Chemical Composition, Antitumor Properties, and Mechanism of the Essential Oil from Plagiomnium acutum T. Kop.

Plagiomnium acutum T. Kop. (P. acutum) has been used as a traditional Chinese medicine for thousands of years to treat cancer but lacks evidence. The objective of this work was to reveal the chemical composition of P. acutum essential oil (PEO) and explore its potential antitumor activity and molecular mechanism. PEO was prepared by the simultaneous distillation-extraction method and characterized by gas chromatography/mass spectroscopy. CCK8 assay, flow cytometry, western blot, and immunofluorescence techniques were used to analyze the effects and mechanism of PEO against cancer cells. A total of 74 constituents of PEO were identified, with diterpenes (26.5%), sesquiterpenes (23.89%), and alcohols (21.81%) being the major constituents. Two terpenoids, selina-6-en-4-ol and dolabella-3,7-dien-18-ol, were detected in PEO for the first time. PEO showed significant cell growth inhibitory activity on HepG2 and A549 cells by blocking the G1 phase and inducing apoptosis, which may be attributed to its upregulation of p21Cip1 and p27Kip1 proteins and interference with mitochondrial membrane potential effect. Dolabella-3,7-dien-18-ol accounts for 25.5% of PEO and is one of the main active components of PEO, with IC50 values in HepG2 and A549 cells of (25.820 ± 0.216) µg/mL and (23.597 ± 1.207) µg/mL, respectively. These results confirmed the antitumor medicinal value of P. acutum and showed great application potential in the pharmaceutical industry.

Characterization of Trace Elements in Atmospheric Deposition Studied by Moss Biomonitoring in Georgia.

The present work was conducted to obtain and highlight the first comprehensive baseline data on atmospheric deposition of trace elements and to evaluate the air quality in Georgia. A total of 120 moss samples were collected over accessible territories in Georgia in the period from 2014 to 2017. Hylocomium splendens (Hedw.) Schimp., Hypnum cupressiforme (Hedw.), and Pleurozium schreberi (Brid.) Mitt. moss species were analyzed by two complementary analytical techniques: instrumental neutron activation analysis and atomic absorption spectrometry. Concentrations of 41 elements in mg/kg were determined. The concentrations were compared with the corresponding values in the literature and are in a good agreement, except for the concentration of Mg, Al, K, Ca, Ti, and Fe, which were higher than those reported for other countries. The principal component and discriminant analyses were implemented to extract information about the similar geochemical features and to decipher the provenance of the studied elements. The analysis showed that a considerable association of crustal elements and the provenance of elements can be considered as a mixture of geogenic and anthropogenic sources. In addition, the influence of different latitudinal climate zones on the distribution of elements in the atmospheric deposition was observed. The enrichment factor shows considerable values for Th and Zr. The spatial distribution of the pollution load index identifies four zones (#12, 38, 53, and 64). The potential ecological risk index and the risk index were calculated and it does not pose significant risk except As and Cd. The data obtained can be used as the first dataset of metal characterization of air pollution in Georgia.

Agarose/κ-carrageenan-based hydrogel film enriched with natural plant extracts for the treatment of cutaneous wounds.

Hydrogels for complex and chronic wound dressings must be conformable, absorb and retain wound exudates and maintain hydration. They can incorporate and release bioactive molecules that can accelerate the healing process. Wound dressings have to be in contact with the wound and epidermis, even for long periods, without causing adverse effects. Hydrogel dressing formulations based on biopolymers derived from terrestrial or marine flora can be relatively inexpensive and well tolerated. In the present article hydrogel films composed by agarose (1.0 wt%), κ-carrageenan at three different concentrations (0.5, 1.0 and 1.5 wt%) and glycerol (3.0 wt%) were prepared without recourse to crosslinking agents, and characterized for their mechanical properties, morphology, swelling and erosion behavior. The films resulted highly elastic and able to absorb and retain large amounts of fluids without losing their integrity. One of the films was loaded with the aqueous extract from Cryphaea heteromalla (Hedw.) D. Mohr for its antioxidant properties. Absence of cytotoxicity and ability to reduce the oxidative stress were demonstrated on NIH-3T3 fibroblast cell cultures. These results encourage further biological evaluations to assess their impact on the healing process.

