Ispitivanje estrogene aktivnosti u vodama pomoću kvasaca

The presence of endocrine-disrupting compounds in wastewater, surface water, groundwater and even drinking water has become a major concern worldwide, since they negatively affect wildlife and humans. Therefore, these substances should be effectively removed from effluents before they are discharged into surface water to prevent pollution of groundwater, which can be a source of drinking water. Furthermore, an efficient control of endocrine-disrupting compounds in wastewater based on biological and analytical techniques is required. In this study, a yeast estrogen screen (YES) bioassay has been introduced and optimized with the aim to assess potential estrogenic activity of waters. First, assay duration, concentration of added substrate to the assay medium and wavelength used to measure the absorbance of the substrate were estimated. Several compounds, such as 17-β-estradiol, 17-α-ethinylestradiol, bisphenol A, nonylphenol, genisteine, hydrocortisone, dieldrin, atrazine, methoxychlor, testosterone and progesterone were used to verify its specificity and sensitivity. The optimized YES assay was sensitive and responded specifically to the selected estrogenic and nonestrogenic compounds in aqueous samples. Potential estrogenicity of influent and effluent samples of two wastewater treatment plants was assessed after the samples had been concentrated by solid-phase extraction (SPE) procedure using Oasis® HLB cartridges and methanol as eluting solvent. Up to 90 % of relative estrogenic activity was detected in concentrated samples of influents to wastewater treatment plants and estrogenic activity was still present in the concentrated effluent samples. We found that the introduced YES assay is a suitable screening tool for monitoring the potential estrogenicity of effluents that are discharged into surface water.Prisutnost spojeva koji imaju estrogeni učinak u otpadnim, površinskim, podzemnim, a čak i pitkim vodama sve više zabrinjava stručnjake diljem svijeta zbog negativnog učinka na zdravlje ljudi i životinja. Stoga takve spojeve treba odstraniti iz otpadnih voda prije njihovog ispuštanja u površinske vode, te na taj način spriječiti zagađivanje podzemnih voda, što se koriste kao izvor pitke vode. Kemijskom analizom i biološkim testovima potrebno je učinkovito pratiti prisutnost spojeva s estrogenim učinkom u otpadnim vodama. U ovom smo radu razvili i optimirali metodu ispitivanja estrogenog učinka spojeva pomoću kvasca (test YES) radi procjene potencijalne estrogene aktivnosti u vodama. Optimirali smo trajanje testa, koncentraciju supstrata dodanog podlozi i valnu duljinu za mjerenje apsorbancije razgrađenog supstrata. Specifičnost i osjetljivost testa YES provjerili smo pomoću sljedećih spojeva: 17-β-estradiol, 17-α-etinilestradiol, bisfenol A, nonilfenol, genistein, hidrokortizon, dieldrin, atrazin, metoksiklor, testosteron i progesteron. Optimiranim testom YES mogu se u uzorcima voda detektirati odabrani spojevi s estrogenim učinkom ili bez njega. Moguća estrogenost pritoka i istoka vode iz dva postrojenja za obradu otpadnih voda određena je prethodnom ekstrakcijom uzoraka vode na čvrstoj fazi (SPE ekstrakcija) pomoću kolone Oasis® HLB i metanola kao otapala. U koncentriranim je uzorcima pritoka u postrojenjima određeno 90 % relativne estrogene aktivnosti. Isto tako je estrogena aktivnost pronađena i u koncentriranim uzorcima istoka iz postrojenja. Iz dobivenih se rezultata može zaključiti da je test YES prikladan za ispitivanje estrogene aktivnosti otpadnih voda, što naknadno dospijevaju u površinske vode

