In vitro Genotoxicity and In silico Docking Analyses of the Essential Oils of Thuja orientalis.

Two main objectives were pursued to assess the reliability of Thuja orientalis essential oils (TOEO). The first objective was to extract TOEO, analyze them by GC-MS, and determine their in vitro genotoxicity against selected plants using the RAPD-PCR method. The second objective was to evaluate the in-silico toxicity of TOEO. The binding sites and energies of each content was calculated against B-DNA. In-silico analyses were performed using a simulation program, AutoDock Vina, and Toxicity Estimation Software Tools. 3-carene, cedrol, and 2-pinene were identified as the predominant components. In vitro studies showed that the TOEO had a more significant impact on reducing genomic stability in wheat compared to the amaranth. The lowest stability was determined as 39.78 % in wheat and 53.58 % in amaranth. Cedrol (-5,7 kcal/mol) and selinene (-5,6 kcal/mol) exhibited the highest binding affinity. The toxicity test indicated that components other than cyclohexene may have toxic effects, none of them were predicted to be mutagenic, and LD50 (mol/kg) values could vary between 1.33 and 1.55.

Chemotaxonomic Variation in Volatile Component Contents in Ancient Platycladus orientalis Leaves with Different Tree Ages in Huangdi Mausoleum.

To gain insight into the differences in the composition and volatile components content in ancient Platycladus orientalis leaves with different tree ages in Huangdi Mausoleum, the volatile components were identified by headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS) method. The volatile components were statistically analyzed by orthogonal partial least squares discriminant analysis and hierarchical cluster analysis, and the characteristic volatile components were screened. The results exhibited that a total of 72 volatile components were isolated and identified in 19 ancient Platycladus orientalis leaves with different tree ages, and 14 common volatile components were screened. Among them, the contents of α-pinene (6.40-16.76%), sabinene (1.11-7.29%), 3-carene (1.14-15.12%), terpinolene (2.17-4.95%), caryophyllene (8.04-13.53%), α-caryophyllene (7.34-14.41%), germacrene D (5.27-12.13%), (+)-Cedrol (2.34-11.30%) and α-terpinyl acetate (1.29-25.68%) were relatively higher (>1%), accounting for 83.40-87.61% of the total volatile components. Nineteen ancient Platycladus orientalis trees were clustered into three groups through the HCA method based on the 14 common volatile components content. Combined with the results of OPLS-DA analysis, (+)-cedrol, germacrene D, α-caryophyllene, α-terpinyl acetate, caryophyllene, ß-myrcene, ß-elemene and epiglobulol were the differential volatile components to distinguish ancient Platycladus orientalis with different tree ages. The results revealed that the composition of the volatile components in ancient Platycladus orientalis leaves with different tree ages was different, showing different aroma characteristics, which provided a theoretical reference for the differential development and application of volatile components in ancient Platycladus orientalis leaves.

Screening of Promising Chemotherapeutic Candidates from Plants against Human Adult T-Cell Leukemia/Lymphoma (VII): Active Principles from Thuja occidentalis L.

During the screening of novel chemotherapeutic candidates from plants against adult T-cell leukemia/lymphoma, we identified that the extracts of Thuja occidentalis (Cupressaceae) showed potent anti-proliferative activity in MT-1 and MT-2 cells. Therefore, we attempted to isolate the active components from this plant. We isolated and identified 32 compounds (1-32; eight lignans, 18 terpenoids, and six flavonoids) from the extracts of the leaves and cones. Their structures were determined by spectroscopic analysis. Several of the isolated compounds inhibited the growth of both cell lines. Lignans showed more potent activity than other classes of compounds. A comparison of the activities of compounds 1-8 revealed that the presence of a trans-lactone (linkage of C-6 to C-7) correlated with increased activity. Diterpenes showed moderate activity, and the presence of a ketone moiety at the C-7 position correlated with increased activity in compounds 12-21. In addition, biflavones showed moderate activity, and the presence of methoxy functions appeared to influence the activity of these compounds. Several lignans were lead compound of anti-cancer reagent (etoposide). In conclusion, not only lignans, but also diterpenes and/or biflavones, may be promising candidates for the treatment of adult T-cell leukemia/lymphoma.