Evaluation of the acute toxicity, phototoxicity and embryotoxicity of a residual aqueous fraction from extract of the Antarctic moss Sanionia uncinata.

BACKGROUND: Ultraviolet (UV) radiation is the main exogenous inductor of skin damage and so photoprotection is important to control skin disorders. The Antarctic moss Sanionia uncinata is an important source of antioxidants and the photoprotective activity of its organic extracts has been investigated. This study aimed to evaluate the potential photoprotection, cytotoxicity and embryotoxicity of residual aqueous fraction (AF) from the moss S. uncinata. METHODS: UV-visible spectrum and SPF (sun protection factor) were determined by spectrophotometry. Embryotoxicity potential was evaluated by Fish embryo-larval toxicity test using zebrafish (Danio rerio) as organism model. Cell death assays by water-soluble tetrazolium salt (WST-1) and lactate dehydrogenase (LDH) were investigated using HaCaT keratinocyte cell line cultured in monolayers and three dimensions (3D). Phototoxicity and association with UV-filters were performed by 3T3 neutral red uptake test. RESULTS: The AF showed sharp absorption bands in the UV region and less pronounced in the visible region. The SPF was low (2.5 ± 0.3), but the SPF values of benzophenone-3 and octyl-methoxycinnamate increased ~ 3 and 4 times more, respectively, in association with AF. The AF did not induce significant lethal and sublethal effects on zebrafish early-life stages. In monolayers, the HaCaT cell viability, evaluated by WST-1, was above 70% by ≤0.4 mg AF/mL after 48 and 72-h exposure, whereas ≤1 mg AF/mL after 24-h exposure. The LDH assay showed that the cell viability was above 70% by ≤0.4 mg AF/mL even after 72-h exposure, but ≤1 mg/mL after 24 and 48-h exposure. In 3D cell culture, an increased cell resistance to toxicity was observed, because cell viability of HaCaT cell by WST-1 and LDH was above ~ 90% when using ≤1 and 4 mg AF/mL, respectively. The AF demonstrated values of photo irritation factor < 2 and of photo effect < 0.1, even though in association with UV-filters. CONCLUSIONS: The residual AF absorbs UV-vis spectrum, increased SPF values of BP-3 and OMC and does not induce embryotoxicity to zebrafish early life-stage. The cell death assays allowed establishing non-toxic doses of AF and phototoxicity was not detected. AF of S. uncinata presents a good potential for skin photoprotection against UV-radiation.

Botrysphones A-C and Botrysphins A-F, Triketides and Diterpenoids from the Fungus Botrysphaeria laricina.

Three new triketides, botrysphones A-C (1-3) and six new isopimarane-type diterpenoids, botrysphins A-F (4-9), together with the known triketides sphaeropsidone (10) and chlorosphaeropsidone (11) and diterpenoids sphaeropsidins A and B (12 and 13), were obtained from culture of the fungus Botrysphaeria laricina associated with the moss Rhodobryum umgiganteum. The structures of the new compounds were established on the basis of extensive spectroscopic techniques including HRMS and 1D and 2D NMR data. Compounds 7 and 12 showed significant quinone reductase inducing activity in Hepa 1c1c7 cells.

A new support material for IgG adsorption: Syntrichia papillosissima (Copp.) Loeske.

In this presented work, Syntrichia papillosissima (Copp.) Loeske (S. papillosissima) was used as a natural phytosorbent for IgG purification. These moss species were collected for the natural habitat and prepared for IgG adsorption studies by cleaning, drying, and grinding to uniform size. Syntrichia papillosissima samples were characterized by using FTIR and SEM studies. Functional groups of S. papillosissima were identified by FTIR analysis, while surface characteristics were determined by SEM studies. A batch system was used for the adsorption of IgG onto S. papillosissima surface and physical conditions of the IgG adsorption medium were investigated by modifying the pH, IgG concentration and temperature. Maximum IgG adsorption onto S. papillosissima was found to be 68.01 mg/g moss by using pH 5.0 buffer system. Adsorption kinetic isotherms were also studied and it was found that, Langmuir adsorption model was appropriate for this adsorption study. Reusability profile of S. papillosissima was also investigated and IgG adsorption capacity did not decrease significantly after 5 reuse studies. Results indicated that S. papillosissima species have the capacity to be used as biosorbent for IgG purification, with its low cost, natural and biodegradable structure.