Contemporary treatment of psoriasis and the care of affected skin

Luskavica je ena izmed najpogostejših kožnih bolezni, ki prizadene okrog 2 % prebivalstva, pri čemer obolevajo tako ženske kot moški. Število znanih primerov v Republiki Sloveniji je povsem primerljivo z drugimi državami. Zaradi različnih tipov bolezni je potrebno vsak primer obravnavati individualno, pri čemer je ključnega pomena, da pred začetkom zdravljenja zdravniki postavijo pravilno diagnozo ter se nato odločijo za terapijo, ki bo najustreznejša za posameznega bolnika. Pri luskavici so zaradi pospešene keratinizacije prizadeta področja kože zelo suha in prekrita z luskami. Zato je poleg zdravljena zelo pomembna tudi nega kože z različnimi negovalnimi izdelki. V diplomski nalogi smo opisali oblike luskavice in dejavnike, ki vplivajo na njen nastanek in potek. Izpostavili smo tudi psihične težave obolelih in nujnost osveščanja splošne populacije o tej bolezni. S pomočjo strokovnjaka, specialista dermatologa-venerologa, ki smo ga intervjuvali, smo raziskali, kako v praksi potekata diagnosticiranje in zdravljenje bolezni. Izvedeli smo, da se v Republiki Sloveniji uporabljajo enake metode zdravljenja in enake zdravilne učinkovine, ki so uveljavljene tudi drugje po svetu in so dokazano najboljše za terapijo luskavice. Tudi za nego z luskavico prizadete kože imamo na slovenskem trgu na voljo precej izdelkov. Informacije o tem, kateri med njimi so najbolj uporabljani oz. katere najbolj priporočajo bolnikom z luskavico, smo pridobili s pogovorom s farmacevtko v eni od lekarn. Vsem izdelkom za nego kože je skupno to, da lajšajo draženje ter vlažijo in pomirjajo suho in srbečo kožo.Psoriasis is one of the most common skin diseases, affecting about 2% of general population, both men and women equally. In Republic of Slovenia its incidence is comparable to other countries in the world. Due to different types of disease, each case should be dealt individually, as it is of utmost importance to set up a proper diagnosis before starting any treatment. In psoriasis, the affected skin areas are very dry and full of scales due to accelerated keratinization. Therefore, in addition to treatment, skin care with various types of products is also important. In this thesis, we described different types of psoriasis and factors influencing its outbreak and course. We also set out psychological problems of psoriatic patients and the necessity to improve awareness of general population regarding this disease. Further on, by interviewing a specialist of dermatology-venereology we investigated how the disease is diagnosed and treated in practice. We found out that in the Republic of Slovenia, the same treatment methods and drugs are used, which are well established elsewhere in the world and are proven to be the best choice for treating psoriasis. There are also many non-prescription skincare products available on the Slovenian market that can be used on psoriasis affected skin. By talking to the pharmacist in one of the community pharmacies we were able to gather information regarding skincare products that are the most suggested and actually the most used by psoriatic patients. All of these products are able to relieve irritation, and hydrate and soothe the dry and itchy skin

Comparison of regulatory requirements for obtaining drug marketing authorisation in the United States of America and selected other countries

