Unravelling Anti-Melanogenic Potency of Edible Mushrooms Laetiporus sulphureus and Agaricus silvaticus In Vivo Using the Zebrafish Model

Severe drawbacks associated with the topical use of depigmenting agents in treatments of skin hyperigmentations impose a great demand for novel, effective, and safe melanogenesis inhibitors. Edible and medicinal mushrooms, known for numerous health-promoting properties, represent a rich reservoir of anti-melanogenic compounds, with the potential to be applied in preventing excessive skin pigmentation. Herein, using zebrafish (Danio rerio) as a preclinical animal model, we have demonstrated that ethanol extract of Laetiporus sulphureus (LSE) and Agaricus silvaticus (ASE) are not toxic at high doses up to 400-500 mu g/mL while effectively inhibit melanogenesis in a dose-dependent manner. At depigmenting doses, the explored extracts showed no adverse effects on zebrafish embryos melanocytes. Even more, they did not provoke inflammation or neutropenia when applied at the highest dose ensuring almost complete the cells depigmentation. Since LSE and ASE have demonstrated significantly higher the therapeutic potential than kojic acid and hydroquinone, two well-known depigmenting agents, overall results of this study strongly suggest that the explored mushrooms extracts could be used as efficient and safe topical agents in treatments of skin hyperpigmentation disorders

Produktivnost golozrnog ječma u organskom i konvencionalnom sistemu gajenja

There is rising interest for hulless barley use in human nutrition and industrial processing. Hulless barley is a relatively new cereal crop; the hulls of it can be separated from grain during threshing. This cereal has been recognized as being more valuable and economic in food industry than covered barley. Several studies show the positive influence of hulless barley food products on human health; it can be utilized in many different food products. Aim of this paper is to investigate possibilities of organic growing of hulless barley, comparing with conventional one, in agroecological conditions of Valjevo hilly region during 2008/2009. Trial was set up in Kotešica village on soil which was not used for agriculture for 7 years. In organic cropping system three combinations of microbiological fertilizer baktofil with zeolite and hydrogel were used prior to sowing. Half of each plot was treated with foliar microbiological fertilizer (Slavol) during crop growing period. In conventional cropping system three variants with mineral fertilizers were included: NPK, NPK+zeolite, NPK+hydrogel. Results of the yield obtaind in the experiment showed no significant differences between control and treatments. Yield average in organic cropping system (4,54 t/ha) was slightly higher than in conventional one (4,48 t/ha), but both of them were lower than in control with no fertilizer (4,65 t/ha). According to yield obtained in separate plots, the highest value gave the treatment NPK+zeolit+slavol. Different microbiological fertilizers combined with NPK fertilizer and zeolite give the maximum results in hulles barly production.Poslednjih desetak godina u svetu raste interes za upotrebom golozrnog ječma u direktnoj ljudskoj ishrani i industrijskoj preradi. Razlog za to je pre svega što golozrni ječam predstavlja bogat izvor rastvorljivih biljnih vlakana, koji povoljno utiču na ljudsko zdravlje. Cilj ovog rada je da se ispita mogućnost organskog gajenja ove biljne vrste, u odnosu na konvencionalni u konkretnim agroekološkim uslovima Valjevskog pobrđa 2008/2009 godine. Ogled je postavljen u selu Kotešica, na zemljištu koje nije korišćeno u konvencionalnoj proizvodnji 7 godina. U organskom sistemu poljoprivredne proizvodnje, korišćene su kombinacije mikrobiološkog đubriva baktofila sa dva poboljšivača zemljišta zeolita i hidrogela, kojima je tretirano zemljište neposredno pred setvu, a u toku vegetacionog perioda neke varijante ogleda su folijarno tretirane mikrobiološkim đubrivom (slavol). U konvencionalnom sistemu poljoprivredne proizvodnje uključene su tri varijante sa kompleksnim mineralnim NPK đubrivom i kombinacija sa zeolitom i hidrogelom, kojima je tretirano zemljište neposredno pred setvu. Dobijeni rezultati prosečnih prinosa golozrnog jećma pokazuju da na parceli, koja nije korišćena dugi niz godina, nisu dobijene značajne razlike između kontrole, bez đubrenja i tretmana sa mikrobiološkim ili mineralnim đubrivima. Prosečni prinos u organskom sistemu gajenja (4,54 t/ha) je bio nešto viši od konvencionalnog (4,48 t/ha), a oba su bila nešto niža od kontrole bez đubrenja (4,65 t/ha). Posmatrajući prinose po pojedinačnim varijantama najveći prinos je postignut u tretmanu NPK+zeolit+slavol. U ovoj varijanti ogleda mikrobiološka đubriva su u kombinaciji sa NPK i poboljšivačem zemljišta dala maksimalne rezultate u gajenju golozrnog ječma