Antibacterial sesquiterpenes from the stems and roots of Thuja sutchuenensis.

Eight new sesquiterpenes with diverse skeletons involving four cuparenes, denominated thujasutchins F-I (1-4), one eudesmane and one cedrol, named thujasutchin J (5) and thujasutchin K (6), as well as two thujopsenes thujasutchins L-M (7-8) together with three known congener compounds (9-11) were isolated from EtOAc soluble fraction of ethanolic extract of the stems and roots of Thuja sutchuenensis. Their structures including absolute configurations were unambiguously established by extensive interpretation of the NMR and mass spectroscopic data, X-ray diffractions, and ECD measurements powered by molecular calculations. The biological assays disclosed that 5 and 9 displayed potent inhibitory effect against Staphylococcus. aureus (CMCC 26003), methicillin-resistant Staphylococcus aureus (JCSC 4744), Bacillus cereus (ATCC 10876), and Staphylococcus epidermidis (ATCC 12228) with MICs ranging from 6.25 to 25 µg/mL.

Thuja occidentalis L. (Cupressaceae): Ethnobotany, Phytochemistry and Biological Activity.

Thuja occidentalis L. (Cupressaceae) has its origins in Eastern North America and is cultivated in Europe and Brazil as an ornamental tree, being known as the "tree of life" or "white cedar". In traditional medicine, it is commonly used to treat liver diseases, bullous bronchitis, psoriasis, enuresis, amenorrhea, cystitis, uterine carcinomas, diarrhea, and rheumatism. The chemical constituents of T. occidentalis have been of research interest for decades, due to their contents of essential oil, coumarins, flavonoids, tannins, and proanthocyanidines. Pharmacology includes antioxidant, anti-inflammatory, antibacterial, antifungal, anticancer, antiviral, protective activity of the gastrointestinal tract, radioprotection, antipyretic, and lipid metabolism regulatory activity. Therefore, the present review represents the synthesis of all the relevant information for T. occidentalis, its ethnobotany, phytochemistry, and a thorough analysis of their pharmacological activities, in order to promote all the biological activities shown so far, rather than the antitumor activity that has promoted it as a medicinal species.

Microbial population dynamics under microdoses of the essential oil arborvitae.

BACKGROUND: With the current concern caused by drug resistant microorganisms, alternatives to traditional antimicrobials are increasingly necessary. Historical holistic treatments involving natural approaches are now of interest as a potential alternative. Many essential oils have antimicrobial properties with the ability to modify bacterial and fungal population dynamics in low concentrations. METHODS: In this study, bacterial and fungal growth in response to varying concentrations of arborvitae oil was assessed using spectrophotometric methods to obtain estimates of population growth parameters including carrying capacity (K) and intrinsic rate of growth (r). Estimates of these parameters were compared among doses within strains using general linear modeling. RESULTS: Results suggest the active component of the essential oil arborvitae is likely of hydrophilic nature and demonstrates the ability to influence both K and r during bacterial and fungal growth in a dose-dependent manner. Highly concentrated doses of arborvitae completely kill Escherichia coli and significantly inhibit Staphylococcus aureus, however these same doses have no effect on Pseudomonas aeruginosa. Accordingly, microdoses of arborvitae demonstrated the ability to inhibit population growth parameters in both prokaryotic and eukaryotic microorganisms. Specifically, K of E. coli, r of Candida auris, and both K and r of Candida albicans were significantly reduced in the presence of microdoses of arborvitae. CONCLUSIONS: Microdoses of essential oils have the ability to inhibit one or both population parameters in both prokaryotic and eukaryotic microorganisms. Some microorganisms appear to be more susceptible to this essential oil arborvitae than other microorganisms. The use of essential oils, such as arborvitae, as novel antimicrobials may prove useful when contending with the current epidemic of multidrug resistant pathogens.

