Content of bioactive constituents and antioxidant potential of Galium L. species

Qualitative phytochemical analyses of eight species from the genus Galium (G. corrudifolium, G. cruciata, G. divaricatum, G. lucidum, G. mollugo, G. palustre, G. parisiense, and G. verum), followed by spectrophotometric evaluation of their total phenolic, flavonoid and iridoid content, as well as antiradical capacity, were conducted. G. cruciata contained the greatest amount of total phenolics (111.00 mg gallic acid equivalents g–1 dry extract), while G. verum had the greatest amount of flavonoids (23.11 mg quercetin equivalents g–1 dry extract) and iridoids (461.30 mg aucubin equivalents g–1 dry extract). The best antioxidant (antiradical) activity was shown by G. cruciata (IC50 (ABTS) = 30.30 µg mL–1; IC50 (DPPH) = 27.62 µg mL–1), followed by G. divaricatum, G. verum and G. palustre. The same species were rich in various bioactive constituents and would be appropriate for inclusion in further investigations considering their biomedical potential. This work is licensed under a Creative Commons Attribution 4.0 International License

Učestalost bakterijemija povezanih s trajnim dijaliznim kateterima Incidence of bacteriemia pertinent to implanted intravascular dialysis catheters

Uvod: Centralni venski kateteri se koriste kao privremeni ili trajni vaskularni pristup za svrhe liječenja hemodijalizom. Nazočnost navedenih intravaskularnih katetera povećava rizik od nastanka bakterijemije. Tunelirani venski kateter je sve češće trajno rješenje u svrhe vaskularnog pristupa liječenja hemodijaliznih pacijenata, ali istovremeno predstavlja i povećanje rizika za nastanak infekcija. Ispitanici i metode: Korišteni podaci dobiveni su uvidom i analizom medicinske dokumentacije pacijenata na hemodijalizi i hospitaliziranih pacijenata na Odjelu nefrologije i endokrinologije od 2014. do 2017. godine u Općoj Bolnici „Dr. T. Bardek“ Koprivnica. Rezultati: Obzirom na broj implantiranih pCVK (ukupno 136) u vremenskom razdoblju od dvije godine (2014.-2015.), i na broj implantiranih TDK (ukupno 60) u vremenskom razdoblju sljedeće dvije godine (2016.-2017.) učestalost bakterijemija povezanih s pCVK iznosi 3,67%, tj. 15,15 na 1000 dana implantiranih intravaskularnih proteza, dok učestalost bakterijemija povezanih s TDK iznosi 5,00%, odnosno 2,35 na 1000 kateter dana. Najčešći uzročnik bakterijemija, neovisno o vrsti primjenjenog katetera, bio je Staphylococcus aureus. Zaključak: Odgovornost je na svim zdravstvenim profesionalcima koji skrbe o bolesnicima s implantiranim intravaskularnim kateterima provoditi higijenu ruku i okoliša, kako bi se učestalost bakterijemija povezanih s dijaliznim kateterima smanjila na najmanji obim

Spirulina - ljekovita cijanobakterija

Species of the genus Arthrospira are multicellular fi lamentous cyanobacteria (blue-green algae). Th ey are commonly known as spirulina. Addition of these microalgae to human diet is an emerging trend. High protein content that reaches up to 70 % of dried biomass along with fi ber, carotenoids, minerals and vitamins is a reason why spirulina is considered as a superfood. Consumption of algal biomass has a long tradition dating back to indigenous people of Africa and Mexico area. Spirulina is very rich in pigments. In particular, C-phycocyanin accounts for more than 10 % of dried biomass. Th is pigment is being widely studied for its anti-infl ammatory and antioxidant properties. Spirulina biomass and C-phycocyanin have a very good safety profi le. Th is article is a review on biomedical applications of spirulina biomass as a chemoprotective agent, lipid lowering agent and metabolic modulator, antiinfl ammatory, immunostimulatory, and anti-cancer agent

Contribution to Globularia phylogeny based on nuclear ribosomal spacer and two chloroplast DNA regions

