Utjecaj medija i temperature na tvorbu gliotoksina u sojeva Aspergillus fumigatus

Gliotoxin is a secondary metabolite of the epipolythiodioxopiperazine family with biologically active internal disulfide bridge. It is produced by many fungal species, including Aspergillus fumigatus and A. terreus. A. fumigatus, which produces gliotoxin and more than twenty other secondary metabolites, is the leading cause of invasive aspergillosis. Gliotoxin production in situ influence the development of aspergillosis. This study investigated the in vitro production of gliotoxin in nine A. fumigatus isolates from the upper respiratory tract of immunocompromised patients. The effects of media composition and incubation temperature were studied. Gliotoxin was extracted from biomass and its concentration was semi-quantitatively analysed using thin-layer chromatography. Gliotoxin production was higher in the yeast-extract liquid medium (YES) than in the synthetic Czapek-Dox liquid medium (CZA). Incubation at 37 °C resulted in higher gliotoxin production than at 25 °C, probably because higher temperatures favour expansive growth of the mycelium. Gliotoxin could be detected after three days of incubation at concentrations 4.06 mg mL-1 (in YES at 37 °C) and 1.07 mg mL-1 (in CZA at 25 °C). YES broth as a medium containing 4 % sucrose and 2 % of yeast extract is a very rich substrate for the production of gliotoxin in vitro.Gliotoksin je sekundarni metabolit iz skupine epipolitiodioksipiperazina s biološki aktivnim internim disulfidnim mostom u molekuli, koji tvore razne plijesni i gljivica Candida albicans. Plijesan Aspergillus fumigatus vodeći je uzročnik invazivnih aspergiloza i također može tvoriti gliotoksin. Pretpostavlja se da in situ tvorba gliotoksina utječe na patogenezu aspergiloze. U ovom radu ispitali smo in vitro tvorbu gliotoksina u devet sojeva A. fumigatus vrste, izoliranih iz imunokompromitiranih pacijenata. Praćen je utjecaj medija i temperature inkubacije na tvorbu gliotoksina. Gliotoksin je ekstrahiran iz biomase i koncentracija mu je utvrđena polukvantitativno tankoslojnom kromatografijom. Tvorba gliotoksina uočena je već nakon trodnevne inkubacije u koncentracijama 4,06 mg mL-1 (u bujonu s kvaščevim ekstraktom – YES na 37 °C) i 1,07 mg mL-1 (u sintetskom Czapek-Dox bujonu – CZA na 25 °C). Tvorba gliotoksina bila je veća u YES bujonu, za razliku od sintetskog CZA bujona. Viša temperatura inkubacije (37 °C) također utječe na jaču tvorbu gliotoksina nego niža temperatura (25 °C). YES bujon s dodatkom 4 % saharoze i 2 % kvaščeva ekstrakta vrlo je bogat supstrat za in vitro tvorbu gliotoksina, kao jednog od lipofilnih sekundardnih metabolita plijesni vrste A. fumigatus

Olive leaf extract activity against Candida albicans and C. dubliniensis – the in vitro viability study

Olive leaf extract is characterized by a high content of polyphenols (oleuropein, hydroxytyrosol and their derivatives), which is associated with its therapeutic properties. The objective of the present research was to evaluate the antifungal activity of olive leaf aqueous extract against Candida albicans ATCC 10231 and C. dubliniensis CBS 7987 strains. Minimum inhibitory concentration (MIC) of the extract was determined by several in vitro assays. The extract showed a concentration dependent effects on the viability of C. albicans with MIC value of 46.875 mg mL–1 and C. dubliniensis with MIC value 62.5 mg mL–1. Most sensitive methods for testing the antifungal effect of the extract were the trypan blue exclusion method and fluorescent dye exclusion method while MIC could not be determined by the method according to the EUCAST recommendation suggesting that herbal preparations contain compounds that may interfere with this susceptibility testing. The fluorescent dye exclusion method was also used for the assessment of morphological changes in the nuclei of the treated cells. According to the obtained results, olive leaf extract is less effective against the tested strains than hydroxytyrosol, an olive plant constituent tested in our previous study

