Fine root production, mortality, and turnover in response to simulated nitrogen deposition in the subtropical Abies georgei (Orr) forest.

Increased nitrogen deposition has important effects on below-ground ecological processes. Fine roots are the most active part of the root system in terms of physiological activity and the main organs for nutrient and water uptake by plants. However, there is still a limited understanding of how nitrogen deposition affects the fine root dynamics in subtropical Abies georgei (Orr) forests. Consequently, a three-year field experiment was conducted to quantify the effects of three forms of nitrogen sources ((NH4)2SO4, NaNO3, and NH4NO3) at four levels (0, 5, 15, and 30 kg N·ha-1·yr-1) on the fine root dynamics in Abies georgei forests using a randomized block-group experimental design and minirhizotron technique. The first year of nitrogen addition did not affect the first-class fine roots (FR1, 0 < diameter < 0.5 mm) and second-class fine roots (FR2, 0.5 < diameter < 1.0 mm). The next two years of nitrogen addition significantly increased the production, mortality, and turnover of FR1 and FR2; the three year of nitrogen addition did not affect the dynamics of the third- class fine roots (FR3, 1.0 < diameter < 1.5 mm) and fourth- class fine roots (FR4,1.5 < diameter < 2.0 mm). Nitrogen addition positively affected the dynamics of FR1, FR2, FR3 and FR4 by positively affecting the carbon, nitrogen, and phosphorus contents of fine roots and indirectly affecting the soil pH. Increased carbon allocation to FR1 and FR2 may represent a phosphorus acquisition strategy when nitrogen is not the limiting factor. The nitrogen addition forms and levels affected the fine root dynamics in the following orde: NH4NO3 > (NH4)2SO4 > NaNO3 and high nitrogen > medium nitrogen > low nitrogen. The results suggest that the different-diameter fine root dynamics respond differently to different nitrogen addition forms and levels, and linking the different-diameter fine roots to nitrogen deposition is crucial.

Structure of KOH-soluble polysaccharides from сoniferous greens of Norway spruce (Picea abies): The pectin-xylan-AGPs complex. Part 1.

Using 7 % KOH, the polysaccharide PAK has been isolated from the coniferous greens of Norway spruce. PAK was found to contain predominantly arabinoglucuronoxylan, xyloglucan and arabinan, but also pectic polysaccharides, glucomannan and arabinogalactan proteins (AGPs), as determined by 1D/2D NMR analysis. It was found that fractionation of PAK on DEAE-cellulose resulted in simultaneous elution of pectins, arabinoglucuronoxylans and AGPs. It was evident that the content of 4-OMe-α-D-GlcpA and xylose, 1,4-ß-D-GlcpA, and T-ß-D-GlcpA increased with an increase in NaCl concentration. However, 1,4-α-D-GalpA content was almost independent of NaCl concentration, indicating unchanged pectic polysaccharide concentration. Interestingly, pectins extracted with 0.1-0.3 M NaCl solutions were richer in rhamnogalacturonan-I (RG-I) than those extracted with water and 0.01 M NaCl. Conclusion: The content of RG-I, AGPs and arabinoglucuronoxylan rises with rising NaCl concentration. An intense signal indicating an intermolecular linkage between the xylan and RG-I domains, i.e. that part of the arabinoglucuronoxylan is covalently bound to RG-I, is observed in the HMBC spectra of the polysaccharides obtained. The discovery here of a new relationship between rhamnogalacturonan I and xylan contradicts the prevailing cell wall model.

Structure of KOH-soluble polysaccharides from сoniferous greens of Norway spruce (Picea abies): The pectin-xylan-AGPs complex. Part 2.

