Magister Pharmaceuticae - MPharm / Diabetes Mellitus, one of the major diseases affecting human population all over the world has caused significant morbidity and mortality. The management of this condition has raised the demand of safe and cost effective remedial measures due to several side effects associated with the present use of modern medicines. Thus, it is crucial to explore other options for diabetes management such as the use of medical plants. Cnicus benedictus L. is one of the known plant species used by traditional medicine practitioners in South Africa for the treatment of various ailments including inflammatory conditions, pain and diabetes. Even though the plant species has been extensively studied, scientifically, no evidence exists in literature to corroborate the claim made by traditional medicine practitioners of its therapeutic success in the treatment of diabetes and pain. Therefore, the objectives of this present study were: to investigate the antidiabetic activity of C. benedictus using leaf methanol extract of the plant species on animal model involving male and female Albino rats; to investigate the antinociceptive activity of the plant species on mice; to determine the safety profile of the plant by investigating the acute toxicity and to carry out HPLC study in order to characterize the plant species. Animals were divided into groups of six each and fasted overnight prior to the induction of diabetes in rats using Streptozocin (STZ). The plasma glucose was measured at intervals of 30 min for 4 hours by means of a glucometer. Cnicus benedictus (100 – 400 mg/kg, i.p.) significantly reduced the blood glucose concentrations of fasted normal rats with percentage maximum reduction ranging from 46 to 79% and chlorpropramide (250 mg/kg, i.p.) significantly reduced the blood glucose concentrations of fasted normal rats by 84%. Cnicus benedictus (100 – 400 mg/kg, i.p.) significantly reduced the blood glucose concentrations of STZinduced diabetic rats with percentage maximum reduction ranging from 44.82 to 66.04% and chlorpropramide (250 mg/kg, i.p.) significantly reduced the blood glucose concentration of STZ-induced diabetic rats by 71.71%. In the oral glucose tolerance test, administration of leaf methanol extract of Cnicus benedictus (100 – 400 mg/kg, i.p.) following oral glucose load on fasted normoglycaemic rats significantly reduced the increased blood glucose concentrations with percentage maximum reduction ranging from 42.45 to 70.75%. Chlorpropramide (250 mg/kg, i.p.) following oral glucose load on fasted normoglycaemic rats significantly reduce the increased blood glucose concentration with a percentage maximum reduction of 79.04%. In acetic acid writhing test, animals were divided into groups of eight per dose. Cnicus benedictus (25-400 mg/kg, i.p.) significantly reduced the number of writhes in mice with percentage inhibition of the writhes ranging from 67.95 to 73.71%. Indomethacin (20 mg/kg, i.p.) and paracetamol (500 mg/kg, i.p.) significantly reduced the number of writhes in mice with percentage inhibition of 75.44 and 69.18% respectively. Combined treatment of lowest and sub-effective doses of C.benedictus (12.5 mg/kg, i.p.) and indomethacin (10 mg/kg, i.p.) significantly reduced the writhes with a percentage inhibition of 58.32%. In hot plat test, animals were divided into groups of eight per dose. Cnicus benedictus (25-400 mg/kg, i.p.) significantly delayed the reaction times of the mice to hot-plate thermal stimulation. Morphine (10 mg/kg, i.p.) significantly delayed the reaction time of the mice to the hot-plate stimulation. The no-adverse-effect-level (NOAEL) of leaf methanol extract of Cnicus benedictus was obtained at 3200 mg/kg (p.o.) and the LD50 value of the plant species was found to be 4000 mg/kg (p.o.). The HPLC fingerprint of the leaf methanol extract of Cnicusbenedictus showed distinct peaks at the following retention times of 6.387, 14.628, 18.431, 23.228 and 29.829 min. In conclusion, the data obtained showed that leaf methanol extract of Cnicus benedictus possesses both antidiabetic and antinocipetive activities. / National Research Foundation (NRF)

Syzygium cordatum is a medicinal plant indigenous to South Africa and Mozambique, commonly used to treat stomach aches, diabetes, respiratory problems and tuberculosis. In spite of the folklore use, adequate scientific data to credit its widespread traditional use is lacking. The objectives of this study were: to evaluate and validate scientifically the successful therapeutic claims by traditional medicine practitioners that Syzygium cordatum is effective in treating diarrhoea and diabetes / to determine the effects of the plant extract on gastrointestinal transit of a charcoal meal in mice / to determine the effects on castor oilinduced intestinal fluid accumulation / to determine the safety profile of the plant by carrying out acute toxicology study and to carry out preliminary screening of the active compounds present in the plant using standard phytochemical analytical procedures. The aqueous leaf extract of Syzygium cordatum (3.125 -50mg/kg, p.o) significantly reduced the faecal output caused by castor oil (0.7ml). All the doses used, reduced faecal output from 100% produced by castor oil to between 40 and 61%. S.cordatum (6.25 &ndash / 50mg/kg, p.o) significantly and in a dose dependent manner, delayed the onset of castor oil-induced diarrhoea.

