Biosynthesis of Silver Nanoparticles Using Salvia pratensis L. Aerial Part and Root Extracts: Bioactivity, Biocompatibility, and Catalytic Potential.

The aim of this research was the synthesis of silver nanoparticles (SPA- and SPR-AgNPs) using the aqueous extracts of the aerial (SPA) and the root (SPR) parts of the plant Salvia pratensis L., their characterization, reaction condition optimization, and evaluation of their biological and catalytic activity. UV-Vis spectroscopy, X-ray powder diffraction (XRPD), scanning electron microscopy with EDS analysis (SEM/EDS), and dynamic light scattering (DLS) analysis were utilized to characterize the nanoparticles, while Fourier transform infrared (FTIR) spectroscopy was used to detect some functional groups of compounds present in the plant extracts and nanoparticles. The phenolic and flavonoid contents, as well as the antioxidant activity of the extracts, were determined spectrophotometrically. The synthesized nanoparticles showed twice-higher activity in neutralizing 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) compared with the respective extracts. SPR-AgNPs exhibited strong antimicrobial activity against almost all of the tested bacteria (<0.0039 mg/mL) and fungal strains, especially against the genus Penicillium (<0.0391 mg/mL). Moreover, they were fully biocompatible on all the tested eukaryotic cells, while the hemolysis of erythrocytes was not observed at the highest tested concentration of 150 µg/mL. The catalytic activity of nanoparticles toward Congo Red and 4-nitrophenol was also demonstrated. The obtained results confirm the possibility of the safe application of the synthesized nanoparticles in medicine and as a catalyst in various processes.

Strategies of accumulation of potentially toxic elements in Minuartia recurva and M. bulgarica.

The aim of this study was to determine the concentrations of potentially toxic elements in soil samples and plant tissues of Minuartia recurva and M. bulgarica, predominantly or exclusively calcifuge species. Biological concentration (BCs) and translocation factors (TFs) were used to evaluate their accumulation potential. Considerable differences were observed between M. recurva and M. bulgarica assessions in terms of accumulation strategies of potentially toxic elements (PTEs). In M. recurva, most of the elements analyzed (Mn, Cu, Zn, Cd, and Co) were transported to the shoot, whereas in M. bulgarica, these elements remained predominantly in the roots. The Cu concentrations in the shoot samples of M. recurva from an abandoned iron-copper mine at Mt. Kopaonik were clearly above the notional hyperaccumulation threshold, characterizing this species as a possible Cu hyperaccumulator. Additionally, strong accumulation potential for Cr, Ni, Zn, Pb, and Cd was observed in M. recurva assessions, but without significant accumulation due to the low concentrations of these elements in the soils. The strong accumulation capacity and the different strategies in tolerance to PTEs indicate a potential of the two species for an application in phytoremediation: M. recurva for phytoextraction and M. bulgarica for phytostabilization.

Plantago subulata as indicator of potentially toxic elements in the substrate.

Plantago subulata is a facultative serpentinophyte, with predominantly ultramafic distribution in Serbia and Montenegro. The plant samples were collected from ultramafic and non-ultramafic substrate, including two abandoned mining sites, with the aim to assess the accumulation potential of this species. The samples were collected from 10 sites in Serbia and Montenegro and element concentrations in plants and soils were determined. Particularly high concentrations of Fe, Cu, Mn, Zn, and Cd were found in the soil and plant samples from an abandoned iron mining site, Mt. Kopaonik, Suvo Rudiste. The concentrations of Ni, Cr, Fe, Co, and Cd were statistically different between soil and plant samples from ultramafic and non-ultramafic bedrock. Considering the fact that concentrations of Zn, Ni, and Cr in roots and shoots were positively correlated with their respective contents in the soil, i.e., the chemical composition of the plant and soil samples reflected the characteristics of the substrate, and for most of the elements analyzed, P. subulata acted as indicator species. For Cd and Pb, only the root concentrations were positively correlated with soil content, indicating exclusion and root sequestration as the main tolerance strategies for these elements. Although below the hyperaccumulation threshold, a strong accumulation capacity of P. subulata was found for Ni and Cu.

Application potential of biogenically synthesized silver nanoparticles using Lythrum salicaria L. extracts as pharmaceuticals and catalysts for organic pollutant degradation.

