Testing selected substances for inhibition of modified 6-phosphofructo-1-kinase in metastatic cancer cells

Encim 6-fosfofrukto-1-kinaza je eden od najpomembnejših regulatornih encimov glikolize. Pri ljudeh najdemo 3 tipe encima, ki so različno izraženi v posameznih tkivih. Za številne tipe raka je značilna, povečana ekspresija izoforme Pfk-L, ki smo jo v raziskavi uporabili, za testiranje inhibicije aktivnosti, ob dodatku izbranih inhibitorjev. Z Gibson assembly tehnologijo smo v plazmidnem vektorju p416GPD pripravili rekombinantni humani nPFKL in sfPFKL gen in ju vnesli v PFK-null sev kvasovke (HD114-8D). Encim nPfk-L smo po namnožitvi v kvasovkah izolirali do homogenosti s frakcionirnim obarjanjem in afinitetno kromatografijo, medtem, ko smo zaradi nestabilnosti transformante sfPFKL, encim sfPfk-L le delno prečistili s frakcionirnim obarjanjem z amnosulfatom. Oba očiščena encima smo uporabili za testiranje inhibicije aktivnosti ob dodatku 15 izbranih inhibitorjev. Uspeli smo določiti tiste, ki bolje inhibirajo aktivnost nPfk-L encima, oziroma tiste, ki bolje inhibirajo sfPfk-L encim, kar predstavlja potencial za tarčno zdravljenje tumorjev. Ugotovili smo tudi, da so transformante z vnesenim nPFKL precej stabilne medtem, ko je sam encim nPfk-L v 30 min neprekinjene meritve, praktično izgubil vso aktivnost. Obratno pa so transformante z vnesenim sfPFKL genom izredno nestabilne, saj so precej hitro izgubile plazmid z vnesenim genom, sam encim sfPfk-L, ki smo ga nekaj sicer uspeli izolirati iz sveže pripravljene transformante pa je bil bolj stabilen.Enzyme 6-phosphofructo-1-kinase is one of the most important regulatory enzymes of glycolysis. There are three phosphofructokinase genes in humans, that are diversely expressed in specific human tissue types. Enchanced expression of Pfk-L isoform, that we used for testing of inhibition upon addition of selected inhibitors, is characteristic for many human tumor types. We used Gibson assembly technology and p416GPD plasmid vector to prepare human nPFKL and sfPFKL gene, and inserted them into PFK-null yeast strain (HD114-8D). Enzyme nPfk-L was purified to homogeneity with ammonium sulfate fractional precipitation and affinity chromatography, while we purified sfPfk-L only with fractional precipitation, due to very low level of extracted sfPfk-L protein, from unstable sfPFKL yeast strain. We used both purified enzymes to test inhibition of activity, upon addition of 15 seleted inhibiotrs. We were able to determine those inhibitors, that show better inhibiton for nPfk-L activity, or sfPfk-L activity respectively. We also noticed that transformants with inserted nPFKL were quite stable, while the enzyme nPfk-L practically lost all of it`s activity after 30 min of continuous measurement. On the contrary, transformants with inserted sfPFKL were very unstable, they lost the majority of their inserted plasmids, while sfPfk-L enzyme preserved its activity

JAVNA SVIJEST O JEZIČNIM POREMEĆAJIMA

U ovom članku opisujem jezične poremećaje, specifične jezične poremećaje i probleme iz područja neujednačene terminologije i dijagnostičkih kriterija za definiranje specifičnih jezičnih poremećaja. Neujednačena terminologija i nejednaki dijagnostički kriteriji štetno utječu na područje kliničke prakse i znanstvenih istraživanja, kao i na svijest javnosti o jezičnim poremećajima. Razina svijesti roditelja posebno je važna, jer ima značajan utjecaj na pravovremeno upućivanje djeteta logopedu što zauzvrat utječe na bolji ishod terapije. Smatram da je ključno poboljšati javno informiranje, jer se samo na taj način može poboljšati polje prevencije i pravodobnog reagiranja roditelja

Les aspects éducatifs de l\u27art dramatique dans l\u27enseignement des langues étrangères

