpuštaju iz dentalnih lukova „rematitan“ i „remanium“ te kakva će biti njihova metabolička aktivnost ako su izloženi ovim ionima. Za određivanje metaboličke aktivnosti korišteni su: test stanične proliferacije (XTT test), „Live/Dead Yeast Viability kit“ te AO/PI boja koja se koristi za brojanje stanica na LUNA-FLTM uređaju. Krivulja rasta stanica kvasca, tretiranih i netretiranih, većeg je porasta i pravilnijeg oblika kod stanica uzgajanih na YPD hranjivoj podlozi nego kod stanica uzgajanih u umjetnoj slini. U tom slučaju stanice su imale minimalan, čak i zanemariv porast. Kod stanične proliferacije primijećeno je da promjena medija ima utjecaj na odgovor stanica obaju sojeva kvasca. Divlji soj kvasca razvije određenu toleranciju i povećava se broj stanica bez obzira što u umjetnoj slini nema hranjivih tvari poput šećera. Metabolički aktivnih stanica, određenih FUN-1 bojom, podjednako je kod oba tretmana („rematitan“ i „remanium“), dok je nešto manji udio metabolički poremećenih stanica kod tretmana „rematitan“-om. Stanice tretirane metalnim ionima mijenjaju oblik i veličinu te rastu u nakupinama („klasteri“). Metalni ioni koji su se otpuštali iz ortodontskih lukova 7 dana pokazali su najveću toksičnost., Yeast Saccharomyces cerevisiae has been used for years as a model organism in a number of studies. Thus, it is also used to study the molecular mechanisms of oxidative stress and to study the effects of metal ions on the metabolic activity of cells. Today, there are a variety of technologies and materials used in dentistry. The main characteristics to be considered in material selection are biocompatibility and quality. Dental alloys usually contain nickel, titanium, chromium, molybdenum, cobalt, etc. It is already known that nickel is the most allergenic metal, chromium is often associated with oxidative stress, but other metals found in dental alloys can as well be potentially toxic. Alloys "rematitan" and "remanium" are often used in the manufacture of dental arches due to their super-elasticity, biocompatibility and easy handling when installing an orthodontic appliance. In this study, it was studied how yeast cells will react to metal ions released from the dental arches "rematitan" and "remanium" and what will be their metabolic activity if they are exposed to these ions. To determine metabolic activity, the following were used: cell proliferation test (XTT test), „Live/Dead Yeast Viability kit“ and AOPI dye used to count cells on the LUNA-FLTM device. The growth curve of yeast cells, treated and untreated, is larger and more regular in cells grown on YPD nutrient medium than in cells grown in artificial saliva. In this case, the cells had a minimal, even negligible increase. In cell proliferation, a change in medium has been observed to affect the cell response of both strains of yeast. The wild strain of yeast develops a certain tolerance and increases the number of cells despite the fact that there are no nutrients such as sugar in artificial saliva. Metabolically active cells, determined by FUN-1 color, are the same in both treatments ("rematitan" and "remanium"), while a slightly lower proportion of metabolically disturbed cells in the treatment with "rematitan". Cells treated with metal ions change shape and size and grow in clusters ("clusters"). Metal ions released from orthodontic arches for 7 days showed the greatest toxicity.

High-intensity training is becoming more popular nowadays when people have less time to engage in prolonged physical activity. Expertly led high intensity training is a safe way to achieve desired fitness goals. The aim of the study was to check if there were significant changes in the concentrations of sodium, potassium, calcium, magnesium, zinc, iron and copper in the blood and urine of twelve trainees after a short but intense training. Blood and urine sampling was performed before and after high intensity training where bodyweight exercises and exercises with external load were used. Statistical analysis was performed using paired t-test (2-tailed) with α=0.05 as statistical significance. The results obtained showed that the measured mineral concentrations varied as a result of intense physical activity, but these variations were small and did not have a general trend of increase or decrease of analyzed mineral content. Based on these results, it can be concluded that, from the standpoint of the mineral concentrations loss, short high-intensity training is safe for the trainee’s health., Visoko intenzivni trening postaje sve popularniji u današnje vrijeme kada ljudi imaju sve manje vremena za bavljenje dugotrajnim fizičkim aktivnostima. Stručno vođen trening visokog intenziteta može biti siguran način postizanja fitnes ciljeva. Cilj istraživanja bio je da se provjeri dešavaju li se značajne promjene u koncentracijama natrija, kalija, kalcija, magnezija, cinka, željeza i bakra u krvi i urinu 12 vježbača poslije kratke, ali intenzivne fizičke aktivnosti. Uzorkovanja krvi i urina provođena su prije i poslije treninga visokog intenziteta u kojem su se koristile vježbe s vanjskim opterećenjem i vlastitom težinom tijela. Statistička analiza rezultata je napravljena korištenjem parnog t-testa (2-tailed) te je kao nivo značajnosti uzeta vrijednost α=0.05. Rezultati su pokazali da se koncentracija mjerenih minerala mijenja usljed naporne fizičke aktivnosti, ali promjene su male i nemaju generalni trend povećanja ili smanjenja koncentracije analiziranih minerala. Na osnovu ovih rezultata može se zaključiti da je, sa stanovišta gubitka minerala, kratak trening visokog intenziteta siguran za zdravlje vježbača.

The presence of nickel could modify bacterial behavior and susceptibility to antimicrobial agents. Adhesion and biofilm formation on orthodontic archwires can be a source of bacterial colonization and possible health hazards. Staphylococcus aureus was subjected to exposure and adaptation to various sub-inhibitory concentrations of nickel. Five strains of bacteria adapted to nickel in concentrations of 62.5–1000 μg/mL were tested for adhesion and biofilm formation on nickel-titanium archwires. Archwires were previously incubated in artificial saliva. Bacteria were incubated with orthodontic wire with stirring for 4 h (adhesion) and 24 h (biofilm formation). The number of adherent bacteria was determined after sonication and cultivation on the Muller-Hinton agar. Disk diffusion method was performed on all bacteria to assess the differences in antimicrobial susceptibility. Bacteria adapted to lower concentrations of nickel adhered better to nickel-titanium than strains adapted to higher concentrations of nickel (p &lt, 0.05). Biofilm formation was highest in strains adapted to 250 and 500 μg/mL of nickel (p &lt, 0.05). The highest biofilm biomass was measured for strains adapted to 250 μg/mL, followed by those adapted to 1000 μg/mL. Bacteria adapted to lower concentrations of nickel demonstrated lower inhibition zone diameters in the disk diffusion method (p &lt, 0.05), indicating increased antimicrobial resistance. In conclusion, bacteria adapted to 250 μg/mL of nickel ions adhered better, demonstrated higher biofilm formation and often had higher antimicrobial resistance than other adapted and non-adapted strains.

The treatment with fixed orthodontic appliances could have an important role in the induction of oxidative stress and associated negative consequences. Because of the simultaneous effects of corrosion, deformation, friction, and mechanical stress on fixed orthodontic appliances during treatment, degradation of orthodontic brackets and archwires occurs, causing higher concentrations of metal ions in the oral cavity. Corroded appliances cause the release of metal ions, which may lead to the increased values of reactive oxygen species (ROS) due to metal-catalyzed free radical reactions. Chromium, iron, nickel, cobalt, titanium, and molybdenum all belong to the group of transition metals that can be subjected to redox reactions to form ROS. The estimation of health risk due to the amount of heavy metals released and the level of selected parameters of oxidative stress generated for the time of treatment with fixed orthodontic appliances is presented. Approaches to avoid oxidative stress and recommendations for the preventive use of topical or systemic antioxidants during orthodontic treatment are discussed.

Objectives: The goal of this study was to determine GD3 expression after treatment with the novel thieno[2,3-b]pyridine derivative, by focusing on CD44+/CD24- CSCs, CD44- epithelial cells, as well as CD44+/ CD24+ hybrid cells. Methods: The MDA-MB-231 cell line was treated with 3-amino-N-(3-chloro-2- methylphenyl)-5-oxo-5,6,7,8-tetrahydrothieno[2,3-b]quinoline-2-carboxamide (compound 1). Cells were plated onto six-well plates and treated with 2 M compound 1 for 48 h. Following this, cells were stained with anti-GD3, anti-CD44 and anti-CD24 antibodies, and then analyzed via flow cytometry. The acquisition of data of triple-stained samples was done by a BD Accuri C6 cytometer (BD Biosciences) and analyzed with the FlowLogic Software. Results: Treatment of MDA-MB-231 cells with compound 1 has led to a reduction of the expression of GD3 in CD44+/CD24- CSCs, as well as CD44- epithelial cells. GD3 expression in CD44+/CD24+ hybrid cells was not significantly reduced. Conclusion: Due to its ability to reduce GD3 expression in different breast cancer subpopulations, compound 1 has a potential to be used in the treatment of triple-negative breast cancer., Ciljevi: Cilj ove studije bio je utvrditi ekspresiju GD3 nakon tretmana novim derivatom tieno[2,3-b]piridina, posebice na CD44+/CD24- matičnim stanicama raka (CSC), CD44- epitelnim stanicama, kao i CD44+/CD24+ hibridnim stanicama. Metode: Stanična linija MDA-MB-231 tretirana je s 3-amino-N-(3-kloro-2-metilfenil)-5- okso-5,6,7,8-tetrahidrotienom[2,3-b]kinolin-2-karboksamid (spoj 1). Stanice su nasađene na ploče sa šest jažica i tretirane s 2 M spojem 1 tijekom 48 sati. Nakon toga, stanice su obojane s protu-GD3, protu-CD44 i protu-CD24 protutijelima, a zatim analizirane protočnom citometrijom. Prikupljanje podataka trostruko obojenih stanica rađeno je citometrom BD Accuri C6 (BD Biosciences) i analizirano softverom FlowLogic. Rezultati: Tretman MDA-MB-231 stanica spojem 1 doveo je do smanjenja ekspresije GD3 na CD44+/CD24- CSC, kao i CD44- epitelnim stanicama. Ekspresija GD3 na hibridnim stanicama CD44+/CD24+ nije bila značajno smanjena. Zaključak: Zbog svoje sposobnosti smanjenja ekspresije GD3 u različitim subpopulacijama raka dojke, spoj 1 ima potencijal za upotrebu u liječenju trostruko-negativnog raka dojke.

Biljne infuzije ubrajaju se u najčešće konzumirane napitke diljem svijeta. Njihova popularnost je u porastu, ponajprije zbog potencijalnih zdravstvenih dobrobiti koje se pripisuju fenolnim sastojcima u tim pripravcima i njihovim antioksidacijskim svojstvima. Premda je u brojnim istraživanjima zabilježen visok antioksidacijski kapacitet biljnih infuzija primjenom metoda koje se oslanjaju na postupak vodene ekstrakcije biljnog materijala, većina ih zanemaruje fiziološke parametre, poput fizikalno-kemijskih uvjeta u probavnom sustavu koji značajno utječu na bioraspoloživost, a time i na biološku aktivnost fenolnih sastojaka. Stoga je u ovom radu usporedno prikazan i antioksidacijski kapacitet biljnih infuzija koji je određen nakon probave in vitro, u uvjetima koji oponašaju one u gastrointestinalnom traktu. Dostupni literaturni podaci ukazuju da bi se procjeni antioksidacijskog kapaciteta biljnih infuzija trebalo dati prednost metodama utemljenim na fiziološkom pristupu nad metodama koje ovaj parametar određuju u vodenim ekstraktima biljnog materijala, obzirom da promjene koje se događaju tijekom probave in vitro mogu doprinijeti poznavanju stvarne bioaktivne snage biljnih infuzija., Herbal infusions are among the most commonly consumed beverages around the world. Their popularity is rising, primarily due to the potential health benefits attributed to the phenolic compounds in these preparations and their antioxidant properties. Although a number of studies have reported high antioxidant capacity of herbal infusions using methods that rely on the process of aqueous extraction of herbal material, most of them neglect physiological parameters, such as physicochemical conditions in the digestive system that significantly affect the bioavailability and thus the biological activity of phenolic compounds.Therefore, in this paper, the antioxidant capacity of herbal infusions, which was determined after in vitro digestion, in conditions that mimic those in the gastrointestinal tract, is presented in parallel. Available literature data suggests that the assessment of antioxidant capacity of herbal infusions should be preferred to methods based on a physiological approach over methods that determine this parameter in aqueous extracts of herbal material, as changes occurring during in vitro digestion may contribute to knowledge of actual bioactive power of herbal infusions.

