Provider: - Institution: University of Zagreb. Faculty of Pharmacy and Biochemistry. Department of microbiology. - Data provided by Europeana Collections- Učinak sekundarnog metabolita masline (Olea ...europaea L.) oleuropeina, njegovog razgradnog produkta hidroksitirosola i vodenog ekstrakta maslinova lista na staničnu vijabilnost medicinski značajne gljivične vrste Candida albicans ispitan je in vitro testovima. Metodom mikrodilucije određene su minimalne inhibitorne koncentracije (MIK) koje uzrokuju 80 % smanjenja stanične vijabilnosti i koje su iznosile 12,5 mg/ml za oleuropein, 6,25 mg/ml za hidroksitirosol i 25 mg/ml za vodeni ekstrakt maslinova lista. Kvantitativna analiza s fluorescentnim bojanjem upućuje na zaključak da je apoptoza primarni način stanične smrti u uzorcima tretiranim sa sub-MIK koncentracijama ovih tvari. Budući da je patogenost vrste određena virulentnim čimbenicima, ispitani su učinci oleuropeina i hidroksitirosola na najznačajnije virulentne čimbenike vrste C. albicans. Utvrđen je inhibitorni učinak ovih fenolnih tvari na promjenu u hifalni oblik rasta kod vrste C. albicans induciranu u uvjetima staničnog gladovanja. Pri sub-MIK koncentracijama, oleuropein i hidroksitirosol nisu pokazali inhibitorno djelovanje na stvaranje biofilma ove vrste. Pri subinhibitornim koncentracijama nakon 24 sata inkubacije, oleuropein i hidroksitirosol su uzrokovali modulaciju stanične površinske hidrofobnosti koja se dovodi u svezu s adherencijom vrste C. albicans na biomaterijale. Uzgoj vrste C. albicans u prisutnosti oleuropeina i hidroksitirosola je uzrokovao inhibiciju aktivnosti kandidinih hidrolitičkih enzima, aspartil-proteaza (Sap) i α-glukozidaza. Tretiranjem vrste C. albicans s oleuropeinom i hidroksitirosolom utvrđeno je višestruko djelovanje ovih tvari uključujući oštećenje stanične stijenke, stanične membrane i otpuštanje citoplazmatskog sadržaja (DNA i proteina). S obzirom da neki antimikotici djeluju kao inhibitori biosinteze ergosterola, provedeno je ispitivanje učinka oleuropeina i hidroksitirosola na modulaciju ergosterola kojima je utvrđeno smanjenje ovog sterola u ovisnosti o primijenjenoj koncentraciji. Uz testove određivanja djelovanja na gljivičnu vrstu, ispitani su mogući citotoksični i genotoksični učinci na stanice nositelja. Navedene tvari nisu pokazale hemolitičku aktivnost na humanim eritrocitima pri testiranim koncentracijama u rasponu 3-60 μM. Ispitivanje učinka na humanim limfocitima periferne krvi provedeno kometnim testom pokazalo je protektivno djelovanje ovih tvari na oštećenja DNA inducirana s mutagenom H2O2, pri čemu je hidroksitirosol imao bolji protektivni učinak u usporedbi s oleuropeinom nakon 120 min predtretmana s ispitivanim tvarima u koncentracijama 1, 5 i 10 μM. Navedeni genoprotektivni učinci su posljedica snažnog antioksidativnog djelovanja ovih fenolnih spojeva utvrđenih ABTS, FRAP i CUPRAC metodom.- Activity of oleuropein, a phenol secoiridoid present in olive tree products, its derivative hydroxytyrosol and water extract of olive leaf were investigated using in vitro tests against opportunistic fungal pathogen Candida albicans. Antifungal susceptibility testing was used to estimate the drop of viability up to 80 % in comparison to the control (untreated cells). Minimal inhibitory concentration (MIC) against C. albicans for oleuropein was 12,5 mg/ml, for hydroxytyrosol was 6,25 mg/ml and for water extract of olive leaf was 25 mg/ml. Morphological changes in the nuclei after staining with fluorescent DNA-binding dyes revealed that apoptosis was a primary mode of cell death in analyzed samples treated with sub-MIC concentrations of tested compounds. In order to understand mode of action of oleuropein and hydroxytyrosol, their effect on C. albicans virulence factors essential for development of infection in the host have been investigated. Both oleuropein and hydroxytyrosol modulate morphogenetic coversion and inhibit filamentation of C. albicans induced in the conditions of reduced nutrient availability. On the other hand, treatment with sub-MIC concentractions of these phenolic compounds did not affect C. albicans biofilm formation. The hydrophobicity assay showed that both compounds, in sub-MIC values, have decreased the cellular surface hydrophobicity, a factor associated with adhesion to epithelial cells. Cultivation of C. albicans in the presence of oleuropein and hydroxytyrosol also inhibits, in the dose-dependent manner, acitivity of aspartyl-proteases (Sap), hydrolytic enzymes important in adherence, tissue penetration, invasion and destruction of host tissue. Most therapies for fungal infections target the ergosterol biosynthesis pathway or its end product ergosterol which is necessary for growth and normal function of fungal cells. At subinhibitory concentrations tested compounds have altered sterol content and subsequently affected the cell membrane of C. albicans. Both compounds induced dose-dependent loss of intracellular material to outer space (DNA, protein leakage) and caused membrane depolarisation indicating their direct effect on the membrane. In addition, cytotoxicity and genotoxicity assays were performed to elucidate the effect of oleuropein and hydroxytyrosol on the host cells. Oleuropein and hydroxytyrosol exhibited no hemolytic activity on human erythrocytes at tested concentrations (3-60 μM). Antigenotoxic effects of oleuropein and hydroxytyrosol against H2O2-induced damage in human lymphocytes were evaluated in vitro by alkaline comet assay. Pretreatment of human lymphocytes with each of the substance for 120 min produced dose-dependent reduction of primary DNA damage in the tested cell type. At tested concentrations (1, 5, 10 μM), hydroxytyrosol showed better protective effect than oleuropein against H2O2-induced DNA breaks. Our results suggest that genoprotective effects of these phenolic compounds could be attributed to their radical scavenging and metal ions chelating activity which were determined using ABTS, FRAP and CUPRAC assays.- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Polimeri su već neko vrijeme dio farmacijskoga oblikovanja lijekova, bilo da se koriste kao pomoćna sredstva ili kao ambalaža za lijekove. Polimeri imaju jedinstvena fizika!no-kemijska svojstva kao ...što su molekulska masa, viskoznost, geliranje ili topljivost, koja uvelike utječu na njihovu primjenu u farmaciji i medicini. Sve je veća primjena polimera u naprednim terapeutskim sustavima kao što su matrični i spremišni sustavi, osmotske pumpe, degradacijski sustavi te sustavi osjetljivi na okolinu. Poznavanje odnosa struktura-svojstvo polimemih tvari te njihova karakterizacija jedan je od preduvjeta za dizajniranje lijekova pomoću polimemih tvari.