Mikotoksinogenost kliničkih i okolišnih Aspergillus fumigatus i A. flavus sojeva

Clinical isolates of fifty strains of A. fumigatus and 30 strains of A. flavus from immmunocompromised patients from the hematological unit were analyzed for mycotoxin production and compared with the same number of environmental isolates (from soil, compost, and air). Only 9 (18%) strains of A. fumigatus produced gliotoxin in a mean concentration 2.22 mg mL-1 (range 0.5-5 mg mL-1). Aflatoxin B1 was detected in 7 (23%) isolates (range from 0.02 to 1.2 mg L-1) and aflatoxin G1 in one (3%) of clinical A. flavus isolates (0.12 mg L-1). In the group of environmental isolates, 11 (37%) were positive for aflatoxin B1 production (range from 0.02 to 1.2 mg L-1) and one for aflatoxin G1 (0.02 mg L-1). Bioautoantibiogram ("bioassay in situ") on TLC plates against Bacillus subtilis NCTC 8236 showed that only gliotoxin-producing strains have bactericidal activity of Rf values corresponding to gliotoxin. The secondary-metabolite profiles of clinical and environmental A. fumigatus and A. flavus isolates were homogenous, except for gliotoxin production, which was detected only in the group of clinical isolates of A. fumigatus (18%).Analizirana je mikotoksinogenost pedeset kliničkih sojeva vrste A.fumigatus i trideset sojeva vrste A. flavus izoliranih iz imunokompromitiranih ispitanika u hematološkom odjelu. Kao usporedba, izoliran je isti broj okolišnih sojeva iz zemlje, komposta i zraka. Utvrđeno je da samo 9 sojeva (18%) vrste A. fumigatus tvori gliotoksin sa srednjom vrijednošću 2,22 mg mL–1 (u rasponu od 0,5 do 5 mg mL–1). Tvorba aflatoksina B1 utvrđena je u 7 sojeva (23%) (raspon od 0,024 do 1,2 mg L–1) i u jednog kliničkog izolata (3%) A.flavus vrste (0,12 mg L–1). U skupini okolišnih sojeva utvrđena je tvorba aflatoksina B1u 11 sojeva (37%) (u rasponu od 0,024 do 1,2 mg L–1) i u jednog soja tvorba aflatoksina G1(0,024 mg L–1). Bioautoantibiogramom na tankoslojnoj kromatografskoj ploči s bakterijskom vrstom Bacillus subtilis NCTC 8236 utvrđeno je da samo gliotoksinogeni sojevi pokazuju baktericidnu aktivnost oko Rf vrijednosti koja odgovara gliotoksinu. Profili sekundarnih metabolita kliničkih i okološnih sojeva A. funmigatus i A. flavus vrsta su bili homogeni, osim za tvorbu gliotoksina koji je utvrđen samo u skupini kliničkih izolata A. fumigatus sojeva (18%)

Onečišćenja u ljekovitom bilju i biljnim proizvodima

Medicinal plants have a long history of use in therapy throughout the world and still make an important part of traditional medicine. Thus, medicinal plants and herbal products must be safe for the patient (consumer). This review addresses biological contaminants (microbes and other organisms) and chemical contaminants (mycotoxins, toxic elements such as heavy metals, and pesticide residues) as major common contaminants of medicinal herbs and herbal products. To prevent and screen for contamination and ensure safety and conformity to quality standards, medicinal herbs and herbal products should be included in appropriate regulatory framework.Ljekovito bilje i biljni proizvodi već tisućljećima nalaze široku primjenu u različitim sustavima tradicionalnog liječenja. Stoga je njihova neškodljivost, ponajprije uvjetovana kakvoćom biljne sirovine, od izuzetne važnosti za javno zdravstvo. Od mogućih čimbenika koji utječu na kakvoću ljekovitog bilja i biljnih proizvoda ovaj pregledni rad osvrće se na najčešće prisutna biološka (mikroorganizmi) i kemijska onečišćenja (mikotoksini, toksični elementi poput teških metala te ostaci pesticida). S ciljem postizanja ujednačenih standarda kakvoće te osiguranja sigurnosti primjene biljnih proizvoda od vitalne su važnosti zakonski propisi koji moraju u odgovarajućim regulatornim okvirima obuhvatiti ovu skupinu proizvoda s naglaskom na sprječavanju i ispitivanju njihovih mogućih onečišćenja