Ciljevi: Keratoakantom je keratinizirajući kožni tumor s centralnim kraterastim udubljenjem, nejasnog uzroka. Obilježavaju ga brzi rast i česta spontana regresija. Protein povezan s citotoksičnim limfocitima T ili CTLA-4 inhibicijski je receptor iz obitelji CD28 koji zaustavlja aktivaciju reagirajućih T-stanica te posreduje suprimirajuće funkcije regulacijskih T-stanica. Cilj studije je utvrditi izražaj CTLA-4 u centru, bazi keratoakantoma i zdravom lateralnom rubu kože kao kontroli te ispitati ovisi li izražaj o stadiju keratoakantoma, spolu i dobi. Materijali i metode: Istraživanje je obuhvatilo 30 uzoraka tkiva uklopljenog u parafinski blok, dobivenih nakon ekscizije keratoakantoma. U imunofluorescencijskom bojenju korišteno je anti–CTLA-4 zečje antitijelo (ab237712, Abcam, UK) te sekundarno antitijelo obilježeno fluoresceinom, a preparati su analizirani fluorescentnom mikroskopijom. Broj stanica pozitivnih na CTLA-4 izražen je u postotku u odnosu na ukupan broj stanica po vidnom polju (400 x). Rezultati: Protein CTLA-4 izražen je u centru tumora u 28 (93.33%) slučajeva, na bazi tumora u 28 (93.33%) slučajeva, a značajno manje na zdravom lateralnom resekcijskom rubu, u 17 (56.67%) uzoraka. U centralnom dijelu tumora postoji statistički značajna razlika u izražaju CTLA-4 među stadijima keratoakantoma (P = 0.020). Postoji statistički značajna razlika između skupine s proliferativnnim stadijem i skupine s neodređenim stadijem keratoakantoma (P = 0.037) te između skupine s proliferativnim stadijem i skupine s regresijskim stadijem keratoakantoma (P = 0.022). Statistički značajna razlika prema spolu nije pronađena u ekspresiji CTLA-4 u centru tumora (P = 0.546), bazi tumora (P = 0.151), niti na zdravom lateralnom resekcijskom rubu (P = 0.914). Nije utvrđena statistički značajna razlika među dobnim skupinama s obzirom na ekspresiju CTLA-4 u centru tumora (P = 0.592), na bazi tumora (P = 0.888) te na zdravom resekcijskom rubu (P = 0.821). Zaključci: Izražaj CTLA-4 je značajno veći u tumoru (centar i baza) nego na zdravom lateralnom resekcijskom rubu. Regresijski stadij keratoakantoma ima veći izražaj CTLA-4 nego proliferativni stadij. Spol i dob nisu povezani s izražajem CTLA-4. Za dodatna pojašnjenja uloge CTLA-4 u razvoju i/ili regresiji keratoakantoma, potrebna su daljnja istraživanja u smijeru fenotipizacije CTLA-4 pozitivnih stanica., Objectives: Keratoacanthoma is a dome-shaped skin tumor with a centralized keratinous plug, characterized by initial rapid growth and often spontaneous regression. Cytotoxic T-lymphocyte-associated protein 4 or CTLA-4 is an inhibitory receptor, belonging to the CD28 immunoglobulin subfamily, that inhibits T cell activation and mediates regulatory T-cell suppressive activity. The aim of this study was to determine whether there is a difference in CTLA-4 expression in the keratoacanthoma center, at the base and in the healthy lateral resection margin as a control, as well as to determine whether it is dependent on keratoacanthoma stage, sex, and age. Materials and methods: The study included 30 samples of paraffin-embedded tissue, obtained after KA excision. Anti–CTLA-4 rabbit antibody (ab237712, Abcam, UK) and a secondary antibody labeled with fluorescein were used for immunofluorescence staining. The slides were analyzed by fluorescence microscopy. The number of CTLA-4-positive cells was expressed as a percentage of the total number of cells per field of view (400 x). Results: CTLA-4 protein was expressed in the tumor center in 28 (93.33%) cases, at the tumor base in 28 (93.33%) cases, and significantly less on the healthy lateral resection margin, in 17 (56.67%) samples. Statistically significant difference in CTLA-4 expression between the stages of KA was found in the central part of the tumor (P = 0.020). Also, statistically significant difference was found between the group with the proliferative stage and the group without exact stage of keratoacanthoma (P = 0.037), as well as between the group with the proliferative stage and the group with the regression stage of KA (P = 0.022). No statistically significant gender difference was found in CTLA-4 expression in the tumor center (P = 0.546), tumor base (P = 0.151), or at the healthy lateral resection margin (P = 0.914). No statistically significant difference was found between age groups with respect to CTLA-4 expression in the tumor center (P = 0.592), at the tumor base (P = 0.888), and at the healthy resection margin (P = 0.821). Conclusions: CTLA-4 expression is significantly higher in the tumor (center and base) than on the healthy lateral resection margin. The regression stage of keratoacanthoma has a higher expression of CTLA-4 than the proliferative stage. Gender and age were not associated with CTLA-4 expression. To further clarify the role of CTLA-4 in the development and/or in the regression of keratoacanthoma, additional research is needed to determine the phenotype of CTLA-4 positive cells.

The goal of each professional athlete is to reach maximal performance. In order to achieve that, physical as well as mental training needs to be part of a regular training routine. Anxiety significantly affects athletic performance while extreme anxiety levels are associated with reduced athletic achievements. Therefore, controlling sport-related anxiety seems to be one of the most significant aspects for elite athletes. In order to understand the biology behind sport-related anxiety, this review is focused on the biochemical responses to such a stressor. Knowing the biochemistry of stress and its limiting factors on athletic performance enables us a greater understanding of the influence of anxiety on professional athletes. It might help to better understand the importance of sport psychology and how our mindset influences our performance. Furthermore, the correlation between the placebo effect and the manner in which the mind controls the body are considered as it might prove an important tool for future athletic training and sport psychology., Cilj svakog profesionalnog sportaša jest postizanje najviše razine sportskih performansi. U tu svrhu fizička i mentalna priprema trebaju biti dio redovitih treninga. Anksioznost značajno utječe na sportsku izvedbu a izrazita tjeskoba povezana je sa smanjenim atletskim dostignućima. Stoga je za sportaše jedna od vrlo važnih značajki sposobnost kontroliranja anksioznosti. Kako bismo razumjeli biološke odgovore povezane s anksioznošću, ovaj je pregledni rad usmjeren na razjašnjavanje biokemijskih odgovora navedenog stresora. Poznavanje biokemijskog odgovora organizma uzrokovanog stresom te njegovih ograničavajućih učinaka na sportsku izvedbu omogućuje nam veće razumijevanje utjecaja tjeskobe na profesionalne sportaše. Navedeno nam daje uvid u bolje razumijevanje važnosti sportske psihologije te pojašnjava na koji način mentalno stanje utječe na fizičku izvedbu. U radu se opisuje i korelacija između placebo učinka i načina na koji um kontrolira fizičku snagu, s obzirom na to da ova saznanja mogu predstavljati važnu komponentu sportske psihologije, te daje mogućnost poboljšanja načina pripreme sportaša.

Sve veća potreba za hranom u svijetu uslijed rasta populacije dovela je do sve veće uporabe pesticida koji svakodnevno predstavljaju potencijalnu opasnost, kako za okoliš, tako i za čovjeka. Imajući to na umu, provedeno je istraživanje koje je za cilj imalo analizirati pokretnost insekticida acetamiprida na tlima Republike Hrvatske. Provedene su analize četiriju uzoraka tla (Š, J, O, Kt) uzorkovanih s područja zapadne Slavonije te uzorka bogatog organskom tvari (Ku) kupljenog u trgovini. Izvršene su analize fizikalno-kemijskih svojstava tla, od mehaničkog sastava (teksture) tla, kiselosti, kapaciteta izmjene kationa (KIK) do sadržaja, sastava te prirode građe humusa, nakon čega je pristupljeno analizi kinetike sorpcijsko-desorpcijskih procesa acetamiprida u tlu čiji ostaci su se analizirali tekućinskim kromatografom s masenim spektrometrom (LC-MS) koristeći pritom Lagergrenov model pseudo-prvog reda, bifazni model kinetike prvog reda te Weber-Morrisov model unutar-čestične difuzije. Analizom mehaničkog sastava (teksture) tla, uzorci Š, J, O i Kt klasificirani su kao glinaste ilovače (srednje do fino teksturirana tla), dok je uzorak Ku klasificiran kao pjeskulja (grubo teksturirano tlo). Reakcija tla okarakterizirana je kao kisela (pH 5,59-6,04), a prema udjelu humusa (organske tvari) u tlu, uzorci Š, J, O i Kt pripadaju slabo humoznim tlima (1,78-2,95 % C), dok uzorak Ku pripada jako humoznim tlima (62,37 % C). Bifazni model kinetike prvog reda pokazan je kao najprikladniji model za opis kinetike sorpcijsko-desorpcijskih procesa acetamiprida u tlu. Uzorak Ku imao je najveće sorbirane i desorbirane količine acetamiprida u tlu, dok je najveće vrijednosti k1 i k2 imao uzorak J. Organska tvar tla imala je najveći utjecaj na spomenute procese, ali analiza zajedničkog utjecaja svih karakteristika tla zajedno najbolje opisuje kinetiku sorpcijsko-desorpcijskih procesa acetamiprida u tlu., The growing need for food in the world due to the population growth has led to the increasing use of pesticides that pose a potential risk to both the environment and humans on a daily basis. With this in mind, this study was conducted to analyze the mobility of the insecticide acetamiprid in the soils of the Republic of Croatia. Analysis of the four soil samples (Š, J, O, Kt) sampled from the area of western Slavonia and a sample rich in organic matter (Ku) purchased from the shop were performed. Analysis of physicochemical soil properties, such as mechanical soil composition (texture), acidity, cation exchange capacity (CEC), composition and structure of soil humus were performed, while kinetics analysis of acetamiprid sorption-desorption processes in soils was conducted using the Lagergren model of the pseudo-first order, Two-site model and Weber-Morris intra-particle diffusion model. Residue of acetamiprid was determined by liquid chromatograph with a mass spectrometer (LC-MS). According to the mechanical composition (texture) of the soil, samples Š, J, O and Kt were classified as clay loams (medium to finely textured soils), while sample Ku was classified as sandy (coarsely textured soil). The soil reaction was acidic (pH 5.59-6.04), while samples Š, J, O and Kt belong to the low humus soils (1.78-2.95% C), and the Ku sample to soils with high humus content (62.37% C). Two-site model has been shown to be the most suitable model for describing the kinetics of sorption-desorption processes of acetamiprid in soil. Sample Ku sorbed and desorbed the highest amount of acetamiprid in the soil, while the sample J had the highest value of k1 and k2. Soil organic matter had the greatest influence on the mentioned processes, but the multiple linear regression analysis that combined soil characteristics best describes the kinetics of sorption-desorption processes of acetamiprid in soil.