Provider: - Institution: University of Zagreb. Faculty of Pharmacy and Biochemistry. Department of pharmacognosy. - Data provided by Europeana Collections- U okviru ovog doktorskog rada po prvi put ...je provedeno opsežno usporedno istraživanje bioloških učinaka
14 prirodnih fenolnih kiselina, prevalidacija metode određivanja ukupnih hidroksicimetnih derivata u biljnim
drogama, fitokemijska karakterizacija fenolnih kiselina i flavonoida te biološki učinci odabranih vrsta roda
Micromeria Bentham (Lamiaceae). Antioksidacijski učinak fenolnih kiselina ispitan je primjenom četiri različite
spektrofotometrijske metode. Hidroksicimetne kiseline su pokazale bolja antioksidacijska svojstva u odnosu na
većinu hidroksibenzojevih kiselina, a iznimno dobar antioksidacijski učinak utvrđen je za ružmarinsku, kavenu i
galnu kiselinu. Citotoksični učinak fenolnih kiselina ispitan je na HepG2 i THP 1 staničnim kulturama primjenom
MTS testa, pri čemu većina testiranih kiselina nije pokazala značajnije smanjenje vijabilnosti stanica. Istraživanje
protuupalnog djelovanja fenolnih kiselina istaknulo je galnu, ružmarinsku, klorogensku, kavenu i sinapinsku kiselinu
kao dobre inhibitore interleukina-6. Komet testom na uzorku humane krvi nakon tretiranja bovericinom, dokazan je
dobar antigenotoksični učinak svih ispitanih fenolnih kiselina.Fitokemijska analiza fenolnih kiselina i flavonoida u
biljnim vrstama porodice Lamiaceae provedena je primjenom kromatografskih (HPTLC, RP-HPLC-DAD, LC-ESIMS/
MS) i spektrofotometrijskih metoda. U etanolnim ekstraktima vrsta M. croatica, M. juliana i M. thymifolia
identificirano je jedanaest fenolnih kiselina i petnaest flavonoida. Kriptoklorogenska, neoklorogenska, 3-p- i 4-pkumaroilkina,
3-(3,4- dihidroksifenil)mliječna, salvianolna kiselina B i J te dimer i trimer ružmarinske kiseline po
prvi put su identificirane u navedenim vrstama. Izuzev luteolinskih i akacetinskih derivata u vrstama M. juliana i M.
thymifolia, ostali su flavonoidi također prvi put identificirani u ispitivanim vrstama roda Micromeria. Ustanovljena su
dobra antioksidacijska, protuupalna i antigenotoksična svojstva odabranih biljnih vrsta, pri čemu se istaknula
endemična vrsta M. croatica.
Dobiveni rezultati ukazali su na veliki biomedicinski potencijal ispitanih fenolnih kiselina i vrsta roda Micromeria.- A comprehensive comparative study of the biological effects of 14 natural phenolic acids, prevalidation of
total hydroxycinnamic assays, identification of phenolic acids and flavonoids as well as biological activities of
selected Micromeria species were conducted for the first time in this doctoral thesis.
Antioxidant activity of phenolic acids was tested using four different spectrophotometric methods. Hydroxycinnamic
acids had better antioxidant properties than most hydroxybenzoic acids. Evaluation of antioxidant activity highlighted
rosmarinic, caffeic and gallic acids as the best ones. Cytotoxic effect of phenolic acids was studied on HepG2 and
THP-1 cell cultures using MTS test, and phenolic acids did not show significant reduction of cell viability.
Investigation of phenolic acids' anti-inflammatory activity revealed gallic, rosmarinic, chlorogenic, caffeic and
sinapinic acids as good inhibitors of interleukin-6 production. Comet assay on human blood cells treated with
beauvericin proved good antigenotoxic effects of all tested phenolic acids.
Phytochemical analysis of polyphenols was carried out using various chromatographic (HPTLC, RP-HPLC-DAD, LCESI-
MS/MS) and spectrophotometric methods. Eleven phenolic acids and fifteen flavonoids were identified in the
ethanolic extracts of M. croatica, M. juliana and M. thymifolia. Chriptochlorogenic, neochlorogenic, 3-p- and 4-pcoumaroylquinic,
3-(3,4- dihydroxyphenil)lactic, salvianolic acid B and J as well as dimer and trimer of rosmarinic
acid were identified in these species for the first time. With exception of luteolin and acacetin derivatives in M. juliana
and M. thymifolia, flavonoids were identified for the first time in the tested species. Good antioxidant, antiinflammatory
and antigenotoxic properties were established for investigated Micromeria species with special emphasis
on endemic M. croatica.
