Anticancer assessment and antibiofilm potential of Laetiporus sulphureus mushroom originated from Serbia.

Laetiporus sulphureus (Bull.) Murrill is a well-known edible mushroom consumed in nutrition as delicacy. It has been used in traditional medicine because of its beneficial effects on human wellness, such as antimicrobial, antioxidant, and anticancer potential. The present study determined the phenolic profile of Laetiporus sulphureus ethanolic extract (LSE) by high-performance liquid chromatographic method. Tolerance of two probiotic bacterial strains Lactiplantibacillus plantarum 229v, Bifidobacterium animalis subsp. lactis and probiotic yeast Saccharomyces boulardii on LSE was analyzed in terms of viability and biofilm formation. Effects of extract on colorectal (HCT-116) and cervical (HeLa) cancer cells viability was determined using MTT test in concentration range: 1-500 µg/mL after 24 and 72 h. Redox parameters (superoxide anion radicals, nitrites, and reduced glutathione) were evaluated using NBT, Griess, and GSH assays in the concentration range of 1-500 µg/mL after 24 and 72 h. Antimigratory activity was determined by wound healing method using selected concentrations of 10 and 50 µg/mL after 24 h. Untreated cells were considered as control. As control cell line, we used healthy fibroblasts (MRC-5). Our results demonstrated abundance of LSE in phenolics, with rosmarinic acid as the main component. LSE induced low tolerance of tested planktonic probiotic strains, with no affection on their ability to form biofilm. No significant cytotoxicity on tested cancer cells was observed, with prooxidative and antimigratory effects noticed. Extract exerted significant antimigratory activity on cancer cells without effect on planktonic and probiotic cultures in biofilm. These results indicate potential application of Laetiporus sulphureus ethanolic extract as natural protector of probiotics with prominent ability to suppress cancer cell motility.

In Silico Investigation of Selected Pesticides and Their Determination in Agricultural Products Using QuEChERS Methodology and HPLC-DAD.

In this study, we considered some pesticides as active substances within formulations for the protection of plant-based food in the Republic of Serbia in silico, because these pesticides have not often been investigated in this way previously, and in an analytical way, because there are not very many available fast, cheap, and easy methods for their determination in real agricultural samples. Seven pesticides were detected in selected agricultural products (tomatoes, cucumbers, peppers, and grapes) using the QuEChERS methodology and HPLC-DAD. Standard curves for the investigated pesticides (chlorantraniliprole, methomyl, metalaxyl, thiacloprid, acetamiprid, emamectin benzoate, and cymoxanil) show good linearity, with R2 values from 0.9785 to 0.9996. The HPLC-DAD method is fast, and these pesticides can be determined in real spiked samples in less than 15 min. We further characterized the pesticides we found in food based on physicochemical properties and molecular descriptors to predict the absorption, distribution, metabolism, elimination, and toxicity (ADMET) of the compounds. We summarized the data supporting their effects on humans using various computational tools to determine their potential adverse effects. The results of our prediction study show that all of the selected pesticides considered in this study have good oral bioavailability, and those with high toxicity, therefore, could be harmful to human health. Chlorantraniliprole was shown in a molecular docking study as a good starting point for a new Alzheimer's disease drug candidate.

Wild-Growing and Conventionally or Organically Cultivated Sambucus nigra Germplasm: Fruit Phytochemical Profile, Total Phenolic Content, Antioxidant Activity, and Leaf Elements.

European elder or elderberry (Sambucus nigra L., Viburnaceae) is a plant species with known high pharmaceutical and nutritional value. However, the Greek native germplasm of S. nigra has not been adequately utilized to date as in other regions. This study evaluates the fruit antioxidant potential (total phenolic content and radical scavenging activity) of wild-growing and cultivated germplasm of Greek S. nigra. In addition, nine cultivated Greek S. nigra genotypes were evaluated regarding the effects of fertilization (conventional and organic) on fruit phytochemical and physicochemical potential (total flavonoids, ascorbic acid content, pH, total soluble solids, and total acidity), as well as on the antioxidant potential (total phenolic content and radical scavenging activity) of fruits and leaves. Additionally, an analysis of macro- and micro-elements in the leaves of the cultivated germplasm was performed. The results demonstrated comparatively higher total phenolic contents of fruits of cultivated germplasm. The genotype was the decisive factor in the fruits' phytochemical potential and leaves' total phenolic content of cultivated S. nigra germplasm. Similarly, fertilization regime effects were found to be genotype-dependent, affecting fruit phytochemical and physicochemical attributes. The trace element analysis results were similar, with genotypes varying significantly in their concentrations of macro- and micro-elements. The current work builds on previous domestication attempts for Greek S. nigra, providing new data on the phytochemical potential of this important nutraceutical species.

Chemometric Approach to the Composition of Flavonoid Compounds and Phenolic Acids and Antioxidant Potential of Artemisia Species from Different Habitats.