Syzygium cordatum is a medicinal plant indigenous to South Africa and Mozambique, commonly used to treat stomach aches, diabetes, respiratory problems and tuberculosis. In spite of the folklore use, adequate scientific data to credit its widespread traditional use is lacking. The objectives of this study were: to evaluate and validate scientifically the successful therapeutic claims by traditional medicine practitioners that Syzygium cordatum is effective in treating diarrhoea and diabetes / to determine the effects of the plant extract on gastrointestinal transit of a charcoal meal in mice / to determine the effects on castor oilinduced intestinal fluid accumulation / to determine the safety profile of the plant by carrying out acute toxicology study and to carry out preliminary screening of the active compounds present in the plant using standard phytochemical analytical procedures. The aqueous leaf extract of Syzygium cordatum (3.125 -50mg/kg, p.o) significantly reduced the faecal output caused by castor oil (0.7ml). All the doses used, reduced faecal output from 100% produced by castor oil to between 40 and 61%. S.cordatum (6.25 &ndash / 50mg/kg, p.o) significantly and in a dose dependent manner, delayed the onset of castor oil-induced diarrhoea.

<p>Glavni cilj ove doktorske disertacije je određivanje hemijskog sastava, biolo&scaron;kih i funkcionalnih karakteristika novih proizvoda od zove. Dobijanje proizvoda na bazi zove zasniva se na primeni tradicionalnih i savremenih tehnolo&scaron;kih procesa proizvodnje. Iskori&scaron;ćenje prirodnog potencijala zove započeto je primenom tradicionalne i savremene (liofilizacija) tehnike su&scaron;enja. U cilju dobijanja visoko-vrednih ekstrakata ploda i cveta zove primenjene su tradicionalna (maceracija) i savremene (ultrazvučna i mikrotalasna) ekstrakcione tehnike sa dva ekstragensa (50% etanol i voda). Dobijanje matičnog soka od plodova zove podrazumevalo je primenu tradicionalnog načina ceđenja, dok je vino od plodova zove dobijeno po standardnom postupku proizvodnje vina. Vino je izloženo različitim temperaturnim tretmanima u različitom vremenskom periodu (60 &deg;C u toku 5 minuta, 60 &deg;C u toku 10 minuta, 70 &deg;C u toku 5 minuta i bez toplotnog tretmana) u cilju evaluacije biolo&scaron;ke aktivnosti dobijenog proizvoda. Etarsko ulje ploda i cveta zove je dobijeno hidrodestilacijom. Ispitivanje efikasnosti primenjenih tehnolo&scaron;kih postupaka su&scaron;enja i ekstrakcije je zasnovano na određivanju biolo&scaron;kih i funkcionalnih karakteristika dobijenih ekstrakata ploda i cveta zove. U ispitivanim ekstraktima ploda i cveta zove dominantne fenolne kiseline su hlorogenska i protokatehinska kiselina, a rutin i kvercetin-3-O-heksozid su dominantna flavonoidna jedinjenja. Biolo&scaron;ke i funkcionalne karakteristike su ispitane primenom različitih in vitro antioksidativnih, neuroprotektivnih, antitirozinaznih i antidijabetogenih testova. Primenom liofilizacije kao savremene tehnike su&scaron;enja i mikrotalasne ekstrakcije kao savremene ekstrakcione tehnike povećava se biopotencijal ispitivanih ekstrakata. Matični sok od plodova zove kao potencijalno novi funkcionalni proizvod je analiziran u cilju definisanja hemijskog, fitohemijskog i nutritivnog sastava, biolo&scaron;kog potencijala i senzorskih