U današnje vrijeme ljudi su sve više opsjednuti “hollywoodskim” osmijehom kojega svi oblici medija svakodnevno serviraju na ovaj ili onaj način (filmovi, TV serije/emisije, socijalne mreže i sl.). Stoga, sve veći broj ljudi počinje koristiti ortodontske naprave, neznajući potencijalne štetne učinke koje njegove komponente mogu imati na čovjekov organizam. Otpuštanje iona metala iz ortodontskih materijala postaje sve veća zabrinutost ortodontskoga svijeta jer, osim uzrokovanja kontaktnoga dermatitisa, moguća je i pojava akutnog ili kroničnog otrovanja. Osim toga, ioni metala mogu uzrokovati oksidativni stres pomoću kojega će se manifestirati njihovi genotoksični i citotoksični učinci. Cilj istraživanja bio je ispitati učinak otpuštenih iona (prvenstveno Ni2+, Mo4+ i Cr3+) na vitalnost i vijabilnost dva soja kvasca: Saccharomyces cerevisiae W303 i divlji tip. Istraživanje je odrađeno u dva dijela – na hranidbenoj podlozi (YPD) i umjetnoj slini. U svakoj su metodi korištene hranidbene podloge i uzorci umjetne sline s onom količinom iona otpuštenih iz ortodonskih bravica i prstenova kroz 3, 7 i 14 dana. Vijabilnost je praćena promjenom broja stanica oba soja kvasca kroz 24 sata na tretiranim hranidbenim podlogama i uzorcima umjetne sline. Rezultati su iskazani preko krivulja rasta te prebrojenih stanica pomoću LUNA-FLTM Dual fluorescence cell counter-a. S druge strane, vitalnost je praćena promjenom metaboličke aktivnosti stanica dvjema metodama: prva – bojanjem stanica LIVE/DEAD Yeast Viability Kit-om i promatranjem stanica fluorescentnim mikroskopom, i druga – korištenjem XTT testa. Također, fluorimetrijskim mjerenjem unutarstanične oksidacije određena je i količina ROS-ova stvorenih u stanicama kvasaca kao posljedica aktivnog metabolizma i rasta na podlogama s iscrpinama metala. Ioni ortodontskih bravica i prstenova utječu na vitalnost i vijabilnost oba soja kvasca Saccharomyces cerevisiae, pogotovo na soj W303 u svim parametrima., Nowadays, people are getting more and more obsessed with the “Hollywood” smile that all forms of media serve in one way or another (movies, TV shows/series, social networks, etc.). Therefore, an increasing number of people are starting to wear orthodontic appliances, unaware of the potential harmful effects that its components could have on the human organism. The release of metal ions from orthodontic materials is becoming a growing concern of the orthodontic world because, in addition to causing contact dermatitis, acute or chronic poisonings are also possible to occur. In addition, metal ions can cause oxidative stress, by which their genotoxic and cytotoxic effects could be manifested. The aim of this study was to examine the effects of released ions (Ni2+, Mo4+ and Cr3+) on the vitality and viability of two yeast strains: Saccharomyces cerevisiae W303 and wild type. The research was done in two parts – on YPD growth media and in artificial saliva. Growth media and artificial saliva samples with the amount of ions released from orthodontic braces and brackets over 3, 7 and 14 days were used in each method. Viability was observed by the change in the number of the cells of both yeast strains over 24 hour-growth in treated growth media and artificial saliva samples. The results were expressed via growth curves and counted cells using a LUNA-FLTM Dual fluorescence cell counter. On the other hand, vitality was observed by the change in cell's metabolic activity using two method. First – cell staining with the LIVE/DEAD Yeast Viability Kit, afterwhich the cells were microscoped using a fluorescence microscopy, and the second – XTT test. Also, fluorimetric measures of intracellular oxidation determined the amount of ROS generated in yeast cells as a result of active metabolism and growth on depleted media. The ions of orthodontic braces and rings affect the vitality and viability of both strains of the yeast Saccharomyces cerevisiae, especially the W303 strain in all parameters.

Titanij se široko primjenjuje zbog svoje biokompatibilnosti i dobrih biomehaničkih svojstava. Pokazuje dobru sposobnost vezanja za živo tkivo i kosti pa se često upotrebljava za zubne, ortopedske i srčane implantate. Upotrebljava se kao čisti titanij ili kao legura na bazi titanija, gdje dodatak drugog elementa titaniju omogućuje dobivanje boljih svojstava od samoga titanija. Unatoč inertnosti, titanij je i dalje podložan koroziji i otpuštanju štetnih metalnih iona u organizam. Većina studija pokazuje da je titanij izrazito biokompatibilni materijal, međutim određene studije ukazuju da materijali od titanija mogu postati nekompatibilni i prouzročiti štetne učinke. S obzirom na široku upotrebu titanija, javljaju se pitanja o mogućoj štetnosti titanija. Stoga je važno i dalje provoditi toksikološka ispitivanja, pomno procijeniti i ispitati potencijalnu štetnost titanija te provjeriti potencijalne zdravstvenerizike za ljude i za ekosustav., Titanium is widely used due to its biocompatibility and good biomechanical properties. Often it is the material of choice for making dental, orthopaedic and heart implants because it has good binding ability to living tissue and bones. It is used as pure titanium or as a titanium alloys, where the addition of another element or more of them allows to obtain better properties than titanium itself. Despite its inertness, titanium is still susceptible to corrosion and the release of harmful metal ions into the body. Most studies show that titanium is a highly biocompatible material, however there are certain studies that show that titanium materials can become incompatible and cause harmful effects. Given the widespread use of titanium, questions arise about its possible harmfulness. It is therefore important to continue to conduct toxicological tests, to carefully evaluate and examine the potential harmfulness of titanium, and to check for potential health risks to humans and the ecosystem.

Dimethoate is an organophosphorus insecticide still used in Croatia and worldwide, with polar structure and high water solubility that make it prone to leaching. This study analyzed how physico-chemical properties of soils affected dimethoate sorption and mobility. For that purpose, five soil samples were collected from three Croatian regions (two coastal and one mountain region). Dimethoate sorption process was analyzed using the batch procedure while its mobility and leaching potential was investigated by column experiment. The results showed that dimethoate sorption can be adequately described by Freundlich model. All isotherms were of L-type with varying degrees of non-linearity, indicating different sorption efficiencies and distribution of sorption sites energies among the soils. Energy distribution was broader in soils richer in organic matter (OM). KF values indicated relatively low sorption efficiency for all soils, with an increase of KF values proportional to OM content. Mechanisms involved in dimethoate sorption and mobility were analyzed by fitting the breakthrough curves (BTCs) with two mathematical models, namely one-site equilibrium (ELM) and two-site nonequilibrium sorption model (NELM). Correlations were quantified by Kendall-Tau test, which revealed the strongest correlation of KF value with OM content, cation exchange capacity and the humic acid content, while correlations with pH, clay content and A465nm/A665nm ratio were negative and insignificant. Based on these findings, a model for prediction of leaching potential was formed. A simplified model for dimethoate sorption/transport was proposed.

Cilj: Utvrditi mogući sinergizam antioksidansa iz prirodnih uzoraka soka aronije i crnog vina te ispitati kinetiku reakcija između DPPH radikala i antioksidansa prisutnih u uzorcima. Materijali i metode: Antioksidativna aktivnost u pet metanolnih uzoraka (ARONIJA, VINO, MIX 1 (50 % soka aronije i 50 % crnog vina), MIX 2 (75 % soka aronije i 25 % crnog vina), MIX 3 (25 % soka aronije i 75 % crnog vina)) određena je pomoću DPPH• i TEAC testova i analizirana spektrofotometrijski, dok se kinetika kemijskih reakcija pratila trima monofaznim i jednim bifaznim matematičkim modelom. Rezultati: Prema kvantifikaciji TEAC testa, najjači antioksidacijski kapacitet smjesa, a ujedno i najbolji sinergizam antioksidansa, pokazuje MIX 1 (75 mmol/kg), zatim MIX 3 i MIX 2 (67 i 62 mmol/kg). Gubitak signala DPPH radikala najjači je u uzorku ARONIJA (81 %), dok je najslabiji u uzorku VINO (21 %). Kombinacijom napitaka, kemijske reakcije s radikalom odvijaju se dvjema vrstama antioksidansa čiji se mehanizam djelovanja odvija u dvjema fazama brzine kemijske reakcije. Zaključak: Dodatak soka aronije u crno vino uzrokuje porast njegove antioksidativne aktivnosti. Odnosno, ukoliko se priprema smjesa aronijinog soka i crnog vina, komponente moraju biti u jednakim udjelima (50 : 50) kako bi se postigao što bolji sinergizam ovih dviju vrsta antioksidansa, a time i ostvario jači zdravstveni učinak., Aim: To determine the synergism of antioxidants from natural samples of chokeberry juice and red wine as well as to examine the reaction kinetics between DPPH radical and antioxidants presented in the samples. Materials and Methods: Antioxidant activity in five methanolic samples (ARONIJA, VINO, MIX 1 (50% chokeberry juice and 50% red wine), MIX 2 (75% chokeberry juice and 25% red wine), MIX 3 (25% chokeberry juice and 75% red wine)) was measured spectrophotometrically using DPPH• and TEAC assays, while the chemical reaction of kinetics was analyzed with three monophasic and one biphasic mathematical models. Results: TEAC test shows the best antioxidants synergism of, as well as the strongest antioxidant capacity for mixture, MIX 1 (75 mmol/kg), followed by MIX 3 and MIX 2 (67 and 62 mmol/kg). The inhibition of DPPH radical signal was the strongest in the ARONIJA sample (81%), while it was the weakest in the VINO sample (21%). By combining beverages, two different types of antioxidants inhibited the DPPH radical causing two phases of reaction with different chemical reaction rates. Conclusion: The addition of chokeberry juice to red wine causes an increase in its antioxidant activity. Furthermore, if the chokeberry juice and red wine mixture is prepared, the samples should be in equal amount (50:50) in order to achieve the best antioxidants synergism and also strengthened the health effect.

Antinuclear antibodies (ANA) represent a family of autoantibodies targeting ubiquitous cellular constituents and are a hallmark of systemic inflammatory autoimmune rheumatic diseases named connective tissue diseases (CTD). The gold standard method for ANA determination is indirect immunofluorescence (IIF) on the human laryngeal epidermoid carcinoma cell line type 2 substrate (HEp-2), but with increasing demand for ANA testing, novel methods eased for automation emerged, which allows testing by staff less experienced in this specific field of laboratory diagnostic. In 2016 The working group (WG) for laboratory diagnostics of autoimmune diseases as part of the Committee for the Scientific Professional Development of the Croatian Society of Medical Biochemistry and Laboratory Medicine (CSMBLM) published the data of a survey regarding general practice in laboratory diagnostics of autoimmune diseases in Croatia. Results indicated high diversity in the performance of autoantibody testing as well as reporting of the results and indicated the need of creating recommendations for the assessment of ANA that would help harmonize diagnostics of systemic autoimmune rheumatic diseases in Croatia. This document encompasses twenty-seven recommendations for ANA testing created concerning indications for ANA testing, preanalytical, analytical, and postanalytical issues, including rational algorithm and quality control assurance. These recommendations are based on the relevant international recommendations and guidelines for the assessment of ANA testing and relevant literature search and should help to harmonize the approach in ANA testing and clarify differences in interpretation of the results obtained using different methods of determination.

Ulcerative colitis (UC) is one of the major forms of inflammatory bowel disease with an increasing incidence and prevalence worldwide. It is a chronic remitting and relapsing disease that occurs spontaneously characterized by gastrointestinal symptoms, and significantly affects the quality of life1. Currently used treatments for UC are unsatisfactory due to various side effects, failure in the induction of remission and prevention of relapse, as well as high economic costs2. Recently, the use of natural compounds has become increasingly attractive in the prevention and treatment of various diseases, including UC. Luteolin (3′, 4′, 5, 7- tetrahydroxyflavone ; Lut) is a common flavonoid present in many edible plants that has been shown to have beneficial effects on various diseases3, however, its effect on UC has been poorly studied. In this study, we investigated the effect of Lut in the posttreatment and cotreatment of experimental colitis induced by dextran sulfate sodium (DSS) in mice. Lut attenuated clinical symptoms of DSS-induced colitis in mice by diminishing body weight loss and improving disease activity index, alleviated colon tissue damage, and reduced inflammation, apoptosis and autophagy. This effect was more pronounced when Lut was applied simultaneously with DSS. Additionally, the role of extracellular signal-regulated kinases 1/2 (ERK1/2) in the mechanism of action of Lut in experimental colitis was investigated as it was shown that Lut increased the activity of ERK1/2. Inhibition of ERK1/2 exacerbated the symptoms of DSS-induced colitis and diminished the protective effects of Lut as evidenced by worsening of inflammation and increased apoptosis and autophagy. Overall, the results of this study contributed to the elucidation of the underlying mechanism of action of Lut in experimental colitis and provided new mechanistic details underlying the antiinflammatory, antiapoptotic and antiautophagic effects of Lut through the activation of the ERK signaling pathway. In conclusion, the results suggest that Lut could be an effective therapeutic candidate for the treatment of UC, or a supplement to conventional therapy, although further clinical studies are required.