Obtained results revealed the great biomedical potential of tested phenolic acids and Micromeria species.- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Provider: - Institution: - Data provided by Europeana Collections- Neravnoteža između toksičnih i esencijalnih elemenata i sustava antioksidacijske zaštite u organizmu, zajedno s ostalim štetnim ...čimbenicima iz čovjekovog okoliša i načina života, može dovesti do povećane osjetljivosti i doprinijeti razvoju poremećaja muškog reprodukcijskog sustava kao što su rak testisa i prostate. Cilj ovog istraživanja bio je odrediti povezanost koncentracije esencijalnih i toksičnih metala i polumetala i aktivnosti antioksidacijskih enzima s razinama tumorskih biljega u uzorcima krvi muškaraca s rakom testisa i prostate, uzevši u obzir dob, indeks tjelesne mase, te naviku pušenja i konzumiranja alkohola. Također, određena je koncentracija metalotioneina u serumu ispitanika s rakom testisa, kao i učinak međudjelovanja cinka, selenija i kadmija s platinom na aktivnost superoksid dismutaze i glutation peroksidaze te primarno oštećenje DNA u
kontrolnih muškaraca u in vitro uvjetima. U ispitanika s novo dijagnosticiranim rakom testisa izmjerene su značajno niže koncentracije esencijalnih elemenata bakra i selenija i značajno viša koncentracija olova u odnosu na kontrolne ispitanike dok je u skupini ispitanika s novo dijagnosticiranim rakom prostate pronađena značajno viša koncentracija olova i niža koncentracija cinka. U obje skupine ispitanika s malignim oboljenjem uočena je značajno viša aktivnost antioksidacijskih enzima bakar-cink superoksid dismutaze i glutation peroksidaze. U ispitanika s rakom testisa, porastom koncentracije platine nakon primljene kemoterapije, snižene su koncentracije većine mjerenih metala i polumetala, a uočen je porast aktivnosti glutation peroksidaze. U ispitanika s rakom prostate nakon radioterapije, uočen je značajan porast koncentracije bakra i selenija te značajno sniženje koncentracije kadmija i cinka. Dobiveni rezultati pokazali su kako je smanjenje izloženosti
toksičnim elementima uz održavanje homeostaze esencijalnih elemenata povezano sa sniženjem razina serumskih tumorskih biljega laktat dehidrogenaze i prostatnog specifičnog antigena. U serumu ispitanika s novo dijagnosticiranim seminomima testisa uočena je viša koncentracija metalotioneina u odnosu na kontrolne ispitanike. Rezultati upućuju na važnost metalotioneina kod germinativnih tumora testisa. U kombinaciji s postojećim biljezima, određivanje metalotioneina moglo bi biti korisno kod dijagnoze raka testisa te u praćenju oboljelih nakon kemoterapije. Na temelju distribucijskog profila platine u serumu ispitanika s germinativnim tumorima testisa možemo zaključiti kako metalotionein ne doprinosi otpornosti na cisplatinu
budući da je ~ 95 % slobodne platine vezano za proteine srednje molekulske mase, dok se tek manji dio (~ 3 %) veže za metalotionein. Rezultati ispitivanja međudjelovanja kadmija, platine, selenija i cinka u in vitro uvjetima pokazuju kako
istovremeno djelovanje platine i kadmija smanjuje aktivnost bakar-cink superoksid dismutaze (za 16 %) i glutation peroksidaze (za 10 %) te izaziva oštećenje DNA (porast % DNA u repu za 106 % u odnosu na kontrolu), dok je dodatak selenija i cinka rezultirao značajnim povišenjem aktivnosti enzima, odnosno popravkom oštećenja DNA. Ovi rezultati naglašavaju važnost što manje izloženosti kadmiju, pogotovo kod ljudi oboljelih od malignih bolesti koji su, tijekom liječenja, a i kasnije, izloženi spojevima poput cisplatine.- The imbalance between toxic and essential elements and mechanisms of antioxidant defence in the organism,
along with other harmful factors from human environment and lifestyle, can lead to increased sensitivity and
contribute to the development of the male reproductive system disorders such as testicular and prostate cancer.
The aim of this study was to determine the association between the concentration of essential and toxic metals
and metalloids and activity of antioxidant enzymes, and levels of tumour markers in blood samples of men with
testicular and prostate cancer, taking into account the age, body mass index, smoking habits and alcohol consumption. In addition, we determined the concentration of serum metallothionein in men with testicular cancer, as well as the in vitro interaction of zinc, selenium, cadmium and platinum on the activity of copper-zinc superoxide dismutase and glutathione peroxidase and induction of DNA strand-breaks. In subjects with newly diagnosed testicular cancer we found significantly lower concentrations of essential elements copper and selenium, and higher concentration of lead compared to control subjects, while in subjects with newly diagnosed prostate cancer we found significantly higher concentration of lead and lower concentration of zinc. In both groups of subjects with malignant disease significantly higher activity of
antioxidant enzymes copper-zinc superoxide dismutase and glutathione peroxidase was observed. The results
show how reducing the exposure to toxic elements while maintaining the homeostasis of essential elements is
associated with decrease in serum levels of tumour markers lactate dehydrogenase and prostate specific antigen.
Increased concentration of platinum in patients with testicular cancer after chemotherapy reduced the
bioavailability of most metals and metalloids and activated the antioxidant defence mechanisms. In patients with
prostate cancer, concentration of copper and selenium significantly increased after radiotherapy while there was
a decrease in concentration of cadmium and zinc. In subjects with newly diagnosed testicular seminoma we found significantly higher concentration of metallothionein compared to control. In combination with the existing markers, the determination of serum metallothionein levels could be useful in diagnosis of testicular cancer and in monitoring patients after chemotherapy. Based on the distribution profile of platinum in the serum of patients with testicular cancer, we
can conclude that metallothionein does not contribute to cisplatin resistance since ~ 95% of serum platinum
binds to medium molecular mass proteins while only ~ 3% is bound to metallothionein. The results of in vitro experiment regarding interactions of cadmium, platinum, selenium and zinc in blood show that simultaneous addition of cadmium and platinum reduces the activity of copper-zinc superoxide dismutase (by 16%) and glutathione peroxidase (by 10%) and induces DNA damage. However, addition of selenium and zinc recovered enzyme activity and repaired the DNA damage. These results emphasize the importance of the less exposure to cadmium, especially for people with malignant disease who are, during treatment and later, exposed to compounds such as cisplatin.- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Provider: - Institution: - Data provided by Europeana Collections- U okviru ovog doktorskog rada, proučavani su mehanizmi pregrađivanja psihofarmaka haloperidola, aripiprazola, paroksetina, ...karbamazepina te lijekovi iz skupine hidantoina i barbiturata. Korištenjem kvantno-kemijskih metoda, izračunate su strukture i reakcijski mehanizmi relevantni za kemijsku sudbinu psihofarmaka u okolišu. Proučavane su reakcije kloriranja, epoksidacije i
hidroksilacije modelnih amina, amida i metabolita psihofarmaka inducirane hipoklorastom kiselinom. Opisani su različiti mehanizmi pregrađivanja (eliminacija, supstitucija, hidridni pomak, ciklizacija), koji uključuju radikalske i ionske međuprodukte. Dobiveni teorijski rezultati odgovaraju eksperimentalnim podacima (konstante brzine reakcija pregrađivanja, spektroskopski podaci o produktima) te ih upotpunjuju s novim fizikalno-kemijskim parametrima.- In this thesis, study of rearrangement reactions of psychopharmaceuticals aripiprazole, haloperidol, paroxetine, and drugs from hydantoin and barbiturate group, is presented. Using quantumchemical methods, structures and reaction mechanisms relevant to chemical fate of psychopharmaceuticals in the environment were calculated. Chlorination, epoxidation, and
hydroxylation reactions for model amides, amines, and psychopharmaceutical metabolites, induced by hypochlorous acid, were studied. Different rearrangement mechanisms (elimination, substitution, hydride shift, cyclization), involving radical and ionic intermediates, were described. Our theoretical results match experimental values (reaction rate constants, spectroscopic product data), and supplement them with additional physicochemical parameters.- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Provider: - Institution: - Data provided by Europeana Collections- Glycans constitute the most abundant and diverse form of the post-translational modifications. While genes
unequivocally determine ...the structure of each polypeptide, there is no genetic template for the glycan part.