Utjecaj dramskih metoda Zvjezdane Ladike na odgoj i obrazovanje djece

Ovaj završni rad obrađuje utjecaj dramskih metoda Zvjezdane Ladike na odgoj i obrazovanje djece. Sadržaj rada sastoji se od 7 poglavlja, uvod, životopis Zvjezdane Ladike, procesna drama, dramske vježbe, uloga igre i dramskih vježbi s obzirom na dječju dob, dramske igre u vrtiću – primjer provedene aktivnosti te zaključak na kraju. U uvodu se objašnjava da je ova tema izabrana zato što je važno unaprijediti dramski odgoj u odgojno obrazovnim institucijama, naročito u predškolskim ustanovama. Zvjezdana Ladika se osvrće na to da scenska umjetnost na dijete ostavi pozitivan utisak, da djetetu razvije ljubav prema kazalištu, da mu pomogne u razvijanju osjećajnosti, empatije, razvoju govornih i komunikacijskih vještina. U idućem poglavlju, prikazana je i pojašnjenja procesna drama. To je vrsta rada s djecom u kojem se isprepliću učenje i umjetnost kroz dramski izričaj. 12 dramskih vježbi je opisano u 4. poglavlju. Ladika je vježbe započinjala na način da je prije svega radila na izgradnji spontanosti svakoga djeteta zato što je smatrala da je spontanost put prema kreativnosti, a da svaka spontanost ne podrazumijeva kreativnost. U 5. poglavlju prikazuje se što je prikladno za koju dječju dob. Igra u životu čovjeka započinje već nakon 3. mjeseca života. Tada je igra naravno nesvjesna i motorička. U dobi između 5. i 6. godina djetetu je u igri važno planiranje igre isto koliko i samo igranje. Iduće poglavlje opisuje dramsku vježbu odnosno dramsku igru koja je provedena u skupini djece u dobi od 4. do 5. godine života. Skupina je pripremala predstavu za svoje roditelje. Predstava se zove “Tko je rekao mijau?”. Na kraju rada se donosi zaključak. Kroz kratke i jednostavne igre dijete nesvjesno ulazi u zamišljeni svijet. Tako se polako nadograđuje nova vježba za vježbom. Dramskih aktivnosti u vrtiću nije ih dovoljno i ne provode se često. Trebalo bi više inzistirati na provođenju dramskih vježbi u vrtićima jer djeca to mogu raditi, odnosno igrati se na takav način, učiti na takav način a pritom uživati.This paper deals with the impact of the Zvjezdana Ladika’s dramatic methods on child upbringing and education. The content of the work consists of 7 chapters, an introduction, a resume of Zvjezdana Ladika, a process drama, drama exercises, the role of games and dramatic exercises with regard to children's age, dramatic games in kindergarten - an example of the activity carried out and a conclusion at the end. The introduction explains that this topic was chosen because it is important to improve dramatic education in educational institutions, especially in preschool institutions. Zvjezdana Ladika points out that stage art gives a child a positive impression, developing a child's love for theater, helping her develop emotional, empathy, developing speech and communication skills. In the next chapter, a process drama is described. This is a kind of work with children that combines learning and art through dramatic expression. 12 drama exercises are described in chapter 4. Ladika began her exercises in a way that she worked primarily to build the spontaneity of every child because she felt that spontaneity was a way to creativity, and that any spontaneity does not imply creativity. Chapter 5 shows what is appropriate for the child's age. Playing in a child's life begins after 3 months of life. Then the game is naturally unconscious and motoric. At age 5 and 6, it is important for a child to plan and organize the game as much as playing. The next chapter describes a dramatic exercise or a dramatic play that was conducted in a group of children ages 4 to 5. The group was preparing a show for their parents. The play is called "Who said the mijau?". Conclusion is at the end of the paper. Through a short and simple game, the child goes unconsciously into the imaginative world. So, a new exercise is slowly being upgraded. Drama activities in kindergarten are not enough and are not often conducted. It should be more insistent on performing drama exercises in kindergartens because children can do it, that is to play in such a way, learn in such a way and enjoy it