The Effect of Ten Essential Oils on Several Cutaneous Drug-Resistant Microorganisms and Their Cyto/Genotoxic and Antioxidant Properties.

In this study, we determined the antimicrobial activity of ten essential oils (EOs)-oregano, thyme, clove, arborvitae, cassia, lemongrass, melaleuca, eucalyptus, lavender, and clary sage-against drug-resistant microorganisms previously isolated from patients with skin infections. The essential oil compositions were determined using gas chromatography coupled to mass spectrometry (GC/MS). The assayed bacteria included Pseudomonas aeruginosa, Proteus vulgaris, Citrobacter koseri, and Klebsiella pneumoniae. Two drug-resistant yeasts (Candida albicans and Candida parapsilosis) were also involved in our survey. Oregano, thyme, cassia, lemongrass and arborvitae showed very strong antibacterial and antifungal activity against all tested strains. These results show that these essential oils may be effective in preventing the growth of the drug-resistant microorganisms responsible for wound infections. In this study, the genotoxic effects of tested essential oils on healthy human keratinocytes HaCaT were evaluated using the comet assay for the first time. These results revealed that none of the essential oils induced significant DNA damage in vitro after 24 h. Moreover, the treatment of HaCaT cells with essential oils increased the total antioxidant status (TAS) level. The obtained results indicate that EOs could be used as a potential source of safe and potent natural antimicrobial and antioxidant agents in the pharmaceutical and food industries.

Botanicals With Dermatologic Properties Derived From First Nations Healing: Part 1-Trees.

INTRODUCTION: First Nations people have a long history of working with medicinal plants used to treat skin diseases. The purpose was to assess the dermatologic therapeutic potential of western red cedar, white spruce, birch, balsam poplar, and black spruce. METHODS: Based on expert recommendations, 5 trees were selected that were used in First Nations medicine for cutaneous healing and have potential and/or current application to dermatology today. We searched several databases up to June 12, 2014. RESULTS: Western red cedar's known active principal compound, ß-thujaplicin, has been studied in atopic dermatitis. White spruce's known active principal compound, 7-hydroxymatairesinol, has anti-inflammatory activity, while phase II clinical trials have been completed on a birch bark emulsion for the treatment of actinic keratoses, epidermolysis bullosa, and the healing of split thickness graft donor sites. Balsam poplar has been used clinically as an anti-aging remedy. Black spruce bark contains higher amounts of the anti-oxidant trans-resveratrol than red wine. DISCUSSION: North American traditional medicine has identified important botanical agents that are potentially relevant to both cosmetic and medical dermatology. This study is limited by the lack of good quality evidence contributing to the review. The article is limited to 5 trees, a fraction of those used by First Nations with dermatological properties.

Oral supplementation of Thuja oreintalis leaf extract moderately affects the behavior in albino mice in a gender specific manner.

Present study was designed to demonstrate the effect ethanolic leaf extract of Thuja orientalis on the learning and memory formation in albino mice. Four week old albino mice were either orally supplemented with 50mg/ml solvent/Kg body weight of Thuja orientalis or with equal volume of saline solution (0.9% NaCl) for seven days. A battery of neurological tests (Elevated plus maze, Rota Rod, light and dark box, open field and novel object recognition test) were applied at the end of dose supplementation. Thuja orientalis leaf extract treated female mice spent significantly les time on rotating rod (P = 0.02) than male treated with saline solution [indicating poor neuromuscular coordination]. While all other neurological test performances varied non significantly (P > 0.05) when compared between Thuja orientalis treated albino mice (of both genders) with their respective control groups indicating a moderate effect of 50mg/ml solvent/Kg body weight of Thuja orientalis on the behavior of young albino mice.

Adsorption characteristics of Pb(II) from aqueous solutions onto a natural biosorbent, fallen arborvitae leaves.

