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Rak glave in vratu predstavlja heterogeno skupino malignih tumorjev. Mednje uvrščamo malignome nosu, obnosnih votlin, ustne votline, žrela, grla in žlez slinavk. Ti tumorji se med seboj razlikujejo ne le po mestu, od koder izvirajo, temveč tudi po bolezenskih znakih, histološki sliki, načinu in pogostosti metastaziranja, hitrosti napredovanja ter ne nazadanje tudi po načinu zdravljenja in izidu bolezni. Resno ogrožajo bolnikovo življenje, hkrati pa prizadenejo del telesa z izredno pomembnimi fiziološkimi funkcijami, ki so bistvenega pomena v človekovem psihičnem, emocionalnem in socialnem življenju. V nastanek raka sta vpleteni predvsem dve skupini genov, in sicer protoonkogeni in tumor supresorski geni. Prvi so med procesom karcenogeneze spodbujeni k čim večjemu izražanju, medtem ko so drugi spremenjeni tako, da je njihovo izražanje zmanjšano ali da je njihov produkt funkcionalno neaktiven. Na sam proces karcenogeneze vplivajo tudi drugi dejavniki, ki ne povzročajo sprememb v nukleotidnem zaporedju DNA in jih imenujemo epigenetski dejavniki. Epigenetski dejavniki se izražajo predvsem tako, da celico vzpodbujajo in jo stalno silijo v izražanje določenih genov. Med karcenogenezo se spremeni metilacija protoonkogenov, tumor supresorskih genov ter genov, ki so odgovorni za popravljalne mehanizme DNA v celici. Hipermetilirajo in s tem se utišajo tumor supresorski geni ter geni, odgovorni za popravljanje nastalih napak na DNA. Hipometilirajo in s tem se aktivirajo protoonkogeni in geni, odgovorni za tumorsko metastaziranje. Namen magistrske naloge je analiza mutacij in metilacije tumor supresorskih genov pri raku glave in vratu. Določali smo tudi prisotnost humanih papiloma virusov (HPV) v tumorskih vzorcih, saj je predvideno, da je status metilacije povezan s HPV povezanimi tumorji raka glave in vratu. Uporabili smo referenčne vzorce zdravega tkiva in testne vzorce tumorskega tkiva. V študijo je bilo vključenih 59 bolnikov, starih med 46 in 80 let. Okužbo s HPV smo odkrili pri petih bolnikih. Mutacije v tumor supresorskem genu predstavljajo povečano verjetnost za nastanek raka v ORL področju. Za testiranje prisotnosti mutacij v tumor supresorskem genu smo uporabili metodo metilacije specifičnega hkratnega pomnoževanja od ligacije odvisnih prob, MS – MLPA. Pri 49 bolnikih je bila ugotovoljena prisotnost metilacije v več kot enem genu. Najpogosteje hipermetilirani geni so RARB, TIMP3, DAPK1 in CDH13, in jih lahko povežemo z razvojem raka glave in vratu. Pravočasno odkrivanje mutacij v genih, povezanih z nastankom raka, je za nosilce mutacij pomembno, saj je dokazana mutacija razlog za prilagojeno klinično spremljanje in preventivne ukrepe pri nosilcih mutacije. Cancer of the head and neck is characterized by a heterogeneous group of malignant tumours. These include malignancies of the nose, the paranasal sinuses, the oral cavity, the pharynx, the larynx and the salivary glands. The tumors can be differentiated not only by place of origin, but also by their symptoms, histological slides, method and frequency of metastasis, progression rate, method of treatment and the outcome of the disease. These tumors pose a serious threat to the patient's life, while affecting a part of the body that controls important physiological functions, which are an essential part of our psychological, emotional and social well-being. Two groups of genes have been shown to be involved in the development of cancer: proto-oncogenes and tumor suppressor genes. During carcinogenesis, the first type of gene increases activity, while the second type is modified to decrease activity so that its product is functionally inactive. The process of carcinogenesis is also impacted by other factors that do not cause changes within the nucleotide sequence of DNA these compounds are called epigenetic factors. Epigenetic factors assert themselves by stimulating cells and forcing them to express certain genes. During carcinogenesis, the methylation of proto-octogens, tumor suppressor genes and the genes responsible for DNA reparative mechanisms in cells, is changed. This leads to hypermethylation, which in turn suppresses the tumor suppressor genes and the genes responsible for correcting any DNA errors. It also leads to hypomethylation and the activation of proto-oncogenes and the genes responsible for tumor metastasis. The purpose of this master's thesis is to analyse the mutations and methylation of tumor suppressor genes in cancer of the head and neck. We also determined the presence of human papilloma virus (HPV) in tumor samples, since a connection between the status of methylation and HPV-associated head and neck tumors has been predicted. We used reference samples of healthy tissue and test samples of tumor tissue. The study included 59 patients aged between 46 and 80. We discovered five instances of HPV infection in our samples. Mutations in the tumor suppressor gene suggest a greater likelihood of developing cancer in the ORL region. We used the method of methylation-specific multiplex ligation-dependent probe amplification, MS – MLPA, to test for the presence of mutations in the tumor suppressor genes. In 49 cases we found methylation in more than one gene. The most common genes to be hypermethylated are RARB, TIMP3, DAPK1 and CDH13, and these can be connected to the development of head and neck cancer. Discovering gene mutations is important for the carriers of mutations, since this provides sufficient grounds for personalized clinical monitoring and other preventive measures.