CILJEVI ISTRAŽIVANJA: Ciljevi ovog istraživanja bili su odrediti izražaj CD15s na epitelnim (CD44-CD24+ i CD44-CD24-) i mezenhimalnim (CD44+CD24- tj. CSC) stanicama MDA-MB-231 stanične linije trostruko negativnog karcinoma dojke te odrediti izražaj CD44 kod CSC CD15s+ i CSC CD15s- stanica nakon primjene tienopiridinskog derivata. MATERIJALI I METODE: Novosintetizirani tienopiridinski derivat, 3-amino-N- (3-kloro-2-metilfenil)- 5 -okso -5,6,7,8-tetrahidrotieno [2,3-b] kinolin- 2-karboksamid, primijenjen je u 2 μM koncentraciji u trajanju od 48 sati na MDA-MB-231 staničnoj liniji. Potom su stanice imunobojene s anti-CD15s odnosno sekundarnim antitijelom na anti-CD15s konjugiranim s eFluor 660 fluorokromom, s anti-CD44-FITC i anti-CD24-PE te je fluorescencija pojedinih kromogena konačno mjerena protočnim citometrom. REZULTATI: U izražaju CD15s u CSC subpopulaciji nije bilo značajne razlike prije i nakon tretmana tienopiridinskim derivatom. Za razliku od CSC subpopulacije, ovaj je tretman rezultirao gotovo dvostrukim smanjenjem izražaja CD15s u subpopulaciji CD44- (P=0,004). Tretman nije utjecao na ekspresiju CD44 u CD15s+ CSC, ali je smanjio izražaj CD44 (P=0,008) u CD15s-CSC. ZAKLJUČAK: Nalazi sniženog izražaja CD15s kod epitelnih stanica i CD44 kod CD15- mezenhimalnih stanica ukazuju na potencijal novosintetiziranog tienopiridinskog derivata kao terapijskog sredstva za trostruko negativni rak dojke., Objectives: The aims of this study were to analyse the expression of CD15s in epithelial (CD44-CD24+) and mesenchymal (CD44+CD24-, CSC) cells in MDA-MB-231 triple-negative breast cancer cell line and to measure expression of CD44 in CSC CD15s+ and CSC CD15s- cells after being treated with thieno[2,3-b]pyridine derivative. Materials and methods: Newly synthesized thieno[2,3-b]pyridine derivative of 3-amino-N-(3-chloro-2-methylphenyl)-5-oxo-5,6,7,8-tetrahydrothieno[2,3-b]quinoline-2-carboxamide was used in 2 μM concentration for 48 hours on MDA-MB-231 cell line. Thereafter, the cells were stained with anti-CD15s secondary antibody conjugated with eFluor 660 on CD15s fluorochrome, anti-CD44-FITC, anti-CD24-PE, and then chromogen fluorescence was analyzed by flow cytometry. Results: There was no difference in expression of CD15s in CSC subpopulation before and after treatment with thieno[2,3-b]pyridine derivative. Unlike CSC subpopulation, this treatment has resulted in almost two times reduction of CD15 expression in CD44- subpopulation (P=0.004). The treatment didn’t have any influence on the expression of CD44 in CD15s+CSC but it had reduced expression of CD44 (P=0.008) in CD15-CSC. Conclusion: The finding of reduced expression of CD15s in epithelial cells and CD44- within mesenchymal cells show the potential of newly synthesized thieno[2,3-b]pyridine derivative as a potential treatment option for triple-negative breast cancer.

Uvod: Ulcerozni kolitis (UK) je kronična bolest koja nastaje kao posljedica neodgovarajućeg imunološkog odgovora. Rak debelog crijeva treći je najčešće dijagnosticirani zloćudni tumor i drugi uzrok smrtnosti. Dosadašnje terapije ovih bolesti nisu se pokazale odgovarajućim. Klorogenična kiselina i luteolin su polifenolni spojevi s pokazanim značajnim farmakološkim učincima u tretmanu različitih bolesti. Cilj istraživanja: Cilj rada je ispitati i predložiti mehanizam djelovanja klorogenične kiseline i luteolina u terapiji UK i raka debelog crijeva. Materijal i metode: Istraživanje je provedeno na in vivo modelu kolitisa i na in vitro modelu raka debelog crijeva. Tijekom razvoja kolitisa praćene su kliničke promjene, a stupanj oštećenja tkiva debeloga crijeva određen je histopatološkom analizom. Izražaj istraživanih proteina određen je Western blot metodom i imunohistokemijski. Uloga signalnog puta izvanstaničnim signalima reguliranih kinaza (ERK) istražena je primjenom inhibitora. Citotoksičnost ispitivanih spojeva određena je testom stanične vijabilnosti i sposobnosti stvaranja kolonija. Autofagija i apoptoza analizirane su fluorescencijski. Uloga autofagije ispitana je primjenom inhibitora autofagije. Rezultati: Klorogenična kiselina i luteolin ublažavaju kliničke simptome i smanjuju intenzitet upale. Klorogenična kiselina smanjuje aktivaciju proteina signalnih puteva proteinskih kinaza aktiviranih mitogenom (MAPK), proteinske kinaze B (PKB (Akt)) i pretvarača signala i aktivatora transkripcije 3 (STAT3) te djeluje antioksidacijski. Luteolin aktivira ERK protein. Klorogenična kiselina ne utječe na vijabilnost stanica, dok luteolin smanjuje njihovu vijabilnost i proliferaciju te djeluje proapoptotski, proautofagično i antimetastatski. Zaključak: Istraživanjem je pokazan protuupalni i antiapoptotski učinak klorogenične kiseline u UK te protuupalni, antiapoptotski i antiautofagični učinak luteolina koji je posredovan ERK signalnim putem. Antikarcinogena aktivnost luteolina se temelji na ERK/FOXO3a ovisnom mehanizmu i antimetastatskom potencijalu Introduction: Ulcerative colitis (UC) is a chronic disease that results from an inadequate immune response. Colon cancer is the third most commonly diagnosed malignancy and the second leading cause of death. Previous therapies for these diseases have not been shown to be adequate. Chlorogenic acid and luteolin are polyphenolic compounds with a shown significant pharmacological effects in the treatment of various diseases. Aim of the study: The aim of this study was to investigate and propose the mechanism of action of chlorogenic acid and luteolin in the treatment of UC and colon cancer. Matherial and methods: The study was performed in an in vivo model of colitis and in an in vitro model of colon cancer. During the development of colitis, clinical changes were monitored and the degree colon tissue damage was determined by histopathological analysis. The expression of the investigated proteins was determined by the Western blot method and immunohistochemically. The role of the extracellular regulated kinases (ERK) signaling pathway was investigated using inhibitor. The cytotoxicity of the test compounds was determined by a cell viability and colony formation assay. Autophagy and apoptosis were analyzed by fluorescence. The role of autophagy was investigated using autophagy inhibitor. Results: Chlorogenic acid and luteolin alleviate clinical symptoms and reduce the intensity of inflammation. Chlorogenic acid reduces the activation of mitogen-activated protein kinases (MAPK), protein kinase B (PKB (Akt)) and signal transducer and transcription activator 3 (STAT3) signaling pathway proteins and has an antioxidant effect. Luteolin activates ERK protein. Chlorogenic acid does not affect cell viability, while luteolin reduces their viability and proliferation and has proaptotic, proautophagic and antimetastatic effect. Conclusion: The study demonstrated the antiinflammatory and antiapoptotic effect of chlorogenic acid in the UC and the antiinflammatory, antiapoptotic and antiautophagic effect of luteolin mediated by the ERK signaling pathway. The anticarcinogenic activity of luteolin is based on the ERK/FOXO3a dependent mechanism and antimetastatic potential.

Nitroksidni radikali su organske molekule koje imaju jedinstvena antioksidacijska svojstva te mogu suzbiti rast tumora. U in vivo uvjetima, ovi radikali ne mogu se učinkovito koristiti jer se nespecifično akumuliraju u zdravom tkivu te se brzo izlučuju iz organizma. U ovom radu istraživane su nanočestice temeljene na poli[N-(2-hidroksipropil)metakrilamidu] (poli(HPMA)) i kolesterolu koje sadrže TEMPONE radikal vezan hidrazonskom (RNP-H) i peptidnom vezom (RNP-P). Veličina i oblik nanočestica određeni su metodom krio-transmisijske elektronske mikroskopije. Utvrđeno je da su dobivene nanočestice sferičnog oblika prosječne veličine 25−50 nm. Otpuštanje radikala s nanočestica praćeno je metodom elektronske spinske rezonancije (ESR) u rasponu pH vrijednosti od 7,4 do 5,0. Pokazano je da u kiseloj sredini koja je karakteristična za tumorske stanice dolazi do brzog otpuštanja radikala vezanih hidrazonskom vezom dok nanočestice koje sadrže radikale vezane peptidnom vezom ostaju stabilne. Nasuprot tome, u fiziološkim uvjetima ne dolazi do značajnog oslobađanja radikala ni sa jedne vrste nanočestica. Ovim istraživanjem pokazano je da se polimerne nanočestice mogu koristiti kao nosači nitroksidnih radikala u novom pristupu terapiji tumora kada se žele izbjeći neželjene nuspojave slobodnih radikala i produljiti njihovo zadržavanje u organizmu., Nitroxide radicals are organic molecules that possess unique antioxidant properties and can suppress tumor growth. Under in vivo conditions, these radicals can not be used effectively because they accumulate nonspecifically in healthy tissue and are rapidly removed from the body. In this work, we investigated nanoparticles based on poly[N-(2-hydroxypropyl)methacrylamide] (poly(HPMA)) and cholesterol containing a TEMPONE radical bound by a hydrazone (RNP-H) and peptide bond (RNP-P) respectively. The size and shape of the nanoparticles were determined by cryo-transmission electron microscopy. Spherical nanoparticles with an average size of 25–50 nm were obtained. The release of radicals from the nanoparticles was investigated by electron spin resonance (ESR) spectroscopy in the pH range from 7.4 to 5.0. It was shown that the radicals bound by hydrazone bond are rapidly released in an acidic environment which is characteristic for tumor cells, while the nanoparticles containing radicals bound by peptide bond remain stable. In contrast, there is no significant release of radicals from both types of nanoparticles under physiological conditions. This research has shown that polymer nanoparticles can be used as a carriers of nitroxide radicals in a new approach to tumor therapy when we want to avoid unwanted side effects of free radicals and prolong their retention in the body.