Polymers containing arabinoglucuronoxylan, fucogalactoxyglucan, pectin and arabinogalactan proteins were obtained from PAK isolated from Norway spruce with 7 % KOH. The pectin core of PAK-I2-F-1 and PAK-I2-F-2 was dominated by RG-I, as treatment with 1,4-α-D-polygalacturonase resulted in almost complete removal of homogalacturonan. Interestingly, the above has not affected the co-fractionation of arabinoglucuronoxylan (AGX), arabinogalactan proteins and rhamnogalacturonan I (RG-I). Since pectin was mainly represented by RG-I, we concluded that xylan is specifically associated with RG-I. Correlations in the HMBC spectrum demonstrate intermolecular interactions between the α-L-Rhap (RG-I) and the Xyl (xylan), indicating a covalently bound AGX:RG-I complex via the Xyl-(1→4)-Rha bond: …→2)-[(2,4-ß-D-Xylp)-(1→4)]-[(α-D-GalpA-(1→2)]-α-L-Rhap-(1→4)-α-D-GalpA-(1→…. In PAK-H1-1-F-1 and PAK-H1-1-F-2, parts of RG-I and xylan were removed by enzymolysis. Part of the xylan was probably attached to the above-mentioned RG-I blocks. The removal of part of RG-I, xylan and the disappearance of the signal in the HMBC spectrum indicating the bond between RG-I and xylan confirms that part of the arabinoglucuronoxylan is covalently bound to RG-I. The observed glycosidic linkage contradicts the dominant PCW model in which pectin and hemicellulose polysaccharide networks are considered as independent components. It can be concluded that alkali-soluble xylan from Norway spruce was detected both in the free state and covalently bound to pectin.

Fir (Abies balsamea) (Pinales: Pinaceae) needle essential oil enhances the knockdown activity of select insecticides.

Because of the increased interest in plant essential oils (PEO) for both home pest control and personal bite protection, the ability of fir needle (Abies balsamea) oil to synergize the 1-h knockdown and 24-h toxicity of 9 different synthetic insecticides was evaluated. Fir needle oil strongly synergized knockdown of the neonicotinoids, clothianidin, and thiamethoxam (between 16- and 24-fold), as well as natural pyrethrins (12-fold), but had less effect with organophosphates and fipronil. For 24-h mortality, only pirimiphos-methyl was strongly synergized by fir needle oil pretreatment (18-fold). Chemical analysis and testing identified delta-3-carene is the most bioactive constituent, producing synergism similar to that of the whole oil. In fact, this constituent synergized the 24-h mortality of clothianidin to a higher degree than fir needle oil itself (4.9-fold vs. 2.4-fold). Synergism is unlikely to be mediated by effects on the nervous system, as fir needle oil caused no change in mosquito central nervous system firing at 100 ppm and did not synergize an inactive concentration of natural pyrethrins (10 nM). To better understand fir needle oil effects, we evaluated the ability of pretreatment with this oil to impact Aedes aegypti monooxygenase degradation of a model substrate, 7-ethoxycoumarin. Interestingly, both fir needle oil and delta-3-carene caused a significant increase in metabolic degradation of 7-ethoxycoumarin, perhaps indicating they upregulate oxidative metabolic processes. Such an action would explain why fir needle oil enhances knockdown, but not 24-h mortality for most of the insecticides studied here, whereas increased bioactivation would explain the synergism of pirimiphos-methyl toxicity.

Gymnosperms of Idaho: Chemical Compositions and Enantiomeric Distributions of Essential Oils of Abies lasiocarpa, Picea engelmannii, Pinus contorta, Pseudotsuga menziesii, and Thuja plicata.