Trace elements in native and transplanted Fontinalis antipyretica and Platyhypnidium riparioides from rivers polluted by uranium mining.

The past uranium/polymetallic mining activities in the Sudety (SW Poland) left abandoned mines, pits, and dumps of waste rocks with trace elements and radionuclides which may erode or leach out and create a potential risk for the aquatic ecosystem, among others. In the present work four rivers affected by effluents from such mines were selected to evaluate the application of aquatic mosses for the bioindication of 56 elements. Naturally growing F. antipyretica and P. riparioides were compared with transplanted samples of the same species. The results demonstrate serious pollution of the examined rivers, especially with As, Ba, Fe, Mn, Pb, Ti, U and Zn, reaching extremely high concentrations in native moss samples. In the most polluted rivers native F. antipyretica and P. riparioides samples showed significantly higher concentrations of As, Ba, Cu, Fe, La, Nd, Ni, Pb, U and Zn than corresponding transplanted samples, whereas at less polluted sites a reverse situation was sometimes observed. Transplanted moss moved from clean to extremely polluted rivers probably protects itself against the accumulation of toxic elements by reducing their uptake. Selection of native or transplanted F. antipyretica and P. riparioides depended on the pollution load.

Accumulation of uranium by aquatic plants in field conditions: prospects for phytoremediation.

A study was undertaken to determine Uranium concentrations in water and aquatic plants in the uraniferous region of Beiras, Central Portugal. Samples were collected from running water (n=200) at places where aquatic species were observed. Plant samples were collected from 28 species of submerged, free-floating and rooted emergent plants including 2 bryophytes and 1 pteridophyte. Uranium concentrations in surface waters ranged from 0.23 to 1,217 µg L(-1). The aquatic plant species studied, including several previously untested species, exhibited the ability to accumulate U in concentrations many times that of the ambient water. In general submerged plants exhibited higher U content followed by rooted emergent and free floating species. The highest U concentrations were observed in the bryophyte Fontinalis antipyretica (up to 4,979 mg kg(-1)) followed by Callitriche stagnalis (1963mgkg(-1)), Callitriche hamulata (379 mg kg(-1)), Ranunculus peltatus subsp. saniculifolius (243 mg kg(-1)), Callitriche lusitanica (218 mg kg(-1)), and Ranunculus trichophyllus (65.8 mg kg(-1)). In two out of three rooted emergent species U seemed to be preferentially partitioned in rhizome/roots with highest rhizome U content recorded in Typha latifolia (380 mg kg(-1)). Among the free-floating species, the highest U content (42.5 mg kg(-1)) was seen in Lemna minor. The bryophyte F. antipyretica and Callitrichaceae members seem to be promising candidates for the development of phytofiltration methodologies based on U accumulation, abundance and biomass production.

Headspace volatile oxylipins of Eastern Himalayan moss Cyathophorella adiantum extracted by sample enrichment probe.

Cyathophorella adiantum (Griff.) M. Fleisch. (Division-Bryophyta, Family-Daltoniaceae), an Eastern Himalayan moss was studied for the first time to identify the volatiles derived from cellular and membrane bound fatty acids. A high capacity sample enrichment probe (SEP) was used for extraction of headspace volatile (HSV) molecules followed by GC-MS analysis. Different short-chain oxylipins like alkenes, alkanes, saturated and unsaturated alcohols, saturated and unsaturated aldehydes, ketones were identified along with free and esterified fatty acids, cyclo compounds and some by-products of secondary metabolites. Fatty acid analysis of neutral lipids (NL) and phospholipids (PL) of this plant exhibits the predominance of C16 and C18 fatty acids. It also reveals some interesting information that might indicate the possible fatty acid precursors for volatile generation and their sources in this plant.

Analysis of selected biomonitors to evaluate the suitability for their complementary use in monitoring trace element atmospheric deposition.