Cilj diplomskega dela je izdelava analize trenutne opremljenosti slovenskih predorov na avtocestnem križu in podajanje predlogov za izboljšavo trenutnega stanja. Vožnji skozi predore se posveča dodatna pozornost z varnostnega vidika, zaradi občutka utesnjenosti, ki ga na odprti trasi ni. Slovenski avtocestni križ predstavlja križišče evropskih transportnih koridorjev in je kot tak pomemben del pan-evropskega transportnega omrežja. Leta 2004 je v veljavo stopila Direktiva evropskega parlamenta in sveta 2004/54/ES, ki ureja minimalne varnostne zahteve za predore znotraj članic EU. Opremljenost dotičnih predorov v Republiki Sloveniji se je po uvedbi direktive izkazala za dobro. Kljub temu pa so razlike med tipi opreme in prostorsko umestitvijo opreme v območju predorov in v samih predorih velike in nedopustne za vzpostavitev željenega višjega nivoja varnosti. Najvplivnejši dejavnik v prometnih nesrečah je seveda človek, poleg prometnega okolja in motornega vozila pa med dejavnike, ki vplivajo na prometne nesreče, zagotovo sodi tudi cestna infrastruktura. Za zagotovitev večje varnosti pri vožnji skozi predore je nujno potrebno poenotenje opreme v območju predorov, ki se uporablja za upravljanje in vodenje prometa, pri običajnem delovanju predora in ob izrednih dogodkih. Dogaja se namreč, da se v različnih predorih uporabljajo različni tipi opreme, njihova prostorska umestitev pa ni pogojena z zakonodajo in je zato prepuščena projektantom in izvajalcem del. Prvi korak v pravo smer bi bil vzpostavitev smernic, ki bi do potankosti urejala področje opremljenosti predorov. Na nacionalnem nivoju bi morale biti urejene tehnične specifikacije in navodila za opremo predorov, s katerimi bi uskladili tudi najmanjše, a še zdaleč ne najmanj pomembne razlike med predori. The goal of this thesis is the analysis of current state of tunnel equipment in Slovenian road tunnels and suggestions for its improvements. From safety point of view an extra thoughtfulness needs to be applied to the matter of driving through road tunnels because of the feeling of constriction while driving through them. Slovenian highway cross consists of two main European transport corridors and represents an important part of Pan-european transport network. That is why European Directive for minimal safety requirements for road tunnels 2004/54/ES came into effect in 2004. Although the status of current tunnel equipment in Slovenian road tunnels is acceptable, it differs among them. For reaching a higher goal in maintaining a satisfactory level of safety in road tunnels, it is necessary to eliminate the differences between types of equipment and its positioning on site. Taken into consideration that human behaviour plays the biggest role in road tunnel safety we also need to consider the effects of type of the vehicle, traffic surroundings and definitely of road infrastructure for safety in road tunnels. Only with unification of different types of equipment and its positioning in tunnels will we achieve the proper safety level in our road tunnels. Often the installation of road tunnel equipment and its positioning is left to the preference of the design engineers and contractors, because there are no guidelines to follow. The first step towards achieving a total unification on this field would be the establishment of technical specifications for traffic management equipment in road tunnels on a national scale.