Background and Purpose: Molecular approach has a major impact on phylogenetic studies of plants, considering that it gives useful information about evolutionary events and relations on all taxonomic levels. The sequence data of the nuclear ITS and of two chloroplast regions, trnL-trnF spacer and rbcL gene, obtained from thirteen Globularia L. taxa, including five Anatolian endemics, representing six sections altogether, were analyzed in order to determine the relations between the European and the Anatolian species and get a better insight into the phylogeny of several closely related Globularia taxa.  Materials and Methods: Total cellular DNA was extracted from fresh or frozen leaf tissue of thirteen Globularia samples. The ITS regions of nuclear DNA and two chloroplast DNA regions were amplified and sequenced. Obtained nuclear and combined plastid data matrices were subjected to Maximum Parsimony analyses. Results and Conclusions: Molecular data that were obtained in this study indicate the existence of separate centers of diversification for the European and the Anatolian Globularia. The results provide support for relationships among the studied Anatolian endemic species and indications for a redefinition of affinities of some of the European species. The results presented herein are discussed along with available morphological, karyological, phytogeographical and molecular data. Keywords: Globularia, ITS, trnL-trnF, rbcL, Maximum Parsimony, Quaternary</p

Morphometric and Physiological Characteristics of Brown Trout (Salmo trutta) from the Ponor River

This paper presents the data related to morphometric and physiological (hematological) characteristics of brown trout (Salmo trutta) from the Ponor River. This river rises near the village Podrašnica (municipality of Mrkonjić Grad), sinks above the ground and after a while, near the settlement of Krupa na Vrbasu, appears as a source of the Krupa River (tributary of the Vrbas River). Fish sampling was performed during the summer of 2015 and during this period 22 Salmo trutta units were caught (11 females and 11 males). On that occasion, several morphometric (total and standard body length) and hematological traits (Hct-hematocrit, Hb-hemoglobin and MCHC - mean corpuscular hemoglobin concentration), body mass and Fulton’s condition factor were analyzed. The average value for total body length was 18.85 cm and for standard body length it was 16.85 cm; the mean value for body mass was 80.38 g, and for Fulton’s condition factor 1.41. In case of hematological parameters, the average value for Hct was 0.39 l/l, Hb 70.62 g/l and MCHC 180.64 g/l erythrocytes. The estimated parameters can serve as indicators for the condition of these aquatic organisms, and indirectly, the state of their environment

Diterpenes and Phenolic Compounds from Salvia brachyodon Vandas

Salvia brachyodon, the short-tooth sage, is one of the rarest plant species and endemic in the Adriatic area of the Balkan Peninsula. As aside from its essential oil, only limited information on its phytochemical composition is known, a more detailed study of the leaves was undertaken. From its leaves two diterpenes, agastanol (2), and a new natural compound 1, i.e., 3-methyl-4-methylen-11,12,14-trihydroxy-8,11,13-abietatrien-7-one, were isolated and identified by NMR spectroscopy and mass spectrometry. In addition, caffeic acid, isoquercitrin, luteolin 7-O-glucoside and rosmarinic acid were identfied by comparison with reference compounds. The fraction containg the diterpenes as well as the isolated compound 1 showed significant antimycobacterial activity against Mycobacterium smegmatis. The diterpenes of S. brachyodon represent promising antimycobacterial substances for further evaluation. Due to the endangered nature of the plant, the wide use of S. brachyodon and its bioactive compounds could be achieved by growing the plants in culture

The Mastic Tree (Pistacia lentiscus L.) Leaves as Source of BACs: Effect of Growing Location, Phenological Stage and Extraction Solvent on Phenolic Content