Instead, hundreds of genes and their products interact in the very complex pathway of glycan biosynthesis
which is further complicated by environmental influences. Therefore, the aim of this thesis was to
determine the extent to which individual differences in immunoglobulin G and total plasma proteins
glycosylation patterns reflect genetic versus environmental influences. A twin study design was used and
study subjects were twins enrolled in the TwinsUK registry, a national register of adult twins. More than
4500 samples were analyzed by HILIC-UPLC (Hydrophilic Interaction Ultra Performance Liquid
Chromatography). A high contribution of the genetic component to N-glycome composition was found.
Variation in levels of 51 of the 76 IgG glycan traits studied was at least 50% heritable and only a small
proportion of N-glycan traits had a low genetic contribution. Heritability of plasma N-glycome was also
high, with half of the plasma glycan traits being at least 50% heritable. Further, epigenome-wide association
(EWA) analysis showed that methylation levels at some genes are also implicated in glycome composition,
both in those with high heritability and those with a lower genetic contribution. The study to investigate the
potential role of glycosylation in kidney function was also conducted. Fourteen IgG glycan traits were
associated with renal function in discovery population and remained significant after validation in an
independent subset of monozygotic twins discordant for renal disease, reflecting difference in
galactosylation, sialylation, and level of bisecting N-acetylglucosamine. Using the weighted correlation
network analysis (WGCNA) for IgG glycan traits, a correlation between low back pain (LBP) and glycan
modules was established. There was a weak positive correlation between pain phenotypes and "proantibody-
dependent cell-mediated cytotoxicity (ADCC)" WGCNA glycan modules (high bisecting Nacetylglucosamine
and low core fucose) and a weak negative correlation between pain phenotypes and
"anti-ADCC" module (high core fucose, no bisecting N-acetylglucosamine). This suggests that glycans are
promising candidates for biomarkers in many different diseases.- Glikani predstavljaju najzastupljeniji i najraznolikiji oblik posttranslacijske modifikacije. Dok geni
nedvosmisleno određuju strukturu svakog polipeptida, za sintezu glikana ne postoji genski predložak.
Umjesto toga, stotine gena i njihovih produkata sudjeluju u vrlo kompleksnoj biosintezi glikana koju
okolišni utjecaji čine još složenijom. Stoga je cilj ovog doktorskog rada odrediti razmjer kojim genski i
okolišni čimbenici utječu na N-glikane imunoglobulina G (IgG) i ukupnih glikoproteina plazme. Da bi se
postigao navedeni cilj, upotrijebljena je studija na blizancima. Ispitanici su regrutirani TwinsUK registrom,
najvećim registrom blizanaca u Velikoj Britaniji. Više od 4500 uzoraka analizirano je HILIC-UPLC
metodom (kromatografijom vrlo visoke djelotvornosti temeljenoj na hidrofilnim interakcijama, eng.
Hydrophilic Interaction Ultra Performance Liquid Chromatography). Velika genska komponenta
(heritabilnost ≥ 50%) pokazana je za 51 od 76 glikanskih svojstava IgG-a. Nasuprot tome, samo je 12
glikanskih svojstava IgG-a pokazalo malu gensku komponentu. Heritabilnost plazmatskog N-glikoma
također se pokazala velikom. Polovica plazmatskih glikanskih svojstava bila je barem 50% heritabilna.
Epigenomska asocijacijska analiza pokazala je da razina metilacije na nekim genima također utječe na
sastav glikoma, i to na visokoheritabilne i niskoheritabilne glikane. Također je provedeno prvo istraživanje
potencijalne uloge IgG glikozilacije u funkciji bubrega. U prvoj je analizi pronađena značajna povezanost s
bubrežnom funkcijom za 14 glikanskih svojstava, a ostala je značajna i nakon validacije na neovisnoj
podskupini monozigotnih blizanaca diskordantnih za bubrežnu funkciju. Ta glikanska svojstva pripadaju
trima glavnim glikozilacijskim karakteristikama IgG-a: galaktozilaciji, sijalinizaciji i razini račvajućeg Nacetilglukozamina.
