Trace Element Contents in the Edible Mushroom Boletus edulis Bull. ex Fries

Study was carried out on the trace element concentrations of edible mushroom Boletus edulis from Gorski kotar, Croatia. Samples were collected between March and December 2013. The analyses were carried out by Inductively Coupled Plasma – Optical Emission Spectrometry (ICP-OES). The results showed that the values of the studied elements decreased in the order: Cu (13.01) > Se (12.25) > Mn (9.19) > Al (4.62) > Ag (2.34) > Hg (2.18) > Cd (1.22) > Ni (0.99) > Pb (0.79) > Sb (0.60) > As (0.41) > Ba (0.38) > Cr (0.22) > Co (0.07). The essential elements in fruiting bodies Boletus edulis were a considerably higher than those of toxic elements. The consumption of wild edible mushroom Boletus edulis from Gorski kotar is safe for human health

Influence of essential oil Helichrysum italicum (Roth) G. Don on the formation of non-tuberculous mycobacterial biofilm

Cilj: Ispitati antimikrobni i antioksidacijski učinak te sposobnost inhibicije biofilma eteričnog ulja (EU) smilja [Helichrysum italicum (Roth) G. Don (H. italicum)] prema netuberkuloznim mikobakterijama M. avium i M. intracellulare. Materijali i metode: Za određivanje minimalne inhibicijske koncentracije (MIK) i minimalne baktericidne koncentracije (MBK) EU H. italicum prema M. avium i M. intracellulare korištena je metoda mikrodilucije u bujonu. Preživljavanje mikobakterija u Middlebrook 7H9 bujonu pod utjecajem EU smilja praćeno je subkultivacijom suspenzija 0, 1, 4 i 8 dan. Za određivanje inhibicije stvaranja biofilma u vodi primjenjena je metoda bojenja biofilma s kristal violetom. Antioksidacijski učinak MIK koncentracije EU određen je DPPH (2,2-difenil-1-pikril-hidrazil) metodom. Rezultati: MIK i MBK vrijednost EU smilja iznosila je 3,2 mg/ml. Koncentracija ½ MIK vrijednosti dovela je nakon prvog dana do redukcije vijabilnosti M. avium za približno 2 log10 odnosno 2,5 log10 za M. intracellulare, dok je četvrti dan zabilježena potpuna inhibicija porasta obje vrste mikobakterija. M. avium je pokazao veću tendenciju stvaranja biofilma. Koncentracija od ¼ i ½ MIK vrijednosti dovela je do statistički značajne inhibicije stvaranja biofilma ispitivanih mikobakterija. MIK koncentracija EU H. italicum je nakon 60 minuta inhibirala 38,2 % DPPH radikala. Zaključak: Subinhibitorne koncentracije EU smilja značajno reduciraju vijabilnost M. avium i M. intracellulare i dovode do značajne inhibicije stvaranja biofilma ispitivanih mikobakterija u vodi što bi omogućilo primjenu niskih efektivnih koncentracija ovog EU, koje nemaju citotoksični učinak, kao prirodnog dezinficijensa u vodi.Objective: To investigate the antimicrobial and antioxidative effect of the essential oil (EO) of immortelle [Helichrysum italicum (Roth) G. Don (H. italicum)] and its ability to inhibit biofilm of nontuberculous mycobacteria, M. avium and M. intracellulare. Materials and methods: a broth microdilution method was used to determine the minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the H. italicum EO. Survival of mycobacteria in Middlebrook 7H9 broth under the influence of H. italicum EO was monitored by subcultivation of suspension of 0, 1, 4 and 8 days. For determination of the inhibition of biofilm formation in water, the biofilm coloring method with crystal violet was applied. The antioxidant effect of MIC concentrations of H. italicum EO was determined by DPPH (2,2-diphenyl-1-picrylhydrazil) method. Results: MIC and MBC value of immortelle EO was 3.2 mg/ml. Treatment of mycobacteria with H. italicum EO at one half times the MIC resulted in approximately 2 log10 reduction of M. avium and 2.5 log10 M. intracellulare respectively over 24 h, and complete reduction of both mycobacteria on the fourth day. M. avium showed a greater tendency to form biofilms. The concentration of one-quarter and one-half of the MIC values resulted in statistically significant inhibition of biofilm production of the tested mycobacteria. MIC concentrations of H. italicum EU after 60 minutes inhibited 38.2% of DPPH radicals. Conclusion: The subinhibitory concentrations of H. italicum EO significantly reduce the viability of M. avium and M. intracellulare and result in substantial inhibition of biofilm production of the tested mycobacteria in water, which would allow the application of low effective concentrations of this EO that do not have a cytotoxic effect as a natural disinfectant in water