Pozadina istraživanja. Tršlja (Pistacia lentiscus L.) je zimzeleni grm iz porodice Anacardiaceae, rasprostranjen u mediteranskim zemljama, gdje se koristi u narodnoj medicini. Dosadašnja istraživanja ekstrakata lista, stapke, ploda i korijena P. lentiscus pokazuju da biljka sadržava veliku koncentraciju sekundarnih metabolita (flavonoida, fenolnih kiselina i tanina), što upućuje na zaključak da ima izvrstan biološki potencijal. Stoga je svrha ovoga rada bila procijeniti utjecaj okolišnih čimbenika i vrste ekstrakcijskog otapala na koncentraciju fenola u listu tršlje uzgojene na četiri različite lokacije duž Jadranske obale (Barbariga, Lun, Hvar i Vela Luka) tijekom tri fenološke faze (faze cvatnje, ranog ploda i zrelog ploda). Eksperimentalni pristup. Budući da listovi tršlje sadržavaju fenolne spojeve koji se razlikuju po strukturi i kemijskim svojstvima, ispitivanje etanolnih i metanolnih ekstrakata provedeno je visokodjelotvornom tekućinskom kromatografijom (HPLC) s UV-Vis PDA detektorom. Fenolni spojevi su identificirani usporedbom retencijskih vremena i dobivenih spektara s odgovarajućim standardima pri 280 i 340 nm. Rezultati i zaključci. U svim su uzorcima određene koncentracije fenolnih kiselina i flavonol glikozida, dok je koncentracija katehina određena samo u metanolnim ekstraktima. Prevladavajući fenolni spoj bila je 5-O-galoilkininska kiselina, zatim su slijedili monogaloil-glukoza, 3,5-di-O-galoilkininska kiselina, 3,4,5-tri-O-galoilkininska kiselina i galna kiselina. Dominantni flavonol glikozid bio je miricetin-3-O-ramnozid, zatim miricetin-3-O-glukozid, miricetin-3-O-glukuronid, kvercetin-3-O-ramnozid i derivat flavonol glikozida. Rezultati pokazuju da masene koncentracije navedenih spojeva variraju ovisno o fenološkoj fazi, lokaciji i korištenom otapalu. Najveće masene koncentracije fenola utvrđene su u uzorcima lista tršlje ubranim na lokaciji Hvar te ekstrahiranim s 80 %-tnom otopinom metanola. Najveće masene koncentracije ukupnih fenola pronađene su u uzorcima lista tršlje ubranim tijekom fenološke faze cvatnje, a ukupnih flavonoida u uzorcima lista tršlje ubranim tijekom fenološke faze ranog ploda. Novina i znanstveni doprinos. Dobiveni rezultati omogućuju bolje razumijevanje fenolnog sadržaja vrste P. lentiscus, što može potaknuti daljnja ispitivanja valorizacije lista tršlje kao farmaceutskog pripravka ili hrane s dodanom vrijednošću.Research background. Mastic tree (Pistacia lentiscus L.) of the Anacardiaceae family is an evergreen shrub from Mediterranean countries where it is used in traditional medicine. Analysis of P. lentiscus leaf, stem, fruit and root extracts showed high concentrations of principal groups of secondary metabolites (flavonoids, phenolic acids and tannins), suggesting the plant possesses great biological potential. Therefore, the aim of this research is to evaluate the impact of environmental parameters and the extraction solvent type on the concentration of phenols in mastic tree leaf extracts grown at four different locations along the Adriatic coast (Barbariga, Lun, Hvar and Vela Luka) during three phenological stages (early flowering, early fruiting and late fruiting). Experimental approach. Since mastic tree plant has phenolic compounds with different structures and chemical properties, ethanolic and methanolic leaf extracts were analysed using high-performance liquid chromatography (HPLC) coupled with UV/Vis PDA detector. Phenolic compounds were identified by comparing the retention times and spectral data with those of standards at 280 and 340 nm. Results and conclusions. In all samples, phenolic acids and flavonol glycosides were quantified, while catechin was quantified only in methanolic extracts. The 5-O-galloylquinic acid was determined as a predominant phenolic compound in all samples followed by monogalloyl glucose, 3,5-di-O-galloylquinic acid, 3,4,5-tri-O-galloylquinic acid and gallic acid, respectively. Myricetin-3-O-rhamnoside was found to be the predominant flavonol glycoside followed by myricetin-3-O-glucoside, myricetin-3-O-glucuronide, quercetin-3-O-rhamnoside and derivative of flavonol glycoside. The mass concentration of these compounds significantly varied during different phenological stages, at different growing locations and used extraction solvents. The highest phenolic mass concentration was determined in the samples harvested at Hvar growing location and extracted in 80 % methanol. The highest total phenolic acid mass concentration was obtained in the samples harvested during the flowering phenological stage and the highest total flavonoid mass concentration in the samples harvested during the early fruiting stage. Novelty and scientific contribution. The obtained data provide a better understanding of the P. lentiscus species phenolic concentration, which can lead to further investigations regarding the valorisation of mastic tree leaves as pharmaceutical products or as food products with added value

Botanical characteristics, phytochemical profile and biological effects of Globularia L. species