Koristeći se WGCNA (eng. weighted correlation network analysis) metodologijom,
provedena je mrežna analiza razina IgG glikana kod blizanaca, da bi se uspostavili klasteri koreliranih
glikana. Pronađene su povezanosti između tih klastera i fenotipova boli kod blizanaca s LBP-om. Opažena
je pozitivna korelacija između fenotipova boli i „pro-stanična citotoksičnost ovisna o protutijelima (eng.
antibody-dependent cell-mediated cytotoxicity, ADCC)“ WGCNA glikanskog modula (visoka razina
račvajućeg N-acetilglukozamina i niska razina sržne fukoze) te negativna korelacija između fenotipova boli
i „anti-ADCC“ modula (visoka razina sržne fukoze, bez račvajućeg N-acetilglukozamina). Rezultati
pokazuju da su glikani obećavajući biomarkeri za mnoge bolesti.- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Provider: - Institution: University of Zagreb. Faculty of Pharmacy and Biochemistry. Department of pharmacognosy. - Data provided by Europeana Collections- U okviru ovog doktorskog rada po prvi put ...je provedeno opsežno usporedno istraživanje fitokemijskog sastava i bioloških učinaka odabranih vrsta roda Thymus hrvatske flore (T. longicaulis C. Presl, T. praecox Opiz subsp. polytrichus (A. Kerner ex Borbás) Jalas, T. pulegioides L., T. serpyllum L. subsp. serpyllum i T. striatus Vahl). Kvalitativna i kvantitativna analiza biološki aktivnih sastavnica provedena je primjenom kromatografskih tehnika (TLC, RP-HPLC-DAD, LC-ESI-MS/MS i GC-MS) i spektrofotometrijskih metoda. Ružmarinska kiselina je bila najzastupljenija polifenolna sastavnica svih istraživanih vrsta. Većina
identificiranih flavonoida bila je iz skupine flavona. Najveći broj pripadao je derivatima luteolina, dok se udjelom isticao skutelarin. Istraživane Thymus vrste značajno su se razlikovale prema sadržaju i sastavu eteričnog ulja. Vrste T. longicaulis i T. pulegioides bile su bogate timolom i/ili karvakrolom, dok je 1,8-cineol bio glavna sastavnica vrste T. serpyllum subsp. serpyllum. U eteričnom ulju planinskih vrsta T. praecox subsp. polytrichus i T. striatus prevladavali su seskviterpenski spojevi. Biološki učinci eteričnih ulja, biljnih ekstrakata i fenolnih sastavnica ispitani su u prikladnim in vitro eksperimentalnim sustavima.
Eterična ulja odabranih Thymus vrsta inhibirala su rast bakterija S. aureus, E. faecalis i E. coli. Ustanovljeno je da se uklapanjem eteričnih ulja u liposome zadržava isti ili se može postići bolji antibakterijski učinak. Istraživanim biljnim ekstraktima dokazana su snažna antioksidacijska svojstva temeljena na sposobnosti hvatanja slobodnih radikala, redukciji i keliranju iona željeza te sposobnost inhibicije lipidne peroksidacije. Thymus ekstrakti su pokazali sposobnost inhibicije Src tirozin kinaze te su
smanjili produkciju proupalnog citokina IL-6 u splenocitima Balb/C miševa. Rezultati provedenih fitokemijskih i bioloških istraživanja pokazali su da, među Thymus vrstama koje rastu u Hrvatskoj, tankolisna majčina dušica (T. longicaulis) ima najveći potencijal za primjenu u farmaciji, usporediv s farmakopejskom vrstom T. vulgaris.- In this doctoral thesis, a comprehensive comparative study of the phytochemical composition and biological effects of selected Thymus species (T. longicaulis C. Presl, T. praecox Opiz subsp. polytrichus (A. Kerner ex Borbás) Jalas, T. pulegioides L., T. serpyllum L. subsp. serpyllum i T. striatus Vahl) from Croatian flora was conducted for the first time. Qualitative and quantitative chemical analyses of bioactive compounds were carried out using various chromatographic (TLC, RP-HPLC-DAD, LC-ESI-MS/MS and GC-MS) and spectrophotometric methods. Rosmarinic acid was found to be the most abundant polyphenolic constituent of all investigated species. Most of the identified flavonoids were from the group of flavones, the largest number belonged to luteolin derivatives, while scutellarin was present with the highest concentration. The investigated Thymus species were significantly different according to the content and composition of
essential oils. Thymol and/or carvacrol were found to be the main components of essential oils of T. longicaulis and T. pulegioides, while T. serpyllum subsp. serpyllum was rich in 1,8-cineole. The mountain species T. praecox subsp. polytrichus and T. striatus were characterised by a high percentage of sesquiterpenes. The biological effects of essential oils, plant extracts and phenolic constituents were studied using different in vitro assays. Selected Thymus essential oils inhibited growth of bacterial strains S. aureus, E. faecalis and E. coli. It was found that encapsulation of essential oils in liposomes can maintain or even improve their antibacterial efficiency. The studied plant extracts displayed strong antioxidant properties by acting as free radical scavengers and reducing agents and, additionally, by chelating transition metals and inhibiting lipid peroxidation. The Thymus extracts showed Src tyrosine kinase inhibitory activity and they