Synergistic potential of Juniperus communis and Helichrysum italicum essential oils against nontuberculous mycobacteria

Objective. The present study evaluated the possible synergistic antimycobacterial interactions of Juniperus communis (J. communis) and Helichrysum italicum (H. italicum) essential oils (EO). Methods. Antimycobacterial potential was tested against Mycobacterium avium and Mycobacterium intracellulare using broth and water dilution method and checkerboard synergy method. Antiadhesion and antibiofilm effect of EOs was evaluated on biotic (HeLa cells) and abiotic surface (polystyrene). To evaluate the possible mechanisms of action, cellular leakage of proteins and DNA was tested and structural changes were visualized with a transmission electron microscope. Results. MIC, MBC and MEC were 1.6 mg/ml for J. communis EO and 3.2 mg/ml for H. italicum EO against both mycobacteria. All combinations of EOs in checkerboard synergy method produced fractional inhibitory concentration index values ranging from 0.501 to 1.5, corresponding to synergistic, additive or indifferent effects. Mycobacterium avium showed a greater tendency to create biofilm but these EO at subinhibitory concentrations (sMIC) effectively blocked the adhesion and the establishment of biofilm. The exposure of both mycobacteria to MICs and sMICs lead to significant morphological changes: acquired a swollen form, ghost-like cell, disorganized cytoplasm detached from the cell wall. OD value of supernatant for both mycobacteria exposed to EOs have confirmed that there is a leakage of cellular material. Conclusion. The leakage of the cellular material is noticeably higher in sMIC, which is probably due to cell wall damage. sMIC of both EOs have an additive or synergistic effect, reducing MICs, limiting adhesion and preventing the formation of biofilms

Evaluation of the Antioxidant Capacity, Antimicrobial and Antiproliferative Potential of Fir (Abies alba Mill.) Honeydew Honey Collected from Gorski kotar (Croatia)