U radu je praćen utjecaj termičke obrade maslinovog i suncokretovog ulja u atmosferi zraka na njegovu antioksidacijsku aktivnost. Varirana su dva parametra: vrijeme i temperatura termičkog tretiranja. Pri tome su odabrani parametri koji odgovaraju realnim parametrima pripreme namirnica i to temperatura od 150 °C i 180 °C te vrijeme 10 min i 20 min. Antioksidacijsko djelovanje praćeno je mjerenjem relativne koncentracije radikala 2,2-difenill- pikrilhidrazil (DPPH) elektronskom spinskom rezonancijom (ESR) tijekom 20 min po dodatku ulja otopini radikala. Ispitivana su četiri uzorka maslinovog ulja i jedan uzorak suncokretovog ulja. Pokazalo se da kod svih uzoraka termička obrada, bez obzira na vrijeme i temperaturu tretiranja, dovodi do porasta antioksidacijske moći prema DPPH radikalu. To se može objasniti prisutnošću radikala nastalih tijekom procesa termičke obrade kao posljedica lipidne peroksidacije. Ti radikali vjerojatno doprinose bržoj redukciji DPPH. Različito antioksidacijsko djelovanje termički tretiranih uzoraka najvjerojatnije potječe od različitog kemijskog sastava ulja. Neki su uzorci za vrijeme tretiranja tijekom 10 min pri 150 °C pokazali bolju aktivnost nego kada su bili tretirani u istom vremenskom intervalu pri višoj temperaturi, dok su drugi pokazali jače antioksidacijsko djelovanje nakon tretiranja tijekom 10 min pri 180 °C. Za dulje vrijeme tretiranja (20 min) pri 180 °C, većini je uzoraka oslabila antioksidacijska moć, u usporedbi s kraćim vremenskim tretiranjem ili tretiranjem pri nižoj temperaturi. U suncokretovom ulju su tokoferoli odgovorni za njegovo antioksidacijsko djelovanje. Njihova uloga u neutraliziranju novonastalih radikala čini se različitom u odnosu na ulogu polifenola kod maslinovog ulja. Može se pretpostaviti da tokoferoli slabije reagiraju s novonastalim radikalima od polifenola pa je stoga uloga tih radikala u neutralizaciji DPPH znatno izraženija kod suncokretovog ulja u usporedbi s maslinovim uljem, The influence of thermal treatment of olive oil and sunflower oil in the presence of air on their antioxidant activity was studied. Two parameters were varied: time and temperature of thermal treatment. The parameters that correspond to the real parameters of food preparation were selected, namely the temperature of 150 ° C and 180 ° C and the time of 10 min and 20 min. Antioxidant activity was monitored by measuring the relative concentration of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical by electron spin resonance (ESR) during 20 min after the addition of oil to the radical solution. Four samples of olive oil and one sample of sunflower oil were investigated. It was shown that for all samples the heat treatment, regardless of the time and temperature of treatment, leads to an increase in antioxidant power against the DPPH radical. This can be explained by the presence of radicals formed during the heat treatment process as a consequence of lipid peroxidation. These radicals probably contribute to a faster reduction of DPPH. The different antioxidant activity of thermally treated samples most likely originates from the different chemical composition of the oil samples. Some samples showed better activity during treatment for 10 min at 150 ° C than when treated during the same time interval at higher temperature, while others showed stronger antioxidant activity after treatment for 10 min at 180 ° C. For a longer treatment time (20 min) at 180 ° C, antioxidant power decreased for most samples, compared to a shorter time treatment or treatment at a lower temperature. In sunflower oil, tocopherols are responsible for its antioxidant activity. Their role in neutralizing newly formed radicals appears to be different from the role of polyphenols in olive oil. It can be assumed that tocopherols react less well with newly formed radicals than polyphenols and therefore the role of these radicals in neutralizing DPPH is significantly more pronounced in sunflower oil compared to olive oil

Polimeri su makromolekule koji danas „grade“ većinu predmeta s kojima se čovjek susreće. Polimeri se koriste najviše za izradu ambalaže, u graditeljstvu, u automobilskoj industriji, za izradu elektronskih uređaja, igračka, namještaja, odjeće, obuće, u medicini ali i poljoprivredi i kućanstvu. Cilj ovog istraživanja bio je ispitati utjecaj alumosilikata (zeolita) kao punila na dinamiku lanaca polimera poli(dimetil-siloksana). Za ispitivanje su se koristile metode elektronske spinske rezonancije (ESR) i razlikovne pretražne kalorimetrije (DSC). Kao proba za ispitivanje elektronskom spinskom rezonancijom korišten je nitroksilni radikal 4-okso-2,2,6,6-tetrametilpiperidin-1-oksil (TEMPONE). Rezultati ispitivanja pokazuju da je pri niskim temperaturama ispod staklišta (Tg) dinamika lanaca spora što se vidi po širokim linijama spektra. Blizu staklišta pojavljuju se složeni, tzv. bimodalni spektri dok se povišenjem temperature pojavljuju uske linije karakteristične za brzo gibanje lanaca. Oblik i koncentracija zeolita također utječu na dinamiku lanaca polimera. Veće koncentracije zeolita i kubični oblik zeolita više usporavaju dinamiku lanaca od manjih koncentracija i bipiramidnog i okruglog oblika zeolita. Isto tako, istraživanjem je pokazano da je elektronska spinska rezonancija osjetljivija metoda za ispitivanje lokalne dinamike lanaca od razlikovne pretražne kalorimetrije., Polymers are macromolecules that today "build" most of the objects that man encounters. Polymers are mostly used for the production of packaging, in construction, in the car industry, for the manufacture of electronic devices, toys, furniture, clothing, footwear, in medicine, but also in agriculture and household. The aim of this study was to examine the influence of aluminosilicates, zeolites, as fillers on the dynamics of poly(dimethyl-siloxane) polymer chains. Investigations were performed using electron spin resonance (ESR) and differential scanning calorimetry (DSC). The nitroxide radical 4-oxo-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPONE) was used as a probe for electron spin resonance. Results of this study show that at low temperatures, below the glass transition temperature (Tg), the dynamics of the chains is slow, as indicated by the presence of the broad lines in ESR spectra. In the vicinity of the glass transition temperature, composite, so-called bimodal spectra appear, while increasing the temperature narrow lines appear, characteristic of fast chain motions. The shape and concentration of the zeolite also affect the dynamics of polymer chains. Higher concentrations of zeolite and the cubic shape of zeolite slow down the chain dynamics more than lower concentrations of both, bipyramidal and round forms of zeolite. Likewise, research has shown that electron spin resonance is a more sensitive method for examining local chain dynamics than differential scanning calorimetry.

Šećerna bolest ili dijabetes predstavlja skup poremećaja metabolizma ugljikohidrata, masti i bjelančevina u organizmu. Riječ je o značajnom javnozdravstvenom problemu koji predstavlja jedan od deset vodećih uzroka smrti u svijetu. Bolest se dijeli na tip I, tip II, gestacijski dijabetes te ostale, manje zastupljene vrste dijabetesa. Dijabetes prate mnoge komplikacije, poput otežanog cijeljenja rana. Mnogi znanstveni radovi dokazuju i naglašavaju važnost proteaza u procesu cijeljenja rana koji je u dijabetesu otežan zbog brojnih čimbenika. Cijeljenje rana je proces koji se odvija u više međusobno povezanih faza. Dipeptdil-peptidaza IV, odnosno molekula CD26 multifunkcionalan je protein koji ima vrlo važnu ulogu u održavanju homeostaze glukoze u krvi. Također, ima vrlo značaju ulogu u patofiziološkim i fiziološkim procesima. Molekula DPP IV/CD26 izražena je na zrelim timocitima, aktiviranim limfocitima T, B, NK-stanicama te makrofagima ali i brojim stanicama kao što su epitelne, endotelne stanice različitih organa i tkiva. Poznato je da DPP IV/CD26 sudjeluje u adheziji, migraciji i apoptozi stanica, razgradnji ekstracelularnog matriksa stanica te ključnim procesima u cijeljenju rana, gdje djeluje kao upalni medijator te potiče reepitalizaciju ozlijeđenog tkiva. Funkcija DPP IV/CD26 je dvostruka; svojom proteolitičkom funkcijom utječe na različite supstrate koji sudjeluju u procesu cijeljenja rana te sudjeluje u modulaciji i stimulaciji imunološkog odgovora u procesu cijeljenja rana., Diabetes mellitus or diabetes is a set of disorders of the metabolism of carbohydrates, fats and proteins in the body. It is a significant public health problem because it is one of the ten leading causes of death in the world. The disease is divided into type I, type II, gestational diabetes and other, less common types of diabetes. Diabetes is accompanied by many complications, such as delayed wound healing. Many scientific papers prove and emphasize the importance of proteases in the wound healing process. The wound healing process in diabetes is hampered by a number of factors. Wound healing is a process that takes place in several interrelated stages. Dipeptidyl peptidase IV, or CD26 molecule, is a multifunctional protein that plays a very important role in maintaining blood glucose homeostasis. It also has a very important role in pathophysiological and physiological processes. The DPP IV / CD26 molecule is expressed on mature thymocytes, active T, B lymphocytes, NK cells, and macrophages and a number of cells such as epithelial, endothelial cells of various organs and tissues. DPP IV / CD26 is known to participate in cell adhesion, migration and apoptosis, degradation of the extracellular matrix of cells and key processes in wound healing, where it acts as an inflammatory mediator and promotes reepitalization of injured tissue. The function of DPP IV / CD26 is twofold; its proteolytic function affects various substrates involved in the wound healing process and participates in the modulation and stimulation of the immune response in the wound healing process.

UVOD: Ulcerozni kolitis (UK) je jedan od dva patohistološka oblika upalne bolesti crijeva. Predstavlja imunološki posredovanu bolest obilježenu ponavljajućim upalnim procesima koji rezultiraju oštećenjem i gubitkom funkcije crijevne sluznice. Pokazano je da su proteini obitelji dipeptidil-peptidaze (DP)4, posebice, nedovoljno istraženi članovi, DP8 i 9 putem svojih enzimskih i kostimulacijskih učinaka uključeni u modulaciju imunoloških mehanizama te proliferaciju i aktivaciju stanica koji vode k razvoju upalne bolesti crijeva. CILJ: Cilj je istraživanja ispitati promjene transkripcijske aktivnosti DP8 i 9, signalnog pretvarača i aktivatora transkripcije (STAT)3 i 6 te inducibilne dušične sintetaze (iNOS) na eksperimentalnom modelu UK izazvanog primjenom natrijevog dekstran sulfata (DSS). MATERIJAL I METODE: Eksperimentalni akutni kolitis je izazvan je u C57BL/6 miševa primjenom 3 % otopine DSS tijekom sedam dana u vodi za piće. Tijekom pokusnog perioda pratio se razvoj i intenzitet kolitisa temeljem promjena kliničkih manifestacija, indeksa aktivnosti bolesti, te lokalnih promjena. Nadalje, pratile su se promjene izražaja DP8 i 9, iNOS te STAT3 i STAT6 na transkripcijskoj razini u tkivu debeloga crijeva tijekom razvoja DSS kolitisa u miša. REZULTATI: Razvoj upalnog procesa na sistemskoj razini rezultirao je pojavom kliničkih manifestacija, porastom indeksa aktivnosti bolesti i hepatosomatskog indeksa. Na lokalnoj razini, razvoj kolitisa praćen je porastom mikroskopskog indeksa oštećenja sluznice debelog crijeva i skraćenjem debelog crijeva. Trećeg dana pokusnog perioda zabilježen je značajan porast transkripcijske aktivnosti DP8, DP9, STAT3 i iNOS. Značajan porast transkripcijske aktivnosti STAT3 i iNOs zabilježen je i sedmog dana. Izražaj STAT6 gena se tijekom razvoja kolitisa nije mijenjao. ZAKLJUČAK: Rezultati ukazuju na ulogu ispitivanih proteaza i signalnih molekula u razvoju i progresiji DSS kolitisa., Introduction. Ulcerative colitis (UC) is one of the two major pathohistological forms of inflammatory bowel disease. It represents an immune mediated disease which is characterized by recurrent episodes of the colon inflammation resulting in damage and loss of the intestinal mucosal function. It has been shown that the proteases dipeptidyl peptidase (DP)4 and recently discovered DP8 and 9 are involved in the modulation of immune mechanisms leading to colitis development as well as in the proliferation and activation of immune cells. Aim. Considering our previous results and experience, we hypothesized that changes in the expression of DP8 and 9 influence on the colitis development. Therefore, the aim of the research was to investigate the role of DP8 and 9, STAT3 and STAT6, and inducible nitric oxide synthase (iNOS) in the induction and progression of dextran sulfate sodium (DSS)-induced experimental colitis in mice. Material and Methods. Colitis was induced in C57BL/6 mice by oral administration of 3% solution of DSS. The health status of animals, changes in body weight, and development of clinical symptoms of colitis were monitored during the experimental period. Animals were sacrificed on day 7. Transcriptional expression of the DP8, DP9, STAT3, STAT6, and iNOS in the colonic tissue were evaluated by RT-qPCR. Results. The development of the colonic inflammation resulted in the appearance of clinical manifestations, an increase in the disease activity index, microscopic damage score and hepatosomatic index. A significant increase in transcriptional activity of DP8, DP9 in the colon was determined on the third day of the experimental period. Likewise, the increase in transcriptional activity of STAT3 and iNOS were determined on the day 3 and 7. Interestingly, development of the colon inflammation has not influenced on the STAT6 gene expression. Conclusion. The results indicate the importance of investigated proteases and signal molecules in the DSS colitis induction and progression.

