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Jedna od alternativa klasičnim, unutarstaničnim, bioprocesima su izvanstanični bioprocesi temeljeni na sojevima kvasca koji izlažu biotehnološki zanimljive enzime na svojoj staničnoj površini. Takvo vezivanje proteina može se, na primjer, postići fuzijom enzima od interesa s proteinom Pir2. Međutim, nije poznato gdje je unutar proteina Pir2 najbolje umetnuti enzim, a da se što bolje sačuva i sposobnost vezanja proteina Pir2 i aktivnost izloženog enzima. U ovom radu konstruiran je plazmid pRSII-pir2bla_CJ koji kodira za fuzijski protein Pir2bla_CJ, spoj proteina Pir2 i enzima β-laktamaze. Nadalje, izmjerena je aktivnost β-laktamaze izložene na staničnoj stijenci kvasca Saccharomyces cerevisiae koji eksprimira ovaj rekombinantni protein i uspoređena s aktivnosti četiri prethodno konstruirane inačice proteina Pir2bla. Iako protein Pir2bla_CJ kombinira odlike dvije prethodno razvijene vrlo efikasne inačice proteina Pir2bla, njegova ekspresija rezultira zanemarivom aktivnošću β-laktamaze, sugerirajući da pravila koja diktiraju efikasno vezanje proteina na staničnu stijenku, uz očuvanje biotehnološki zanimljive enzimske aktivnosti, nisu aditivna. Some classical, intracellular bioprocesses could be replaced with extracellular ones, i.e. bioprocesses based on yeast strains displaying biotechnologically interesting enzymes on their cell surface. We can bind such enzymes to the cell wall, for example, by fusing them with Pir2 proteins. However, it remains unclear where in the Pir2 protein we should insert the enzyme to preserve both the enzymatic activity and the Pir2 binding ability. In this work, we constructed a plasmid encoding protein Pir2bla_CJ, a fusion of the protein Pir2 and the enzyme β-lactamase. Furthermore, we measured the β-lactamase activity of Pir2bla_CJ on the Saccharomyces cerevisiae cell surface and compared it to four previously constructed variations of Pir2bla protein. Although Pir2bla_CJ combined characteristics of two high-performing Pir2bla variations, its β-lactamase activity was negligible, suggesting that the rules allowing for the robust binding of proteins on the cell wall, and preservation of biotechnologically relevant enzymatic activity, are not additive.

After synthesizing proteins, yeast Saccharomyces cerevisiae can retain them within the cell, secrete them into the medium, or bind them to the cell surface. In the third case, yeast anchors proteins by covalently binding them to β-glucan residues, thus allowing for surface display, a cell-engineering technique that can remake the entire cell surface into catalytically active living material. To immobilize proteins of interest closer to their N-terminus, the proteins are regularly fused with one of five Pir proteins (proteins with internal repeats), which covalently bind to β-1, 3-glucan. However, beyond the requirement for glutamine in their characteristic internal repeats, the mechanism of attaching these proteins to the yeast cell wall remains unclear. Thus, the Pir-based surface display continues to rely on guesswork and intuition, which is less than efficient. To address this issue, we inserted β-lactamase at five positions in Pir2 (Hsp150) protein and followed its activity and Pir2-binding efficiency through enzymatic and immunochemical methods. Moreover, we constructed and tested additional six truncated Pir2-β-lactamase variants, thus determining the minimal portion of Pir2 required for efficient binding to the yeast cell wall. Finally, to enrich our experimental results with structural insights, we used a deep- learning Alphafold2-based algorithm to in silico predict the structure of Pir2-β-lactamase fusions. Therefore, we present and rationalize a novel set of practical guidelines for a reproducible, straightforward, and efficient Pir-based yeast surface display.