U radu su ispitani antiproliferativni, antimikrobni i antioksidacijski učinci medljikovaca jele (Abies alba Mill.) prikupljenih s planinskog područja Hrvatske (Gorski kotar) a u svrhu potencijalne primjene umjesto standardnih antibiotika i kemoterapeutika. Preživljavanje stanica, annexin V test i protočna citometrijska analiza poslužili su za procjenu antiproliferativnog učinka na indukciju apoptoze i smrti humanih tumorskih staničnih linija: HeLa, MCF-7, SW620, CFPAC-1, MIA PaCa-2 i normalnih diploidnih humanih fibroblasta (BJ). Antimikrobna je aktivnost testirana na različitim sojevima bakterija Staphylococcus i Acinetobacter uporabom difuzijske i mikrodilucijske metode. DPPH˙ testom određena je sposobnost uklanjanja radikala, dok je matematičkim modelima proučavana kinetika inhibicije DPPH˙. Uočen je antiproliferativni učinak na sve ispitivane stanične linije kao i na normalne diploidne fibroblaste (BJ), kolorektalni adenokarcinom (SW620, metastatski) i adenokarcinom epitela dojke (MCF-7, metastatski). Mehanizmi antiproliferativnog učinka uključuju nakupljanje stanica u sub-G1 fazi u svim ispitivanim stanicama te indukciju apoptoze osobito u SW620 i MCF-7 stanicama. Antibakterijski testovi pokazuju da su rezistentni sojevi obaju bakterija, uključujući i multirezistentni soj A. baumannii ATCC® BAA-1605™, bili osjetljivi na sve ispitane uzorke meda. Proučavanjem kinetike uklanjanja radikala dokazano je da antioksidansi prisutni u medu posjeduju različite sposobnosti hvatanja radikala, da reagiraju s njima različitim brzinama i time uzrokuju dva stupnja reakcije. Rezultati istraživanja pokazuju da hrvatski medljikovac ima terapeutski potencijal zbog snažne biološke aktivnosti te može služiti u zaštiti ljudskog zdravlja.The paper examines the antiproliferative, antimicrobial and antioxidative effects of fir (Abies alba Mill.) honeydew honey from mountain region of Croatia (Gorski kotar) as a potential replacement for standard antibiotics and chemotherapeutic agents. Cell viability, annexin V assay and flow cytometry analysis served to analyse the antiproliferative effect on, apoptosis induction in and cell death of cancer cell lines: HeLa, MCF-7, SW620, CFPAC-1, MIA PaCa-2 and normal diploid human fibroblasts (BJ). Antimicrobial activity was tested against Staphylococcus and Acinetobacter strains by agar well diffusion and microdilution assays. The DPPH˙ assay determined the radical scavenging activity, while mathematical models helped to evaluate the kinetic data of DPPH˙ inhibition. Antiproliferative effect on all tested cell lines and the prominent effect on normal diploid human fibroblasts (BJ), colorectal adenocarcinoma (SW620, metastatic) and breast epithelial adenocarcinoma (MCF-7, metastatic) was observable. The mechanisms of antiproliferative effect included accumulation of cells in the sub-G1 phase in all tested cells and induction of apoptosis in SW620 and MCF-7 cells predominantly. The antibacterial assays showed that antibiotic resistant strains of both bacteria, including multi-resistant strain A. baumannii ATCC® BAA-1605™, were sensitive to all tested honey samples. Radical scavenging assay suggests that antioxidants present in the honey possess different radical suppressing abilities and that they react at different rates with radicals, thereby causing two steps of reaction. The results of the study indicate that Croatian fir honeydew honey has a therapeutic potential due to the strong biological activity and can serve to protect human health

Antimikobakterijski potencijal eteričnog ulja plodova borovice (Juniperus communis) u pitkoj vodi