Formation of biofilms allows bacterial cells to survive in adverse environ- ments. Nontuberculous mycobacteria are ubiquitous in aqueous environ- ments, where they adhere to surfaces and create a biofilm. This has led to the emergence of healthcare-associated infections and the use of bioma- terials in medicine. Essential oils (EO) are substances of natural origin whose effect on microorganisms has been the subject of numerous studies. Here, we investigated the effect of Juniperus communis EO on nontuberculous mycobacteria and their early and mature biofilm formation in sterilised tap water. The combination of Juniperus communis EO and increasing ambient temperature showed a synergistic effect on the reduction of biofilm forma- tion of Mycobacterium avium and Mycobacterium intracellulare on a polystyrene surface. A significant antibiofilm effect of Juniperus communis EO was also found at subinhibitory concentrations, suggesting a potential role for it as an alternative disinfectant of natural water.

Određivanje antioksidacijskog kapaciteta popularan je cilj istraživanja znanosti o hrani, budući je ovo pitanje vezana za rok trajanja, senzorsku kvalitetu kao i uz potencijalne zdravstvene dobrobiti hrane, uključujući maslinovo i arganovo ulje koji su istaknut izvor fenolnih antioksidansa u prehrani. U ovom radu, za određivanje antioksidacijskog kapaciteta maslinovog i arganovog ulja primjenjena je ABTS metoda. Kao bi se ovaj parametar relevantnije procijenio, antioksidacijski kapacitet ulja određen je u (a) vodeno-organskim ekstraktima izdvojenim iz ulja kemijskom ekstrakcijom i (b) u topljivoj bioraspoloživoj frakciji i netopljivoj rezidualnoj frakciji koje zaostaju nakon procesa probave in vitro. Potonji pristup čini osnovu metode globalnog antioksidacijskog odgovora, utemeljene na primjeni enzima i osmišljene tako da oponaša probavu u gastrointestinalnom traktu. Postupak probave in vitro povećao je sadržaj ukupnih fenolnih sastojaka i antioksidacijski kapacitet određen ABTS metodom u bioraspoloživim frakcijama uzoraka ulja u usporedbi s kemijskim ekstraktima. Nadalje, značajan udio fenola od ukupnog sadržaja fenolnih sastojaka zaostaje u netopljivoj rezidualnoj frakciji analiziranih ulja nakon probave in vitro. Ove frakcije treba uzeti u obzir zajedno s onim topljivim pri procjeni antioksidacijskog kapaciteta ulja, jer zadržavaju značajnu antiradikalsku aktivnost prema ABTS radikalu. Primjena GAR metode za mjerenje antioksidacijskog kapaciteta mogla bi pridonijeti boljem poznavanju stvarne bioaktivne snage i zdravstvenog potencijala maslinovog i arganovog ulja, obzirom da doprinosi potpunijem oslobađanju fenolnih sastojaka iz matriksa ulja te omogućuje mjerenje antiradikalske aktivnost u rezidualnoj netopljivoj frakciji, koja se ne uzima u obzir pri određivanju antioksidacijskog kapaciteta tradicionalnim metodama koje se oslanjaju na primjenu organskih otapala za kemijsku ekstrakciju bioaktivnih sastojaka., The assessment of food antioxidant capacity is a popular objective of food science research, as this issue is related to shelf life, sensory quality and potential health benefits of foods, including olive and argan oil, which are prominent source of phenolic antioxidants in the diet. In this study, antioxidant capacity of oils was determined by the ABTS assay. Within the aim to provide more relevantly assess this parameter, antioxidant capacity was measured in (a) aqueous-organic extracts prepared by traditional chemical method, and (b) in both soluble and insoluble fractions obtained by Global Antioxidant Response (GAR) method, designed as an in vitro enzyme-based approach that mimics digestion in the gastrointestinal tract. The in vitro digestion process increased the content of total phenolic constituents and the antioxidant capacity in the bioavailable fractions of oil samples compared to the initial chemical chemical extracts. Moreover, a significant proportion of phenolics remained in the insoluble fractions of analysed oils after in vitro digestion. These fractions should be taken into account together with the soluble ones when assessing antioxidant capacity of oils, as they are characterized by significant antiradical activity against ABTS radical. GAR method enabled more complete release of phenolics from the oil matrix and permitted the measurement of the antiradical activity in the insoluble fraction, which is excluded from the traditional extraction methods employing organic solvents to extract bioactive compounds. Hence, the application of GAR method for measurement of antioxidant capacity could contribute to a better knowledge of the bioactive power and health potential of olive and argan oil.

Korištenje velikih količina pesticida, kao i njihova visoka topljivost u vodi predstavljaju popriličnu opasnost za okoliš, u vidu onečišćenja voda, zagađenja tla, zraka te narušavanja homeostaze ekosustava. Iz toga razloga, cilj istraživanja bio je ispitati utjecaj karakteristika tla s lokaliteta Grobnik, Matulji, Međimurje i Otočac uz poseban naglasak na utjecaj organske tvari tla na sorpcijsko ponašanje dimetoata. Izvršena su određivanja aktualne, supstitucijske i hidrolitičke kiselosti tla, količine i sastava humusa, određivanja ugljika u ekstraktu, huminskim i fulvinskim kiselinama te spektrofotometrijsko određivanje prirode građe huminskih kiselina pri valnim duljinama od 465 i 665 nm. Određivanje sorpcijskog kapaciteta tla za dimetoat provedeno je pomoću tekućinskog kromatografa visoke djelotvornosti (HPLC). Reakcija tla bila je neutralna i alkalna (pH=6,73-7,40), a udio organske tvari u tlu se nalazio u rasponu od 1,21-3,47% te svrstava tla u slabo humozna i dosta humozna tla. Prema Freundlichovom modelu opisa procesa sorpcije organskih spojeva na tlu dobivene su sorpcijske izoterme C oblika kod uzoraka GR, M2 i L1, L oblika kod uzorka MEĐ te S oblika kod uzorka M1, a vrijednosti koeficijenta KF su bile u rasponu od 0,20-6,91 (mg/kg)(mg/L)1/n. Rezultati ukazuju kako sorpcijsko ponašanje dimetoata u tlu nije uvjetovano samo organskom tvari tla, već zajedničkim učinkom svih karakteristika tla., The use of large pesticide quantities, as well as their high water solubility, represent a significant environmental hazard, in the form of water, soil, air pollution and disruption of the ecosystem homeostasis. For that reason, the aim of this study was to investigate the effect of soil characteristics from following locations: Grobnik, Matulji, Međimurje and Otočac with the main emphasis on the effect of organic carbon soil characteristics on the sorption behavior of dimethoate. Actual, exchangeable and hydrolytic acidity of soil, character and humus content, amount of carbon in extract, humic and fulvic acids and spectrophotometric determination of the natural humic acid structure at wavelengths of 465 and 665 nm were performed. Determination of the soil sorption capacity for dimethoate was performed using a high performance liquid chromatograph (HPLC). The soil reactions were neutral and alkaline (pH=6.73-7.40) and the proportion of organic matter content in soil ranged from 1.21-3.47% classifying soils as poorly and fairly humus soils. According to Freundlich's model for describing the sorption process of organic compounds in soil, sorption isotherms were classified as C type for the samples GR, M2 and L1, L type for the sample MEĐ and S type for the sample M1. Furthermore, the values of the KF coefficient varied from 0.20-6.91 (mg/kg) (mg/L)1/n. The results of the study indicate that the sorption behavior of dimethoate in soil is not only caused by soil organic matter content, than by the combined effect of all soil characteristics.

U ljudskom organizmu pronađene su brojne uloge soka aronije i crnog vina. Ovi napitci poznati su po jakom antioksidativnom učinku, kardioprotektivnom, hepatoprotektivnom, antibakterijskom te neuroprotektivnom djelovanju u kojima antioksidansi reduciraju brojne slobodne radikale koji uzrokuju veliki broj bolesti. Kinetika njihovih reakcija može se opisati matematičkim pristupom, odnosno modeliranjem pomoću jednadžbi. Ovim istraživanjem proučavan je sinergizam antioksidanasa iz soka aronije i crnog vina. Pripremljeno je pet uzoraka (ARONIJA, VINO, MIX 1, MIX 2, MIX 3) kojima se analizirala antioksidativna aktivnost uz DPPH• i TEAC test te kinetika kemijskih reakcija s tri monofazna i jednim bifaznim matematičkim modelom. Ukazano je da smjesa aronije i crnog vina u omjeru 1:1 (MIX 1), pokazuje najjači sinergizam ovih komponenti te najjači antioksidativni kapacitet u odnosu na druge mješavine. Uspoređivanjem statističkih pokazatelja, o slaganju eksperimentalnih i modelnih podataka, utvrđeno je kako bizafni model prvog reda najbolje opisuju kinetiku kemijskih reakcija DPPH radikala i antioksidanasa, dok je najlošiji opis kinetike dao model nultog reda. Time je i dokazano postojanje dvaju vrsta antioksidanasa koji reagiraju s DPPH radikalom različitim brzinama te uzrokuju i dvije faze reakcije. Dodatak soka aronije u crno vino uzrokuje porast njegove antioksidativne aktivnosti. Odnosno, ukoliko se priprema smjesa aronijinog soka i crnog vina, komponente moraju biti u jednakim udjelima kako bi se postigao što bolji sinergizam ovih dvaju vrsta antioksidanasa i što bolji zdravstveni učinak., The antioxidants from aronia juice and red wine are responsible for countless roles in the human body. Besides the antioxidant activity, these beverages are known for their cardioprotective, hepatoprotective, antibacterial and neuroprotective effects, but their main function is to reduce free radicals that can cause a large number of diseases. The kinetics of these reactions could be described by a mathematical approach or by equation modeling. In this reasearch, the synergism of antioxidants from aronia juice and red wine was examined. Five samples (ARONIA, WINE, MIX 1, MIX 2, MIX 3) were prepared in order to analyze the antioxidant activity, using the DPPH• and TEAC assay. Furthermore, the kinetics of chemical reactions between DPPH• and antioxidants were studied using three single-phase and one biphasic mathematical models. Results of the study pointed that mixture of aronia and red wine in a 1:1 ratio (MIX 1) demonstrates the strongest synergism of aronia juice and red wine as well as the strongest antioxidant capacity compared to other mixtures. Statistical parameters which tested differences between experimental and model data, showed that the biphasic first-order model best describes the chemical reaction kinetics between DPPH radical and antioxidants in all mixtures. This confirmed that two types of antioxidans react with the DPPH radical at different rates, making two different reaction phases. The addition of aronia juice to the red wine increases its antioxidant activity. Therefore, mixture od aronia juice and red wine in equal quantity make the most out of their synergism and have a much better health effect.

Nitinol legura, legura nikla i titana, jedan je od materijala koji imaju dobru biokompatibilnost, otporni su na koroziju i sadrže još poneka svojstva zbog kojih se često koriste u izradi implantata. No zbog nepovoljnog fiziološkog stanja okoline i mehaničkih oštećenja moguća je biorazgradnja legure i otpuštanje iona metala u tijelo. Cilj ovog istraživanja je odrediti učinak iona metala koji se otpuštaju iz dentalnih naprava izrađenih od Nitinol legure na stanice kvasaca. Pratio se učinak ekstrakta Nitinol legure, Ni2+ (NiCl2) i Ti4+ (TiO2) iona. Kao radni mikroorganizam korišten je soj W303 kvasca Saccharomyces cerevisiae. Metabolička aktivnost kvasaca određena je dvjema metodama: fluorimetrijski i fluorescentnim mikroskopom uz pomoć „Live/Dead Yeast Viability kit“-a. Povećanje broja stanica nakon 20 sati uzgoja manje je kod tretmana žicom i niklom nego kod kontrolnih, netretiranih stanica. Najveći učinak na metaboličku aktivnost uočen je, i fluorimetrijski i fluorescentnim mikroskopom, kod tretmana žicom pošto je to zapravo objedinjeno tretman niklom i titanom., Nitinol alloy, alloy made of nickel and titanium, is one of the materiala that are used for biomedical applications because of their good biocompatibility, corrosion resistance and other properties. However, it may come to implant biodegradation and ions can be released to peri-implant tissues because of the aggressive physiological environment along with mechanical loading. This research aims to determin the effect of ions that are released from dental implants made of Nitinol alloy on yeast cells. we tracked effects of Nitinol alloy extract, Ni2+ (NiCl2) and Ti4+ (TiO2) ions. Work microorganism used in this research was yeast Saccharomyces cerevisiae (strain W303). Metabolic activity of yeast cells was determined with two methods: in microplate reader and under the fluorescens microscope using „Live/Dead Yeast Viability kit“. The increase in number of yeast cells after 20 h of culturing is lower after treatment with alloy and nickel than in the control sample. The biggest effect on metabolic activity of yeast cells was determined after the treatment with alloy because it is both treatment with nickel and titanium at the same time.