Vrsta Globularia alypum L. široko je primjenjivana biljna vrsta u narodnoj medicini mediteranskih zemalja i najbolje istražena predstavnica roda Globularia L. U okviru ovog doktorskog rada prvi je put provedeno opsežno usporedno istraživanje botaničkih obilježja, fitokemijskog profila i bioloških učinaka četiriju vrsta roda Globularia, koje je, uz vrstu G. alypum, uključivalo tri slabije istražene srodne vrste: G. cordifolia L. (G. cordifolia subsp. cordifolia (L.) Hayek), G. meridionalis (Podp.) O. Schwarz (G. cordifolia subsp. bellidifolia (Ten.) Wettst.) i G. punctata Lapeyr. Istraživanjem su obuhvaćeni uzorci 19 samoniklih populacija, sabrani u vrijeme cvatnje na području Hrvatske i okolice kroz više različitih sezona (do pet). U cilju procjene biomedicinskog potencijala slabije istraženih vrsta provedena je kvalitativna i kvantitativna kemijska analiza biološki aktivnih sastavnica primjenom različitih kromatografskih i spektroskopskih tehnika (TLC, HPLC-DAD-ESI-MS, LC-PDA-ESI-MSn, UV/Vis spektrofotometrija, GC-FID, GC-MS), a prikladnim metodama u uvjetima in vitro ispitani su odabrani biološki učinci prethodno utvrđeni kod vrste G. alypum (antidijabetski, antioksidativni, protuupalni, antimikrobni i citotoksični). Navedenim istraživanjima prethodila su temeljna botanička istraživanja na ekološkoj, morfološkoj, histološko-anatomskoj, citološkoj i molekularnoj razini, kojima se u kombinaciji s fitokemijskim istraživanjima pokušalo pridonijeti rješavanju taksonomskog spora oko razdvojenosti vrsta G. cordifolia i G. meridionalis. Sve četiri istraživane vrste pokazale su antidijabetski, antioksidativni, protuupalni, antimikrobni i citotoksični učinak u uvjetima in vitro, što je bilo moguće povezati s njihovim utvrđenim fitokemijskim profilima. Rezultati usporedbe fitokemijskog profila ukazali su na veću sličnost vrste G. punctata (podrod Globularia) s vrstom G. alypum (podrod Abolaria) od ostalih dviju vrsta (podrod Abolaria). Podjelu na podrodove nije podupirala ni usporedba histološko-anatomskih, molekularnih i fitokemijskih značajki, koja je ukazala na veću sličnost vrsta G. cordifolia i G. meridionalis s vrstom G. punctata. Također, kod uzoraka uključenih u ovo istraživanje nisu uočena karakteristična ekološka, morfološka, histološko-anatomska, citološka, molekularna ili fitokemijska obilježja koja bi podupirala razdvajanje na vrste G. cordifolia i G. meridionalis.Globularia alypum L. is a plant species widely used in folk medicine of Mediterranean countries and the best explored representative of the genus Globularia L. In this doctoral thesis, a comprehensive comparative study of botanical characteristics, phytochemical profile and biological effects of four Globularia species, which, apart from G. alypum, included three less explored related species: G. cordifolia L. (G. cordifolia subsp. cordifolia (L.) Hayek), G. meridionalis (Podp.) O. Schwarz (G. cordifolia subsp. bellidifolia (Ten.) Wettst.) and G. punctata Lapeyr., was conducted for the first time. Samples from 19 natural populations, collected during flowering time in Croatia and surrounding area in several different seasons (up to five), were used in this study. In order to evaluate the biomedical potential of less investigated species, qualitative and quantitative chemical analyses of biologically active constituents were conducted using different chromatographic and spectroscopic techniques (TLC, HPLC-DAD-ESI-MS, LC-PDA-ESI-MSn, UV/Vis spectrophotometry, GC-FID, GC-MS), while suitable in vitro methods were used to evaluate selected biological effects, previously established for G. alypum (antidiabetic, antioxidative, anti-inflammatory, antimicrobial and cytotoxic). The aforementioned research was preceded by basic botanical research on ecological, morphological, histo-anatomical, cytological and molecular level, by which, in combination with the phytochemical research, an attempt was made to contribute to the resolution of the taxonomic dispute over the separation of the species G. cordifolia and G. meridionalis. All four examined species showed antidiabetic, antioxidative, anti-inflammatory, antimicrobial and cytotoxic activity in vitro, which could be linked to their established phytochemical profiles. The results of the comparison of phytochemical profiles showed greater similarity of G. punctata (subgenus Globularia) to G. alypum (subgenus Abolaria) than those of the other two species (subgenus Abolaria). The division into subgenera was also not supported by the comparison of histo-anatomical, molecular and phytochemical characteristics, which indicated greater similarity of G. cordifolia and G. meridionalis to G. punctata. Moreover, no ecological, morphological, histo-anatomical, cytological, molecular or phytochemical characteristics, that would support the separation of G. cordifolia and G. meridionalis into different species, were observed for the samples included in this study

Botanical characteristics, phytochemical profile and biological effects of Globularia L. species