Conifers are of great economic value in terms of lumber production, important for construction and other uses such as pulp and paper. They are also important sources of essential oils. Conifer species have been vital to the ethnobotany and traditional herbal medicine of many different Native American groups. The objective of this work was to obtain and analyze the essential oils of several conifer species (Abies lasiocarpa, Picea engelmannii, Pinus contorta, Pseudotsuga menziesii, and Thuja plicata) growing in Idaho. The foliar essential oils were obtained by hydrodistillation and then analyzed by gas chromatographic methods, including GC-MS, GC-FID, and chiral GC-MS. The essential oils were obtained in varying yields from 0.66% up to 4.70%. The essential oil compositions were largely dominated by monoterpene hydrocarbons and oxygenated monoterpenoids. The chiral monoterpenoids were generally rich in the (-)-enantiomers for members of the Pinaceae, but the (+)-enantiomers predominated in the Cupressaceae. The essential oil compositions obtained in this work are qualitatively similar, but quantitatively different, to previously reported compositions and confirm and complement the previous reports. However, this is the first comprehensive analysis of the chiral terpenoid components in these conifer species. Additional research on essential oils of the Pinaceae and Cupressaceae is needed to describe the chemical profiles, chemical compositions, and enantiomeric distributions more reliably in the various species and infraspecific taxa of these two families.

"Smart Extraction Chain" with Green Solvents: Extraction of Bioactive Compounds from Picea abies Bark Waste for Pharmaceutical, Nutraceutical and Cosmetic Uses.

Secondary metabolites from the sawmill waste Picea abies bark were extracted using an innovative two-step extraction that includes a first step with supercritical CO2 (SCO2) and a second step using green solvents, namely ethanol, water, and water ethanol mixture. Maceration (M), ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE) techniques were applied in the second step. A total of nineteen extract were obtained and yield were compared. Bark extracts were characterized by LC-DAD-MSn and classes of compounds were quantified as abietane derivatives, piceasides, flavonoids, and phenolics to compare different extractions. Obtained extracts were studied by in vitro assay to evaluate potential pharmaceutical, nutraceutical and cosmetic uses assessing the antioxidant activity as well as the inhibitory activity on target enzymes. Results show that the "smart extraction chain" is advantageous in term of yield of extraction and phytoconstituent concentration. SCO2 extract, presenting a unique composition with a large amount of abietane derivatives, exerted the best activity for amylase inhibition compared to the other extracts.

Novel Approaches for Inhibiting the Indoor Allergen Der f 2 Excreted from House Dust Mites by Todomatsu Oil Produced from Woodland Residues.

House dust mite (HDM) is a globally ubiquitous domestic cause of allergic diseases. There is a pressing demand to discover efficient, harmless, and eco-friendly natural extracts to inhibit HDM allergens that are more likely to trigger allergies and challenging to be prevented entirely. This study, therefore, is aimed at assessing the inhibition of the allergenicity of major HDM allergen Der f 2 by todomatsu oil extracted from residues of Abies Sachalinensis. The inhibition was investigated experimentally (using enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)) and in silico using molecular docking. The results showed that todomatsu oil inhibits the allergenicity of Der f 2 by reducing its amount instead of the IgG binding capacity of a single protein. Moreover, the compounds in todomatsu oil bind to Der f 2 via alkyl hydrophobic interactions. Notably, most compounds interact with the hydrophobic amino acids of Der f 2, and seven substances interact with CYS27. Contrarily, the principal compounds fail to attach to the amino acids forming the IgG epitope in Der f 2. Interestingly, chemical components with the lowest relative percentages in todomatsu oil show high-affinity values on Der f 2, especially ß-maaliene (-8.0 kcal/mol). In conclusion, todomatsu oil has been proven in vitro as a potential effective public health strategy to inhibit the allergenicity of Der f 2.

Screening of the antioxidant and antibacterial effects of extracted essential oils from Thunbergia coccinea, Acacia polyacantha, Polygonum micrpcephallum, Abies spectabilis and Clerodendrum colebrookianum.