Teoretiki različnih obdobij in različnih predelov sveta so si v večini enotni v tem, da je človek ustvarjalno bitje. Tako lahko nasprotuje destruktivnemu, napreduje, ustvarja in izumlja. Spodbujanje ustvarjalnosti ima zato v šoli velik pomen tako za razvoj vsakega posameznika, kot tudi celotne družbe. Kljub temu ustvarjalnost ne deluje na ukaz. Eden od načinov, da jo spodbudimo pri otrocih in mladostnikih, je vpeljava umetnosti same v pouk. Nekateri predmeti za to dopuščajo več prostora kot drugi. Tako si bomo v tej magistrski nalogi pogledali vpeljavo gledališke umetnosti, in sicer pri predmetih tujih jezikov, natančneje pri francoščini. Ravno pri teh predmetih je, tako kot pri gledališču, pomembno kreiranje z besedami in nastopanje s celim telesom. Poleg tega imata gledališka dejavnost in učenje tujega jezika še veliko več drugih skupnih točk. Gledališka dejavnost naj bi pozitivno vplivala na razvoj jezikovnih kompetenc in ob tem nudi nabor pozitivnih učinkov na vzgojo. Oboje bomo podrobneje preučili v načrtovanem magistrskem delu. Čeprav Slovenija z raziskovanjem in uvajanjem umetnosti v šoli nekoliko zaostaja za državami kot so Nizozemska, Anglija in ZDA, se zanimanje za to področje povečuje. Ni popolnoma jasno, ali zato ker smo skozi različne raziskave dokazali, da umetnost doprinaša k razvoju človeka, ali zato ker se je pojavila skrb, da bi umetnost izginila iz šolskega prostora. Ta magistrska naloga bo pomagala bolje raziskati in razumeti to področje ter ponuditi nova vprašanja in odgovore v še ne popolnoma raziskanem odnosu med poljema gledališke umetnosti in šolstva. Takšen prispevek bo imel vse večji pomen, saj lahko umetnost spodbuja ustvarjalnost in pozitivno vpliva na učenčevo razumevanje sveta ter samega sebe, na njegovo izražanje, vrednotenje, sodelovanje, razvoj odnosa do lepega, motivacijo, osebnostno rast in celo na telesni razvoj. Močno vlogo lahko ima tudi na samo usvajanje tujega jezika, saj lahko s pomočjo umetnosti, natančneje gledališča, učenci lažje razvijejo svoje govorne spretnosti, izražanje, vživljanje v jezik in vlogo govorca tujega jezika, estetiko, pomnjenje in so bolj motivirani za učenje tujega jezika. Vse te predpostavke, ki narekujejo, da ima umetniška dejavnost v šoli vrsto pozitivnih učinkov, bomo v tem delu poskusili potrditi s pomočjo kvalitativnih raziskovalnih postopkov z opazovalno prakso skupine gimnazijcev med pripravo krajše gledališke predstave v francoščini ter z intervjuji učencev, njihovih mentorjev in enega gledališkega umetnika, ki je skupaj z dijaki pripravljal celovečerno predstavo.Theoreticians of different periods and different parts of the world found a common ground in acknowledging that the human is a creative being. Because of this he can oppose destructionhe can progress, create and invent. This is a key reason why creativity in schools should get a lot of support as it is important for the developement of every individual as well as its society. Creativity however does not work on demand. One of many ways to awaken it in children and adolescents is with introducing art itself in school classes. Some arts offer more space for that than others. Thus, in this Master thesis we will research the introducing of theatre art in the subjects of foreign languages, mostly French. Significant in subjects such as theatre is creativeness with words and performing with the whole body. Beside that, theatre activity and learning a foreign language have many other things in common. Theatre activity should positively influence the development of linguistic competences and at the same time it should offer a long list of positive effects on education in sense of raising up. Both is studied in detail in this Master thesis. Even though Slovenia is slightly behind in researching and introducing arts in school when compared to the Netherlands, United Kingdom or the USA, interest in this field is increasing. It is not evident if the reason for this interest comes from the researches who proved all the qualities that arts bring to the development of a human being, or if it could come from a fear of art disappearing from schools. This Master thesis will try to explore and comprehend this field we are entering and offer new questions and answers as well in the not yet completely researched relationship between field of theatre art and education in schools. The meaning of this contribution is to grow, as arts encourage creativity and they positively influence a learners\u27 understanding of the themselves and the world. It also influences their expressing, valuing, cooperating, the sense for aesthetics, motivation, personal growth and even physical progress. Next to that it can also have a powerful role on learning a foreign language as with help of art, especially theatre art, students can develop their rhetorics, memory, the settling into a language and into the role of a native speaker, and they can also appear more motivated for learning this foreign language. We will try to confirm all these presumptions that indicate that artistic activites in school awaken several positive effects by observing a group of secondary school pupils during their preparation of a short theatre performance in French and with interviews of four students, two professors and one artist that were preparing the theatre pla