In this study, the potential of the oriental arborvitae leaves for the adsorption of Pb(II) from aqueous solutions was evaluated. Brunauer-Emmett-Teller analysis showed that the surface area of arborvitae leaves was 29.52 m(2)/g with pore diameter ranging from 2 to 50 nm. X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy showed C-C or C-H, C-O, and O-C=O were the main groups on the arborvitae leaves, which were the main sites for surface complexation. Finally, effects of adsorbent dose, initial pH, contact time, and coexisting natural organic matters (humic acid (HA)) on the adsorption of Pb(II) were investigated. The results indicated that the pHZPC (adsorbents with zero point charge at this pH) was 5.3 and the adsorption reached equilibrium in 120 min. Isotherm simulations revealed that the natural arborvitae leaves exhibit effective adsorption for Pb(II) in aqueous solution, giving adsorptive affinity and capacity in an order of 'no HA' > 5 mg/L HA > 10 mg/L HA, and according to the Langmuir models, the maximum adsorptions of Pb(II) were 43.67 mg/g, 38.61 mg/g and 35.97 mg/g, respectively. The results demonstrated that the oriental arborvitae leaves showed high potentials for the adsorption of Pb(II) from aqueous solutions.

External Nursing Applications in the Supportive Management of Prolonged Postoperative Ileus: Description of Interventions and Case Report.

Prolonged postoperative ileus is a common but clinically challenging problem that leads to patient discomfort and prolonged hospitalization; the condition is managed through a multimodular program of supportive measures. In anthroposophic nursing, the management of prolonged postoperative ileus involves additional tools, including external abdominal compresses and massages with plant or silver-containing oils and ointments. We describe 3 typical techniques: Oxalis tincture compresses, Thuja/Argentum ointment compresses, and massage with "Wala Melissenöl" (containing Melissa officinalis, Carvum cari, Foeniculum amari, and Origanum majorana). A 61-year-old man with chronic pain from adhesions after multiple abdominal surgical procedures developed a prolonged postoperative ileus after an elective ileostomy reversal. Following slow recovery during the first postoperative days, he began vomiting. A nasogastric tube was inserted, and daily Oxalis tincture compresses and massage with "Wala Melissenöl" and Thuja/Argentum ointment compresses were applied on the abdomen. The patient's symptoms gradually improved over the next 10 days. No prokinetic medications were needed to manage this episode. External abdominal nursing applications with plant substances and silver can be an additional tool in the management of prolonged postoperative ileus.

Short Communication: Evaluation of the chemical composition of essential oil of Thuja occidentalis leaves grown in Peshawar, Pakistan by gas chromatography mass spectrometry.

Essential oil extracted from the fresh leaves of Thuja occidentalis were evaluated for its chemical composition employing GC-MS. Total of twenty nine components were identified and determined quantitatively using the area normalization procedure. Alpha-pinene and (+)-4-carene were found in high amount with a percentage concentration of 54.78 and 11.28 respectively. Other compounds which yielded appreciable amounts are: alpha-cedrol (6.87%), terpinolene (5.88%), p-menth-1-en-8-ol acetate (5.21%), beta-myrcene (4.04%), beta-pinene (2.26%), germacrene D (1.72%), sabinene (1.65%) and D-Limonene (1.62%).

UPLC-MS-ELSD-PDA as a powerful dereplication tool to facilitate compound identification from small-molecule natural product libraries.

