Revealing fungal activity in the presence of ionic liquids

Dissertation presented to obtain the Ph.D degree in BiochemistryIonic liquids constitute a vast and heterogeneous group of chemicals, generally non-volatile and of high solvent quality. They are already used in industrial processes; future applications depend heavily on conscious design of ionic liquids. Given especially the global demand for sustainable chemicals, understanding environmental risks is a priority, necessitating a multidisciplinary research approach, covering a broad range of disciplines from biology to chemistry. Ascomycota fungi are highly suitable model organisms, especially due to their environmental ubiquity and important role in the biotic decay of pollutants. This thesis reports the first ever use of Ascomycota fungi to investigate ionic liquids ecotoxicity and environmental persistence. Fungal strains of Penicillium and Aspergillus were in general found to be more tolerant to ionic liquids containing imidazolium, pyridinium, pyrrolidinium, cholinium or phosphonium cations, than any other microorganism tested to date (Chapters II, III and IV). The capacity of the strains to tolerate the ionic liquids tested was apparently correlated to their phylogeny. Ionic liquid toxicity was evaluated using common parameters, such as growth inhibition and death. Less frequently evaluated parameters were also analysed, including monitoring of the integrity of the cellular boundaries of fungal conidia by microscopy (Chapter IV) and determining the diffusible fungal metabolome by ESI-MS and LC (Chapter II and V). Overall, these data significantly contribute to current understanding of structure-activity relationships in ionic liquids. For example toxicity is apparently a function of alkyl chain length of both anion and cation (Chapters III and IV, respectively). A critical review of current understanding of toxicity and environmental impact of the principal ionic liquid groups made it clear that the common generalisation of ionic liquids being either “green” or “toxic”(...)Financial support provided by Fundação para a Ciência e Tecnologia (fellowship BD 31451/2006). The work was partially supported by a grant from Iceland, Liechtenstein and Norway through the EEA financial mechanism (Project PT015)

Izraženost receptora estrogena i progesterona u ljudskom inače nespecificiranom duktalnom invazivnom karcinomu dojke: postoji li razlika između žena u predmenopauzi i postmenopauzi ?

Determination of hormone receptors is of utmost importance in planning therapy in patients with breast cancer. The aim of the study was to assess the expression of estrogen (ER) and progesterone (PR) receptors in ductal invasive breast carcinoma not otherwise specified (NOS) according to patient menopausal state and tumor histopathology. The study included 549 patients treated at University Department of Surgery, Rijeka University Hospital Center, between January 1, 2000 and January 1, 2005. The patients were diagnosed with breast cancer and underwent mastectomy. ER and PR status was determined by immunohistochemistry. Study results showed no statistically significant differences in the expression of ER and PR, tumor size and grade of histologic differentiation between premenopausal and postmenopausal women. However, tumor size and grade of histologic differentiation differed significantly according to the expression of hormone receptors. Tumors greater than 5 cm in size were mostly ER- in premenopausal (P=0.012) and PRin postmenopausal (P=0.044) patients. Poorly differentiated cancers were associated with ER-PRstatus in both premenopausal and postmenopausal patients (P<0.001). Hormone dependent tumors (ER+PR+) were of smaller diameter and lower histologic grade, while hormone independent tumors (ER-PR-) had greater diameter and higher histologic grade, the difference being statistically significant (P=0.004 and P<0.001, respectively). Study results on the characteristics of ductal invasive carcinoma according to hormone status were consistent with those described in the literature. Considering controversies about the role of steroid receptors in endocrine therapy response, our future objective is assessment of the 5-year prognosis in our patients.Određivanje hormonskih receptora iznimno je važno u planiranju liječenja kod bolesnica s rakom dojke. Cilj ove studije bio je procijeniti izraženost receptora estrogena (ER) i progesterona (PR) u inače nespecificiranom duktalnom invazivnom karcinomu dojke (NOS) u odnosu na menopauzalni status žene i histopatologiju tumora. U studiju je bilo uključeno 549 žena liječenih na Klinici za kirurgiju Kliničkog bolničkog centra Rijeka između 1. siječnja 2000. i 1. siječnja 2005. godine. Kod bolesnica je bio dijagnosticiran rak dojke i one su podvrgnute mastektomiji. Status ER i PR određen je imunohistokemijskim metodama. Rezultati nisu pokazali nikakvih značajnih razlika u izraženosti ER i PR, veličini tumora i stupnju histološke diferencijacije između žena u predmenopauzi i onih u postmenopauzi. Međutim, veličina tumora i stupanj histološke diferencijacije razlikovali su se značajno u odnosu na izraženost hormonskih receptora. Tumori veći od 5 cm uglavnom su bili ER- kod žena u predmenopauzi (P=0,012) i PR- kod onih u postmenopauzi (P=0,044). Slabo diferencirani karcinomi bili su udruženi sa statusom ER-PR- kod bolesnica u predmenopauzi kao i kod onih u postmenopauzi (P<0,001). Hormonski ovisni tumori (ER+PR+) bili su manjeg promjera i nižeg histološkog stupnja, dok su hormonski neovisni tumori (ER-PR-) imali veći promjer i viši histološki stupanj, i ta je razlika bila statistički značajna (P=0,004 odnosno P<0,001). Rezultati dobiveni za značajke duktalnog invazivnog karcinoma prema hormonskom statusu bili su sukladni onima u literaturi. S obzirom na proturječja o ulozi steroidnih receptora u endokrinom odgovoru na terapiju naš je budući cilj procijeniti 5-godišnju prognozu kod naših bolesnica