reduced the production of proinflammatory cytokine IL-6 in Balb/C mice splenocytes. The results of phytochemical and biological investigations suggest that, among Croatian Thymus species, T. longicaulis has the greatest potential for pharmaceutical usage comparable to that of pharmacopoeial species T. vulgaris.- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Provider: - Institution: - Data provided by Europeana Collections- Mikofenolna kiselina (MPA) pokazuje varijabilnu farmakokinetiku (PK). Metabolizira se enzimima UGT u MPA-glukuronid koji podliježe ...enterohepatičkoj recirkulaciji. U izlučivanje putem žuči i bubrega uključen je i protein multirezistencije 2 (MRP2) kodiran genom ABCC2. Ispitali smo PK MPA u stanju ravnoteže u ispitanika s bubrežnim presatkom u odnosu na genotip ABCC2 donora i primatelja, uz druge kliničke kovarijable uključujući komedikaciju s ciklosporinom ili takrolimusom. Bolesnici (n=68, muškarci=53%, dob 15-72 godine) su dobivali mikofenolat-natrij, (EC-MPS) (n= 45, 2x720 mg/dan) ili mikofenolat mofetil (MMF) (n=23, 2x500 do 2x1000 mg/dan) i svi su dobivali ciklosporin (n=43) ili takrolimus (n= 25) te kortikosteroide. Uzorak krvi se uzimao tijekom jednog intervala doziranja (12 sati) u 0, 0.5, 1, 2, 3, 8, 12 h nakon jutarnje doze. MPA je analizirana metodom HPLC. Genotipizacija ABCC2 -24C>T i 1249G>A je provedena metodom PCR u stvarnom vremenu. PK parametri (ln-transformirani) su analizirani prilagodbom generalnog linearnog modela s neovisnim parametrima za dob, spol, indeks tjelesne mase, liječenje (EC-MPS ili MMF), vrstu kalcineurinskog inhibitora te genotip donora i primatelja (-24C>T i 1249G>A). S obzirom na genotip donora ABCC2: alel -24T je neovisno povezan s nižimom Cmin (omjer geometrijskih sredina, GMR=0.61, 90% CI 0.40-0.92), nižom ostatnom koncentracijom Co (GMR=0.54, 0.35-0.83), nižom ostatnom koncentracijom C12 (GMR 0.61, 0.37-0.99) i većim oscilacijama koncentracija MPA (GMR=1.88, 1.09-3.23). S obzirom na genotip primatelja ABCC2: alel 1249A je neovisno povezan s nižom Cmin (GMR=0.55, 0.36-0.66), nižom ostatnom koncentracijom C12 (GMR=0.54, 0.32-0.90) i većim oscilacijama koncentracija MPA (GMR=1.85, 1.05-3.28). Pacijenti liječeni ciklosporinom i koji su bili nositelji alela ABCC2 1249A (niska aktivnost), imali su za 30 % niži AUC MPA u odnosu na pacijenate liječene s takrolimusom, a koji su imali genotip visoke aktivnosti, ABCC2 1249GG. Kombinacija ciklosporina i varijantnih alela ABCC2 (-24T, 1249A) bila je povezana s Cmax/AUC, povišenima za 60%, odnosno 52%. Zaključujemo da je farmakokinetika MPA pod značajnim utjecajem polimorfizama ABCC2 donora i primatelja te ciklosporina u istodobnoj terapiji što može biti klinički značajno.- Mycophenolic acid (MPA) displays variable pharmacokinetics (PK). It is metabolized by UGTs to MPA-glucuronide which is subject to enterohepatic recirculation. Biliary and kidney excretion involves multidrug resistant protein 2 (MRP2) coded by ABCC2. We assessed steady-state PK of MPA in renal allograft recipients in respect to donor and recipient ABCC2 genotypes, along with other clinical covariates including commedication with cyclosporine or tacrolimus. Patients (n=68, men=53%, age 15-72 years) were treated with mycophenolate sodium (EC-MPS) (n= 45, 2x720 mg/day) or mycophenolate mofetil (MMF) (n=23, 2x500 to 2x1000 mg/day), and all received cyclosporine (n=43) or tacrolimus (n= 25) and corticosteroids. Blood samples were taken during one dosing-interval (12 hrs) at 0, 0.5, 1, 2, 3, 8, and 12 h after the morning dose. MPA was analysed by HPLC method. Genotyping of -24C>T and 1249G>A was performed by Real-time PCR method. PK parameters (ln-transformed) were analysed by fitting generalized linear models with independents: age, sex, body mass index, treatment (EC-MPS or MMF), calcineurin inhibitor type, donor and recipient genotypes (-24C>T and 1249G>A). Regarding the donor ABCC2 genotypes: -24T allele was independently associated with lower Cmin (geometric mean ratio, GMR=0.61, 90% CI 0.40-0.92), lower trough Co (GMR=0.54, 0.35-0.83), lower trough C12 (GMR 0.61, 0.37-0.99) and greater MPA concentration oscillations (GMR=1.88, 1.09-3.23). Regarding the recipient genotypes: 1249A allele was independently associated with lower Cmin (GMR=0.55, 0.36-0.66), lower trough C12 (GMR=0.54, 0.32-0.90) and greater % swing (GMR=1.85, 1.05-3.28). Patients treated with cyclosporine and who were carriers of ABCC2 1249A (low activity) , had a 30% lower AUC MPA compared to the patients treated with tacrolimus and who had the high activity genotype, 1249GG. The combination of cyclosporine and variant alleles ABCC2 (-24T, 1249A) was associated with a Cmax / AUC, increased by 60 % and 52 %. In conclusion, the pharmacokinetics of MPA is significantly affected by the donor and recipient ABCC2 polymorphisms and cyclosporine in concomitant therapy, which could be clinically relevant.
The thesis is- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Provider: - Institution: University of Zagreb. Faculty of Pharmacy and Biochemistry. Centre for applied pharmacy. - Data provided by Europeana Collections- Dosadašnja klinička istraživanja upućuju na ...zaključak da američki ginseng (AG) pridonosi regulaciji glikemije na način da, slično djelovanju derivata sulfonilureje, povećava postprandijalnu koncentraciju inzulina u serumu. Stoga je cilj ovog istraživanja bio ispitati učinkovitost i sigurnost odabranog ekstrakta američkog ginsenga kao dopune uobičajenoj terapiji ŠBT2 (prehrana i/ili lijekovi), koristeći randomizirani, dvostruko-slijepi, placebom-kontrolirani, paralelni dizajn studije i posljedično pridonijeti razumijevanju predloženog mehanizma djelovanja ekstrakta američkog ginsenga u regulaciji glikemije u bolesnika sa ŠBT2. Sedamdeset četiri ispitanika s kontroliranom ŠBT2 (spol: 28 M i 46 Ž; dob: 62,9 ± 9,49 godina; ITM: 32 ± 5,3 kg/m2; HbA1c: 7,0 ± 1,3%), randomiziranih u intervencijsku (n = 35) ili kontrolnu skupinu (n = 39), završilo je studiju. Svakom je ispitaniku bio dodijeljen ili odabrani preparat AG-a (etanolni ekstrakt korijena) ili placebo koji sadrži pšenične mekinje (2 kapsule od 500 mg po obroku = 3 g dnevno) u razdoblju od 12 tjedana. Ishodi mjerenja uključivali su mjere učinkovitosti (hemoglobin A1c (HbA1c)-primarni, koncentracija glukoze u plazmi natašte (GUP-NT), koncentracija inzulina u serumu natašte i model procjene homeostaze (HOMA-IR)); sigurnosti primijenjene terapije (jetrena i bubrežna funkcija, koagulacijski parametri, arterijski tlak i arterijska elastičnost) i suradljivosti (broj vraćenih kapsula). Među ispitanim skupinama nije bilo statistički značajne razlike u vrijednostima hemoglobina A1c kao primarnog ishoda