Sesquiterpene Lactones of Amphoricarpos autariatus ssp autariatus from Montenegro - Antifungal Leaf - Surface Constituents

The composition of leaf cuticular neutral lipids of Amphoricarpos autariatus ssp. autariatus collected at canyon of river Tara (North Montenegro) was investigated by GC/MS (nonpolar fraction), LC-ESI TOF MS and H-1 NMR spectroscopy (more polar fraction). The nonpolar fraction (ca. 15% of the whole surface extract) contained C-27 (-) (33) n-alkanes, those with odd-number of carbons predominating. The LC-ESI MS and H-1 NMR of the more polar fraction revealed 13 sesquiterpene lactones, constituting ca. 97.5% of the lactone mixture, identified as the known guaianolides, so-called amphoricarpolides, found previously in the aerial parts of the genus. The lactone fraction exhibited considerable in vitro effect against eight fungi, i.e. Aspergillus ochraceus, A. niger, A. versicolor, Penicillium funiculosum, P. ochrochloron, Trichoderma viride, Fusarium verticillioides and Fulvia fulvum

Comparative evaluation of antimutagenic and antimitotic effects of Morchella esculenta extracts and protocatechuic acid

Morchella esculenta (L.) Pers. methanolic extracts, obtained from fruiting bodies growing wild in Serbia and Portugal, were screened for their antimutagenic properties and compared to protocatechuic acid, previously identified in both species. Salmonella typhimurium TA100 reversion assay was used for the antimutagenic properties. Methanolic extracts expressed important antimutagenic potency towards S. typhimurium, which was documented by index of antimutagenicity (I). A sample from Serbia expressed slightly higher antimutagenic properties with an inhibition rate of 58.7%. A sample from Portugal gave an inhibition rate of 51.7%. Protocatechuic acid had an inhibition rate I of his(+) revertants of 72.4%. Cell viability in the presence of extracts was also documented. M. esculenta samples from Serbia and Portugal possessed novel biological potential for the studied species, as well as its phenolic compound - protocatechuic acid, identified in both samples. Genotoxic effect, regarding mitotic index and chromosomal aberration score, was also assessed by using the Allium cepa L. assay. Protocatechuic acid showed the most significant decrease in mitotic index, as well as decrease in chromosomal aberration score

Gut microbial dysbiosis occurring during pulmonary fungal infection in rats is linked to inflammation and depends on healthy microbiota composition