Plasma electrolytic oxidation of valve metals

Ova doktorska disertacija bavi se proučavanjem fenomena Plazma Elektrolitičke Oksidacije (PEO) ventilnih metala (Al, Mg, Ti, Та) i oksidnih slojeva koji se dobijaju u elektrolitima koji sadrže volfram, pre svega u 12-volframsilicijumskoj kiselini. Primenom metoda optičke emisione spektroskopije i analize procesa u realnom vremenu, omogućeno je određivanje: temperature, elektronske gustine plazme, dimenzionalne distribucije i aktivne površine mikro pražnjenja. Pokazano je da PEO procesi na svim ispitivanim metalima pokazuju tendenciju da se broj mikro pražnjenja smanjuje sa vremenom, a da površine poprečnih preseka mikro pražnjenja rastu. Sa porastom vremena PEO procesa broj kanala za pražnjenje se smanjuje, dok njihov dijametar raste i dolazi do povećanja hrapavosti oksidnih površina. Procenat aktivne površine istovremeno izložen događajima mikro pražnjenja takođe raste u početku PEO procesa, dostiže svoj maksimum, a potom polako opada. Sve spektralne linije detektovane u optičko emisionim spektrima u toku PEO procesa pripadaju ventilnim metalima i jonskim i atomskim elementima prisutnim u elektrolitu. Pored jasno izraženih atomskih i jonskih linija, u optičko emisionom spektru dobijenom tokom PEO procesa na magnezijumu, jasno je vidljiva i vibraciona traka MgO molekula, koja odgovara elektronskom prelazu sa pobuđenog B1Σ+ nivoa na osnovni elektronski nivo X1Σ+. Na sličan način, u spektru dobijenom tokom PEO procesa na aluminijumu opaža se jaka vibraciona traka AlO sistema, koja odgovara elektronskom prelazu sa pobuđenog B2Σ+ nivoa na osnovni elektronski nivo X2Σ+. U slučaju PEO procesa na aluminijumu i magnezijumu, koristeći spektroskopske podatke dobijene za AlO, OH i MgO trake, uz primenu kvantno-mehaničkih proračuna, određene su temperature plazme. Iz vibracionih traka MgO molekula procenjeno je da su temperature plazme (11000 ± 2000) K, dok su u slučaju podataka iz OH trake dobijene nešto niže vrednosti temperature (3500 ± 500) K. Iz analiza vibracione trake AlO molekula u slučaju PEO procesa u limunskoj kiselini procenjena temperatura je (8000 ± 2000) K...This doctoral dissertation is focused on investigation of a relatively new, highvoltage anodization process, called Plasma Electrolytic Oxidation (PEO) on valve metals (such as Al, Mg, Ti and Ta) and their oxide coatings obtained in tungsten containing heteropoly acids. Very useful data concerning temperature, dimensional distribution of the microdischarges, their spatial density, and electron density were obtained using optical emission spectroscopy and real time imaging of PEO events. It is shown that the size of microdischarges becomes larger while their number decreases with increased time of PEO. Spatial density of microdischarges is the highest in the early stage of the PEO process and then slowly decreases. The percentage of oxide coatings’ area simultaneously covered by active discharge sites decreases slowly with extended PEO time. All atomic and ionic lines detected in optical emission spectra originate either from electrolyte or from the metal substrate. During the PEO process on magnesium, wide transition band was also recorded corresponding to the v′−v″=0 band sequence of the B1Σ+ – X1Σ+ emission transition of MgO. During the PEO process on aluminum, visible band of B2Σ+ - X2Σ+ emission of AlO system was also detected. By applying spectroscopic, quantum-chemical and thermodynamical methods to analyze spectra in the range of those bands it was possible to determine the temperature of the microdischarges. In the case of PEO process on magnesium, calculated temperatures are about 11000 K ± 2000 K, while analysis of OH bands in the same sysstem yields sligtly lower value of 3500 K ± 500 K. Analysis of AlO band system gave temperature of 8000 K ± 2000 K. The analysis of hydrogen Balmer Hβ line shape in the case of PEO of tantalum and titanium, indicates the presence of two types of discharges during PEO process and the electron number densities of Ne ≈ 0.8×1015 cm−3 and Ne ≈ 2.1×1016 cm−3 are determined. These two electronic concentrations are similar to those obtained during the PEO process on aluminum..