Four Artemisia species from Serbia were selected for the study: A. annua L., A. absinthium L., A. vulgaris L. and A. scoparia Waldst. et Kit. because of the proven excellent action in treating certain medical conditions and diseases. A. absinthium L., A. vulgaris L., A. annua L. and A. scoparia Waldst. et Kit. collected from different habitats across Serbia (48 samples in total) were studied from the statistical aspect considering the phenolic and flavonoid contents, compositions, and antioxidant activities of methanol extracts in correlation with the soil type. The components were identified using HPLC (High Performance Liquid Chromatography), while antioxidant activities were determined by seven assays (TP (Total phenolic content), TF (Total flavonoid content), DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-based, ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radical cation-based, FRAP (Ferric Reducing Antioxidant Power), TRP (Total Reducing Power), and CUPRAC (Cupric Reducing Antioxidant Capacity). The results were processed using five statistical methods (PCA (Principal Component Analysis), ANOVA (Analysis of Variance), MANOVA (Multivariate analysis of variance), DA (Discriminant analysis), and AHC (Agglomerative Hierarchical Clustering)). Principal component analysis enabled very well separation of the analyzed Artemisia species based on the content of total phenolics, total flavonoids, and the values obtained from antioxidant tests, but not on the individual compounds identified and quantified by HPLC. The MANOVA analyses showed that for A. scoparia and A. annua there was a significant effect of soil type on the total phenolics, total flavonoids, and antioxidant tests. In contrast, for A. vulgaris and A. absinthium, that effect was not significant. Additional MANOVA analyses showed a significant effect of soil type on phenolic and flavonoid compounds in the case of A. vulgaris, A. annua, and A. absinthium. The overall correct classification rate of all samples of four investigated Artemisia species by the discriminant analysis was 81.25 % using the training sample and 72.92 % the cross-validation results based on TP, TF, and antioxidant tests, but a much lower based on selected compounds identified and quantified by HPLC. MANOVA analyses based on particular Artemisia species show that soil is a significant factor affecting the measurable variables.

Comparative in vitro studies of the biological potential and chemical composition of stems, leaves and berries Aronia melanocarpa's extracts obtained by subcritical water extraction.

Preparation of functional products as well as natural-based products requires non-toxic but effective extraction techniques. In this study, subcritical water extraction was used for the extraction of different aronia parts in order to explore their potential. Stems, leaves and berries of Aronia melanocarpa were extracted under the following conditions: temperature 130 °C; pressure 35 bar; time 20 min. The total phenols and flavonoid contents of the produced extracts were evaluated by conventional spectrophotometric methods. Additionally, the main phenolic compounds were also identified and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). The biological potential of the extracts was evaluated by determining their antioxidant (DPPH, ABTS and lipid peroxidation assays), antimicrobial, enzyme inhibitory (cholinesterase and elastase) and cytotoxic effects (HeLa, A-549, LS-174T, MRC-5 cell lines). The results indicate that leaves and berries extracts exhibited stronger antioxidant action when compared with stems. The strongest cholinesterase and elastase inhibitory activity was also found in berries extract. Similarly, the extracts obtained from leaves and berries showed considerable cytotoxic effects against tested cell lines. A moderate antimicrobial effects was observed too. Demonstrated biological potential of all three aronia parts can trace a new road map for developing newly designed functional products.

Biological activity and chemical profile of Lavatera thuringiaca L. extracts obtained by different extraction approaches.

BACKGROUND: Lavatera thuringiaca L. is herbaceous perennial plant from Malvaceae family, which is known for its biological activity and richness in polyphenolic compounds. Despite this, the information regarding the biological activity and chemical profile is still insufficient. PURPOSE: Aim of this study was to investigate biological potential and chemical profile of Lavatera thuringiaca L., as well as influence of applied extraction technique on them. STUDY DESIGN AND METHODS: Two conventional and four non-conventional extraction techniques were applied in order to obtain extracts rich in bioactive compound. Extracts were further tested for total phenolics, flavonoids, condensed tannins, gallotannins and anthocyanins contents using spectrophotometric assays. Polyphenolic profile was established using HPLC-DAD analysis. Biological activity was investigated regarding antioxidant, cytotoxic and antibacterial activities. Four antioxidant assays were applied as well as three different cell lines for cytotoxic and fifteen bacterial strain for antibacterial activity. RESULTS AND CONCLUSION: Results showed that subcritical water extraction (SCW) dominated over the other extraction techniques, where SCW extract exhibited the highest biological activity. Study indicates that plant Lavatera thuringiaca L. may be used as a potential source of biologically compounds.

Phenolic profiles and metal ions analyses of pulp and peel of fruits and seeds of quince (Cydonia oblonga Mill.).