During the previous few decades, it has been seen that there is a rapid emergence of pathogens resistant to multiple antibiotics. This has now become a global crisis. Some unexplored or less explored plants also provide some antibacterial, bactericidal and antioxidant properties. The antibacterial, bactericidal effects of extracted essential oils (EEOs) of Thunbergia coccinea, Acacia polyacantha, Polygonum micrpcephallum, Abies spectabilis and Clerodendrum colebrookianum was tested in comparison with standard antibiotics. The methods chosen were disc diffusion and deduction of minimum inhibitory concentration (MIC) by microbroth dilution assays of the EEOs against the bacterial strains.The antioxidant activity was found out utilizing DPPH free radical scavenging assay, MDA, Hydrogen peroxide radical inhibition assay and Superoxide radical inhibition assay (O 2 -). Some commonly used standard antibiotics (metronidazole, amoxicillin, clarithromycin, rifampicin, clindamycin and oxacillin,) were utilized to compare the EEO antibacterial action. Clerodendrum colebrookianum (85.17 ± 3.06 µg MDA/g extract) had a reasonable MDA. Acacia polyacantha in MIC had values of 3.86 ± 0.25 to 6.20 ± 0.16. Polygonum micrpcephallum had excessive H2O2 (48.27 ± 2.4 5%). The antibacterial actions determined by the paper disc­diffusion technique of the EEO extracted from these plants showed that most had some antibacterial actions. Also, it was seen that the bactericidal action of the EEO extracted from E. alba was most potent against S. pyogenes (4.06 ± 0.15). The extract of the plant at varying concentrations (20, 40, 60, 80 and100 mg/mL) demonstrated noteworthy (P< 0.001) anthelmintic action in an effective change when the dose was adjusted. In conclusion, most of the tested plants contain a medicinal value, which can be utilized in the future to supplement artificial medicines and cure emerging diseases that create havoc for mankind.

Developing Forest Therapy Programmes Based on the Health Benefits of Terpenes in Dominant Tree Species in Tara National Park (Serbia).

Numerous medical studies have shown the positive effects of forests on different aspects of human health. This study deals with the content of major terpenes in dominant coniferous species in Tara National Park, Serbia, in order to explore the potential for the development of a novel health tourism programme based on forest therapy. Main terpenes were analysed using a headspace-sampling technique coupled with gas-chromatography-mass spectrometry (Head-space-GC/MS). Needles of fir and spruce growing in the vicinity of hiking trails were investigated for possibilities to perform such therapy. Major detected terpenes were α-cadinol and spathulenol previously described as antiviral, antitumor, antimicrobial and immunomodulatory agents. The results of the study were favourable and worked well with the existing walking infrastructure in the observed area of the Tara Mountain, as they act as invaluable resources for designing the structured forest bathing walks. The study not only adds to the knowledge in the environmental and public health realm but also to tourism and sustainability studies.

In vitro evaluation of the effects of Viscum album homeopathic preparations on murine melanoma cells

Viscum album extract (VA) is a complementary treatment in cancer, with in vitro and in vivo cytotoxic effects on several tumor types when applied in phytochemical doses. However, highly diluted ethanolic homeopathic preparations' effects and mechanisms need further study. Aims:To assess the in vitro effects of highly diluted VA from the subspecies V. album abietis and V. album album at different potency levels in different dilution ratios on murine melanoma cells. Methodology:The VA mother tinctures (MT)from Abies alba (MTA) and Quercus robur (MTQ) were prepared with summer and winter samples, harvested in Switzerland. They were submitted to homeopathic ethanolic maceration and a subsequent dynamization process. MTA, MTQ and the following respective potencies were tested in B16F10 murine cells: 3x, 12x, 30x, 6cH, 12cH, 200cH, 2LM, 3LM, and 5LM. Dynamized water, dynamized and non-dynamized ethanol, and carboplatin were used as control groups. The mitochondrial activity and cell viability analysis were performed at 1, 24, 48, and 72 hours by in vitro incubation. MTA and MTQ harvested in summer, as well as 12x, 200cH and 5LM potencies were also tested to cell apoptosis and necrosis markers, reactive oxygens species (ROS) production, inflammatory cytokines profile, cell morphology, and migratory capacity. Results and discussion: MTA and MTQ induced a decrease in cell metabolism and higher cytotoxicity within 1 hour, with significant morphological changes and increased production of ROS and inflammatory cytokines. Both homeopathic dilutions 12x and 5LM showed an influence on cell metabolism, cell replication, and oxidative stress modulation with inflammatory cytokines, mitosis, and migration pattern changes. On the other hand, Quercus robur and Abies alba 200cH showed increased on cytotoxicity and ROS levels, respectively. Conclusion:The in vitro effects of Viscum album homeopathic solutions in melanoma cells highlight the promising antitumoral potential and reinforce the need for further research to better understanding their mechanisms of action.