istraživanja. Ipak, u usporedbi s placebom, u skupini koja je uzimala AG uočen je silazni trend vrijednosti hemoglobina A1c za 0,84% (P = 0,836) te su ispitanici za vrijeme studije ostali dobro kontrolirani (HbA1c = 7,12%). Odabrani pripravak AG-a također je smanjio koncentraciju GUP-a u plazmi natašte za 4,42%, koncentraciju inzulina u serumu natašte za 11,54% i vrijednosti HOMA-IR indeksa za 18,84%, bez statistički značajne razlike među uspoređivanim skupinama (P = 0,947, P = 0,373, P = 0,489, za svaki pojedinačno). Razlika početnih i krajnjih vrijednosti vaskularnih pokazatelja između dviju skupina, i to sistoličkog arterijskog tlaka (SAT) i augmentacijskog indeksa (AI), bila je statistički značajna. U usporedbi s placebom, AG u primijenjenoj dozi značajno je smanjio radijalni AI za 5,25% (P = 0,041) i SAT za 11,72% (P < 0,001), dok je izostao učinak na dijastolički arterijski tlak. Ishodi sigurnosti i suradljivosti ostali su također nepromijenjeni. Dobiveni rezultati upućuju na zaključak da nakon 12 tjedana suplementacije ekstrakt AG-a, iako ne smanjuje vrijednost HbA1c, održava zadovoljavajuću kontrolu glikemije u bolesnika s reguliranom ŠBT2, bez nuspojava.- According to previous clinical studies it has been suggested that American ginseng (AG) is effective in improving glycemic control in type 2 diabetes through increasing post-prandial insulin levels, much like the conventional therapy sulphonylurea. The aim of the present study was to test the efficacy and safety of 12 weeks of supplementation with the selected AG extract as an adjunct to usual therapy in people with type 2 diabetes (diet and/or conventional diabetes therapy), using a randomized, double-blind, placebo-controlled design, and subsequently to contribute to better understanding of the proposed mechanism of action of the American ginseng extract in glucose regulation in type 2 diabetes. Seventy-four participants with well-controlled type 2 diabetes (sex: 28 M and 46 F, age: 62.9 ± 9.49, BMI: 32 ± 5.3, HbA1c: 7.0 ± 1.3), randomized to either intervention (n = 35) or control (n = 39) group, completed the study. Participants were randomly assigned to one of the two interventions and received, prior to each of the main meals, three times daily two 500 mg capsules (total 3g/ day), of either American ginseng extract or identical-appearing placebo capsules containing wheat bran. Outcomes included measures of efficacy (glycated hemoglobin (HbA1c)-primary, fasting plasma glucose (FPG), fasting plasma insulin (FPI), homeostasis model assessment (HOMA-IR)); safety (liver and kidney function, haemostatic function, blood pressure and arterial stiffness); and compliance (returned capsules). There was no change in the primary endpoint, HbA1c, between the treatment and placebo groups. Nevertheless, there was a trend toward lower HbA1c by 0.84% (P = 0.836) compared to placebo. Furthermore, the participants remained well-controlled (HbA1c = 7.12%) throughout. The selected AG treatment also decreased FPG, FPI and HOMA by 4.42%, 11.54% and 18.84%, respectively, with no significant between-treatment difference (P = 0.947, P = 0.373, P = 0.489, respectively). Between-treatment end differences of vascular parameters, namely systolic blood pressure (SBP) and augmentation index (AI) were statistically significant. Compared to placebo, 3 g of AG significantly lowered radial AI by 5.3% (P = 0.041) and systolic BP by 11.7% (P< 0.001) at 12 weeks. No effect was observed with diastolic BP. Safety and compliance outcomes remained unchanged. The study results indicate that after 12-weeks of supplementation, even though the AG extract did not decrease HbA1c levels, it maintained good glycemic control in patients with well-controlled type 2 diabetes without adverse effects.- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
Provider: - Institution: University of Zagreb. Faculty of Pharmacy and Biochemistry. Department of pharmaceutical technology. - Data provided by Europeana Collections- Nesanica je jedan od najčešćih ...poremećaja spavanja koji zahvaća oko 30% odrasle populacije, a značajno je
češći kod starije populacije i psihičkih bolesnika. Zahvaljujući svojim farmakološkim svojstvima, zaleplon je hipnotik
izbora za kratkotrajno liječenje nesanice. Međutim, slaba topljivost i razgradnja u jetri ograničavaju mu oralnu
bioraspoloživost na samo 30%. Stoga je cilj ovog rada pripremom inkluzijskih kompleksa s ciklodekstrinima
poboljšati topljivost lijeka u vodi, a uklapanjem u oblik s kontroliranim oslobađanjem razviti novi terapijski sustav
koji bi mogao značajno unaprijediti terapijsku učinkovitost ovog lijeka.
Spektrofluorimetrijska i solubilizacijska ispitivanja su pokazala da β-ciklodekstrin sa zaleplonom stvara
komplekse prikladne stabilnosti, no topljivost nastalog kompleksa je ograničena. Od kemijski modificiranih derivata,
nasumično metilirani β-ciklodekstrin se pokazao najprikladnijim, dok je topljivost i stabilnost inkluzijskih kompleksa
zaleplona sa sulfobutil-β-ciklodekstrinom i hidroksipropil-β-ciklodekstrinom nešto niža. Nastajanje inkuzijskih
kompleksa s navedenim derivatima ciklodekstrina je potvrđeno primjenom 1H NMR spektroskopije, a određena je i
njihova struktura. Sustav karakterizira istovremeno prisustvo dva načina vezanja. Prvi, dominantni način uključuje
inkluziju fenilnog prstena molekule zaleplona u centralnu šupljinu molekule ciklodekstrina, a drugi inkluziju
pirazolo1,5-a pirimidinskog prstena molekule lijeka, dok je nastajanje kompleksa višeg reda isključeno.
Termodinamička ispitivanja su pokazala da je nastajanje kompleksa zaleplona s β-ciklodekstrinom i njegovim
metiliranim i hidroksipropiliranim derivatima entalpijom uvjetovani proces, dok je kompleksiranje lijeka sa sulfobutil-
β-ciklodekstrinom entropijom vođen proces. U oba slučaja riječ je o spontanom procesu.