V težkih časih kot so sedaj je ključnega pomena za gospodarstvo, da se podjetja posvetijo inoviranju. Številni posamezniki generirajo raznorazne inovativne predloge, vendar zaradi nezmožnosti podpornih institucij, da tem podjetnikom nudi celostno podporo ideje ne preidejo iz invencije v inovacijo. Pomislite koliko koristi bi imeli vsi udeleženci v tem procesu, če bi le znali podpirati dobre inovativne ideje posameznikov. Gre se predvsem za finančne težave, saj denarja ki je namenjen raziskovalno-razvojnim dejavnostim ne uporabljamo na pravi način. Kar hočemo povedati je to, da je težava v alokaciji sredstev. Seveda so tudi številne druge težave, ki preprečujejo podjetjem, da bi se razvijala tako v začetni fazi svojega poslovanja kot tudi v končnih fazah. Podpornih institucij je v Sloveniji zelo veliko. Mogoče je tudi to težava, saj bi bilo bolj smiselno, da jih je manj, vendar da so takšne ki nudijo celostno podporo inoviranju podjetjem. Po statističnih podatkih se je v obdobju od 2008 do 2010 delež inovacijsko aktivnih podjetij v Sloveniji zmanjšal glede na obdobje 2006-2008. Kar predstavlja še dodatno težavo je to, da se je v večini EU držav zgodilo ravno nasprotno. Delež inovacijsko aktivnih podjetij se je v obdobju 2008-2010 povečal glede na obdobje 2006-2008. V diplomskem seminarju smo tudi ugotovili, da največ podpore inoviranju nudijo tehnološki parki. Do te odločitve so nas privedli številni dejavniki. Možno je seveda, da bi po drugih dejavnikih katere druge institucije nudile več podpore inoviranju, vendar po naših kriterijih in analizi njihovih dejavnosti smo se odločili da tehnološki parki nudijo največ podpore inoviranju. Pri drugih institucijah je enostavno bilo preveč nepopolnosti pri izpolnjevanju naših kriterijev in tako se nismo odločili za njih. V prihodnjih letih pričakujemo od podpornih institucij predvsem večjo vključitev v celoten proces pomoči podjetjem, ki bi rade vstopile na trg z inovativnim izdelkom/storitvijo. Pričakujemo tudi boljšo porabo sredstev, saj iz leta v leto za raziskovalno-razvojne dejavnosti porabimo več sredstev, kot smo pa prej ugotovili se je stopnja inovacijsko dejavnih podjetij zmanjševala v Sloveniji. Podjetja, podporne institucije in država bodo morali narediti nekaj veliki potez, saj nam bo to pomagalo na dolgi rok, poleg tega pa bomo vsi imeli koristi zaradi tega. In difficult times such as now it is vital for the economy that companies devote to innovation. Many individuals generate all kinds of innovative proposals, but due to the inability of the support institutions, that should offer full support to the entrepreneurs, ideas don’t make a transition from invention to innovation. Think of how much benefit would have all participants have in this process, if only we knew how to support good innovative ideas of individuals. It is primarily for financial problems, because of money that is intended for research and development activities are not used in the right way. What we want to say is that the problem is in the allocation of resources. Of course, there are also many other problems that prevent enterprises to develop both in the initial stages of their business, as well in the final stages. There are a lot of support institutions in Slovenia. Maybe that is also a problem, as it would make more sense to have that fewer, but such that offer comprehensive support for the innovative companies. According to statistics, in the period of 2008 to 2010, the share of innovation active enterprises in Slovenia decreased when comparing it to the period of 2006-2008. What constitutes a further problem is that in most EU countries opposite happened. The share of innovation active enterprises in the period of 2008-2010 increased, comparing it to the period of 2006-2008. In the diploma seminar, we also found that the most support to the innovation is provided by the technology parks. To these decisions we were led by a number of factors. It is possible, of course, that if we compared support institutions by other factors, we would come to the decision that they offered more support to the innovation, but according to our criteria and analysis of their activities we decided that technology parks offer the most support to the innovation. Other institutions that we looked into had just to many imperfections at meeting our criteria, and so we didn’t decided for them. In the years to come we expect for the support institutions in particular, greater involvement in the whole process of assistance to the companies wishing to enter the market with an innovative product/service. We also expect a better use of resources, since every year we spend more and more money for the field of research and development however the rate of innovation active businesses declined in Slovenia. Companies, support institutions and the government will have to make some big moves, because it will help us in the long run, in addition we will all benefit from it.