Administration of spruce bark (Picea abies) extracts in young lambs exhibits anticoccidial effects but reduces milk intake and body weight gain.

BACKGROUND: Eimeria spp. are widespread apicomplexan parasites known to cause coccidiosis in livestock, resulting in reduced animal welfare and productivity, particularly in sheep. The treatment options are limited, and there is an emerging development of resistance against registered pharmaceuticals. Spruce bark is rich in plant secondary metabolites (PSM), such as condensed tannins, which are bioactive compounds previously shown to have antiparasitic activity. Here, we examined the anticoccidial properties of bark extract of Norway spruce (Picea abies) against a field isolate of ovine Eimeria spp. by treating Eimeria-infected pre-ruminant lambs with water-extracted bark daily for 12 days. We hypothesised that the bark extract would reduce the faecal oocyst excretion and, consequently, the severity of diarrhoea. RESULTS: Oral administration of spruce bark extract significantly reduced the excretion of Eimeria oocysts in milk-fed lambs post treatment till the end of the trial 22 days post infection. This difference in oocyst excretion between the treated and the untreated infected animals increased with time. Compared to the untreated and the sham-infected control group, the group treated with bark extract had softer faeces and reduced milk intake during the treatment period. After discontinuing the treatment, the treated animals got a more solid and formed faeces compared to that of the untreated control group, and the milk intake increased to the level of the sham-infected, untreated control group. The bark extract treated animals had a lower body weight and a lower mean daily body weight gain throughout the whole duration of the experiment. CONCLUSIONS: Bark extract from Norway spruce showed marked anticoccidial properties by reducing the faecal oocyst count and associated diarrhoea in young lambs. Simultaneously we experienced detrimental effects of the treatment, displayed as reduced feed intake and daily body weight gain. Therefore, we suggest conducting similar studies with lower bark extract dosage to explore the possibilities of a better trade-off to reduce the negative impact while maintaining the antiparasitic effect.

Abietane diterpenes from Abies spectabilis and their anti-pancreatic cancer activity against the MIA PaCa-2 cell line.

An ethanolic extract of the stem of Abies spectabilis exhibited strong cytotoxicity against MIA PaCa-2 human pancreatic cancer cells preferentially under nutrient-deprived conditions. Therefore, phytochemical investigation of this bioactive extract was carried out, and that led the isolation of ten compounds (1-10) including a new abietane-type diterpene (1). The structure of the new compound (1) was elucidated by combined spectroscopic techniques, including HRFABMS, NMR and quantum ECD calculation. All the isolated compounds were evaluated for their efficacy against MIA PaCa-2 human pancreatic cancer cell line by employing an anti-austerity strategy. Among the tested compounds, dehydroabietinol (5) displayed the most potent activity with a PC50 value of 6.6 µM. Dehydroabietinol (5) was also found to retard the MIA PaCa-2 cell migration under normal nutrient-rich conditions displaying its anti-metastatic potential. Investigation on the mechanism suggested that dehydroabietinol (5) is an inhibitor of the key cancer cell survival Akt/mTOR/autophagy signaling pathway.

Efficacy and mechanism of essential oil from Abies holophylla leaf on airway inflammation in asthma: Network pharmacology and in vivo study.