Six hydroxycinnamic acids were identified and determined quantitatively in methanol and acetone extracts from quince peel and pulp, namely 3-O-caffeoylquinic acid (3-CQA), 4-p-coumaroylquinic acid (HC1), 4-O-caffeoylquinic acid (4-CQA), 5-O-caffeoylquinic acid (5-CQA), derivative of p-coumaroylquinic acid (HC2) and 3,5-dicaffeoylquinic acid (3,5-diCQA). The most abundant hydroxycinnamic acid was 5-CQA (neochlorogenic acid) with 259.12-481.4mg/kgf.w. in peel and 97.33-217.36mg/kg in quince pulp. Six flavonols were determined in the extracts from quince, quercetin-3-galactoside (Q-Ga), quercetin-3-rutinoside (Q-Ru), quercetin-3-glucoside (Q-Glu), kaempferol-3-rutinoside (K-Ru), kaempferol-3-glucoside (K-Glu) and derivative of quercetin produced in the reaction between quercetin-glucoside and p-coumaric acid (Q-Glu-p-CouA). Elemental analysis of quince seeds has not been performed previously. Also, using principal component and cluster analyses, we determined a strong negative relationship between total phenols and flavonoids, and Ni and Pb, specifically higher concentrations of these compounds were associated with lower concentrations of these metals.

Statistical characteristics of selected elements in vegetables from Kosovo.

Zinc, copper, iron, chromium and cobalt are essential elements for human health, showing toxicity only in high concentrations, while lead and cadmium are extremely toxic even as traces. Therefore, it is important to monitor the contents of toxic metals in vegetables. Large number of vegetables is grown and used in nutrition, in Kosovo. The concentrations of selected elements in vegetables (radish, onion, garlic and spinach) from Kosovo were determined using ICP-OES method. Oral intake of metals and health risk index were calculated. Statistical analysis indicated numerous positive correlations between concentrations of selected elements in vegetables. As a result of principal component analysis, 15 new variables were obtained which were characterized by eigenvalues. The sequence of health quotients for the heavy metals followed the decreasing order Zn = Mn > Pb > Cu > Ni > Fe > Cd > Co > Cr. The health quotients for all investigated heavy metals were below 1 (one), which is considered safe. The vegetables from Kosovo are mainly safe for use in everyday diet.

A simple method for the ampicillin determination in pharmaceuticals and human urine.

The new kinetically-based spectrophotometric method for the determination of microquantities of ampicillin is proposed in the present paper. Ampicillin degradation in strong alkaline medium was applied for the method development. The reaction rate was monitored at 265 nm. A differential variation of the tangent method was used to process the kinetic data. The method is valid over the 3.49-55.84 µg/mL ampicillin concentration interval with relative standard deviation (RSD) range 7.79-3.20%. The calculated detection limit was determined at 2.58 µg/mL based on the 3.3S0 criterion. The interference effects of some metal ions, anions, amino acids and other molecules were investigated in order to assess the method selectivity. The method was successfully applied to determining the content of ampicillin in commercial pharmaceutical preparations and human urine. The obtained results were in good correlation with the HPLC method results. The newly developed method is simple, inexpensive and efficient for the analysis of a large number of samples at room temperature in a short time.

Heavy metal content in different types of smoked meat in Serbia.

Concentrations of Fe, Cu, Ni, Cr, Pb, Mn, Zn and Cd in pork, beef, turkey and chicken samples were determined using inductively coupled plasma-optical emission spectrometry (ICP-OES). The mean concentration ranges in milligrams per 100 g of the studied metals in all samples were 0.6924-1.2154 for Fe, 0.6492-0.9831 for Cu, 0.0012-0.0027 for Pb, 0.041-0.0510 for Ni, 0.1186-0.1481 for Mn, 0.7257-5.2726 for Zn and 0.0042-0.0050 for Cd. The levels of analysed elements were in accordance with European standards for all metals except for manganese in all samples and for nickel in a certain number of samples. Zn level in beef was significantly higher compared to other samples, and Pb and Cd were found in concentrations well below the recommended daily intake.

Antioxidant capacities and phenolic levels of different varieties of Serbian white wines.

The biologically active compounds in wine, especially phenolics, are responsible for reduced risk of developing chronic diseases (cardiovascular disease, cancer, diabetes, etc.), due to their antioxidant activities. We determined the contents of total phenolics (TP) and total flavonoids (TF) in selected Serbian white wines by colorimetric methods. Total antioxidant activity (TAA) of the white wines was analyzed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity assay. Medas beli had the highest content of TP, TF and TAA. The radical scavenging capacity (RSC) and total antioxidant activity (TAA) of white wines were 15.30% and 1.055 mM Trolox equivalent, respectively. Total phenolic (TP) and total flavonoid (TF) contents in white wines ranged from 238.3 to 420.6 mg gallic acid equivalent per L of wines and 42.64 to 81.32 mg catechin equivalent per L of wines, respectively. A high and significant correlation between antioxidant activity and total phenolic content was determined in wines (R2 = 0.968, p < 0.01). For the individual polyphenols determination we used a high performance liquid chromatography (HPLC)-diode array detection (DAD) technique. The majority of white wine polyphenols was represent by four hydroxycinnamic acids (HCAs).