Tlo je površinski sloj Zemljine kore koji ima brojne uloge. Jedna od najvažnijih je osiguravanje hrane za ljude i životinje. S obzirom na rastuću populaciju na Zemlji, raste i potreba za hranom. Samim time poljoprivrednici se trude u kraćem vremenu, proizvesti više hrane, te posežu za kemijskim supstancama zvanim pesticidi. Cilj ovog istraživanja bio je istražiti utjecaj organske tvari tla na sorpcijsko/desorpcijske procese imidakloprida. Analizirana su četiri tla s područja Hrvatske, ispitane su njihove fizikalnokemijske karakteristike te udio humusa i njegov sastav, potom je tlima uklonjena organska tvar te su na njima provedeni sorpcijsko/desorpcijski procesi imidakloprida. Ravnotežna koncentracija imidakloprida određena je HPLC analitičkom tehnikom. Za prikaz ravnotežnih procesa sorpcije/desorpcije korišteni su Freundlichov i Langmuirov model izotermi. Freundlichov model pokazao se bolji za opisivanje sorpcijsko/desorpcijskih procesa imidakloprida u odnosu na Langmuirov model, te su dobiveni parametri Freundlichovim modelom korišteni za utvrđivanje utjecaja karakteristika tla na sorpcijsko/desorpcijsko ponašanje imidakloprida u tlu. Dobivene su izoterme L oblika. Kod tala s uklonjenom organskom tvari vrijednosti kapaciteta sorpcije/desorpcije su niži u odnosu na netretirana tla. Srednja vrijednost faktora KOC za netretirano tlo iznosi 193,22 kg/L, a za tretirano 370,80 kg/L što imidakloprid svrstava u kategoriju srednje pokretnih mikrozagađivala. Utvrđena je pozitivna ovisnost između organskog ugljika, udjela gline, kapaciteta izmjene kationa na koeficijent sorpcije (KF) odnosno desorpcije (KFd) kod netretiranih tala, dok je ta ovisnost manja kod tretiranih tala. Višestrukom linearnom regresijom utvrđeno je da je parametar ,,organski ugljik“ dominantan u sorpcijsko/desorpcijskim procesima imidakloprida u tlu, što dovodi do zaključka da organska tvar znatno utječe na procese sorpcije i desorpcije mikrozagađivala., Soil represents the surface layer of the Earth's crust, and it has many roles. One of the most important roles is providing food for humans and animals. With the expansion of the world's population, the need for food also grows. As a results, farmers are trying to produce more food in a shorter time using chemicals called pesticides. The aim of this research was to investigate the influence of organic matter content on the sorption/desorption processes of imidacloprid. For that purpose four soils with different physicochemical properties from coastal regions of Croatia were used. Furthermore, the sorption/desorption processes of imidacloprid in soils with reduced organic matter content were also investigated. The equilibrium concentration of imidacloprid was determined by HPLC analytical technique. To describe the imidacloprid equilibrium sorption/desorption processes, the Freundlich and Langmuir isotherm models were used. The Freundlich model proved to be better for description of the imidacloprid sorption/desorption processes compared to the Langmuir model, so the parameters obtained with the Freundlich model were used to determine the influence of soil characteristics on the sorption/desorption behaviour of imidacloprid. In all analysed soils imidacloprid isotherms were of L-shape. In soils with reduced organic matter, the values of sorption/desorption capacity were lower compared to untreated soils. The mean values of the KOC factor for untreated and treated soils were 193.22 and 370.80 kg/L, classifying imidacloprid in the category of medium mobile micro pollutants. Results of the research indicate a positive dependence between organic carbon, clay content, cation exchange capacity and the sorption (KF) and desorption (KFd) coefficients found in untreated soils, while this dependence was lower in treated soils. Multiple linear regression showed that organic matter content was the predominant soil characteristic influencing the sorption/desorption processes of imidacloprid in the soil, leading to the conclusion that organic matter significantly affects the processes of sorption and desorption of imidacloprid in analyzed soils.

Background: Aucubin is pharmacologically active natural compound which possesses numerous beneficial properties in vitro and in vivo. Purpose: This study aimed to evaluate the protective effect of aucubin against cisplatin (CP)-induced acute kidney injury in mice and the mechanism of its action. In addition, a modulation of CP-induced cyototoxicity was investigated in vitro. Study design: Aucubin was administrated to mice orally (po) and intraperitoneally (ip) (1.5 and 5 mg/kg) for two consecutive days, two days after ip injection of cisplatin (CP), 11 mg/kg. Human cervical cancer HeLa cells were treated with CP 30 µM and aucubin 0–1000 μM. Methods: Hematoxylin and eosin staining, western blot, immunohistochemistry and immunofluorescence were used to evaluate the renoprotective activity of aucubin. The cell viability assay was used to evaluate in vitro cytotoxicity of aucubin. Results: Treatment with aucubin by both routes of administration ameliorated histopathological changes and reduced elevated serum markers of kidney injury. CP administration increased renal expression of HO-1 and 4-HNE, as well as TNF-α, which was dose-dependently ameliorated by the administration of aucubin, indicating the suppression of oxidative stress and inflammation, respectively. Moreover, aucubin reduced increased renal expression of cleaved caspase-3 and -9 and decreased PARP cleavage, suggesting amelioration of CP-induced apoptosis. Mechanistically, aucubin normalized the activation of ERK1/2, PI3K/Akt, FOXO3a, STAT3 and NF-κB signaling in mice kidneys. Aucubin was slightly more successful in the amelioration of kidney injury when administered by ip than by po route. In human cervical cancer cells, aucubin did not modulate the CP cytotoxicity in doses up to 1 mM. Conclusions: The findings of this study show that aucubin acts as a protective agent against CP-induced nephrotoxicity, without interference with the CP anticancer activity in clinically relevant doses.

Water (H2O) is a polar inorganic compound that is almost colorless at room temperature. It is by far the most common substance on Earth and among very few substances, which exists on the Earth’s surface as a solid, liquid and gaseous substance. As a relatively simple chemical compound, it has been described as a “universal solvent” or “solvent of life”. It is also the third most common molecule in the Universe. Water molecules form strong polar hydrogen bonds with each other, which, in turn, define its specific physical and chemical properties. Hydrogen bonds are responsible for many unique properties such as the anomaly of water (the fact that its solid form, the ice, is less dense than the liquid form), a relatively high boiling point (100 °C) and a high heat capacity. Water is an amphoteric molecule, which means that it can show properties of both acids and alkaline solutions, depending on the chemical environment. In this paper, a structural analysis of a water molecule and its most important physical properties was carried out in detail with regard to its anomalies. One of the important physical properties of a liquid that strongly influences our environment is the low density of ice in relation to the liquid phase of water, and the phenomenon of the negative expansion coefficient of cold water. This work also describes the solutions of the nuclear Schrödinger equations, which provide information about the internal motions (vibrations and rotations) of a water molecule., Voda (H2O) je polarni anorganski spoj, koji je na sobnoj temperaturi tekućina bez okusa i mirisa, gotovo bezbojna. Količinski je daleko najzastupljeniji spoj na Zemlji i među rijetkim je tvarima koje na Zemljinoj površini postoje u krutoj, tekućoj i plinovitoj fazi. Kao relativno jednostavan kemijski spoj, opisana je kao „univerzalno otapalo“ ili „otapalo života“. Ujedno je i treća najzastupljenija molekula u Svemiru. Molekule vode međusobno tvore snažne vodikove veze, što joj definira specifična fizikalna i kemijska svojstva. Vodikove veze zaslužne su za mnoga jedinstvena svojstva vode, poput anomalije vode (činjenice da je kruti oblik – led – manje gustoće od tekuće vode), relativno visoke temperature vrenja (100 °C) i velikog toplinskog kapaciteta, što su ključni preduvjeti za očuvanje života na Zemlji. Voda je amfoterna molekula, što znači da može pokazivati svojstva i kiseline i lužine, ovisno o kemijskom okruženju u kojem se nalazi. U radu je provedena strukturna analiza molekule vode i njenih najvažnijih fizikalnih svojstava, s naglaskom na anomaliju vode. Jedno od važnih fizikalnih svojstava tekućine, a koja uvelike utječu na naš okoliš, je niska gustoća leda u odnosu na tekuću fazu vode te fenomen negativnog koeficijenta ekspanzije hladne vode. Također je opisano rješenje nuklearne Schrödingerove jednadžbe, koja daje informacije o unutarnjim gibanjima (vibracijama i rotaciji) molekule vode.

Acanthamoeba is an opportunistic protozoon, widespread in the aquatic environment, where it can be in endosymbiosis with over 30 pathogenic bacteria, including nontuberculous mycobacteria (NTM). Protozoa play a crucial role in mycobacterial pathogenesis and serve as a reservoir of infection. Since the first step in bacteria making contact with amoebae is adhesion, we were interested in investigating whether essential oils (EOs) can affect it. To that end we investigated the effects of juniper (Juniperus communis) and immortelle (Helichrysum italicum) EOs against Mycobacterium avium, M. intracellulare, and M. gordonae in tap water and against their adhesion to Acanthamoeba castellanii by combining them in synergistic EO concentrations. M. avium and M. intracellulare adhered to A. castellanii to a greater extent than M. gordonae. The adhesion of all NTMs was prevented by the subinhibitory concentrations of EOs. When comparing the effect of synergistic combinations of EOs and the effect of a single concentration from a combination, a higher percentage of adhesion inhibition in all synergistic combinations observed, except against M. gordonae. Neither oil was cytotoxic to A. castellanii. Our findings suggest that the EOs or their components weaken the contact of environmental NTMs and free-living amoebae and indirectly diminish their pathogenic potential, which could be of value in developing strategies for maintenance of water supply systems., Akantameba je oportunistička protozoa, široko rasprostranjena u vodenom okružju, gdje može kao endosimbionte sadržavati više od 30 različitih patogenih bakterija, uključujući netuberkulozne mikobakterije (NTM). Protozoe kao rezervoar infekcije imaju ključnu ulogu u mikobakterijskoj patogenezi. Sposobnost mikobakterija da se vežu, ulaze I umnožavaju se unutar protozoa vjerojatno im je omogućila da postanu intracelularni patogeni. Budući da je adhezija prvi korak u uspostavi kontakta bakterija s amebama, zanimalo nas je istražiti mogu li prirodni spojevi poput eteričnih ulja utjecati na taj kontakt. Cilj ove studije bio je istražiti učinak eteričnih ulja borovice (Juniperus communis) i smilja (Helichrysum italicum) prema NTM-ovima (M. avium, M. intracellulare i M. gordonae) u pitkoj vodi. Također, ispitana je mogućnost inhibicije adhezije tih NTM-ova na Acanthamoeba castellanii (A. castellanii) primjenom sinergijskih koncentracija eteričnih ulja. M. avium i M. intracellulare, u odnosu na M. gordonae, u većoj su mjeri adherirali na A. castellanii. Adhezija svih NTM-ova inhibirana je subinhibicijskim koncentracijama eteričnih ulja borovice i smilja. Kad se usporedi učinak sinergijskih kombinacija i učinka pojedine koncentracije iz sinergije, uočen je veći postotak inhibicije adhezije kod primjene svih sinergijskih kombinacija, osim jedne prema M. gordonae. Oba eterična ulja u ispitivanim koncentracijama nisu imala citotoksično djelovanje na stanice A. castellanii. Na taj su način primijenjene subinhibicijske koncentracije eteričnih ulja sa smanjenim toksičnim učinkom, uz zadržan antimikrobni i antiadhezivni učinak. Primjena eteričnih ulja ili njihovih komponenata u umjetnom vodenom okolišu mogla bi oslabiti kontakt NTM-ova i slobodno živućih ameba i posredno dovesti do smanjenja patogenoga potencijala okolišnih NTM-ova.