This study was designed to evaluate the optimal conditions for the eco-friendly synthesis of silver nanoparticles (AgNPs) using Lythrum salicaria L. (Lythraceae) aqueous extracts and their potential application and safe use. AgNPs synthesized using L. salicaria aerial parts (LSA-AgNPs) and root extract (LSR-AgNPs) were characterized by UV-Vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM/EDS), and X-ray powder diffraction (XRPD). Dynamic light scattering (DLS) was used for the determination of the size distribution profiles of the obtained nanoparticles. Both L. salicaria extracts showed high phenolic content, while the flavone C-glucosides orientin, vitexin, and isovitexin were detected in extracts using HPLC. The synthesized AgNPs displayed growth inhibition of the tested bacteria and fungi in concentrations between 0.156 and 1.25 mg mL-1. The studied nanoparticles also showed antioxidant potential and gained selectivity at different concentrations on different cancer cell lines. Concentrations of LSA-AgNPs were found to be 20.5 and 12 µg mL-1 towards A431 and SVT2, respectively, while LSR-AgNPs were effective only against A431 cancer cells (62 µg mL-1). The hemolytic activity of LSA-AgNPs in concentrations up to 150 µg mL-1 was not observed, while LSR-AgNPs in the highest applied concentration hemolyzed 2.8% of erythrocytes. The degradation possibility of Congo red and 4-nitrophenol using LSA-AgNPs and LSR-AgNPs as catalysts was also proven. The results indicate that L. salicaria may be used for the eco-friendly synthesis of AgNPs with possible applications as antimicrobial and selective cytotoxic agents towards cancer cell lines, as well as in catalytic degradation of pollutants.

Accumulation of trace elements in Tussilago farfara colonizing post-flotation tailing sites in Serbia.

The purpose of this study was to examine the accumulation potential of spontaneously developed Tussilago farfara populations colonizing sites with different levels of anthropogenic pollution. Physical characteristics of the soil are presented, together with the concentrations of macroelements and microelements (Ca, Mg, Fe, S, Al, Pb, Zn, Cu, Cd, Mn, As, Sb, Ag, Ti, and Sr) in both soil and plants. The biological concentration, accumulation, and translocation factors were used to assess the potential for heavy metal accumulation. Considerable differences were found among assessions from unevenly contaminated habitats, particularly in comparison with an unpolluted site. In line with the ore's characteristics, substrate samples from polluted sites were heavily contaminated with Pb, Zn, As, and Sb. Increased levels of microelements were also detected in plant samples from flotation tailings. Despite active absorption of Zn, Cu, Cd, Mn, and Sr by the plants from mining sites, the detected quantities of these elements in all samples were below the hyperaccumulation threshold. However, the obtained results indicate that the use of T. farfara from such sites in traditional medicine could pose a risk to human health due to accumulation of several toxic elements in the plant's aboveground tissues. Additionally, as a successful primary colonizer and stabilizer of technogenic substrates, T. farfara has an important role in the initial phases of revegetation of highly contaminated sites.

Assessment of trace element accumulation potential of Noccaea kovatsii from ultramafics of Bosnia and Herzegovina and Serbia.

In this work, we present the results of the investigation of trace elements (Fe, Mg, Ni, Zn, Cu, Cr, Co, Cd, Pb) accumulation potential of Noccaea kovatsii (Heuff.) F. K. Mey., from the Balkan Peninsula. The study included eight populations from ultramafic soils, six from Bosnia and Herzegovina, and two from Serbia. Principal component analysis (PCA) was used to reveal relationships of elements in soil, and Pearson's correlation coefficients for analysing associations of available quantities of elements in soil and those in roots and shoots of N. kovatsii. Uptake and translocation efficiency was assessed by using bioconcentration (BCF) and translocation factors (TF). All the analysed populations of N. kovatsii emerged as strong Ni accumulators, with the highest shoot concentrations of 12,505 mg kg-1. Even thought contents of Zn in plant tissues of N. kovatsii were under the hyperaccumulation level (602 mg kg-1 and 1120 mg kg-1 respectively), BCF was up to 667, indicating that certain surveyed populations have strong accumulative potential for this element.

Analysis of Wild Raspberries (Rubus idaeus L.): Optimization of the Ultrasonic-Assisted Extraction of Phenolics and a New Insight in Phenolics Bioaccessibility.