Vrsta Globularia alypum L. široko je primjenjivana biljna vrsta u narodnoj medicini mediteranskih zemalja i najbolje istražena predstavnica roda Globularia L. U okviru ovog doktorskog rada prvi je put provedeno opsežno usporedno istraživanje botaničkih obilježja, fitokemijskog profila i bioloških učinaka četiriju vrsta roda Globularia, koje je, uz vrstu G. alypum, uključivalo tri slabije istražene srodne vrste: G. cordifolia L. (G. cordifolia subsp. cordifolia (L.) Hayek), G. meridionalis (Podp.) O. Schwarz (G. cordifolia subsp. bellidifolia (Ten.) Wettst.) i G. punctata Lapeyr. Istraživanjem su obuhvaćeni uzorci 19 samoniklih populacija, sabrani u vrijeme cvatnje na području Hrvatske i okolice kroz više različitih sezona (do pet). U cilju procjene biomedicinskog potencijala slabije istraženih vrsta provedena je kvalitativna i kvantitativna kemijska analiza biološki aktivnih sastavnica primjenom različitih kromatografskih i spektroskopskih tehnika (TLC, HPLC-DAD-ESI-MS, LC-PDA-ESI-MSn, UV/Vis spektrofotometrija, GC-FID, GC-MS), a prikladnim metodama u uvjetima in vitro ispitani su odabrani biološki učinci prethodno utvrđeni kod vrste G. alypum (antidijabetski, antioksidativni, protuupalni, antimikrobni i citotoksični). Navedenim istraživanjima prethodila su temeljna botanička istraživanja na ekološkoj, morfološkoj, histološko-anatomskoj, citološkoj i molekularnoj razini, kojima se u kombinaciji s fitokemijskim istraživanjima pokušalo pridonijeti rješavanju taksonomskog spora oko razdvojenosti vrsta G. cordifolia i G. meridionalis. Sve četiri istraživane vrste pokazale su antidijabetski, antioksidativni, protuupalni, antimikrobni i citotoksični učinak u uvjetima in vitro, što je bilo moguće povezati s njihovim utvrđenim fitokemijskim profilima. Rezultati usporedbe fitokemijskog profila ukazali su na veću sličnost vrste G. punctata (podrod Globularia) s vrstom G. alypum (podrod Abolaria) od ostalih dviju vrsta (podrod Abolaria). Podjelu na podrodove nije podupirala ni usporedba histološko-anatomskih, molekularnih i fitokemijskih značajki, koja je ukazala na veću sličnost vrsta G. cordifolia i G. meridionalis s vrstom G. punctata. Također, kod uzoraka uključenih u ovo istraživanje nisu uočena karakteristična ekološka, morfološka, histološko-anatomska, citološka, molekularna ili fitokemijska obilježja koja bi podupirala razdvajanje na vrste G. cordifolia i G. meridionalis.Globularia alypum L. is a plant species widely used in folk medicine of Mediterranean countries and the best explored representative of the genus Globularia L. In this doctoral thesis, a comprehensive comparative study of botanical characteristics, phytochemical profile and biological effects of four Globularia species, which, apart from G. alypum, included three less explored related species: G. cordifolia L. (G. cordifolia subsp. cordifolia (L.) Hayek), G. meridionalis (Podp.) O. Schwarz (G. cordifolia subsp. bellidifolia (Ten.) Wettst.) and G. punctata Lapeyr., was conducted for the first time. Samples from 19 natural populations, collected during flowering time in Croatia and surrounding area in several different seasons (up to five), were used in this study. In order to evaluate the biomedical potential of less investigated species, qualitative and quantitative chemical analyses of biologically active constituents were conducted using different chromatographic and spectroscopic techniques (TLC, HPLC-DAD-ESI-MS, LC-PDA-ESI-MSn, UV/Vis spectrophotometry, GC-FID, GC-MS), while suitable in vitro methods were used to evaluate selected biological effects, previously established for G. alypum (antidiabetic, antioxidative, anti-inflammatory, antimicrobial and cytotoxic). The aforementioned research was preceded by basic botanical research on ecological, morphological, histo-anatomical, cytological and molecular level, by which, in combination with the phytochemical research, an attempt was made to contribute to the resolution of the taxonomic dispute over the separation of the species G. cordifolia and G. meridionalis. All four examined species showed antidiabetic, antioxidative, anti-inflammatory, antimicrobial and cytotoxic activity in vitro, which could be linked to their established phytochemical profiles. The results of the comparison of phytochemical profiles showed greater similarity of G. punctata (subgenus Globularia) to G. alypum (subgenus Abolaria) than those of the other two species (subgenus Abolaria). The division into subgenera was also not supported by the comparison of histo-anatomical, molecular and phytochemical characteristics, which indicated greater similarity of G. cordifolia and G. meridionalis to G. punctata. Moreover, no ecological, morphological, histo-anatomical, cytological, molecular or phytochemical characteristics, that would support the separation of G. cordifolia and G. meridionalis into different species, were observed for the samples included in this study