Provider: - Institution: University of Zagreb. Faculty of Pharmacy and Biochemistry. Department of microbiology. - Data provided by Europeana Collections- Fumonizin B1 (FB1), bovericin (BEA) i ...okratoksin A (OTA) su svjetski rašireni mikotoksini koji izazivaju čitav
niz patoloških promjena in vivo i in vitro, a čije je istodobno pojavljivanje u kukuruzu dokazano na području
Hrvatske. Stoga je u ovom radu ispitan mehanizam djelovanja FB1, BEA i OTA i njihove interakcije u PK 15
stanicama (epitel bubrega svinje), praćenjem preživljavanja stanica, integriteta plazmatske i mitohondrijske
membrane, razine lipidne peroksidacije i antioksidacijskog kapaciteta stanice te stupnja apoptoze i
genotoksičnog učinka niskih koncentracija mikotoksina (0,05 g/mL, 0,5 g/mL i 5 g/mL), tijekom 24 i 48
sati. Fumonizin B1, BEA i OTA su citotoksični za PK 15 stanice što se očituje smanjenjem vijabilnosti i
povećanjem katalitičke aktivnosti laktat dehidrogenaze i glutamat dehidrogenaze ovisno o dozi mikotoksina. Sva
tri mikotoksina u najvećoj koncentraciji značajno stimuliraju lipidnu peroksidaciju, a već pri nižim
koncentracijama smanjuju koncentraciju glutationa u PK 15 stanicama. Fumonizin B1, BEA i OTA uzrokuju
apoptozu PK 15 stanica pri čemu se značajnije morfološke apoptotičke promjene zamjećuju nakon 48-satnog
izlaganja. Okratoksin A ima jače djelovanje na aktivnost kaspaze-3 već nakon 24-satnog tretiranja u
koncentracijama od 0,5 i 5 g/mL, dok BEA i OTA uzrokuju značajnije povećanje aktivnosti ovog enzima
nakon duljeg izlaganja. FB1 i BEA su genotoksični za PK 15 stanice u koncentracijama od 0,5 i 5 g/mL, dok
OTA značajno povećava broj mikronukleusa već kod najmanje koncentracije. Sva tri mikotoksina imaju
klastogeni i aneugeni učinak pri čemu je klastogeno djelovanje OTA i BEA više izraženo. U koncentracijama
0,05 i 0,5 g/mL nakon 24- i 48-satnog izlaganja, kombinacije dva i sva tri mikotoksina mikotoksina pokazuju
uglavnom aditivane a manje sinergističke interakcije što se odražava na smanjenje vijabilnosti, narušavanje
integriteta stanične i mitohondrijske membrane, stimulaciju lipidne peroksidacije i smanjnje antioksidacijskog
kapaciteta te apoptozu i mutagenezu. Nakon 24-satnog izlaganja navedenim kombinacijama u koncentraciji 5
g/mL zapažen je aditivan, sinergistički i antagonistički učinak, a nakon 48-satnog tretmana u istoj koncentraciji
više je izražen antagonizam. Istodobni unos niskih koncentracija ovih mikotoksina putem hrane negativno se
odražava na antioksidacijski sustav te može pridonijeti imunosupresiji, nefrotoksičnosti i karcinogenezi u ljudi i
životinja tijekom dulje izloženosti.- Fumonisin B1 (FB1), beauvericin (BEA) and ochratoxin A are widespread mycotoxins, which cause a great
number of pathological changes in vivo and in vitro. Their co-occurrence was established in maize in Croatia.
The mechanism of action of FB1, BEA and OTA, as well as their interactions, were studied in the PK 15 cells
(porcine kidney epithelial cells). We examined the effects of low concentrations (0.05, 0.5 and 5 g/mL) of
individual and combined mycotoxins on cell viability, mitochondrial and plasma membrane integrity, lipid
peroxidation, antioxidative status, and apoptotic and genotoxic events in PK 15 cells. Decrease of cell viability
and increase of catalytic activity of LDH and GLDH show that FB1, BEA and OTA are cytotoxic to PK 15 cell
in dose dependent manner. Individual treatment of cells with 5 g/mL of FB1, BEA and OTA significantly
stimulated lipid peroxidation, while lower concentrations significantly reduced levels of glutathione. Significant
morphological apoptotic changes were observed after 48 hours of exposure to individual mycotoxins. Ochratoxin
A activated caspase-3 already after 24 hours of treatment with 0.5 and 5 g/mL, while BEA and OTA
significantly affected this enzyme after prolonged exposure. Fumonisn B1 and BEA are genotoxic to PK 15 cells
in concentrations of 0.5 and 5 g/mL, while OTA significantly increased the number of micronuclei already after
treatment with lowest concentrations. All of three mycotoxins mainly act as clastogens. Combined treatment
with two or all of three mycotoxins in concentrations of 0.05 and 0.5 g/mL during 24 and 48 hours, showed
mainly additive and less synergistic interactions on cell viability, mitochondrial and plasma membrane integrity,
stimulation of lipid peroxidation and decrease of antioxidative capacity, apoptotic and genotoxic events in PK 15
cells. After 24 hours of exposure to combined mycotoxins in concentration of 5 g/mL, additive, synergistic and
antagonistic effects were observed, while after 48 hours of exposure to same concentration we observed
antagonism. Simultaneous prolonged exposure to mycotoxins in low dietary concentrations can negatively affect
antioxidant status contributing the immunosuppression, nephrotoxicity and carcinogenicity in animals and
humans.- 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 analysis. - Data provided by Europeana Collections- Provedena je botanička i ...fitokemijska karakterizacija vrsta roda Plantago L. iz zapadnog
dijela Hrvatske: P. altissima L., P. argentea Chaix, P. coronopus L., P. holosteum Scop. subsp.