Dodatkom hipromeloze je značajno povećan solubilizacijski potencijal metiliranog β-ciklodekstrina uslijed
nastajanja ternarnih kompleksa lijek-ciklodekstrin-polimer, dok hidrofilni polimeri nisu utjecali na topljivost i
stabilnost kompleksa zaleplona s β-ciklodekstrinom. Binarni i ternarni kompleksi zaleplona s navedenim
ciklodekstrinima i hidrofilnim polimerima u čvrstom stanju pripremljeni su tehnikom sušenja raspršivanjem.
Diferencijalna pretražna kalorimetrija, difrakcija rentgenskih zraka na prašku te pretražna elektronska mikroskopija su
pokazale djelomičnu amorfizaciju lijeka u binarnim i ternarnim kompleksima s β-ciklodekstrinom, dok su kompleksi s
metiliranim-β-ciklodekstrinom bili u potpunosti amorfne strukture. Binarni kompleksi s metiliranim-β-
ciklodekstrinom te ternarni kompleksi s metiliranim-β-ciklodekstrinom i hipromelozom bili su najučinkovitiji u
povećanju topljivosti zaleplona u odnosu na ostale komplekse. Korištenjem manitola kao topljivog punila te ternarnog
kompleksa zaleplona s metiliranim β-ciklodekstrinom i hipromelozom pripremljene su tablete koje omogućuju
oslobađanje doze lijeka u samo 5 minuta, osiguravajući brz početak djelovanja lijeka. Topljivi kompleksi zaleplona s u
vodi topljivim, polimernim derivatom ciklodekstrina su pripremljeni i tehnikom mljevenja u vibracijskim
mikromlinovima. Nakon 90 minuta mljevenja pri frekvenciji od 24 Hz dobiven je amorfni produkt koji otapanjem u
vodi prelazi u inkluzijski kompleks, što je dokazano 1H-NMR spektroskopijom.
Primjenom tehnike sušenja raspršivanjem pripremljene su mikrosfere s pH ovisnim oslobađanjem zaleplona
kao novi terapijski sustav za liječenje nesanice koju karakterizira prerano buđenje te nemogućnost naknadnog
usnivanja. Matriks koji sadrži 80% Eudragita S100 i 20% glicerolmonostearata pokazao se optimalnim u sprječavanju
neželjenog oslobađanja lijeka pri nižim pH-vrijednostima medija, dok je uklapanjem 50% doze lijeka u obliku
inkluzijskog kompleksa s metiliranim β-ciklodekstrinom osigurano potpuno oslobađanje lijeka u umjetnom crijevnom
mediju, kinetikom nultog reda (k0=155 μg/min).- Insomnia is one of the most common sleep disorders affecting about 30% of adult population, while its
prevalence is even higher among elderly and psychiatric patients. Pharmacological properties makes zaleplon the drug
of choice for the short term treatment of insomnia, but its low aqueous solubility and intensive metabolism in liver are
reducing its oral bioavailability to only 30%. Therefore, the aim of this work was to improve aqueous solubility of
zaleplon through inclusion complexation and to develop a novel drug delivery system able to enhance its therapeutical
efficiency.
Spectrofluorimetric and phase solubility studies showed that parent β-cyclodextrin forms stable inclusion
complexes with zaleplon, but aqueous solubility of the complexes formed was limited. Among chemically modified β-
cyclodextrin derivatives, randomly methylated derivative was the most efficient, while the solubility and stability of
zaleplon complexes with sulphobutyl- and hydroxypropyl-β-cyclodextrin was less pronounced. The inclusion complex
formation in all cases was confirmed by 1H-NMR spectroscopy. The structure of the complexes formed was
characterised by the presence of two different binding modes which existed simultaneously in the solution. The first
and the dominant one occurs through inclusion of phenyl moiety of the drug into central cavity of cyclodextrin
molecule, while the other involves the inclusion of the pyrazolo1,5-apyrimidine ring of the drug. The formation of
higher order complexes was not demonstrated. Thermodynamic studies showed that complexation of zaleplon with β-
cyclodextrin and its randomly methylated and hydroxypropylated derivatives was enthalpy driven, while entropy
driven complexation was observed in case of sulphobutyl-β-cyclodextrin. In all cases, inclusion complex formation
was a spontaneous process.
Addition of hypromellose enhanced significantly the solubilising and complexing potential of randomly
methylated β-cyclodextrin due to ternary complex formation, while hydrophilic polymers had no effect on solubility
and stability of zaleplon complexes with parent β-cyclodextrin. Binary and ternary zaleplon complexes with
cyclodextrins and polymers tested were prepared by spray-drying. Differential scanning calorimetry, X-ray powder
diffractometry and scanning electron microscopy showed only partial drug amorphization of the drug in binary and
ternary complexes with β-cyclodextrin, while all complexes with randomly methylated β-cyclodextrin were
completely amorphous. The inclusion complex formation in spray-dried products was confirmed by the solid state
NMR. Binary and ternary complexes with randomly methylated β-cyclodextrin and hypromellose were the most
efficient in increasing the dissolution rate of zaleplon. The use of mannitol as a soluble filler and ternary inclusion
complex with randomly methylated β-cyclodextrin and hypromellose allowed formulation of tablets which released
the complete drug dose in 5 minutes, providing fast onset of drug action. Highly soluble zaleplon complexes with
soluble, polymeric β-cyclodextrin derivative were also prepared by co-grinding in high energy vibrational micromill.
Co-grinding at frequency of 24 Hz resulted in an amorphous product which is readily soluble in water, forming
inclusion complexes as demonstrated by 1H-NMR spectroscopy.
Spray-drying technique was used to prepare pH-responsive zaleplon microspheres with delayed release
properties as a novel drug delivery system for treatment of specific type of insomnia characterised with premature
awaking and inability to fall asleep again. Matrix containing 80% of Eudragit S100 and 20% of glycerol monostearate
was demonstrated to be optimal, preventing the drug release at high pH-values. The incorporation of 50% of the drug
dose in form of binary inclusion complex with randomly methylated β-cyclodextrin ensured complete drug release in
simulated intestinal media, with the zero-order release rate (k0=155 μg/min).- All metadata published by Europeana are available free of restriction under the Creative Commons CC0 1.0 Universal Public Domain Dedication. However, Europeana requests that you actively acknowledge and give attribution to all metadata sources including Europeana