The generation of natural product libraries containing column fractions, each with only a few small molecules, using a high-throughput, automated fractionation system, has made it possible to implement an improved dereplication strategy for selection and prioritization of leads in a natural product discovery program. Analysis of databased UPLC-MS-ELSD-PDA information of three leads from a biological screen employing the ependymoma cell line EphB2-EPD generated details on the possible structures of active compounds present. The procedure allows the rapid identification of known compounds and guides the isolation of unknown compounds of interest. Three previously known flavanone-type compounds, homoeriodictyol (1), hesperetin (2), and sterubin (3), were identified in a selected fraction derived from the leaves of Eriodictyon angustifolium. The lignan compound deoxypodophyllotoxin (8) was confirmed to be an active constituent in two lead fractions derived from the bark and leaves of Thuja occidentalis. In addition, two new but inactive labdane-type diterpenoids with an uncommon triol side chain were also identified as coexisting with deoxypodophyllotoxin in a lead fraction from the bark of T. occidentalis. Both diterpenoids were isolated in acetylated form, and their structures were determined as 14S,15-diacetoxy-13R-hydroxylabd-8(17)-en-19-oic acid (9) and 14R,15-diacetoxy-13S-hydroxylabd-8(17)-en-19-oic acid (10), respectively, by spectroscopic data interpretation and X-ray crystallography. This work demonstrates that a UPLC-MS-ELSD-PDA database produced during fractionation may be used as a powerful dereplication tool to facilitate compound identification from chromatographically tractable small-molecule natural product libraries.

Bioactive sesquiterpenes from the essential oil of Thuja orientalis.

A phytochemical investigation on the essential oil of Thuja orientalis resulted in the isolation and identification of three new sesquiterpenes, 3α-methoxy-4α-epoxythujopsane (1), Δ³,¹5-4ß-epoxythujopsene (2), and Δ³,4-thujopsen-2,15-diol (3), together with eight known sesquiterpenoids (4-11). The structures of these new compounds were elucidated based on spectroscopic data analyses including extensive 2D-NMR data and HR-ESIMS. The full assignments of ¹H and ¹³C NMR chemical shifts for thujopsadiene (4) were obtained by 2D-NMR for the first time. All compounds (1-11) showed antiproliferative activities against the SK-OV-3 and SK-MEL-2 cell lines with IC50 values of 5.85-28.64 µM. In addition, compounds 1, 3, 4, 7, 8, and 9 significantly inhibited nitric oxide production in lipopolysaccharide-activated BV-2 cells with IC50 values of 3.93-17.85 µM without cell toxicity.

Hair growth-promoting activity of hot water extract of Thuja orientalis.

BACKGROUND: Thuja orientalis has been traditionally used to treat patients who suffer from baldness and hair loss in East Asia. The present study sought to investigate the hair growth-promoting activity of T. orientalis hot water extract and the underlying mechanism of action. METHODS: After T. orientalis extract was topically applied to the shaved dorsal skin of telogenic C57BL/6 N mice, the histomorphometric analysis was employed to study induction of the hair follicle cycle. To determine the effect of T. orientalis extract on the telogen to anagen transition, the protein expression levels of ß-catenin and Sonic hedgehog (Shh) in hair follicles were determined by immunohistochemistry. RESULTS: We observed that T. orientalis extract promoted hair growth by inducing the anagen phase in telogenic C57BL/6 N mice. Specifically, the histomorphometric analysis data indicates that topical application of T. orientalis extract induced an earlier anagen phase and prolonged the mature anagen phase, in contrast to either the control or 1% minoxidil-treated group. We also observed increases in both the number and size of hair follicles of the T. orientalis extract-treated group. Moreover, the immunohistochemical analysis reveals earlier induction of ß-catenin and Shh proteins in hair follicles of the T. orientalis extract-treated group, compared to the control or 1% minoxidil-treated group. CONCLUSION: These results suggest that T. orientalis extract promotes hair growth by inducing the anagen phase in resting hair follicles and might therefore be a potential hair growth-promoting agent.

Diterpene and biflavone derivatives from Thuja koraiensis and their cytotoxicities against A549 cells.