karakteristika. Ispitivanja dobijenog vina su bila usmerena na utvrđivanje optimalnih uslova za proizvodnju voćnog vina. Definisanjem hemijskog i fitohemijskog sastava i evaluacijom biopotencijala vina određen je optimalan temperaturni profil za dobijanje jednog od novih funkcionalnih proizvoda. Na osnovu utvrđenog hemijskog sastava etarsko ulje ploda i cveta zove se pokazalo kao potencijalno novi prirodni agens za održavanje svežine i produženja roka trajanja prehrambenih proizvoda. Zova je nesumnjivo samonikla biljna vrsta koja u budućnosti osnovano može biti polazna sirovina za kreiranje i dobijanje novih prehrmabenih proizvoda na domaćem i inostranom trži&scaron;tu.</p> / <p>The main goal of this doctoral dissertation is to determine the chemical composition, biological and functional characteristics of new elderberry products. Elderberry products were obtained via traditional and modern technological processes. The exploitation of the natural potential of the elderberry started with the application of traditional and modern (lyophilization) drying techniques. To obtain high-value extracts of fruits and flowers, traditional (maceration) and modern (ultrasonic and microwave) extraction techniques with two solvents (50% ethanol and water) were applied. Obtaining the juice from the elderberry fruits implied the application of the traditional cold pressing method, whereas wine from the elderberry fruits was obtained in accordance with the standard procedure of wine production. The wine was exposed to different temperature treatments in different periods (60&deg;C for 5 minutes, 60&deg;C for 10 minutes, 70&deg;C for 5 minutes and without heat treatment) to evaluate the biological activity of the product. The essential oil of the fruit and flower was obtained by hydrodistillation. The examination of the efficiency of the applied technological procedures of drying and extraction is based on observing the biological and functional characteristics of the obtained extracts of the said fruits and flowers. In the examined fruit and flower extracts, the dominant phenolic acids are chlorogenic and protocatechuic acid, while rutin and quercetin-3-O-hexoside are the dominant flavonoid compounds. Biological and functional characteristics were examined using various in vitro antioxidant, neuroprotective, antityrosinase, and antidiabetic tests. The application of lyophilization and microwave extraction (as modern drying and extraction techniques) increased the biopotential of the analyzed extracts. Elderberry juice, a potentially new functional product, was analyzed to define the chemical, phytochemical and nutritional composition, biological potential, and sensory characteristics. The wine was tested in order to determine the optimal conditions for the production of fruit wine. By defining the chemical and phytochemical composition and evaluating the biopotential of wine, the optimal temperature profile for obtaining one of the new functional products was determined. Based on the obtained chemical composition, it is determined that the essential oil of fruits and flowers is a potentially new natural agent for maintaining freshness and extending the shelf life of food products. Without a doubt, elderberry is a wild plant species that could be used in the future as the starting material for creating and obtaining new food products on the domestic and foreign markets.</p>