depauperata, P. holosteum Scop. subsp. holosteum, P. holosteum Scop. subsp. scopulorum, P.
lagopus L. i P. maritima L. Utvrđena su morfološko-anatomska obilježja istraživanih biljnih
taksona i detaljno opisana građa listova i stabljika, te oblik i raspored trihoma. Histokemijskim
reakcijama dokazane su trjeslovine, sluzi, glikozidi, alkaloidi i proteini. Primjenom TLC metode i
taložnih reakcija utvrđena je prisutnost iridoida, flavonoida, fenolnih kiselina, trjeslovina,
triterpenskih kiselina, saponina i sterola. Unaprijeđena je MEKC metoda i provedena
karakterizacija iridoidnih sastavnica. Vodeni ekstrakti listova sadržavali su do 0,27% aukubina i
do 1,81% katalpola (P. argentea). U većine uzoraka detektirane su još dvije iridoidne supstancije.
Spektrofotometrijski određen sadržaj polifenolnih spojeva u nadzemnim biljnim organima najveći
je u taksona P. holosteum subsp. holosteum, a trjeslovinama i flavonoidima najbogatiji su
nadzemni dijelovi vrste P. argentea. Primjenom programskog paketa ESKULAP potvrđena je
valjanost analitičkog postupka za određivanje tanina s Folin-Ciocalteusovim fenolnim reagensom.
Reverzno-faznom HPLC analizom uspješno je provedena karakterizacija flavonoida i fenolnih
kiselina u metanolno-vodenim ekstraktima listova. Maseni udjeli tih tvari znatno su varirali među
taksonima: rutin (0,005-0,024%), hiperozid (0,007-0,020%), izokvercitrin (0,020%, P. argentea)
kvercitrin (0,001-0,013%), kvercetin (0,006-0,028%), klorogenska kiselina (0,002-0,115%) i
kavena kiselina (0,001-0,046%). Utvrđena je statistički značajna varijabilnost u distribuciji
analiziranih bioaktivnih tvari među analiziranim vrstama. Eksperimentalni fitokemijski podaci
obrađeni su metodom multivarijatne statistike. Utvrđeno je da se vrsta P. argentea fitokemijski
najviše razlikuje od ostalih taksona i da posjeduje najveći fitoterapijski potencijal od svih
istraživanih vrsta roda Plantago L.- Plantago altissima L., P. argentea Chaix, P. coronopus L., P. holosteum Scop. (subsp. depauperata,
subsp. holosteum and subsp. scopulorum), P. lagopus L. and P. maritima L. were botanically and
phytochemically investigated. Complete morphological and anatomical characterization of leaves and
stems was carried out. Histochemical reactions revealed the presence of tannins, mucilage, glycosides,
alkaloids, inulin and proteins. The existence of iridoids (aucubin and catalpol), flavonoids (rutin, hyperosid,
isoquercitrin, quercitrin and quercetin), saponin substances, triterpenes (oleanolic and ursolic acids) and
sterols (β-sitosterol and stigmasterol) was confirmed using TLC. Micellar electrokinetic chromatography
(MEKC) was used for characterization of iridoid substances. The yield of aucubin and catalpol was up to
0.27% and 1.81% of the dry mass of the leaves, respectively. Besides aucubin and catalpol, two related
compounds were determined in plant samples. The results of spectrometric measurements showed that P.
argentea and P. holosteum subsp. holosteum generally contained the highest amounts of total polyphenols,
tannins and flavonoids, while the lowest contents were measured in samples of P. coronopus.
Mathematical and statistical program ESKULAP was used for the evaluation of the analytical procedure
for tannin determination with Folin-Ciocalteu's phenol reagent. Characterizations of flavonoids and
phenolic acids were successfully performed by reversed phase HPLC. Yields of analysed substances varied
among examined species: rutin (0,005-0,024%), hyperosid (0,007-0,020%), isoquercitrin (0,020%, P.
argentea), quercitrin (0,001-0,013%), quercetin (0,006-0,028%), chlorogenic acid (0,002-0,115%) and
caffeic acid (0,001-0,046%). Experimental phytochemical data were evaluated by cluster analysis.
Significant differences were observed in contents and distribution of iridoids, flavonoids and phenolic acids
between investigated plant species. These substances might be good chemotaxonomical markers in the
study of genus Plantago. The results of present study showed that P. argentea had the highest
phytotherapeutic potential of all examined Plantago species and it could be a good resource of biologically
active compounds.- 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