During the screening of the cytotoxicity of rare Korean endemic plants, the extract of Thuja koraiensis Nakai displayed potent cytotoxicity against the adenocarcinomic human alveolar basal epithelial A549 cell line. Through a series of separations via column chromatography, three undescribed abietanes, an undescribed labdane along with a labdane, and a biflavonoid were purified from methylene chloride (CH2Cl2) fraction possessing a potent cytotoxic effect. Extensive 1D and 2D NMR spectroscopic data analyses, in combination with quantum chemical calculations were conducted to establish the planar and absolute configurations of thujakoraienes A-C. The chemical structure of thujakoraiene D was elucidated by spectroscopic data analysis and competing enantioselective acylation. Thujakoraienes A and C along with 7,7″-di-O-methylamentoflavone, showed cytotoxic effects on A549 cells, with IC50 values of 64.86, 47.97, and 16.14 µM, respectively. Finally, thujakoraiene C and 7,7″-di-O-methylamentoflavone were identified as potent cytotoxic compounds in A549 cells, followed by an additional cytotoxicity test in the normal human lung fibroblast MRC-5 cell line. This is the first study on the non-volatile chemicals in the extract of T. koraiensis and comparison of chemical profiles of T. orientalis and T. koraiensis.

Pinusolide isolated from Biota orientalis inhibits 5-lipoxygenase dependent leukotriene C4 generation by blocking c-Jun N-terminal kinase pathway in mast cells.

Pinusolide, an herbal medicine isolated from Biota orientalis L. (B. orientalis), inhibited 5-lipoxygenase (5-LO)-dependent leukotriene C4 (LTC4) generation in immunoglobulin E (IgE)/Ag-induced bone marrow-derived mast cells (BMMCs) in a concentration-dependent manner. To clarify the action mechanism of pinusolide on the inhibition of LTC4 generation, we examined the effect of pinusolide on phosphorylation of cytosolic phospholipase A2 (cPLA2), as well as translocation phospho-cPLA2 and 5-LO to nucleus. Inhibition of LTC4 generation by pinusolide was accompanied by a decrease in cPLA2 phosphorylation which occurred via a decrease in intracellular Ca2+ influx and blocking the c-Jun N-terminal kinase (JNK) pathways. However, pinusolide had no effect on extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinas phosphorylation. Taken together, the present results suggest that potent inhibition of 5-LO dependent LTC4 generation by pinusolide requires both suppression of calcium influx and JNK phosphorylation.

Three new diterpenoids from the leaves of Thuja orientalis.

Three new diterpenoids, 18-formyloxy-8 ß-hydroxysandaracopimar-15-ene (1), 15(R)-N-butoxypinusolidic acid (2), and 15,16-dihydro-15,16-dimethoxylambertianic acid (3), along with twelve known compounds (4-15) were isolated from MeOH extracts of leaves of Thuja orientalis L. The structures of the three new compounds were elucidated on the basis of spectroscopic analyses, including extensive 2D-NMR data. The absolute stereochemistry of compound 2 was clarified by a CD spectroscopic study. The isolated compounds were evaluated for cytotoxicity against A549, SK-OV-3, SK-MEL-2, and HCT-15 cell lines in vitro using the sulforhodamin B (SRB) bioassay.

Anti-allergic effect of lambertianic acid from Thuja orientalis in mouse bone marrow-derived mast cells.

Lambertianic acid is a bioactive diterpene found in the leaves of Thuja orientalis. Its effect on the bone marrow-derived mast cell (BMMC) mediated allergy and inflammation mechanism remains unknown. In this study, lambertianic acid was evaluated for its effect on the allergic mediators, including prostaglandin D(2) (PGD(2)), leukotriene C(4) (LTC(4)), ß-hexosaminidase (ß-Hex) and cyclooxygenase-2 (COX-2) protein, in phorbol 12-myristate 13-acetate (PMA) plus calcimycin-stimulated BMMCs. The results revealed that lambertianic acid inhibited the production of interleukin-6 (IL-6), PGD(2) and LTC(4), the expression of COX-2 and the degranulation of ß-hexosaminidase in the PMA plus calcimycin-induced BMMCs. Taken together, these findings implied that lambertianic acid may possess the potential in the treatment of allergy.