Seed quantity and quality in fruit heads of Aster lanceolatus Willd.: Implications for invasion success

Aster lanceolatus Willd. is a herbaceous perennial that is considered invasive in many European countries. In Serbia, this plant inhabits wet habitats and forms widespread monospecific stands. The objective of this research is to determine whether generative reproduction has an important role in the expansion of this species to new areas. In 13 different localities, fruit heads were collected from lateral and terminal parts of infructescence. Seed quantity and germination parameters were determined for seeds in the fruit heads. The results showed that the position of the fruit heads did not have a major impact on germination parameters. However, germination parameters differed among the localities. The findings of this study suggest that A. lanceolatus produces a great amount of viable seeds that germinate in an amount sufficient to ensure a successful spread of this invasive species to new areas. [Projekat Ministarstva nauke Republike Srbije, br. 43007: Studying climate change and its influence on the environment: impacts, adaptation and mitigation

Cytogenetic Follow-Up in Testicular Seminoma Patients Exposed to Adjuvant Radiotherapy

Early stage testicular seminoma is a radiosensitive tumor. Its incidence has significantly increased during the last decade especially in the young population. Although the therapy for testicular seminoma gives very satisfying results, the evaluation of genome damage caused by the therapy is of a great importance in order to recognize possible related health risks. The present study was performed on ten patients diagnosed with seminoma stage I; pT1/2N0M0S0, treated with adjuvant radiotherapy (a radiation dose of 25 Gy divided in 16 fractions) after orchidectomy. To assess the possible existence of an increased baseline DNA/chromosome damage in patients we also selected the appropriate control group of ten healthy men. The levels of primary DNA/chromosome damage in peripheral blood lymphocytes, as well as the dynamics of their repair were studied using the alkaline comet assay, chromosome aberration and cytokinesis-block micronucleus assay. Altogether four blood samples per patient were collected in the course of the therapy: before and after receiving the first dose of radiotherapy, in the middle of the radiotherapy cycle, and after the last dose of radiotherapy. Other two follow-up blood samples were collected six and twelve months after the cessation of therapy. As observed, the administration of the first radiation dose significantly increased the levels of DNA damage in almost all patients compared to their baseline values. Specific patterns of DNA damage were recorded in samples analyzed in the middle of radiotherapy and after receiving the last dose, indicating the possibility of an adaptive response in some patients. The levels of chromosomal aberrations and the incidence of micronuclei also increased in the course of therapy but gradually declined during the follow-up period. Our results confirmed the existence of post-irradiation damage in peripheral blood lymphocytes (and possibly in other non-target cells) of cancer patients which may represent a risk for the secondary cancer development. Considering that the majority of patients with testicular cancer are of a younger age, they represent a population deserving special attention. As cytogenetic screening may detect high-risk individuals, it might be useful in regular medical monitoring of seminoma patients after the successful therapy

Cytogenetic Follow-Up in Testicular Seminoma Patients Exposed to Adjuvant Radiotherapy

Early stage testicular seminoma is a radiosensitive tumor. Its incidence has significantly increased during the last decade especially in the young population. Although the therapy for testicular seminoma gives very satisfying results, the evaluation of genome damage caused by the therapy is of a great importance in order to recognize possible related health risks. The present study was performed on ten patients diagnosed with seminoma stage I; pT1/2N0M0S0, treated with adjuvant radiotherapy (a radiation dose of 25 Gy divided in 16 fractions) after orchidectomy. To assess the possible existence of an increased baseline DNA/chromosome damage in patients we also selected the appropriate control group of ten healthy men. The levels of primary DNA/chromosome damage in peripheral blood lymphocytes, as well as the dynamics of their repair were studied using the alkaline comet assay, chromosome aberration and cytokinesis-block micronucleus assay. Altogether four blood samples per patient were collected in the course of the therapy: before and after receiving the first dose of radiotherapy, in the middle of the radiotherapy cycle, and after the last dose of radiotherapy. Other two follow-up blood samples were collected six and twelve months after the cessation of therapy. As observed, the administration of the first radiation dose significantly increased the levels of DNA damage in almost all patients compared to their baseline values. Specific patterns of DNA damage were recorded in samples analyzed in the middle of radiotherapy and after receiving the last dose, indicating the possibility of an adaptive response in some patients. The levels of chromosomal aberrations and the incidence of micronuclei also increased in the course of therapy but gradually declined during the follow-up period. Our results confirmed the existence of post-irradiation damage in peripheral blood lymphocytes (and possibly in other non-target cells) of cancer patients which may represent a risk for the secondary cancer development. Considering that the majority of patients with testicular cancer are of a younger age, they represent a population deserving special attention. As cytogenetic screening may detect high-risk individuals, it might be useful in regular medical monitoring of seminoma patients after the successful therapy