V magistrski nalogi smo primerjalno vrednotili regulativne zahteve za pridobitev dovoljenja za promet z zdravilom v Združenih državah Amerike (ZDA), Kanadi, Avstraliji in državah Evrazijske unije (EAEU). Proučili smo tudi ključne zahteve in izzive pri pridobivanju dovoljenja za promet z zdravilom v Evropski uniji (EU). Regulativne zahteve so ključne za zagotavljanje varnosti, učinkovitosti in kakovosti zdravil. Namen naloge je bil primerjati regulativne okvirje v različnih državah ter identificirati ključne razlike in podobnosti, s poudarkom na izzivih v EU in pristopih drugih držav za reševanje teh težav. Uporabili smo primerjalno analizo podatkov iz uradnih regulativnih dokumentov za ZDA, Kanado, Avstralijo, EAEU in EU. Analizo smo izvedli na podlagi kriterijev, kot so postopek pridobivanja dovoljenja, trajanje postopkov, zahteve glede varnosti in učinkovitosti ter posebni programi za pospešitev postopkov. V nadaljnjem poglavju smo podrobno obravnavali regulativne zahteve v izbranih državah. V ZDA vodi postopek Uprava Združenih držav Amerike za hrano in zdravila s strogimi zahtevami za klinične študije in programi za pospešitev postopkov za inovativna zdravila. Kanada ima podoben sistem, ki omogoča hitro obravnavo zdravil z visokim terapevtskim potencialom. Avstralija preko Avstralske uprave za zdravila zahteva obsežne dokaze o varnosti in učinkovitosti, EAEU pa stremi k harmonizaciji postopkov, vendar se sooča z izzivi pri implementaciji. Ugotovitve kažejo, da so regulativni postopki v vseh regijah strogi in zahtevajo obsežne dokaze o varnosti in učinkovitosti zdravil. Kljub temu se postopki in trajanje razlikujejo, pri čemer ZDA in EU izstopata s strogimi regulativnimi okviri. Zaključujemo, da primerjalna analiza regulativnih zahtev ponuja dragocen vpogled v različne pristope in izzive pri pridobivanju dovoljenj za promet z zdravili. Raziskava poudarja potrebo po nadaljnji harmonizaciji za izboljšanje učinkovitosti postopkov in dostopnosti zdravil na trgih ter predlaga nadaljnje raziskave na tem področju.In this master’s thesis, we comparatively evaluate the regulatory requirements for obtaining a marketing authorisation for a medicinal product in the United States of America (USA), Canada, Australia, and the countries of the Eurasian Economic Union (EAEU). We also examine the key requirements and challenges for obtaining a marketing authorisation for a medicinal product in the European Union (EU). Regulatory requirements are key to ensuring the safety, efficacy, and quality of medicinal products. The aim of this assignment was to compare the regulatory frameworks in different countries and identify key differences and similarities, focusing on the challenges in the EU and approaches taken by other countries to address these issues. We used a comparative analysis of data from official regulatory documents for the US, Canada, Australia, EAEU and the EU. The analysis was based on criteria such as the authorisation process, duration of procedures, safety and efficacy requirements, and specific programmes to speed up procedures. In the following section, we discuss in detail the regulatory requirements in selected countries. In the US, the process is led by the US Food and Drug Administration with strict requirements for clinical studies and fast-track programmes for innovative medicines. Canada has a similar system that allows for the fast-tracking of drugs with high therapeutic potential. Australia, through the Therapeutic Goods Administration, requires extensive evidence of safety and efficacy, and the EAEU is working towards harmonisation of procedures, but faces challenges in implementation. The findings show that regulatory procedures in all regions are stringent and require extensive evidence on the safety and efficacy of medicines. However, procedures and durations vary, with the US and the EU standing out with their strict regulatory frameworks. We conclude that a comparative analysis of regulatory requirements provides valuable insights into the different approaches and challenges in obtaining marketing authorisations for medicines. The study highlights the need for further harmonisation to improve the efficiency of procedures and the accessibility of medicines on the markets and suggests further research in this area

Morphological traits and phenotypic variability of different genotypes of sweet maize (Zea mays L. saccharata Koern.)