The biomonitoring properties of oak tree bark compared with the epiphytic moss Hypnum cupressiforme and the influence of the tree bark, as its growth substrate, on the content of heavy metals in moss were investigated. Samples of the epiphytic moss H. cupressiforme and oak tree bark (Quercus spp.) were collected in Eastern Romania at a total of 44 sampling sites. Parallel moss and bark samples were collected from the same sides of the trunk circumference. V, Cr, Ni, Cu, Zn, As, Mo, Cd, In, Tl, Sn, Pb, and Bi were determined by ICP-MS. Principal component analysis was used to identify possible sources of metals in bark and moss. Six factors explaining 87 % of the total variance in the data set were chosen. The main factors represent long-range atmospheric transport of elements (Zn, Cd, (Pb), Bi, (Mo), (Tl)), local emissions from industrial sources (As, Cr, Ni, V), road traffic (Pb, Zn) and agricultural activities (Cu, (Zn)). The element concentrations in moss and bark samples are of the same order of magnitude. For almost all the elements, higher concentrations were obtained in moss. Significant correlations between concentrations in moss and bark samples were obtained for 7 of the 13 elements: V, Ni, Cu, Zn, Cd, In, and Bi, all typical anthropogenic pollutants. The use of tree bark for monitoring purposes might be an alternative in areas where there is a scarcity of mosses.

Effect of methanolic and water extract of Leucobryum bowringii Mitt. on growth, migration and invasion of MCF 7 human breast cancer cells in vitro.

Inhibitory effects of methanol and water extract of L. bowringii. on the adhesion, migration, invasion and matrix metalloproteinase (MMP) activities of MCF 7 human breast cancer cell line are reported. Cells were cultured with 10, 25, 50 microg/mL methanolic or water extract of L. bowringii. Culture medium containing 0.1% DMSO was used as a solvent control. Ultra structural analysis by electron microscopy revealed typical features of apoptosis. A remarkable dose-response parallelism was observed between methanolic extract with growth, migration and invasion of breast cancer cells. Fractionation of methanolic extract by RP-HPLC revealed a pool of phenolic acids. Hoechst 33342 staining assay reveals massive chromatin condensation and subsequent cleavage of structural components of nucleus. The results indicate that methanol extracts inhibit the growth of human breast cancer cells partially through the inhibition of metallo proteinases MMP-2 and MMP-9 activities. Methanolic extract has more anti-metastatic effects in cell based assay than water extract. Clinical application of L. bowringii extract as a bioactive chemopreventive compound may be helpful in limiting breast carcinoma invasion and metastasis.

pH induced protein-scaffold biosynthesis of tunable shape gold nanoparticles.

In this paper, a pH-inductive protein-scaffold biosynthesis of shape-tunable crystalline gold nanoparticles at room temperature has been developed. By simple manipulation of the reaction solution's pH, anisotropic gold nanoparticles including spheres, triangles and cubes could be produced by incubating an aqueous solution of sodium tetrachloroaurate with Dolichomitriopsis diversiformis biomasses after immersion in ultrapure Millipore water overnight. A moss protein with molecular weight of about 71 kDa and pI of 4.9 was the primary biomolecule involved in the biosynthesis of gold nanoparticles. The secondary configuration of the proteins by CD spectrum implied that the moss protein could display different secondary configurations including random coil, α-helix and intermediate conformations between random coil and α-helix for the experimental pH solution. The growth process of gold nanoparticles further showed that the moss protein with different configurations provided the template scaffold for the shape-controlled biosynthesis of gold nanoparticles. The constrained shape of the gold nanoparticles, however, disappeared in boiled moss extract. The gold nanoparticles with designed morphology were successfully reconstructed using the moss protein purified from the gold nanoparticles. Structural characterizations by SEM, TEM and SAED showed that the triangular and cubic gold nanoparticles were single crystalline.

PAH concentrations in the moss species Hylocomium splendens (Hedw.) B.S.G. and Pleurozium schreberi (Brid.) Mitt. from the Kielce area (south-central Poland).