BACKGROUND: Asthma is one of the most common chronic inflammatory diseases of the airways. Essential oil from Abies holophylla leaf (EOA) has been reported to have anti-inflammatory property. This study aimed to predict the inhibitory effect of EOA against asthma by network analysis and to confirm the underlying mechanism of EOA on airway inflammation. PURPOSE AND STUDY DESIGN: The effects and underlying mechanisms of EOA on asthma were investigated by in silico network pharmacology and an experimental in vivo study. METHODS: To define the effectiveness of EOA on asthma, the network pharmacology was constructed using major components of EOA. EOA (0.0003 and, 0.03 v/v%) was aerosolized by nebulizer 3 times a week for 5 min for 7 weeks. After 3 weeks of treating the mice with EOA, asthma was induced by sensitizing them with ovalbumin (OVA) and PM10. The effects of EOA on the IL-17 related signaling pathway was confirmed using an asthmatic model. RESULTS: The network analysis showed that EOA is highly associated with the IL-17-related signaling pathway. EOA inhibited respiratory epithelium hyperplasia, collagen deposition and goblet cell activation in the lung and trachea tissues. In addition, EOA reduced the number of eosinophils, lymphocytes and macrophages in BALF. Furthermore, in the asthmatic model of mice, we showed that EOA inhibited IL-17-related cytokines, increased Treg-related cytokines and decreased the TRAF6 and MAPK and, suppressed the nuclear transcriptional activities of NF-kB. CONCLUSIONS: The network pharmacology and in vivo study indicated that EOA may have an inhibitory effect on airway inflammation in asthma exposure through the IL-17-related signaling pathway.

Abieshanesides A and B, two unique ent-18,19-dinoricetexane diterpenoid glycosides from Abies beshanzuensis M.H. Wu.

Two unprecedented ent-18,19-dinoricetexane diterpenoid glycosides, named abieshanesides A (1) and B (2), together with seven known compounds, have been isolated from the dead trunks and branches of Abies beshanzuensis M.H. Wu. To our knowledge, abieshanesides A and B represent the first ent-18,19-dinoricetexane diterpenoid glycosides found in natural sources. Their structures and absolute configurations were elucidated by using a combination of spectroscopic techniques and comparison of experimental and calculated electronic circular dichroism (ECD) data. The MTT experiments showed that (E)-resveratrol (7) could inhibit viability of MH7A cells with the IC50 value of 12.5 µM. Compound 7 was able to block MH7A cell proliferation and was associated with G0/G1-phase cell cycle arrest. Flow cytometric analysis showed that the treatment by 7 significantly induced the proliferation of MH7A cells in a dose-dependent manner.

Comparison of the Content of Extractives in the Bark of the Trunk and the Bark of the Branches of Silver Fir (Abies alba Mill.).

The main objective of our study was to investigate the possible differences in the chemical composition of extractives from the bark of silver fir (Abies alba) with respect to the location of the bark sample on the tree, viz. differences in extract composition between stem bark and branch bark samples. Extractives in the bark samples from branches, depending on the distance of the sample from the trunk, were also analysed, and the stem bark samples were analysed with respect to their inner and outer parts. The results of the chemical analysis of extractives were supported by information about their antifungal and antioxidant effects. After felling and sampling silver fir trees, the collected bark samples were ground and freeze-dried. Extraction of bark samples was followed by a system of accelerated extraction using only water as a solvent. The extracts were analysed chemically using gravimetry, spectrophotometry and chromatography. Free-radical-scavenging activity was measured using the DPPH method, and the antifungal effect towards three moulds and three wood-decaying fungi was investigated with antifungal assay using the agar well diffusion method. It was found that the moisture content in bark samples decreased intensively just after the bark samples were peeled off the stem. Detailed chromatographic analysis showed that the bark extracts contained 14 compounds, among which phenolic acids, flavonoids and lignans were found to be the characteristic ones. The content of hydrophilic extractives in the branch bark samples decreased with increasing distance of the sample location from the tree stem. The largest amounts of phenolic extractives were measured in stem bark, followed by branch bark sampled at the point at which the branch entered the tree. Analysis of the separated parts of the bark showed that the outer layers of stem bark contained larger amounts of phenolic extractives, as well catechin and epicatechin, compared to the inner layers. Concentrated extracts of branch bark showed the largest free-radical-scavenging activity among the investigated samples, while strong antifungal effects of the bark extract were not found.