Mycobacterium avium complex-related diseases are often associated with poorly maintained hot water systems. This calls for the development of new control strategies. The aim of this study was to investigate the activity of essential oils (EOs) from the Mediterranean plants, common juniper, immortelle, sage, lavandin, laurel, and white cedar against Mycobacterium avium ssp. avium, Mycobacterium intracellulare, and Mycobacterium gordonae in culturing broth and freshwater as their most common habitat. To do that, we developed a new method of water microdilution to determine their minimal effective concentrations (MEC). The most active EO was the one from the common juniper with the MEC of 1.6 mg mL-1. Gas chromatography / mass spectrometry the juniper EO identified monoterpenes (70.54 %) and sesquiterpenes (25.9 %) as dominant component groups. The main monoterpene hydrocarbons were α-pinene, sabinene, and β-pinene. The juniper EO significantly reduced the cell viability of M. intracellulare and M. gordonae at MEC, and of M. avium at 2xMEC. Microscopic analysis confirmed its inhibitory effect by revealing significant morphological changes in the cell membrane and cytoplasm of all three bacteria. The mode of action of the juniper EO on the cell membrane was confirmed by a marked leakage of intracellular material. Juniper EO has a great practical potential as a complementary or alternative water disinfectant in hot water systems such as baths, swimming pools, spa pools, hot tubs, or even foot baths/whirlpools.Ispitali smo aktivnost eteričnih ulja borovice [Juniperus communis (J. communis)], smilja (Helichrysum italicum), kadulje (Salvia officinalis), lavandina (Lavandula hybrida), lovora (Laurus nobilis) i tuje (Thuja occidentalis) prema sojevima Mycobacterium avium ssp. avium (M. avium), Mycobacterium intracellulare (M. intracellulare) i Mycobacterium gordonae (M. gordonae). Za određivanje minimalnih inhibicijskih koncentracija (MIK) i minimalnih baktericidnih koncentracija (MBK) testiranih eteričnih ulja koristili smo se metodom mikrodiluacije u tekućem bujonu (Middlebrook 7H9), a za određivanje minimalne efektivne koncentracije (MEK) umjesto bujona korištena je sterilna voda iz slavine kojoj je dodano 0,05 % Tweena 80. Dvostruka serijska razrjeđenja eteričnih ulja, počevši od 0,1 do 51,2 mg mL-1, u tekućem bujonu ili u sterilnoj vodi iz slavine inokulirana su s mikobakterijskom suspenzijom uz dodatak resazurina. Najučinkovitije eterično ulje prema svim ispitivanim sojevima mikobakterija bilo je eterično ulje J. communis s MBK / MIK / MEK vrijednošću od 1,6 mg mL-1. Metodom plinske kromatografije i masene spektrometrije analizirano je eterično ulje J. communis. Udio monoterpena iznosio je 70,54 %, a seskviterpena 25,9 %. Glavni monoterpenski ugljikovodici bili su α-pinen, sabinen i β-pinen. Eterično ulje J. communis pokazalo je značajan inhibicijski učinak na M. intracellulare i M. gordonae pri MEK i na M. avium pri dvostrukim MEK. Elektronskom mikroskopijom kod svih triju sojeva mikobakterija nakon izlaganja djelovanju eteričnog ulja J. communis otkrivene su značajne morfološke promjene stanične membrane i citoplazme. Učinak eteričnog ulja J. communis na destrukciju stanične membrane mikobakterija potvrđen je značajnim otpuštanjem unutarstaničnog materijala mjerenjem apsorbancije supernatanta pri 260 nm i 280 nm. Zaključno, u našem smo radu razvili novu metodu za ispitivanje antimikrobnog učinka eteričnih ulja ili drugih prirodnih tvari na netuberkulozne mikobakterije koja oponaša uvjete kao u vodenim sustavima. Prirodni proizvodi, osobito eterična ulja, imaju ne samo velik potencijal kao antimikrobni agensi nego i moguću praktičnu primjenu kao alternativni dezinficijensi

Juniperus communis essential oil limit the biofilm formation of Mycobacterium avium and Mycobacterium intracellulare on polystyrene in a temperature-dependent manner

Formation of biofilms allows bacterial cells to survive in adverse environments. Nontuberculous mycobacteria are ubiquitous in aqueous environments, where they adhere to surfaces and create a biofilm. This has led to the emergence of healthcare-associated infections and the use of biomaterials in medicine. Essential oils (EO) are substances of natural origin whose effect on microorganisms has been the subject of numerous studies. Here, we investigated the effect of Juniperus communis EO on nontuberculous mycobacteria and their early and mature biofilm formation in sterilised tap water. The combination of Juniperus communis EO and increasing ambient temperature showed a synergistic effect on the reduction of biofilm formation of Mycobacterium avium and Mycobacterium intracellulare on a polystyrene surface. A significant antibiofilm effect of Juniperus communis EO was also found at subinhibitory concentrations, suggesting a potential role for it as an alternative disinfectant of natural water

In vitro Antiproliferative and Antimicrobial Activity of the Essential Oil from the Flowers and Leaves of Helichrysum italicum (Roth) G. Don Growing in Central Dalmatia (Croatia)