Forty eight samples of mosses Hylocomium splendens and Pleurozium schreberi from the Kielce area (south-central Poland) were analyzed for seventeen polycyclic aromatic hydrocarbons. The PAH concentrations varied from 558.00 µg/kg (ppb) to 4457.00 µg/kg in H. splendens and from 643.00 to 3086.00 µg/kg in P. schreberi. In both moss species phenanthrene (75.00-732.00 µg/kg), fluoranthene (101.00-577.00 µg/kg) and pyrene (68.00-568.00 µg/kg) predominated, but H. splendens accumulated more PAHs than P. schreberi at eighteen different sites. The highest PAH concentrations were noted at sites located close to the housing estates (sites 1-10) and potential emission sources (e.g. heat and power generating plant "Kielce S.A.") (sites 19-21). The moss samples displayed the diverse ring sequence, but the most prevalent was four>five>three>six. The diagnostic ratios (three+four ring/total PAHs, Flu/Py, Phen/Ant, Phen/Phen+Ant, Flu/Flu+Py, IndPy/IndPy+BghiPe and BaA/BaA+Chr) indicated that coal and petroleum combustion was a principal PAH source in the study area.

Radioactivity measurements in moss (Hypnum cupressiforme) and lichen (Cladonia rangiformis) samples collected from Marmara region of Turkey.

The present study was conducted to compare the (137)Cs, (40)K, (232)Th, and (238)U activity concentrations in epigeic moss (Hypnum cupressiforme) and lichen (Cladonia rangiformis). The activity levels in 37 moss and 38 lichen samples collected from the Marmara region of Turkey were measured using a gamma spectrometer equipped with a high purity germanium (HPGe) detector. The activity concentrations of (137)Cs, (40)K, (232)Th, and (238)U in the moss samples were found to be in the range of 0.36-8.13, 17.1-181.1, 1.51-6.17, and 0.87-6.70 Bq kg(-1) respectively, while these values were below detection limit (BDL)-4.32, 16.6-240.0, 1.32-6.47, and BDL-3.57 Bq kg(-1) respectively in lichen. The average moss/lichen activity ratios of (137)Cs, (40)K, (232)Th, and (238)U were found to be 1.32 +/- 0.57, 2.79 +/- 1.67, 2.11 +/- 0.82, and 2.19 +/- 1.02, respectively. Very low (137)Cs concentrations were observed in moss and lichen samples compared to soil samples collected from the same locations in a previous study. Seasonal variations of the measured radionuclide activities were also examined in the three sampling stations.

Metal-tolerant moss Scopelophila cataractae accumulates copper in the cell wall pectin of the protonema.

The growth kinetics in the presence of copper (Cu) of the protonema of the moss Scopelophila cataractae and the matrix polysaccharides of its cell walls have been analyzed in this study. Protonemal cells cultured in a medium containing 0.2mM CuSO(4) showed a rapid accumulation of Cu, reaching a maximum between 30 and 60d at approximately 65 micromolg(-1) DW. Uronic acids were found in similar amounts in cell walls of both control and Cu-treated cells, whereas arabinose and galactose decreased to 61-67% in the presence of Cu. Cell wall polysaccharides were determined after successive extraction with 50mM CDTA, 50mM Na(2)CO(3), 1M KOH, and 4M KOH. The pectic fractions (CDTA- and Na(2)CO(3)-soluble) decreased to 47% and the hemicellulosic fractions (1M KOH- and 4M KOH-soluble) to 86% under Cu application. Approximately 43% of the Cu taken into cell walls was released following endo-pectate lyase treatment, suggesting that two-fifths of the total Cu accumulation was tightly bound to the homogalacturonan of the cell wall pectin.

Auxin-binding proteins without KDEL sequence in the moss Funaria hygrometrica.

Whereas the important plant growth regulator auxin has multiple effects in flowering plants, it induces a specific cell differentiation step in the filamentous moss protonema. Here, we analyse the presence of classical auxin-binding protein (ABP1) homologues in the moss Funaria hygrometrica. Microsomal membranes isolated from protonemata of F. hygrometrica have specific indole acetic acid-binding sites, estimated to be about 3-5 pmol/mg protein with an apparent dissociation constant (K (d)) between 3 and 5 microM. Western analyses with anti-ABP1 antiserum detected the canonical endoplasmic reticulum (ER)-localised 22-24 kDa ABP1 in Zea mays, but not in F. hygrometrica. Instead, polypeptides of 31-33 and 46 kDa were labelled in the moss as well as in maize. In F. hygrometrica these proteins were found exclusively in microsomal membrane fractions and were confirmed as ABPs by photo-affinity labelling with 5-azido-[7-(3)H]-indole-3-acetic acid. Unlike the classical corn ABP1, these moss ABPs did not contain the KDEL ER retention sequence. Consistently, the fully sequenced genome of the moss Physcomitrella patens, a close relative of F. hygrometrica, encodes an ABP1-homologue without KDEL sequence. Our study suggests the presence of putative ABPs in F. hygrometrica that share immunological epitopes with ABP1 and bind auxin but are different from the classical corn ABP1.