Osnovna dejavnost podjetja TDR Ferolegure d.o.o. je proizvodnja in prodaja legur, med katere spadajo nodulatorji, cepiva in polnjene žice. V nalogi bomo izpostavili nodulatorje z besedo ferosiliko magnezij, FeSiMg. Nodulatorji so po sestavi visokokakovostne kompleksne zlitine, katerih osnovna naloga je vnos magnezija v sivo litino. FeSiMg delajo iz osnovne zlitine FeSi, ki mu dodajajo določen delež magnezija, ki je glavni element v zlitinah. Zlitine FeSiMg vsebujejo 5 do 35 % magnezija. Poleg magnezija zlitina vsebuje še naslednje osnovne elemente: Si, Al, Ca, Fe in redke zemlje, predvsem Ce, La, Pr, Nd in druge lantanide – pod skupnim imenom ''Mischmetall'' (MM). Namen diplomske naloge je certificiranje vrednosti masnega deleža magnezija v FeSiMg 25 – 5 – 1. Število 25 pomeni, da je masni delež magnezija v zlitini 25%, kar prišteva ta nodulator k skupini predlegur tipa FeSiMg z visoko vsebnostjo magnezija (5 za 5% Ca, 1 za 1% CeMM). Za zagotavljanje zanesljive vsebnosti magnezija je potreben ustrezen analizni postopek, pri katerem sta pomembna parametra ponovljivost postopka in pravilnost rezultatov. Zaradi specifičnih karakteristik magnezija se v nalogi osredotočamo na določanje magnezija v vzorcu z optično emisijsko spektrometrijo z induktivno sklopljeno plazmo, ICP–OES. Za primerjavo rezultatov smo vzorce vzporedno analizirali z že uveljavljeno metodo kot referenčno metodo, s kompleksometrično titracijo, ki pa je v nalogi ne bomo statistično obravnavali. Študirali smo tudi vpliv načina vzorčenja na analizo zlitine in s tem povezano homogenost vzorca. Obdelali smo pripravo vzorcev za analizo in ugotovili potrebne eksperimentalne pogoje za magnezij. V zlitini ima vsak element svojo določeno vlogo. Zato smo obdelali medelementne vplive, ki smo jih primerjali na podlagi rezultatov, dobljenih z analizo multielementnih vzorcev in vzorcev, pripravljenih s postopkom vakuumske filtracije, da smo izločili druge osnovne elemente kot možne interference. Kakovost FeSiMg je glede zrnatosti in kemijske sestave območij vsebnosti Si, Al, Ca, Fe, Mg in redkih zemelj prilagojena zahtevam kupca, zato mora biti v ozkem koncentracijskem območju zanesljivo nadzorovana. The main activity of the company TDR Ferolegure d.o.o., is the production and sale of alloys, including nodularises, inoculants and cored wires. This thesis refers to nodularises with the term Ferrosilicon Magnesium (FeSiMg). Nodularises are structurally high quality complex alloys mainly used for magnesium addition into grey cast iron. FeSiMg is produced from the basic alloy FeSi by adding a certain amount of magnesium as the alloy’s main element. FeSiMg alloys contain between 5 – 35% of magnesium. In addition to magnesium, the alloy contains the following elements: Si, Al, Ca, Fe and rare earths, especially Ce, La, Pr, Nd and others lanthanides – under the common name ‘’Mischmetall’’ (MM). Purpose of this thesis is the certification of the mass fraction of magnesium in FeSiMg 25 – 5 – 1. The number 25 indicates that the mass fraction of magnesium in the alloy is 25%, this classifies this nodulariser as a magnesium-rich master alloy (5% Ca, 1% CeMM). To ensure the required amount of magnesium we need an appropriate analytical procedure, with a focus on repeatability and accuracy. Due to the specific characteristics of magnesium the thesis focuses on two analytical methods for determination of magnesium content, which are also compared. A study of how the sampling method affects the analysis of the alloy and the related sample homogeneity is included. We did a sample preparation and determined the required experimental conditions for magnesium. Within the alloy each element has a certain role. Therefore, we studied the inter - elemental effects, and compared them based on the results from the analysis of multi - elemental samples and samples prepared through vacuum filtration to eliminate other elements as possible interferences. The samples are analyzed with inductively coupled plasma optic emission spectrometry (ICP – OES). We determine the mass fraction of calcium and magnesium in the filtered samples for the determination of magnesium through complexometric titration. The quality of FeSiMg is regarding the granularity and chemical composition of content ranges of Si, Al, Ca, Fe, Mg and lanthanides determi adjusted to the client’s needs, therefore we need to ensure reliable control within a narrow concentration range.