A simple and efficient ultrasonic-assisted extraction (UAE) technique was developed in order to find optimal conditions for the extraction of total phenolic compounds, flavonoids and anthocyanins in wild raspberry (Rubus idaeus L.) fruits. Several extraction variables, including methanol composition (v/v, %), solid-solvent ratio (g/mL), time (min) and extraction temperature (°C) were optimized using response surface methodology (RSM). Under optimal conditions for extraction, the total phenolics were found in the concentration of 383 mg GAE/100 g of fresh fruit weight, while HPLC-PDA analysis of the optimized extract showed the presence of cyanidin-3-glucoside, cyanidin-3-sophoroside, catechin, gallic and ellagic acid. The experimental values of DPPH and ABTS radical scavenging activities were 29.0 and 39.5 µmol Trolox/g of fresh fruit weight, respectively. In vitro simulated gastrointestinal digestion showed great raspberry phenolics stability. Our study assessed the bioaccessible phenolics in wild raspberry fruits and showed optimal conditions for the effective extraction of bioactive compounds for their analysis.

Potential of Equisetum ramosissimum Desf. for remediation of antimony flotation tailings: a case study.

A flooding event caused collapsing of the Stolice flotation tailing dam and spilling of large volumes of sludge into environment. Urgent remediation measures have not been applied due to the lack of financial resources. Remediation values for Sb, Zn, and Pb in the flotation tailing samples were exceeded 20.5, 4.2, and 1.15 times, respectively, emphasizing the need for remediation. Plants growing on mine spoils represent useful tools for environmental monitoring and soil remediation. The appearance of Equisetum ramosissimum as a dominant colonizer on the flotation tailings indicates that biological reclamation of the site is possible. Equisetum ramosissimum shows the ability to phytostabilize and immobilize available fractions of Fe, Mn, Zn, Pb, and Sb. Transfer rate of metals from roots to shoots reveals exclusion of elements from the shoots, preventing their further spreading through the food chain. The results of this study show that E. ramosissimum can be an additional tool for environmental monitoring and remediation of flotation tailings after hazardous events.

Atmospheric Solids Analysis Probe with Mass Spectrometry for Chlorpyrifos and Chlorpyrifos-Oxon Determination in Apples.

Chlorpyrifos (CPS) is a toxic pesticide present in several pesticide formulations, with low degradability by natural processes. The degradation leads to the toxic metabolite chlorpyrifos-oxon (CPO). The analytical techniques used for the CPS and CPO analysis, like UPLC-PDA and GC-MS, are accurate but also expensive and time consuming, and they need sample pretreatment. In the search of a more rapid and simple analytical procedure, atmospheric solids analysis probe with mass spectrometry (ASAP-MS) was optimized for the determination of CPS and CPO in apples (Malus domestica "Idared"). The identification of the analytes was based on protonated ion and isotopic pattern, while the quantification was based on peak intensities. The obtained results were confirmed by re-validated UPLC-PDA and GC-MS techniques. CPS and CPO concentrations determined by ASAP-MS and UPLC-PDA showed moderate discrepancies (on average by 10-20%), thus demonstrating that ASAP-MS can be a semiquantitative tool for the quantification of these compounds. As additional goal of this work, the efficiency of a gamma irradiation treatment to remove CPS and CPO from apples was tested by analyzing their content before and after the irradiation: 89-99% of CPS and CPO were degraded with doses of 3.5-3.8 kGy and 66-72 h of irradiation per sample. Identical degradation results were obtained by UPLC-PDA and ASAP-MS, indicating that the latter technique is well suitable to rapidly check pesticide degradation in apples.

Micro-edaphic factors affect intra-specific variations in trace element profiles of Noccaea praecox on ultramafic soils.

The aim of this study was to compare trace element profiles of Noccaea praecox (Wulfen) F. K. Mey. growing on ultramafic soils in different habitat types and to observe differences in uptake and translocation of trace elements. Physico-chemical characteristics of the soil and concentrations of P2O5, K2O, Ca, Fe, Mn, Zn, Cu, Ni, Cr, Pb, Cd, and Co in plant samples were presented. Biological concentration, accumulation, and translocation factors were calculated to estimate accumulation potential of different N. praecox accessions. All of the studied accessions were Ni hyperaccumulators (with shoot concentrations up to 14,593 mg kg-1), but with notable differences in accumulation and translocation rates. Significant differences in accumulation and translocation patterns of trace elements were observed among accessions from habitats characterized as serpentine steppes on dry, shallow soils in contrast to the accessions from habitats with higher soil moisture, and higher content of organic matter.