In the present paper, chemical composition, in vitro antimicrobial and antiproliferative activity of the essential oil from the flowers and leaves of Helichrysum italicum (Roth) G. Don (Central Dalmatia, Croatia) as a potential replacement for standard antibiotics and chemotherapeutic agents was analyzed. Essential oil was isolated by steam distillation and analyzed by GC/MS. Antimicrobial activity was carried out by agar-well diffusion and microdilution assays with Gram (+), Gram (-) bacteria and one yeast. Antiproliferative effect, apoptosis induction and cell death on cancer cell lines: HeLa, MCF-7, SW620, CFPAC-1 and MIA PaCa-2 were analyzed by cell viability and Annexin-V assay as well as by flow cytometric analysis. In essential oil, -pinene, -curcumene and neryl acetate was found as major compounds. The antimicrobial assays, showed that essential oil had weak to moderate antimicrobial potential with S. aureus and S. epidermidis as the most sensitive bacterial strains. Essential oil treatment possessed moderate antiproliferative impact on MCF-7 and HeLa cell lines, while analyzing cell cycle treatment had no significant effect on tested cells except on MIA PaCa-2 with the highest cells increase in sub G1 phase cell cycle. However, treatment caused significantly induction of apoptosis in MCF-7 and HeLa cells, but not in MIA PaCa-2 cells. In this cell line, multiple cell death mechanism existed with involving apoptosis, senescence or necrosis. Results of the study provide a promising basis for the evaluation of the potential use of essential oil from H. italicum (Roth) G. Don as a source of alternative nature antibiotics or anticancer agents for the prevention or treatment of different disease

Evaluation of the Antioxidant Capacity, Antimicrobial and Antiproliferative Potential of Fir (Abies alba Mill.) Honeydew Honey Collected from Gorski kotar (Croatia)

The paper examines the antiproliferative, antimicrobial and antioxidative effects of fir (Abies alba Mill.) honeydew honey from mountain region of Croatia (Gorski kotar) as a potential replacement for standard antibiotics and chemotherapeutic agents. Cell viability, annexin V assay and flow cytometry analysis served to analyse the antiproliferative effect on, apoptosis induction in and cell death of cancer cell lines: HeLa, MCF-7, SW620, CFPAC-1, MIA PaCa-2 and normal diploid human fibroblasts (BJ). Antimicrobial activity was tested against Staphylococcus and Acinetobacter strains by agar well diffusion and microdilution assays. The DPPH˙ assay determined the radical scavenging activity, while mathematical models helped to evaluate the kinetic data of DPPH˙ inhibition. Antiproliferative effect on all tested cell lines and the prominent effect on normal diploid human fibroblasts (BJ), colorectal adenocarcinoma (SW620, metastatic) and breast epithelial adenocarcinoma (MCF-7, metastatic) was observable. The mechanisms of antiproliferative effect included accumulation of cells in the sub-G1 phase in all tested cells and induction of apoptosis in SW620 and MCF-7 cells predominantly. The antibacterial assays showed that antibiotic resistant strains of both bacteria, including multi-resistant strain A. baumannii ATCC® BAA-1605™, were sensitive to all tested honey samples. Radical scavenging assay suggests that antioxidants present in the honey possess different radical suppressing abilities and that they react at different rates with radicals, thereby causing two steps of reaction. The results of the study indicate that Croatian fir honeydew honey has a therapeutic potential due to the strong biological activity and can serve to protect human health

ERA industrial technology roadmap for low-carbon technologies in energy intensive industries

The EU has to drastically accelerate the clean energy transition and increase Europe's energy independence from fossil fuels – and from Russia. This focus is not new: decarbonisation of industry is a key element on the EU’s path to achieving the objective of climate neutrality by 2050 and an intermediate target of reducing greenhouse gas emissions by at least 55% by 2030, as laid down in the European Climate Law. However, bringing innovative low-carbon industrial technologies quickly to the market has become more urgent than ever. The European Research Area (ERA) industrial technology roadmap sketches out the key technologies and the means to transfer them to the industrial ecosystem for energy-intensive industries at EU and national level.Scaling up and deploying the – manageable – number of innovative low-carbon technologies currently at high technology readiness is needed to reach the 2030 emission objectives and to further reduce industry dependence on gas.In order to make best use of the public toolbox to leverage private R&I investment, to increase cross-sector cooperation and accelerate deployment, a number of opportunities for actions were identified, such as to intensify the cooperation with European standardisation organisations (e.g. CEN, CENELEC) and industrial partnerships to identify and fill main standardisation gaps for innovative low-carbon industrial technologies.https://data.europa.eu/doi/10.2777/92567https://dx.doi.org/10.2777/92567KI-01-21-501-EN-NISBN 978-92-76-44692-7EUR 2021.5872 E