Autofagijom se uklanjaju štetne ili nepotrebne tvari iz stanice od makromolekularnih nakupina do čitavih organela poput mitohondrija. Procesom selektivnog uklanjanja mitohondrija, tzv. mitofagijom, uklanjaju se nefunkcionalni mitohondriji čije nakupljanje može dovesti do razvoja neurodegenerativnih bolesti i tumora. Osim toga, mitofagija je neophodna prilikom diferencijacije i razvoja pojedinih stanica, poput eritrocita, kada se ovim procesom uklanjanju svi zdravi i funkcionalni mitohondriji iz eritrocitnih prekursora. Svi oblici selektivne autofagije izrazito su specifični prema citoplazmatskom sadržaju koji se pakira u autofagosom zahvaljujući receptorima selektivne autofagije iako mehanizmi njihovog djelovanja nisu u potpunosti poznati. U ovoj doktorskoj disertaciji proučavani su molekularni mehanizmi regulacije uklanjanja mitohondrija putem receptora selektivne autofagije proteina NIX, smještenog u vanjskoj mitohondrijskoj membrani. Fosforilacija serina, uzvodno od domene LIR proteina NIX, doprinosi stvaranju novih i stabilnih veza između receptora i proteina LC3B, većoj regrutaciji autofagosoma i učinkovitijem uklanjanju mitohondrija iz stanice. Dimerizacija receptora drugi je mehanizam aktivacije, a ostvaruje se specifičnom defosforilacijom S212 smještenog u međumembranskom mitohondrijskom dijelu te ima isti učinak na započinjanje i napredovanje mitofagije kao i fosforilacija domene LIR. Konačno, kombinirani mehanizmi LIR-ovisne regrutacije autofagosoma i dimerizacije receptora pokazuju kumulativni učinak na aktivaciju receptora NIX. Poznavanje molekularne osnove regulacije NIX-ovisne mitofagije ključno je za bolje razumijevanje mehanizama diferencijacije pojedinih stanica te određenih patoloških stanja koji u svojoj podlozi imaju poremećeni proces mitofagije., Autophagy is important for removal of harmful and unnecessary components in the cells, from macromolecular aggregates to entire organelles such as mitochondria. Mitophagy, a process of selective removal of mitochondria, is crucial for elimination of dysfunctional mitochondria whose accumulation can lead to the development of neurodegenerative diseases and tumors. Furthermore, mitophagy is essential for differentiation and development of particular cell types, such as erythrocytes, for elimination of complete healthy and functional mitochondrial population from the erythrocyte`s precursors. All forms of selective autophagy are highly specific to the cytoplasmic content packaged inside the autophagosomes due to the selective autophagy receptors whose mechanisms of action are not fully identified. This doctoral thesis studies the molecular mechanisms of mitochondrial removal via selective autophagy receptor NIX located in the outer mitochondrial membrane. Phosphorylation of two serines, juxtaposed to NIX LIR domain, provide the formation of new and stable interactions between the receptor and LC3B protein, also more robust recruitment of autophagosomes and efficient removal of mitochondria. The second mechanism of NIX activation, dimerization, is achieved by specific S212 dephosphorylation located in the intermembrane mitochondrial space and has the same effect on mitophagy initiation and progression as LIR phosphorylation. Finally, the combined mechanisms of LIR-dependent autophagosomal recruitment and receptor dimerization show a cumulative effect on NIX receptor activation. The knowledge of the molecular basis of NIX-dependent mitophagy regulation is crucial for better understanding the mechanisms of individual cell`s differentiation and the development of pathological conditions that underlie the disturbed process of mitophagy.

Osim antioksidacijskog djelovanja, fitokemikalije pokazuju i druge farmakološke učinke, uključujući modulaciju aktivacije različitih signalnih putova uključenih u oštećenje tkiva. Cisplatin (CP) je protutumorski lijek koji, pored željenog učinka, dovodi i do neželjenih nuspojava, uključujući nefrotoksičnost. Cilj ovih istraživanja bio je utvrditi mehanizam djelovanja nekih od uobičajenih fitokemikalija, sekoiridoida oleuropeina, diterpenoidnog glikozida steviozida, monoterpenoida karvakrola i triterpenoida oleanolične kiseline, kod toksičnog oštećenja bubrega CP-om u miševa. Mužjaci BALB/cN miševa tretirani su oralno s nekoliko prethodno odabranih doza fitokemikalija tijekom 2 uzastopna dana, 48 sati nakon intraperitonealne primjene CPa (13 mg/kg). Dva dana kasnije, primjena CP-a rezultirala je histopatološkim promjenama koje ukazuju na oštećenja bubrega. Sve primjenjivane fitokemikalije, ovisno o dozi, ublažile su promjene koje ukazuju na indukciju oksidacijskog stresa, apoptozu, upalu ili autofagiju u oštećenim bubrezima. Rezultati trenutnih studija ukazuju na to da fitokemikalije mogu smanjiti akutno oštećenje bubrega uzrokovano CP-om potiskujući one procese koji dovode do oštećenja bubrega izazvanih CP-om modulacijom različitih signalnih putova. Dodatno, istražili smo učinak karvakrola i oleanolične kiseline na stanice raka vrata maternice (HeLa) te učinak istih prilikom primjene CP-a. Karvakrol je prilikom kotretmana inducirao rezistenciju na CP u HeLa stanicama suzbijanjem apoptoze i modulacijom autofagije. Zanimljivo je da je oleanolična kiselina povećala citotoksičnost CP-a u HeLa stanicama inducirajući citotoksičnu autofagiju. Kemosenzitacija HeLa stanica na CP oleanoličnom kiselinom ukazuje na njen potencijalni blagotvorni učinak za pacijente liječene CP-om kako bi se u terapiji mogle koristiti i manje doze od uobičajenih. In addition to their antioxidant activity, phytochemicals also show other pharmacological effects, including modulation of the activation of various signaling pathways involved in tissue damage. Cisplatin (CP) is a chemotherapeutic agent which, in addition to its desired effect, leads to unwanted side effects, including nephrotoxicity. The aim of this study is to determine the mechanism of action of some common phytochemicals, secoiridoid oleuropein, diterpenoid stevioside glycoside, monoterpenoid carvacrol, and triterpenoid oleanolic acid in a mouse model of toxic kidney injury induced by CP. Male BALB/cN mice were orally administered with several previously picked doses of phytochemicals for 2 days, 48 h after intraperitoneal administration of CP (13 mg/kg). Two days later, CP treatment resulted in histopathological changes showing kidney injury. All tested phytochemicals dosedependently restored changes that indicate the induction of oxidative stress, apoptosis, inflammation or autophagy in damaged kidneys. The results of the current study suggest that phytochemicals can attenuate CP-induced acute renal injury by suppressing those processes that lead to the kidney damage induced by CP through modulation of various signaling pathways. Additionally, we tested the effect of carvacrol and oleanolic acid on human cervical cancer cells, HeLa, and the effect on their cotreatment with CP. Carvacrol induced CP resistance in HeLa cells through suppression of apoptosis and modulation of autophagy. Interestingly, oleanolic acid increased CP cytotoxicity in HeLa cervical cancer cells by inducing cytotoxic autophagy. The chemosensitation of HeLa cells to CP suggests a potential beneficial effect of oleanolic acid in cervical cancer patients due to reduced CP dosage requirements

UVOD: Ulcerozni kolitis (UK) je idiopatska kronična upalna bolest gastrointestinalnog sustava praćena mnogim komplikacijama. Upala zahvaća sluznicu debelog crijeva i to najčešće završni dio debelog crijeva, a etiopatogeneza nije u potpunosti razjašnjena. Istraživanja ukazuju kako upala nastaje u interakciji niza čimbenika uključujući genetsku predispoziciju, imunološki sustav, mikroorganizme i okoliš. U organizmu, morfološki razlikujemo tri vrste stanične smrti, jedna od njih je apoptoza ili programirana stanična smrt. Apoptoza je proces koji sudjeluje u regulaciji održavanja tkivne homeostaze, a njezini poremećaj može dovesti razvoja mnogih oboljenja ukjučujući imunološke i degenerativne te tumora. Pokazano je da stupanj apoptoze, kako upalnih tako i epitelnih stanica, utječe na brzinu razvoja i intenzitet upalnog procesa. Brojna istraživanja ukazuju da hrana bogata klorogenim kiselinama pogoduje ljudskom zdravlju te da imaju antioksidativni i protuupalni učinak. CILJ: Cilj predloženog istraživanja je ispitati izražaj pro- i protuapoptotskih molekula u kolitisu te utjecaj klorogenične kiseline (KGK) na razvoj upale i aktivaciju procesa apoptoze. MATERIJALI I METODE: U sklopu provedbe istraživanja korišteni su C57BL/6 miševi u kojih je primjenom 2,5% otopine natrijevog dekstran sulfata (DSS) kroz sedam dana izazvan pokusni model kolitisa. Procjena uspostave i intenziteta kolitisa praćena je na dnevnoj bazi putem pojavnosti i intenziteta markera upalnog procesa te izračunom indeksom aktivnosti bolesti. Na proteinskoj razini, u tkivu debeloga crijeva, pratila se promjena izražaja pro- i protuapoptotskih molekula. REZULTATI: Tijekom primjene DSS otopine zabilježen je pad tjelesne mase, porast indeksa aktivnosti bolesti te oštećenje sluznice kod svih ispitivanih skupina životinja. Međutim, u skupini miševa kod kojih se je uz DSS primijenila KGK gubitak tjelesne mase značajno je manji, a također i indeks aktivnosti bolesti. U upalno promijenjenom tkivu životinja kod kojih je primjenjena samo DSS otopina, u odnosu na kontrolnu skupinu, pojačan je izražaj Bax, kaspase 9, cijepane kaspaze 8, a smanjen izražaj Bcl-2 i kaspaze 8. Primjena KGK dovodi do obrata izražaj istih, a stupanj promjene ovisan je o primjenjenoj dozi. ZAKLJUČAK: Primjena KGK značajno utječe na intenzitet kolitisa te na izražaj pro- i protuapoptotskih molekula. Stoga, rezultati studije ukazuju da bi se KGK mogla koristiti u terapiji upalom posredovanih bolesti. Ključne riječi: Ulcerozni kolitis, apoptoza, klorogenična kiselina, animalni modeli., INTRODUCTION: Ulcerative colitis (UC) is an idiopathic chronic inflammation of gastrointestinal system, accompanied with wide variety of extraintestinal manifestations. Inflammation affects colon's mucous, most often final portion of the large intestine and etiopathogenesis of inflammation is still not fully understood. There are 3 types of cell death in multicellular organism, one of them is being apoptosis well known as programmed cell death. Apoptosis plays a fundamental role in human development and tissue homeostasis. Aberrant regulation of apoptosis is linked with a various disease, including immunological and developmental disorders, and cancer. According to vast number of studies, food rich with chlorogenic acids favours human health and has antioxidative and anti-inflammatory impact. AIM: The aim of the proposed study was to investigate the expression of pro- and anti-apoptotic molecules in DSS colitis and the influence of chlorogenic acid (CHA) on the development of inflammation and the activation of the apoptosis. MATERIALS AND METHODS: In the current study male C57BL/6 mice were used. Colitis was induced by by administration of 2,5%. DSS in driking water for seven days. Evaluation of colitis development and severity was monitored daily through occurence and intensity of clinical markers and by calculation of disease activity index. Analysis of results established disease activity indeks. At the protein level, changes in the expression of pro- and anti-apoptotic proteins were observed in the colon tissue. RESULTS: During the administration of the DSS solution, a decrease in body weight, and increase in the disease activity index, and mucosal damage in all groups of animals were observed. However, CHA supplementation attenuated weight loss significantly and conequently disese activity index was lower. A increased expression of Bax, caspase 9, cleaved caspase 8 and decreased expression of Bcl-2 and caspase 8 was determined in mice treated only with DSS in comparison to the control group. Treatment with CHA dose-dependently influenced on the expression of proapoptotic and antiapoptotic molecules. CONCLUSION: The appliance of KGK significanlty affects colitis development as well as the expression of pro and antiapoptosis molecules. The results of the study suggest that CHA could be usful for the tretment of inflammation.