Metal accumulation in populations of Calamagrostis epigejos (L.) Roth from diverse anthropogenically degraded sites (SE Europe, Serbia).

Heavy metal accumulation is recognized as a very important global pollution problem in the last decades. Plant species have been recognized as natural bioindicators of environmental pollution, especially the amount of heavy metals in soils. Moreover, only a limited number of plant species can survive in highly contaminated soils. It is also known that metal accumulation can vary greatly among different populations of the same species. This study examines the chemical composition and accumulation potential of the expansive clonal grass Calamagrostis epigejos at five localities exposed to different levels of anthropogenic pressure. Considerable differences were observed between uptake, translocation, and accumulation of total and available heavy metals, such differences corresponding to soil physico-chemical characteristics and the level of site pollution. The results indicate that Calamagrostis epigejos uptakes a significant portion of the available fraction of heavy metals in the soil and stores it in the roots, thereby exhibiting a certain potential for metal phytostabilization.

Strategies of heavy metal uptake by three Armeria species growing on different geological substrates in Serbia.

This study surveyed three species of the genus Armeria Willd. from five ultramafic outcrops, two non-ultramafic (schist) soils, and one tailing heap of an abandoned iron-copper mine from Serbia. Similarities and differences among the three Armeria species growing on different geological substrates in the ability to control uptake and translocate nine metals were examined. Chemical characteristics of the soil and plant samples (concentrations of P2O5, K2O, Ca, Fe, Mn, Ni, Zn, Cu, Cr, Co, Cd, and Pb) are presented. In order to assess accumulative potential of these three Armeria species, biological concentration, accumulation, as well as translocation factors were used. Three investigated Armeria species growing on eight different localities showed large differences in heavy metal uptake, translocation, and accumulation. The differences were present among the plant samples of the same species and even more among three different Armeria species and were primarily the result of the different contents of available heavy metals in the investigated soils. Additionally, differences might be the consequence of diverse responses and possible presence of supplementary resistance mechanisms in the plants from the ultramafic soils. None of the three Armeria species showed shoot hyperaccumulative potential for any of the investigated heavy metals and they could be considered as root accumulators, considering their potential to accumulate medium to large amounts of Zn (BCF up to 134), Cr (BCF up to 148), and Cd (BCF up to 9) in their roots.

Synthesis, antioxidant and antiproliferative activities of 1,3,4-thiadiazoles derived from phenolic acids.

Two 2-amino-1,3,4-thiadiazoles containing phenolic hydroxyl groups were combined with different carboxylic acid chlorides giving sixteen amide derivatives with good antioxidant and antiproliferative potential. The compound 3'c with an adamantane ring displayed excellent DPPH radical scavenging activity and good cytotoxic activity against human acute promyelocytic leukemia HL-60 cells, while 1,3,4-thiadiazole 3'h with 4-chlorophenyl moiety was found to be the most effective in inhibition of survival of lung carcinoma A549 cells. All examined thiadiazoles except 3a and 3'a exerted higher cytotoxic activities on A549 and HL-60 cancer cells when compared with normal fibroblasts MRC-5, pointing to selectivity in their antiproliferative action. Some of the most active novel compounds 3c, 3'c, 3'g and 3'h induced significant increase in the percentage of HL-60 cells in the subG1 cell cycle phase in comparison with the control cells. The induction of cell death in HL-60 cells by these compounds was at least partially dependent on activation of caspase-3 and caspase-8. The compounds 3c and 3'c exerted strong antiangiogenic activity. Furthermore, compounds 3c, 3'c, 3'g and 3'h showed the ability to down-regulate the MMP2 and VEGFA expression levels in the treated HL-60 cells when compared with the control cell samples.

Protein molecular forms of insulin-like growth factor binding protein-2 change with aging.

Aging is considered to be an adaptive mechanism to altered needs of an organism and/or to altered stimuli. Plasma concentrations of insulin-like growth factor binding protein-2 (IGFBP-2) increase with age and it is generally assumed that IGFBP-2 is a negative predictor of healthy aging. The aim of this study was to examine the distribution of IGFBP-2 molecular forms in different age groups and, specifically, the relationship between IGFBP-2 and alpha-2-macroglobulin (α2M). The relative amount of monomer IGFBP-2 was the highest in young persons, making up approximately 2/3 of the total circulating IGFBP-2. This gradually decreased with age down to 1/3 of total IGFBP-2 in elderly individuals. Fragmented IGFBP-2 increased with age and contributed almost 60% to the total immunoreactive IGFBP-2 in the age group 61-80 years. IGFBP-2/α2M complexes represented 10-12% of the total IGFBP-2 in the two younger groups but half this level in the oldest group. The significance of these changes and whether they affect more IGF-dependent or independent interactions are unknown. Due to drastic proteolysis of IGFBP-2, it may be postulated that either over-release of IGFBP-2-bound IGFs causes unwanted events or IGFBP-2 fragments are able to over-stimulate cellular processes.