Begov Most – Staničenje, zaštitna arheološka istraživanja u 2015. godini na trasi autoputa E80, Koridor 10 – istočni krak

U junu i julu 2015. godine nastavljena su aštitna istraživanja Arheološkog instituta na lo kalitetu Begov ost – taničenje u oni ekspropria ije na trasi i gradnje autoputa 0 oridor 10 – istočni krak. aštitna arheološka iskopavanja apočeta su 201 . godine kada je istražena površina od 1 2 sonde 1 1 višeslojnog arheološkog nala išta Begov ost etković et al. u šta pi . ada je otkriven deo kasnoantičke hrišćanske nekropole i ver ena . veka – istraženo je 5 gro ova kao i kulturni slojevi i akarnog do a eneolita sa o jekti a i nogo rojni nala i a kulturne grupe Bu anj alkua rivodol. a iskopavanja su ila prekinuta na ahtev investitora oridora r ije d.o.o. Beograd. eđuti s o iro da je projekto trase autoputa 0 planirano da čitav deo nala išta u oni ekspropria ije ude negiran e ljani radovi a pri i gradnji sao raćajni e nadležni avod a aštitu spo enika kulture u Nišu doneo je rešenje o tehnički uslovi a aštite ovog lokaliteta koji uključuju i potpuno istraživanje ugroženog dela nala išta

Nekropola Slog – Timacum Minus, zaštitna iskopavanja 2014. godine

Zaštitna iskopavanja nekropole Slog u Ravni kod Knjaževca u okviru projekta “Arheološka istraživanja, prezentacija i promocija rimskog utvrđenja i grada Timacum Minus u Ravni kod Knjaževca”, koji realizuju Arheološki institut u Beogradu i Zavičajni muzej u Knjaževcu, započeta su 2013. godine u cilju očuvanja podataka i nalaza sa ove višeslojne nekropole iz kasnoantičkog i ranosrednjovekovnog perioda. Nekropola je istraživana u okviru zaštitnih iskopavanja prilikom rekonstrukcije puta Ravna - Debelica 1994-1996. godine, a rezultati su publikovani u monografiji koju je objavio Arheološki institut (Petković et al. 2005). Godine 2012, ekipa Arheološkog instituta koja je radila na obradi i analizi arheološke građe sa iskopavanja lokaliteta Timacum Minus 1975-1998. godine, radi pripreme monografije, rekognosciranjem je utvrdila da je nekropola Slog ugrožena vertikalnim usecanjem puta Ravna – Debelica u istoimeno brdo, odnosno da je zapadna padina tog uzvišenja izložena eroziji. Na taj način, više grobova je otkriveno prilikom odrona zemlje, što je omogućilo nelegalnim kopačima da pljačkanjem nalaza uz pomoć metal-detektora dodatno devastiraju nekropolu Slog. Tada je bio istražen jedan takav grob (G. 141), već opljačkan i sa dislociranim skeletom, a u njemu je nađena srebrna oplata pojasne garniture i bronzani novac Konstantina, koje su pljačkaši prevideli. Naredne, 2013. godine Arheološki institut je započeo zaštitna istraživanja najugroženijeg dela nekropole, zapadno od rimskih zidanih grobnica. U ovoj kampanji, koja je trajala tri nedelje, otvorena je sonda većih dimenzija, u kojoj je otkriveno 17, a istraženo 15 grobova kasnoantičke i ranosrednjovekovne nekropole (Петковић, Радиновић, Илијић 2014; Petković, Ilijić, Radinović 2014)

Prognostic Values of Morphological and Clinical Parameters in pT2 – pT3 Prostate Cancer in Elderly People

Prostate cancer is a disease of elderly men, hthe incidence of whic increases in an age dependent manner. This study presents the correlation of clinical and morphological parameters in locally confined (pT2) and locally advanced (pT3) prostate cancer. We analyzed a group of elderly men treated with radical prostatectomy in the period 1999–2008 in the University Hospital Rijeka. We found no statistical association between pT stage and age categories, preoperative prostate-specific antigen, digitorectal examination and biopsy Gleason score. There was a significant correlation of higher Gleason score in prostate specimens after radical prostatectomy and a higher frequency of a positive surgical margin in tumors with pT3 than in pT2 stage (p=0.003; p=0.011 respectively). Recurrence-free survival was shorter in patients with tumors with positive surgical margins as well as in patients with pT3 stage (p=0.030; p=0.001 respectively). We conclude that higher tumor grade and positive surgical margins are indicators of a worse prognosis in our patients