<p>&nbsp;У оквиру ове докторске дисертације испитан је хемијски састав и биолошке активности ЕtOH, H₂Oи CHCl_3&nbsp; екстраката четири врсте гљива рода<em> Ganoderma</em>&nbsp; (Basidiomycota):&nbsp;<em> G. applanatum,&nbsp; G. lucidum,G. pfeifferi,&nbsp; G. resinaceum</em>&nbsp; са територије Војводине.Хемијски састав анализираних врста је одређен<br />применом: ААЅ методе (састав макро-&nbsp; имикроелемената у сувим остацима гљива) и LC-MS/MS технике (квантитативни састав фенолних једињења и флавоноида) при чему је детектовано 12 једињења. Спектрофотометријским методама је одређен садржај протеина, шећера, укупних фенола и флавоноида, код којих је највећи садржај протеина утврђен за ЕtOH екстракте&nbsp;<em> G. applanatum&nbsp; </em>и&nbsp; <em>G. pfeifferi</em>. Испитивања биолошких активности екстраката обухватила су: одређивање<span id="cke_bm_779S" style="display: none;">&nbsp;</span><em>&nbsp; in vitro</em><span id="cke_bm_779E" style="display: none;">&nbsp;</span>&nbsp; и&nbsp;<em> in vivo</em> антиоксидантне, антимикробне, антиинфламаторне, антипролиферативне и антијабето<span id="cke_bm_780E" style="display: none;">&nbsp;</span>гене&nbsp;&nbsp; aктивности.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</p><p>Антиоксидантна активност (способност неутрализације слободних радикала и редукциони потенцијал) је одређена спектрофотометријским методама, при којој су најбољу активност остварили Н₂О екстракти&nbsp;<em> G</em>. <em>applanatum.</em> Антимикробнa активност&nbsp; анализираних екстраката одређена је испитивањем антибактеријског, антифунгалног и антивиралног потенцијала где се издвојила<em> G. pfeifferi</em> врста. Антиинфламаторни потенцијал EtOH и&nbsp; CHCl₃екстраката одређен је<em>&nbsp; ex vivo&nbsp;</em> методом мерењем способности инхибиције продукције медијатора инфламације&nbsp; (продукти&nbsp; метаболизма арахидонске киселине) при којој су бољу активност испољили CHCl₃ екстракти.</p><p>Ефекат EtOH и H₂O екстраката врста рода Ganoderma&nbsp;&nbsp; на раст MCF ћелијске линије испитан је MTT тестом, а посебно су се издвојили&nbsp; EtOH&nbsp; екстракти врста после 72h.</p><p>Остварена антидијабетогена активност EtOH и Н₂О екстраката врста&nbsp; <span id="cke_bm_795S" style="display: none;">&nbsp;</span><em>G. pfeifferi&nbsp; </em><span id="cke_bm_795E" style="display: none;">&nbsp;</span>и&nbsp;<em> G. resinaceum</em>&nbsp; код алоксан-индукованог&nbsp;<em> D. mell<span id="cke_bm_796E" style="display: none;">&nbsp;</span>itus-a&nbsp;</em> на&nbsp; експерименталним&nbsp; животињама&nbsp; праћена je регенерацијом&nbsp; &szlig;- ћелија&nbsp; Лангерхансових острваца панкреаса. Као потенцијални нефро-&nbsp; и&nbsp; хепатопротективни агенси се издвајају екстракти<em>&nbsp; G. resinaceum.</em></p><p>Сумарно, укупни биопотенцијал анализираних врста рода&nbsp; Ganoderma&nbsp; на основу спроведених анализа хемијске&nbsp;&nbsp; kарактеризације и биолошке активности упућује&nbsp; на&nbsp; могућност њихове потенцијалне примене као нутрацеутика и додатака исхрани, у будућности уз неопходност додатних микохемијских истраживања ових врста, посебно терпеноида и полисахарида, као и других биолошких активности као што је неуропротективна.