Public awareness of developmental language disorders in Slovenia

Jezikovne motnje so motnje v razumevanju in/ali uporabi govorjenega, pisnega ali drugega simbolnega sistema (npr. znakovni jezik). Specifična jezikovna motnja je del razvojnih jezikovnih motenj, nima znanega vzroka in zajema različna področja jezikovnega funkcioniranja. Za poimenovanje jezikovnih motenj neznanega vzroka so v uporabi tudi drugi različni termini, kot so: govorno-jezikovna motnja, razvojna disfazija, specifični jezikovni primanjkljaji ipd. Prav tako so v uporabi različni diagnostični kriteriji, ki skupaj z neenotno terminologijo vplivajo na poznavanje jezikovnih motenj, javno ozaveščenost in dostop do logopedskih storitev. Pogosto pravilne informacije o jezikovnih motnjah v javnosti niso dovolj razširjene ali pa so celo nedosledne. V teoretičnih izhodiščih magistrskega dela smo najprej predstavili govorno-jezikovne motnje, nato pa smo natančno opisali razvojne jezikovne motnje, specifično jezikovno motnjo, zgodovino poimenovanja oziroma terminologije jezikovnih motenj ter diagnostične kriterije za opredelitev specifične jezikovne motnje. Opisali smo tudi pomen javne ozaveščenosti na področju logopedije in predstavili izsledke do sedaj narejenih raziskav. Z raziskavo smo želeli ugotoviti stopnjo javne ozaveščenosti o jezikovnih motnjah v Sloveniji. Prav tako je bil osrednji namen ugotoviti, ali obstajajo statistično pomembne razlike v stopnji javne ozaveščenosti o jezikovnih motnjah v Sloveniji glede na starost, spol, izobrazbo in socioekonomski status. Vzorec je zajemal 90 oseb z območja celotne Slovenije in je bil sestavljen iz treh starostnih skupin18−39 let, 40−59 let ter 60 in več let. Vse starostne skupine so bile enako zastopane. Vprašani so izpolnili vprašalnik o javni ozaveščenosti o jezikovnih motnjah. Z raziskavo smo ugotovili, da je stopnja javne ozaveščenosti v Sloveniji vprašljiva. Večina vprašanih pozna izraz jezikovna motnja, hkrati pa ga ne zna pravilno opredeliti in ne pozna značilnosti motnje. Prav tako smo ugotovili, da obstajajo nekatere statistično pomembne razlike v javni ozaveščenosti o jezikovnih motnjah med različnimi starostnimi skupinami in glede na spol, izobrazbo ter socioekonomski status.Language disorders are disorders in understanding and/or use of spoken, written, or other symbolic systems (e.g. sign language). A specific language impairment is a part of developmental language disorders. It has no known cause and includes different fields of language functioning. To name the language disorders of unknown cause, other different terms are also in use, such as speech-language disorder, developmental dysphasia, specific language deficits, etc. Different diagnostic criteria are also in use. Together with non-uniform terminology, they influence the knowledge of language disorders, public awareness, and access to speech-and-language-therapist services. Usually, accurate information on language disorders is not spread enough in public or is even inconsistent. In the theoretical starting points of the master’s thesis, we first presented speech-language disorders. Then, we accurately described developmental language disorders, the specific language impairment, the history of naming and the terminology of language disorders, and diagnostic criteria to define the specific language impairment. We also described the significance of public awareness in the field of speech and language therapy and presented the findings of recent research. By research, we wished to ascertain the level of public awareness of language disorders in Slovenia. The central purpose was also to ascertain whether there are statistically significant differences in the level of public awareness in language disorders in Slovenia with regards to age, gender, education, and socioeconomic status. The sample included 90 persons from the area of entire Slovenia and was composed of three age brackets: 18−39 years, 40−59 years, and 60 years and more. All the age brackets were represented equally. The participants filled in the questionnaire on public awareness of language disorders. By the research, we ascertained that the level of public awareness in Slovenia is questionable. Most of the participants know the expression language disorder. At the same time, they cannot define it correctly and do not know the characteristics of the disorder. We also ascertained that there are some statistically significant differences in public awareness of language disorders among different age brackets and with regards to gender, education, and socioeconomic status

Promoting hygiene in the preschool child

In my dissertation, I researched how incentives affect the hygienic condition in a kindergarten. In the theoretical part, I wrote about the history of hygiene, that is, from the first attempts of a soap to the first guidelines for hand washing. I focused on hand hygiene, when and how to wash our hands, and on hand disinfection. I also summarized some research in which there were examples of activities that we can do in kindergarten. I also wrote about the hygiene of sneezing/coughing and wiping a nose, and there I also mentioned some activities for wiping a nose and learning to blow a nose. Finally, I write about the hygiene in indoor areas. Empirical part involved 34 pre-school teachers and 18 children from the age group 2—4 years. Pre-school teachers participated in an anonymous survey questionnaire containing 17 closed, open and combined questions. The children participated in the activity; before and after the activity, I observed the children washing their hands, sneezing/coughing and wiping their noses, and entered the data in observation sheets. Research has shown that pre-school teachers support hand washing, proper sneezing/coughing and wiping the nose. It is important for them to wash their hands before lunch, after use or. assistance in toilets and after arrival from outside premises. They also find it important to throw the handkerchief in the bin after use. However, they do not find it so important to wash their hands every time they sneeze/cough. Pre-school teachers plan activities from various fields that promote hygiene. They allow children to be independent in taking care of their hygiene, but pre-school teachers are still checking how well the children are doing. Pre-school teachers often remind children to wash their hands, but children, depending on their age, are in the minority or in most cases independent in taking care of hygiene. The motivation for learning hygiene was provided by a puppet, which the children were enthusiastic about and expressed great interest in the content. With just one day of activity, I noticed great advances in child hygiene, both in theory and in practice. The results showed that hand hygiene, sneezing/coughing and wiping the nose increased after the performed activity