Utjecaj dramskih metoda Zvjezdane Ladike na odgoj i obrazovanje djece

Ovaj završni rad obrađuje utjecaj dramskih metoda Zvjezdane Ladike na odgoj i obrazovanje djece. Sadržaj rada sastoji se od 7 poglavlja, uvod, životopis Zvjezdane Ladike, procesna drama, dramske vježbe, uloga igre i dramskih vježbi s obzirom na dječju dob, dramske igre u vrtiću – primjer provedene aktivnosti te zaključak na kraju. U uvodu se objašnjava da je ova tema izabrana zato što je važno unaprijediti dramski odgoj u odgojno obrazovnim institucijama, naročito u predškolskim ustanovama. Zvjezdana Ladika se osvrće na to da scenska umjetnost na dijete ostavi pozitivan utisak, da djetetu razvije ljubav prema kazalištu, da mu pomogne u razvijanju osjećajnosti, empatije, razvoju govornih i komunikacijskih vještina. U idućem poglavlju, prikazana je i pojašnjenja procesna drama. To je vrsta rada s djecom u kojem se isprepliću učenje i umjetnost kroz dramski izričaj. 12 dramskih vježbi je opisano u 4. poglavlju. Ladika je vježbe započinjala na način da je prije svega radila na izgradnji spontanosti svakoga djeteta zato što je smatrala da je spontanost put prema kreativnosti, a da svaka spontanost ne podrazumijeva kreativnost. U 5. poglavlju prikazuje se što je prikladno za koju dječju dob. Igra u životu čovjeka započinje već nakon 3. mjeseca života. Tada je igra naravno nesvjesna i motorička. U dobi između 5. i 6. godina djetetu je u igri važno planiranje igre isto koliko i samo igranje. Iduće poglavlje opisuje dramsku vježbu odnosno dramsku igru koja je provedena u skupini djece u dobi od 4. do 5. godine života. Skupina je pripremala predstavu za svoje roditelje. Predstava se zove “Tko je rekao mijau?”. Na kraju rada se donosi zaključak. Kroz kratke i jednostavne igre dijete nesvjesno ulazi u zamišljeni svijet. Tako se polako nadograđuje nova vježba za vježbom. Dramskih aktivnosti u vrtiću nije ih dovoljno i ne provode se često. Trebalo bi više inzistirati na provođenju dramskih vježbi u vrtićima jer djeca to mogu raditi, odnosno igrati se na takav način, učiti na takav način a pritom uživati.This paper deals with the impact of the Zvjezdana Ladika’s dramatic methods on child upbringing and education. The content of the work consists of 7 chapters, an introduction, a resume of Zvjezdana Ladika, a process drama, drama exercises, the role of games and dramatic exercises with regard to children's age, dramatic games in kindergarten - an example of the activity carried out and a conclusion at the end. The introduction explains that this topic was chosen because it is important to improve dramatic education in educational institutions, especially in preschool institutions. Zvjezdana Ladika points out that stage art gives a child a positive impression, developing a child's love for theater, helping her develop emotional, empathy, developing speech and communication skills. In the next chapter, a process drama is described. This is a kind of work with children that combines learning and art through dramatic expression. 12 drama exercises are described in chapter 4. Ladika began her exercises in a way that she worked primarily to build the spontaneity of every child because she felt that spontaneity was a way to creativity, and that any spontaneity does not imply creativity. Chapter 5 shows what is appropriate for the child's age. Playing in a child's life begins after 3 months of life. Then the game is naturally unconscious and motoric. At age 5 and 6, it is important for a child to plan and organize the game as much as playing. The next chapter describes a dramatic exercise or a dramatic play that was conducted in a group of children ages 4 to 5. The group was preparing a show for their parents. The play is called "Who said the mijau?". Conclusion is at the end of the paper. Through a short and simple game, the child goes unconsciously into the imaginative world. So, a new exercise is slowly being upgraded. Drama activities in kindergarten are not enough and are not often conducted. It should be more insistent on performing drama exercises in kindergartens because children can do it, that is to play in such a way, learn in such a way and enjoy it

Welcome to the emerging third era of antimicrobial therapy

Problem otpornosti mikroba na antibiotike može se sagledati kao globalni, kompleksan i sistemski problem s društvenim, kulturnim, socijalnim i političkim posljedicama. Nekoliko je strategija kako prevladati otpornost mikroba na antibiotike, no danas postoji potreba za globalnom strategijom u solidarnoj borbi protiv ove sporo-razvijajuće i progresivne nepogode. Budući da na sve uključene dionike u ovom dinamičnom procesu postoji javni pritisak, ujedno je taj pritisak usmjeren i na znanstvenu zajednicu. Odgovor je na povratak na specifičnu terapiju putem primjene terapijskih monoklonalnih protutijela, ali i primjene inhibitora mikrobne virulencije, bakteriofagnih lizina, antimikrobnih peptida, potencijatora učinka antibiotika, imunomodulatora i dr. Ti pristupi, od kojih je većina orjentirana mikrobno-specifično, uvode nas u treće doba antimikrobne terapije koje se polako rađa napuštajući dominaciju antibiotske molekularne paradigme.The problem of antimicrobial resistance (AMR) could be seen as a global, complex and systemic problem with social, cultural and political consequences. There are several strategies how to overcome AMR, but today we need a unique global and solidary strategy to combat this slowly emerging and progressive disaster. Because all stakeholders in this dynamic challenging process are involved, scientists are also under public pressure to resolve this problem. Nowadays, scientific approach to combat AMR is focused on the need for a specific therapy. This approach includes the use of therapeutic monoclonal antibodies, microbial virulence inhibitors, bacteriophage lysins, antimicrobial proteins, potentiators of current antibiotics, immunomodulators, etc. These approaches, most of which are microbe-specific oriented, introduce us to the third era of antimicrobial therapy, which slowly arises from the domination of molecular paradigmatic era of antibiotics