Glycosylated constituents isolated from the trunk of Abies holophylla and their anti-inflammatory and neurotrophic activity.

Eleven previously undescribed glycosylated compounds with phenolic (abeoside A-F), monoterpenyl (abeoside G and H), or 2-heptanyl (abeoside I-K) aglycone, and twenty one reported compounds were isolated from the trunk of Abies holophylla. The structures of the previously undescribed compounds were elucidated on the basis of the conventional NMR and HRMS data analysis, and the absolute configuration of sugar units were assigned by chiral derivatization and LC-MS analysis. All the isolated compounds were evaluated for their anti-neuroinflammatory and neurotrophic activities. Among the evaluated compounds, twelve compounds including abeoside A, B, E, G, H, J, and K exhibited strong anti-neuroinflammatory activities with IC50 values of 4.6-18.2 µM by inhibiting production of LPS-induced NO levels, and abeoside C and 1-O-[(S)-oleuropeyl]-ß-D-glucoside showed powerful effects on the stimulation of NGF secretion levels with 157.09 ± 8.53% and 154.74 ± 1.24%, respectively.

A new approach for analysis of polyprenols by a combination of thin-film chemical deposition and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

RATIONALE: The polyprenols are involved in some essential biosynthetic pathways and serve as ubiquitous components of cellular membranes, so their fingerprinting in natural samples is of great interest. Previous studies indicate that due to the high hydrophobicity of polyprenols their direct analysis by mass spectrometry with soft ionization techniques may be difficult and require preliminary off-line derivatization. Hence, a method for rapid and sensitive screening of polyprenols is required. METHODS: A combination of thin-film chemical deposition and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was used for analysis of the polyprenol profile of Abies sibirica L. extract. Polyprenol-based monolayers were formed at the interphase of aqueous barium acetate solution, supplemented with 2,5-dihydroxybenzoic acid, and an n-hexane solution of polyprenols directly on a MALDI target plate. RESULTS: Peaks corresponding to [M - H + Ba]+ ions were observed in the MALDI-TOF mass spectra of polyprenols. A total of nine polyprenol homologues were identified with a polyprenol of 16 isoprene units dominating. The limit of detection was established at the level of 6 pg. Possible mechanisms of formation of [M - H + Ba]+ ions of polyprenols were discussed. CONCLUSIONS: The proposed approach can be suitable for high-throughput screening of polyprenols in biological samples of different origin due to easy sample preparation and high sensitivity.

Effects of Siberian fir terpenes extract Abisil on antioxidant activity, autophagy, transcriptome and proteome of human fibroblasts.

BACKGROUND: Abisil is an extract of Siberian fir terpenes with antimicrobial and wound healing activities. Previous studies revealed that Abisil has geroprotective, anti-tumorigenic, and anti-angiogenic effects. Abisil decreased the expression of cyclin D1, E1, A2, and increased the phosphorylation rate of AMPK. OBJECTIVE: In the present study, we analyzed the effect of Abisil on autophagy, the mitochondrial potential of embryonic human lung fibroblasts. We evaluated its antioxidant activity and analyzed the transcriptomic and proteomic effects of Abisil treatment. RESULTS: Abisil treatment resulted in activation of autophagy, reversal of rotenone-induced elevation of reactive oxygen species (ROS) levels and several-fold decrease of mitochondrial potential. Lower doses of Abisil (25 µg/ml) showed a better oxidative effect than high doses (50 or 125 µg/ml). Estimation of metabolic changes after treatment with 50 µg/ml has not shown any changes in oxygen consumption rate, but extracellular acidification rate decreased significantly. Abisil treatment (5 and 50 µg/ml) of MRC5-SV40 cells induced a strong transcriptomic shift spanning several thousand genes (predominantly, expression decrease). Among down-regulated genes, we noticed an over-representation of genes involved in cell cycle progression, oxidative phosphorylation, and fatty acid biosynthesis. Additionally, we observed predominant downregulation of genes encoding for kinases. Proteome profiling also revealed that the content of hundreds of proteins is altered after Abisil treatment (mainly, decreased). These proteins were involved in cell cycle regulation, intracellular transport, RNA processing, translation, mitochondrial organization. CONCLUSIONS: Abisil demonstrated antioxidant and autophagy stimulating activity. Treatment with Abisil results in the predominant downregulation of genes involved in the cell cycle and oxidative phosphorylation.