</p> / <p>&nbsp;U okviru ove doktorske disertacije ispitan je hemijski sastav i biološke aktivnosti EtOH, H₂Oi CHCl_3&nbsp; ekstrakata četiri vrste gljiva roda<em> Ganoderma</em>&nbsp; (Basidiomycota):&nbsp;<em> G. applanatum,&nbsp; G. lucidum,G. pfeifferi,&nbsp; G. resinaceum</em>&nbsp; sa teritorije Vojvodine.Hemijski sastav analiziranih vrsta je određen<br />primenom: AAЅ metode (sastav makro-&nbsp; imikroelemenata u suvim ostacima gljiva) i LC-MS/MS tehnike (kvantitativni sastav fenolnih jedinjenja i flavonoida) pri čemu je detektovano 12 jedinjenja. Spektrofotometrijskim metodama je određen sadržaj proteina, šećera, ukupnih fenola i flavonoida, kod kojih je najveći sadržaj proteina utvrđen za EtOH ekstrakte&nbsp;<em> G. applanatum&nbsp; </em>i&nbsp; <em>G. pfeifferi</em>. Ispitivanja bioloških aktivnosti ekstrakata obuhvatila su: određivanje<span id="cke_bm_779S" style="display: none;">&nbsp;</span><em>&nbsp; in vitro</em><span id="cke_bm_779E" style="display: none;">&nbsp;</span>&nbsp; i&nbsp;<em> in vivo</em> antioksidantne, antimikrobne, antiinflamatorne, antiproliferativne i antijabeto<span id="cke_bm_780E" style="display: none;">&nbsp;</span>gene&nbsp;&nbsp; aktivnosti.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</p><p>Antioksidantna aktivnost (sposobnost neutralizacije slobodnih radikala i redukcioni potencijal) je određena spektrofotometrijskim metodama, pri kojoj su najbolju aktivnost ostvarili N₂O ekstrakti&nbsp;<em> G</em>. <em>applanatum.</em> Antimikrobna aktivnost&nbsp; analiziranih ekstrakata određena je ispitivanjem antibakterijskog, antifungalnog i antiviralnog potencijala gde se izdvojila<em> G. pfeifferi</em> vrsta. Antiinflamatorni potencijal EtOH i&nbsp; CHCl₃ekstrakata određen je<em>&nbsp; ex vivo&nbsp;</em> metodom merenjem sposobnosti inhibicije produkcije medijatora inflamacije&nbsp; (produkti&nbsp; metabolizma arahidonske kiseline) pri kojoj su bolju aktivnost ispoljili CHCl₃ ekstrakti.</p><p>Efekat EtOH i H₂O ekstrakata vrsta roda Ganoderma&nbsp;&nbsp; na rast MCF ćelijske linije ispitan je MTT testom, a posebno su se izdvojili&nbsp; EtOH&nbsp; ekstrakti vrsta posle 72h.</p><p>Ostvarena antidijabetogena aktivnost EtOH i N₂O ekstrakata vrsta&nbsp; <span id="cke_bm_795S" style="display: none;">&nbsp;</span><em>G. pfeifferi&nbsp; </em><span id="cke_bm_795E" style="display: none;">&nbsp;</span>i&nbsp;<em> G. resinaceum</em>&nbsp; kod aloksan-indukovanog&nbsp;<em> D. mell<span id="cke_bm_796E" style="display: none;">&nbsp;</span>itus-a&nbsp;</em> na&nbsp; eksperimentalnim&nbsp; životinjama&nbsp; praćena je regeneracijom&nbsp; &szlig;- ćelija&nbsp; Langerhansovih ostrvaca pankreasa. Kao potencijalni nefro-&nbsp; i&nbsp; hepatoprotektivni agensi se izdvajaju ekstrakti<em>&nbsp; G. resinaceum.</em></p><p>Sumarno, ukupni biopotencijal analiziranih vrsta roda&nbsp; Ganoderma&nbsp; na osnovu sprovedenih analiza hemijske&nbsp;&nbsp; karakterizacije i biološke aktivnosti upućuje&nbsp; na&nbsp; mogućnost njihove potencijalne primene kao nutraceutika i dodataka ishrani, u budućnosti uz neophodnost dodatnih mikohemijskih istraživanja ovih vrsta, posebno terpenoida i polisaharida, kao i drugih bioloških aktivnosti kao što je neuroprotektivna.