Namen naloge je bil s poljskim poskusom preučiti 7 različnih genotipov sladke koruze in sicer križancev \u27Zarja\u27 in \u27Gold Cup\u27 ter 5 populacij sladke koruze \u27True Gold Sweet Corn\u27, \u27Golden Bantam\u27, \u27Ashworth\u27, \u27Extrasusser Zuckermais Damaun\u27 in \u27Mezdi KS\u27 na pridelek, čas cvetenja in čas mlečne zrelosti storžev, nekaterih lastnosti rastline in storža ter izenačenost posameznih genotipov v preučevanih lastnostih. Poskus je bil izveden na poskusnem polju Biotehniške fakultete v Jabljah pri Trzinu po metodiki naključnega bloka v 2 ponovitvah. V času rastne dobe smo beležili čas cvetenja ter izmerili višine rastlin in višine vrhnjih storžev. Storže smo pobirali v času mlečne zrelosti v 3 rokih. Storže smo prešteli, sveže storže stehtali, izmerili dolžino in premer storžev in prešteli vrste zrnja na storžih ter izračunali povprečne vrednosti, koeficient variacije in variacijsko širino. Glede na čas cvetenja sta bili najzgodnejši sorti \u27Ashworth\u27 in \u27Zarja\u27, najpoznejši pa \u27Gold Cup\u27 in \u27True Gold Sweet Corn\u27. Najzgodnejši sorti glede na čas mlečne zrelosti sta \u27Ashworth\u27 in \u27Extrasusser Zuckermais Damaun\u27, od 50 % svilanja do pobiranja sta potrebovali 25 oziroma 21 dninajpoznejši sorti \u27True Gold Sweet Corn\u27 in \u27Gold Cup\u27 pa 26 oziroma 29 dni. Skupno število dni od polnega cvetenja do pobiranja mlečno zrelih storžev se je gibalo med 12 in 39 dnevi, v odvisnosti od sorte, časa pobiranja in enakomernosti dozorevanja posamezne sorte. Sorta \u27Mezdi KS\u27 ima največjo povprečno maso storža, najdaljše in najdebelejše storže, z največjim številom vrst zrnja ter najvišjimi rastlinami. Sorta \u27Ashworth\u27 je najzgodnejša sorta, a ima najmanjše vrednosti za maso in dolžino storža ter višino rastline. Sorta \u27Zarja\u27 ima za večino lastnosti povprečne vrednosti, a je v vseh lastnostih najbolj izenačena.The purpose of our study was to investigate the 7 genotypes (varieties) of sweet maize, namely 2 hybrids (\u27Zarja\u27 and \u27Gold Cup\u27) and 5 populations (\u27True Gold Sweet Corn\u27, \u27Golden Bantam\u27, \u27Ashworth\u27, \u27Extrasusser Zuckermais Damaun\u27 and \u27Mezdi KS\u27). The study included flowering and milk maturity time, lenght, diameter and weight of ears, plant height, height of the uppermost ear and homogeneity of investigated traits. Fot all traits, the minimum, maximum, mean and CV (%) values were calculated. The trial took place on the experimental fields of the Biotechnical Faculty at Jablje near Ljubljana. Regarding the flowering time, the earliest were \u27Ashworth\u27 and \u27Zarja\u27, while the latest were \u27Gold Cup\u27 and \u27True Gold Sweet Corn\u27. Considering the milk maturity time, the earliest varieties were \u27Ashworth\u27 and \u27Extrasusser Zuckermais Damaun\u27, and the period from 50 % silking to milk maturity was 25 and 21 days, respectivelywhile the latest varieties, \u27True Gold Sweet Corn\u27 and \u27Gold Cup\u27, needed 26 and 29 days, respectively. The total number of days from 50 % silking to milk maturity varied between 12 and 39 days, depending on variety, harvest time and the homogeneity of milk ripening time of individual variety. \u27Mezdi KS\u27 was characterised by the highest average weight of ears, and the longest and the thickest ears. \u27Ashworth\u27 was the earliest variety and it also exhibited the lowest values of weight and length of ears. \u27Zarja\u27 was characterised by average values for most traits and was the most homogeneous in all studied traits.

Yeast Estrogen Screen Assay as a Tool for Detecting Estrogenic Activity in Water Bodies

The presence of endocrine-disrupting compounds in wastewater, surface water, groundwater and even drinking water has become a major concern worldwide, since they negatively affect wildlife and humans. Therefore, these substances should be effectively removed from effluents before they are discharged into surface water to prevent pollution of groundwater, which can be a source of drinking water. Furthermore, an efficient control of endocrine-disrupting compounds in wastewater based on biological and analytical techniques is required. In this study, a yeast estrogen screen (YES) bioassay has been introduced and optimized with the aim to assess potential estrogenic activity of waters. First, assay duration, concentration of added substrate to the assay medium and wavelength used to measure the absorbance of the substrate were estimated. Several compounds, such as 17-β-estradiol, 17-α-ethinylestradiol, bisphenol A, nonylphenol, genisteine, hydrocortisone, dieldrin, atrazine, methoxychlor, testosterone and progesterone were used to verify its specificity and sensitivity. The optimized YES assay was sensitive and responded specifically to the selected estrogenic and nonestrogenic compounds in aqueous samples. Potential estrogenicity of influent and effluent samples of two wastewater treatment plants was assessed after the samples had been concentrated by solid-phase extraction (SPE) procedure using Oasis® HLB cartridges and methanol as eluting solvent. Up to 90 % of relative estrogenic activity was detected in concentrated samples of influents to wastewater treatment plants and estrogenic activity was still present in the concentrated effluent samples. We found that the introduced YES assay is a suitable screening tool for monitoring the potential estrogenicity of effluents that are discharged into surface water