While the effect of gut microbiota and/or inflammation on a distant body site, including the lungs (gut–lung axis), has been well characterized, data about the influence of lung microbiota and lung inflammation on gut homeostasis (lung–gut axis) are scarce. Using a well-characterized model of pulmonary infection with the fungus Aspergillus fumigatus, we investigated alterations in the lung and gut microbiota by next-generation sequencing of the V3–V4 regions of total bacterial DNA. Pulmo- nary inflammation due to the fungus A. fumigatus caused bacterial dysbiosis in both lungs and gut, but with different characteristics. While increased alpha diversity and unchanged bacterial composition were noted in the lungs, dysbiosis in the gut was characterized by decreased alpha diversity indices and modified bacterial composition. The altered homeostasis in the lungs allows the immigration of new bacterial species of which 41.8% were found in the feces, indicating that some degree of bacterial migration from the gut to the lungs occurs. On the contrary, the dysbiosis occurring in the gut during pulmonary infection was a consequence of the local activity of the immune system. In addition, the alteration of gut microbiota in response to pulmonary infection depends on the bacterial composition before infection, as no changes in gut bacterial microbiota were detected in a rat strain with diverse gut bacteria. The data presented support the existence of the lung–gut axis and provide additional insight into this mechanism. IMPORTANCE Data regarding the impact of lung inflammation and lung microbiota on GIT are scarce, and the mechanisms of this interaction are still unknown. Using a well-characterized model of pulmonary infection caused by the opportunistic fungus Aspergillus fumigatus, we observed bacterial dysbiosis in both the lungs and gut that supports the existence of the lung–gut axis. KEYWORDS fungal lung infection, gastrointestinal microbiota, lung microbiota, lung-gut axis, rats B acteria inhabit every part of the human body, but most of them are found in the gut. Gut microbiota are responsible for many functions, including nutrient metabolism, immunomodulation, maintenance of host physiology, and protection against pathogen overgrowth (1). To date, numerous scientific studies confirm the important role of gut bacteria in health and disease. This microbial community impacts not only local immunity but also a distant body site, such as the lungs. Disturbances in gut bacterial composition have been linked to asthma (2), chronic obstructive pulmonary disease (3), cystic fibrosis (4), and lung cancer (5). Furthermore, pulmonary involvement was noted in inflammatory gastrointestinal disease characterized by microbial dysbiosis (6), Month XXXX Volume 0 Issue 0 10.1128/spectrum.01990-23 1 Editor Agostinho Carvalho, University of Minho, Braga, Portugal Address correspondence to Maja Tolinacki, [email protected]. The authors declare no conflict of interest. See the funding table on p. 15. Received 11 May 2023 Accepted 25 July 2023 Published 25 August 2023 Copyright © 2023 Popovic et al. This is an open- access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Downloaded from https://journals.asm.org/journal/spectrum on 09 October 2023 by 147.91.199.205. supporting the existence of a gut–lung axis. The gut bacterial microbiota or some of their constituents impact the immune response in the lungs against viruses (7–9), bacteria (10–13), fungi (14), and allergic airway inflammation (15) mainly through the effect of the gut microbiota (or their metabolites) on the immune cell activity. While the gut–lung axis is well characterized, the influence of the lung microbiota as well as lung inflammation on gut homeostasis has attracted much more attention in recent years. The first indication of the lung–gut axis was a higher prevalence (compared to healthy subjects) of gastrointestinal symptoms in patients with asthma (16) and chronic obstructive pulmonary disease (17). The existence of gastrointestinal symptoms in patients with pulmonary virus infection has also been documented (18). Gastrointesti- nal symptoms (abdominal pain, nausea, vomiting, and diarrhea) were noted in 11.6% of children with influenza infection (18), and a later study showed a decrease in alpha diversity in the feces of influenza-infected patients compared to healthy controls (19). Fecal bacterial samples from patients with COVID-19 infection were shown to cluster separately from those in healthy controls as well, but in the majority of these patients, SARS-Cov-2 could be detected in the feces (20). Experimental studies in mice confirmed the occurrence of gut dysbiosis following respiratory influenza virus infection (21–25) and respiratory syncytial virus infection (24), despite the fact that the virus has not been detected in the gut (21, 22, 24, 25). It has been shown that the alteration of gut microbiota is a consequence of infection with live virus particles, as administration of an attenuated influenza vaccine had no effect on the microbiota (24). Bacterial dysbiosis in the gut also occurs following pulmonary bacterial infection. A decrease in alpha diversity indices and differential relative abundance of fecal microbiota were noted in patients with pulmonary tuberculosis (26, 27) and in mice infected with Mycobacterium tuberculosis (28) and Klebsiella pneumoniae (29). Even administration of the major component of the outer membrane of Gram-negative bacteria, lipopolysac- charide, to the lungs caused gut bacteria dysbiosis (30). In addition to pulmonary infections caused by viruses or bacteria, alteration of the gut microbiota was noted in mice exposed to hyperoxia (31) and in patients with lung cancer (compared to healthy individuals) (32) indicating that pulmonary inflammation/injury affects the gut microbiota regardless of its origin. Despite a growing body of evidence for interaction between the lungs and gut, there is still a lot of work to be done to understand this crosstalk. There are virtually no data regarding gut microbiota changes during pulmonary infection caused by fungi. Our previous study showed an alteration in immune-mediated homeostasis of the gut in a rat model of sublethal pulmonary infection with A. fumigatus (33). Using the same experimental model of infection in Dark Agouti (DA) rat strain, we aimed to investigate changes in the lung and gut microbiota by next-generation sequencing of the V3–V4 regions of total bacterial DNA in these two organs. Possible mechanisms of lung–gut communication were also investigated. In addition, to examine whether gut dysbiosis is a general characteristic during pulmonary fungal infection, we analyzed feces from infected Albino Oxford (AO) rats, a strain that develop quantitatively different immune response to fungus A. fumigatus (34) and whose gut microbiota was previously shown to respond differently to oral cadmium administra- tion (35) compared to DA rats

Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014

Supplementary material for: [https://doi.org/10.1016/j.ijantimicag.2016.08.014]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2364

Apigenin-7-o-glucoside versus apigenin: insight into the modes of anticandidal and cytotoxic actions

Bioactive potential of apigenin derivative apigenin-7-O-glucoside related to its antifungal activity on Candida spp. and cytotoxic effect on colon cancer cells was studied and compared with bioactive potential of apigenin. Antifungal activity was tested on 14 different isolates of Candida spp. using membrane permeability assay, measuring inhibition of reactive oxidative species and inhibition of CYP51 C. albicans enzyme. Cytotoxic potential of apigenin- 7-O-glucoside was tested on colon cancer HCT116 cells by measuring cell viability, apoptosis rate and apoptosis- and colon cancer-related gene expression. Obtained results indicated considerable antifungal activity of apigenin-7-O-glucoside towards all Candida isolates. Breakdown of C. albicans plasma membrane was achieved upon treatment with apigenin-7-O-glucoside for shorter period of time then with apigenin. Reduction of intra-and extracellular reactive oxidative species was achieved with minimum inhibitory concentrations of both compounds, suggesting that reactive oxidative species inhibition could be a mechanism of antifungal action. None of the compounds exhibited binding affinity to C. albicans CYP51 protein. Besides, apigenin-7-O-glucoside was more effective compared to apigenin in reduction of cell's viability and induction of cell death of HCT116 cells. Treatment with both compounds resulted in chromatin condensation, apoptotic bodies formation and apoptotic genes expression in HCT116 cells, but the apigenin-7-O-glucoside required a lower concentration to achieve the same effect. Compounds apigenin-7-O-glucoside and apigenin displayed prominent antifungal potential and cytotoxic effect on HCT116 cells. However, our results showed that apigenin-7-O-glucoside has more potent activity compared to apigenin in all assays that we used

Apigenin-7-O-glucoside versus apigenin

Bioactive potential of apigenin derivative apigenin-7-O-glucoside related to its antifungal activity on Candida spp. and cytotoxic effect on colon cancer cells was studied and compared with bioactive potential of apigenin. Antifungal activity was tested on 14 different isolates of Candida spp. using membrane permeability assay, measuring inhibition of reactive oxidative species and inhibition of CYP51 C. albicans enzyme. Cytotoxic potential of apigenin-7-O-glucoside was tested on colon cancer HCT116 cells by measuring cell viability, apoptosis rate and apoptosis- and colon cancer-related gene expression. Obtained results indicated considerable antifungal activity of apigenin-7-O-glucoside towards all Candida isolates. Breakdown of C. albicans plasma membrane was achieved upon treatment with apigenin-7-O-glucoside for shorter period of time then with apigenin. Reduction of intra- and extracellular reactive oxidative species was achieved with minimum inhibitory concentrations of both compounds, suggesting that reactive oxidative species inhibition could be a mechanism of antifungal action. None of the compounds exhibited binding affinity to C. albicans CYP51 protein. Besides, apigenin-7-O-glucoside was more effective compared to apigenin in reduction of cell’s viability and induction of cell death of HCT116 cells. Treatment with both compounds resulted in chromatin condensation, apoptotic bodies formation and apoptotic genes expression in HCT116 cells, but the apigenin-7-O-glucoside required a lower concentration to achieve the same effect. Compounds apigenin-7-O-glucoside and apigenin displayed prominent antifungal potential and cytotoxic effect on HCT116 cells. However, our results showed that apigenin-7-O-glucoside has more potent activity compared to apigenin in all assays that we used

Antimicrobial Activity of the Freshwater Bryozoan Hyalinella Punctata (Hancock, 1850)

The antimicrobial activity of the freshwater bryozoan Hyalinella punctata (Hancock, 1850) was tested by microdilution method against eight bacteria and eight fungi for the first time. All five crude extracts (hexane, acetone, dimethyl sulfoxide, methanol and water) showed good antibacterial and antifungal potential in vitro wherein the acetone extract was the most active (MICs 0.50-7.00 mu g/ml and MBCs 2.50-10.00 mu g/ml)