</p> / <p>Whitin this doctoral thesis the chemical composition and biological activity of EtOH, H<span id="cke_bm_699S" style="display: none;">&nbsp;</span>₂<span id="cke_bm_699E" style="display: none;">&nbsp;</span>O and CHCl₃extracts of four fungal species which belong to genus<em> Ganoderma&nbsp;</em> (phylum Basidiomycota)<span id="cke_bm_700E" style="display: none;">&nbsp;</span>:&nbsp;<em> G. applanatum,&nbsp; G. lucidum,&nbsp; G. pfeifferi,&nbsp; G. resinaceum</em>&nbsp; were determinated. The samples were collected from different localities in Vojvodina. Chemical characterization included: AAS methods (compositon of macro- and&nbsp; microelements in d.w. of fungi) and LC-MS/MS technique (quantitative analysis of phenolic compounds and flavonoids) wherein the 12 selected phenolic compounds were detected. The total proteins, sugars, phenolics and flavonoids content were&nbsp;&nbsp;&nbsp; determined using spectrophotometric methods. The highest protein content was determined in EtOH extracts of<span id="cke_bm_707S" style="display: none;">&nbsp;</span><em> G. applanatum&nbsp; </em><span id="cke_bm_707E" style="display: none;">&nbsp;</span>and <em>G. pfeifferi</em>&nbsp; species. In order to assess the biological potential, the in vitro&nbsp; and in vivo antioxidant, antimicrobial, anti-inflammatory, antiproliferative and antidiabetic activities of the extracts were investigated.<span id="cke_bm_708E" style="display: none;">&nbsp;</span><br />&nbsp;&nbsp; The antioxidant activity (the ability of neutralizing free radicals and reduction potential) estimated byspectrophotometric methods. The highest&nbsp;&nbsp; antioxidant potential was noticed in H₂O extracts of&nbsp; <em>G. applanatum.</em> Evaluation of antimicrobial activity included the estimation of antibacterial, antifungal and antiviral activity, whereby the&nbsp; species&nbsp;<em> G. pfeifferi&nbsp;</em> showed the highest potential The anti-inflammatory activity of EtOH and&nbsp; CHCl₃&nbsp; extracts was determined by&nbsp; ex vivo&nbsp; method measuring the ability of production inhibition of inflammation mediators&nbsp; (products of arachidonic acid metabolism), where the CHCl₃&nbsp; extracts were exhibited better activity.<br />&nbsp;&nbsp; The effect of EtOH and H₂O extracts of<em>&nbsp; Ganoderma</em> species on the growth of the cell line MCF-7, has been examined using MTT assay (stand out ethanolic extracts of analyzed species after 72h incubation period).<br />&nbsp;&nbsp; Achieved antidiabetic activity of EtOH and H₂O extracts o<em>f<span id="cke_bm_725S" style="display: none;">&nbsp;</span> G. pfeifferi<span id="cke_bm_725E" style="display: none;">&nbsp;</span>&nbsp; </em>an<em>d G. resinaceum</em>&nbsp; at alloxan-i<span id="cke_bm_726E" style="display: none;">&nbsp;</span>nduced D. mellitus in experimental animals was followed by regeneration of&nbsp; cells of Langerhans pancreatic islets. Extracts of&nbsp;<em> G.&nbsp;&nbsp; resinaceum&nbsp;</em> were allocated as a potential nephro- and&nbsp; hepatoprotective agents.<br />In summary, the overall biological potential of the analyzed species of the genus&nbsp; <em>Ganoderma</em>&nbsp; based on results for chemical and biological characterization indicate that they could be used&nbsp; as a nutraceuticals and food supplements in the future, with further the necessity of additional mycochemical investigation (especially terpenoids and polysaccharides) and other biological activity such as neuroprotective.</p>