Sport kao fenomen modernog doba postaje sve masovniji i samim time se povećava njegov utjecaj na svakodnevni život. Dostupnost sporta svima čini ga univerzalnim. Kako sport utječe na živote sve većeg broja ljudi tako raste i njegov kulturni i društveni utjecaj. Velike sportske priredbe poput Olimpijskih igara, svjetskih i europskih prvenstava imaju i značajan utjecaj na ekonomske prilike. Stoga ne čudi da se sport i njegov utjecaj nalazi u središtu istraživanja brojnih znanosti. Također sport se vrlo često povezuje sa religijom. Još od antičkih vremena sportska natjecanja poput Olimpijskih igara imala su religijsku poveznicu jer su održavana u čast bogovima. Moderni sport predstavlja natjecanje sportaša koje naglasak pretežito stavlja na izvedbu i koje u svojem središtu nema religijskih značajki. Ipak, sportaši svoju religioznost pokazuju na brojne načine, a molitva je jedan od najčešćih kao i meditacija i specifični rituali prije natjecanja. U ovom radu navode se objašnjenja pojmova vjere i religije kao i načini njihova mjerenja. Prikazuje se povezanost između religije i sporta te kako religija utječe na ponašanja povezana sa zdravljem. Sport se prikazuje iz perspektive kršćanstva te se opisuje crkveno učenje o sportu i stav pojedinih kršćanskih autoriteta o istome. U posljednjem dijelu rada navodi se kako sportaši iskazuju vlastitu religioznost na terenu i izvan njega te iz kojeg razloga i s kojim ciljem., Sport as a phenomenon of the modern age is becoming more and more widespread and thus its impact on everyday life is increasing. The availability of sports to everyone makes it universal. As sport affects the lives of an increasing number of people, so does its cultural and social impact. Major sporting events such as the Olympic Games, World and European Championships also have a significant economical impact. It is therefore not surprising that sport and its impact are at the center of research of many sciences. Also, sport is very often associated with religion. Since ancient times, sports competitions such as the Olympic Games have had a religious connection because they were held in the honor of the gods. Modern sport is a competition of athletes that focuses mainly on performance and that has no religious features at its core. Still, athletes show their religiosity in a number of ways, and prayer is one of the most common as are meditation and specific pre-competition rituals. This papre provides explanations of the concepts of faith and religion as well as ways to measure them. The connection between religion and sport nad how religion influences health-related behaviours are presented. Sport is presented from the perspective of Christianity and describes the church's teachinh on sport as well as the attitude of individual Christian authorities about it. The last part of the paper states how athletes express their own religiosity on and off the field, for what reason and for what purpose. In this paper, we will analyze the definition of faith and religiosity that will facilitate our further understanding of the connection between religion and sport and their similarities and differences. Given that Christianity and sport nurture similar Christian values such as wisdom, loyalty, courage, equality (...) it can be concluded that they represent what is at the heart of modern sport. We will also analyze Christianity and its impact on behavior and its connection to sport. Furthermore, the influence of the church on sport was contributed by Christian authorities who spoke of sport and physical exercise. Finally, we will show religiosity and the expression of religiosity in athletes.

Cilj istraživanja bilo je utvrditi mogućnost predikcije rezultata u testu na 2000 metara na veslačkom ergometru, kao pokazatelju veslačke uspješnosti, testom vršne izlazne snage. U istraživanju je sudjelovalo 8 veslačica i 12 veslača Veslačkoga kluba Iktus iz Osijeka (AS ± SD: dob 15,6 ± 0,6 god; visina 179,0 ± 10,3 cm; masa 74,0 ± 11,7 kg). Uvjet za sudjelovanje bilo je minimalno 12 mjeseci aktivnoga treniranja veslanja. Svi ispitanici imali su jednaku pripremu za testiranje koja se sastojala od četiriju faza. Prva faza jest pedaliranje na statičnom biciklu, druga dinamičko istezanje, treća aktivacija i četvrta veslanje. Testiranje se sastojalo od 12 zaveslaja na veslačkom ergometru; 6 zaveslaja bilo je uvodnih nakon kojih je slijedilo 6 maksimalnih zaveslaja. Prosjek tih 6 maksimalnih zaveslaja (u W) korišten je kao rezultat u testu vršnog izlaza snage. Svi ispitanici obavili su testiranje na veslačkom ergometru Concept II (Model D). Korišten je Pearsonov koeficijent korelacije za utvrđivanje povezanosti između rezultata u testu na veslačkom ergometru na 2000 metara i rezultata u testu vršne izlazne snage te regresijska analiza za definiranje regresijske jednadžbe tj. predikcijskog modela. Rezultati su pokazali da je regresijski model statistički značajan (p < 0,001), koeficijent multiple korelacije iznosi R = 0,959; a koeficijent determinacije R2 = 0,920. Jednadžba kojom se može prognozirati veslačka uspješnost, odnosno rezultat u testu na 2000 metara temeljem rezultata u testu vršne izlazne snage, glasi: Rezultat na 2000 m = 23,27 + 0,42 x (prosjek 6 maksimalnih zaveslaja). Test vršne izlazne snage jest vremenski i energetski ekonomičniji od testa na 2000 m te se može predložiti kao korisna metoda za provjeru treniranosti veslačica i veslača mlađih dobnih kategorija. The aim of the study was to determine the possibility of predicting the results in the test at 2000 meters on a rowing ergometer, as an indicator of rowing success, with the test of peak power output. The study involved 8 female rowers and 12 male rowers of the Iktus Rowing Club aged 14 to 16, average age 15.6 +/- 0.6, height 179 +/- 10.3 cm and weight 73.95 +/- 11.7 kg. The condition for participation was a minimum of 12 months of active rowing training. All subjects had the same preparation which consisted of 4 phases. The first phase was pedaling on a static bicycle, the second phase dynamic stretching, the third phase activation and the fourth phase rowing. Testing consisted of 12 strokes, 6 strokes were introductory, followed by 6 strokes performed maximally strongly on a rowing ergometer. The average of these 6 maximum strokes (in W) was used as a result in the peak power output test. All subjects performed the test on a rowing ergometer Concept II (Model D). Pearson's correlation coefficient was used to determine the relationship between the results in the test on a rowing ergometer at 2000 meters and the results in the test of peak power output and regression analysis to define a regression equation i.e. a prediction model. The results showed that the regression model was statistically significant (p