Relation between edaphic factors and vegetation development on copper mine wastes: a case study from Bor (Serbia, SE Europe).

The relationship between edaphic characteristics and vegetation growing on mine wastes in the Bor region (East Serbia, SE Europe) was studied using multivariate statistical analysis. The influence of edaphic factors on the composition of plant life-forms was also investigated, since it could reflect strategies for the avoidance of or tolerance to disturbances of ecosystems. The goal was to provide potential models for the restoration and management of this and similar mine waste areas. The results of this study imply that soil textures, nitrogen contents, reclamation technology and the presence of hydrothermally altered andesite as the type of bedrock significantly influenced plant colonization and vegetation composition of the Bor mine wastes. These edaphic factors explained 30.3 % of the total variation in the vegetation data set. It was also revealed that the pattern of plant life-forms found on the considered site groups corresponded to the soil texture. Based on their relative abundance on the investigated sites and relationships with soil properties it is concluded that therophytes and geophytes are unsuccessful primary colonizers of the Bor mine wastes. Hemicryptophytes of psammophytic character were the most successful primary colonizers and therefore potential candidates for anthropogenically-assisted natural recovery. This study suggested that an assessment of edaphic factors should be widely used in the characterization of mine wastes prior to reclamation. Estimation of their role in the development of existing mine vegetation should predate reclamation procedures. Thus, approaches based on adequate plant life-forms should have a more prominent role in future mine reclamation schemes.

Edaphic factors affecting the vertical distribution of radionuclides in the different soil types of Belgrade, Serbia.

The specific activities of natural radionuclides ((40)K, (226)Ra and (232)Th) and Chernobyl-derived (137)Cs were measured in soil profiles representing typical soil types of Belgrade (Serbia): chernozems, fluvisols, humic gleysols, eutric cambisols, vertisols and gleyic fluvisols. The influence of soil properties and content of stable elements on radionuclide distribution down the soil profiles (at 5 cm intervals up to 50 cm depth) was analysed. Correlation analysis identified associations of (40)K, (226)Ra and (137)Cs with fine-grained soil fractions. Significant positive correlations were found between (137)Cs specific activity and both organic matter content and cation exchange capacity. Saturated hydraulic conductivity and specific electrical conductivity were also positively correlated with the specific activity of (137)Cs. The strong positive correlations between (226)Ra and (232)Th specific activities and Fe and Mn indicate an association with oxides of these elements in soil. The correlations observed between (40)K and Cr, Ni, Pb and Zn and also between (137)Cs and Cd, Cr, Pb and Zn could be attributed to their common affinity for clay minerals. These results provide insight into the main factors that affect radionuclide migration in the soil, which contributes to knowledge about radionuclide behaviour in the environment and factors governing their mobility within terrestrial ecosystems.

Transfer of natural and anthropogenic radionuclides to ants, bryophytes and lichen in a semi-natural ecosystem.

Few data are available to quantify the transfer of both natural and anthropogenic radionuclides to detritivorous invertebrates to facilitate estimation of the internal dose to such biota in models used to assess radiation exposure. To enhance the available data, activity concentrations of (137)Cs, (40)K, (90)Sr, (239 + 240)Pu, (241)Am, (235)U and (238)U were measured in ants (Formicidae) and corresponding undisturbed soil collected from the Zlatibor mountain in Serbia and ant/soil concentration ratios (CR) calculated. The (241)Am concentration ratios for ants were fourfold higher than those calculated for ants in a previous study whereas they are similar to the more numerous data previously reported for a range of detritivorous invertebrates in other studies. CR values for (137)Cs in ants were similar to the few other reported values and slightly lower than those for a range of detritivorous invertebrates. Those for (239 + 240)Pu were slightly higher than those for ants in two other studies but they were close to upper limit of a range of data reported for detritivorous invertebrates. All the CR values will be included in a future revision of the ERICA Tool database and will particularly improve the information available for uranium.