Antioxidant activity of Sanionia uncinata, a polar moss species from King George Island, Antarctica.

Antioxidant agents counter reactive oxygen species (ROS) and can be used in cosmetic and medicinal applications. The goal of this study was to evaluate the antioxidant activity of an Antarctic moss species from King George Island (Antarctica), tentatively designated as KSJ-M5. On the basis of morphological characteristics, KSJ-M5 was identified as Sanionia uncinata (Hedw.) Loeske (Amblystegiaceae). The identification was confirmed by comparing the partial sequence of the ITS (internal transcribed spacer) region with that in GenBank. The antioxidant activity of an ethanol extract of KSJ-M5 was evaluated by analyzing its reducing power, superoxide scavenging activity, ABTS [2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)] cation scavenging activity, and DPPH (1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity. The reducing power of 1 mg of KSJ-M5 extract was equivalent to 31.9 +/- 0.9 microg (Mean +/- SD, n = 3) of the commercial standard, BHT (butylated hydroxytoluene). IC(50) values of the KSJ-M5 extract for DPPH free-radical scavenging activity, superoxide scavenging activity, and ABTS cation scavenging activity were found as 356 +/- 26.8 microg/mL, 466.2 +/- 43.4 microg/mL, and 181.3 +/- 12.2 microg/mL, respectively. The total phenolic content in 1 mg of KSJM5 extract was equivalent to 12.7 +/- 2.7 microg of pyrocatechol. These results clearly showed that KSJ-M5 could be an important source of natural antioxidant agents for improved medicinal and cosmetic applications.

[Studies on chemical constituents from moss Rhodobryum roseum II].

OBJECTIVE: To study the chemical constituents from Rhodobryum roseum. METHOD: The compounds were isolated and purified by recrystallization and chromatography with silica gel and sephadex LH-20 column. Their structures were identified by physicochemical properties and spectral analysis. RESULT: Eight compounds were isolated and identified as apigenin (1), quercetin (2), oleanolic acid (3), 2alpha-hydroxyoleanolic acid (4), 2alpha-hydroxyursolic acid (5), protocatechuic acid (6), p-hydroxybenzoic acid (7), p-hydroxycinnamic acid (8). CONCLUSION: Compounds 1-8 were isolated from R. roseum for the first time.

Antimicrobial activity of Bryum argenteum.

The antimicrobial activity of Bryum argenteum ethanol extracts was evaluated by microdilution method against four bacterial (Escherichia coli, Bacillus subtilis, Micrococcus luteus and Staphilococcus aureus) and four fungal species (Aspergillus niger, Penicillium ochrochloron, Candida albicans and Trichophyton mentagrophyes). All the investigated ethanol extracts have been proved to be active against all bacteria and fungi tested.

Differences in concentration of heavy metals between native and transplanted Plagiothecium denticulatum: a case study of soils contaminated by oil well exudates in South East Poland.

For a period of 60 days, the terrestrial moss Plagiothecium denticulatum was transplanted from a clean control site to a soil contaminated with crude oil exudates (Krosno, South East Poland). Native P. denticulatum growing on this contaminated soil was collected during the same period. Concentrations of the metals Al, Ba, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, V, and Zn as well as the macroelements N, P, K, Ca, Mg, and S were determined in P. denticulatum and in soils from the contaminated and control sites. Contaminated soil was the main contributor of Ba, Co, Cr, Cu, Fe, Mn, Ni, Pb, V, and Zn to native and transplanted P. denticulatum. Other sources of contamination apart from the soil existed for Cd and Cu. Transplanted P. denticulatum accumulated significantly more Al, Cr, Cu, Ni, Pb, and Zn, and native P. denticulatum accumulated significantly more N, P, K, and Ca. Higher levels of N, P, K, and Ca in native P. denticulatum indicates a possible mechanism to prevent the loss of these elements as observed in the transplanted P. denticulatum.