Gas Chromatographic-Mass Spectrometric Analysis, Antibacterial, Antioxidant and Antiproliferative Activities of the Needle Essential Oil of Abies pindrow growing wild in Kashmir, India.

The essential oil composition of the leaves of Abies pindrow, growing in Kashmir, India, along with its antioxidant, antibacterial and anticancer activity is reported for the first time. Gas chromatography coupled with mass spectrometry (GC-MS) revealed the presence of 12 constituents, representing 99.9% of the total oil. The major constituents of the oil were limonene (38.9%), α-pinene (36.5%), ß-pinene (6.9%), and α-selinene (4.4%). The essential oil was dominated by the presence of monoterpene hydrocarbons (90.2%), followed by sesquiterpene hydrocarbons (6.761%), oxygenated sesquiterpenes (2.096%) and oxygenated monoterpenes (0.942%). The monoterpene rich essential oil was subjected to antibacterial activity against 4 Gram negative bacteria and 2 Gram positive bacteria at three different concentrations using Agar Well Diffusion Method taking streptomycin sulphate as reference. The oil displayed significant and broad spectrum antibacterial activity against different bacteria used. The minimum inhibitory concentration (MIC) of the active essential oil was determined using Agar Dilution Method. Highest antibacterial activity was shown by the oil against E. Coli (25 mm), and the lowest by Bacillus subtilis (14 mm) and Pseudomonas aeruginosa (14 mm). The oil was subjected to cytotoxic activity by MTT assay against human mammary carcinoma (MCF), human ductal breast epithelial tumour (T47D), human lung adeno-carcinoma epithelial (A549) and rat glial (C6) cell lines at three different concentrations. The results revealed significant sensitization of the cell lines with highest inhibition against human ductal breast epithelial cell line (51%) and the lowest against rat glial cell line (33%) at a concentartion of 50 µg/mL. The oil displayed a significant free radical scavenging activity with DPPH.

Characterization of pectin-xylan-glucan-arabinogalactan proteins complex from Siberian fir Abies sibirica Ledeb.

Polysaccharide ASK was isolated from the Abies sibirica foliage by extraction with an aqueous KOH solution. ASK was shown to contain structurally different polymers such as arabinoglucuronoxylans, xyloglucans, glucomannans, arabinogalactan-proteins (AGPs). The pectic polysaccharides were also found in the alkaline extract of ASK and were represented by regions of homogalactorunan and rhamnogalactouronan-I whose side sugar chains were made up chiefly of highly branched 1,5-α-l-arabinan. The potential couplings between those polysaccharides were examined. Our studies showed simultaneous elution of pectin, xyloglucans, arabinoglucuronoxylans and AGPs, indicating that pectins can be covalently bound to the other cell-wall polysaccharides. NMR spectroscopy results revealed that the polysaccharides obtained by ion-exchange chromatography almost had no free reducing ends. These findings corroborate the conclusion that pectin, AGPs, glucan and xylan are bound together. The existence of the covalently bound complex of pectin-xylan-xyloglucan-AGP is suggested herein. Pectin and xylan are hypothesized to be covalently linked through RG-I regions.