Cinnamic acid derivatives from welsh onion (Allium fistulosum) and their antibacterial and cytotoxic activities.

INTRODUCTION: Cinnamic acids are a class of compounds based on phenyl propanoid backbone (C6-C3) isolated from plants and microorganisms, exhibiting interesting biological activities. OBJECTIVE: To characterise cinnamic acids through the phytochemical study of welsh onion, Allium fistulosum, and to evaluate their antibacterial and cytotoxic properties. MATERIAL AND METHODS: The phytochemical study of A. fistulosum was performed through chromatographic techniques, including reversed phase medium-pressure liquid chromatography (MPLC) and high-pressure liquid chromatography (HPLC). Preliminary analysis of crude chromatographic fractions from the organic extracts was carried out by proton nuclear magnetic resonance (1 H-NMR) in order to prioritise the study of those having phenyl propanoid skeleton. The structural identification of the isolated compounds was performed through analysis of spectroscopic data, mainly one-dimensional (1D) and two-dimensional (2D) NMR. The antibacterial activity was assessed against gram negative (Escherichia coli) and gram positive (Staphylococcus aureus) bacteria while the cytotoxic property was evaluated on breast cancer cell line (MCF-7). RESULTS: The 1 H-NMR study of crude fractions and application of a straightforward method to purify the phenyl propanoid compounds by reversed phase MPLC and HPLC, allowed the effortless isolation of several cinnamic acids, including two new rare phenolic imidates (1 and 2). The use of an entirely NMR approach for structural elucidation of the isolated metabolites allowed the isolated material to be kept for further pharmacological tests. CONCLUSION: These results corroborate the importance of the use of 1D and 2D NMR to the identification of new phenyl propanoids, potential lead compounds against bacteria and cancer cells.

Molecular Networking-Based Analysis of Cytotoxic Saponins from Sea Cucumber Holothuria atra.

The saponin composition of a specimen of black sea cucumber, Holothuria atra collected in the Persian Gulf was studied by a combined approach including LC-MS/MS, Molecular Networking, pure compound isolation, and NMR spectroscopy. The saponin composition of Holothuria atra turned out to be more complex than previously reported. The most abundant saponins in the extract (1⁻4) were isolated and characterized by 1D- and 2D-NMR experiments. Compound 1 was identified as a new triterpene glycoside saponin, holothurin A5. The side chain of the new saponin 1, unprecedented among triterpene glycosides, is characterized by an electrophilic enone function, which can undergo slow water or methanol addition under neutral conditions. The cytotoxic activity of compounds 1⁻4, evaluated on the human cervix carcinoma HeLa cell line, was remarkable, with IC50 values ranging from 1.2 to 2.5 µg/mL.

Aspects of quality of life affected in morbidly obese patients who decided to undergo bariatric surgery: A qualitative study to design a native questionnaire.

BACKGROUND: Obesity is a known prevalent major health issue. The aim of this study is to assay Iranian patients' problems with obesity and their expectations of bariatric surgery. MATERIALS AND METHODS: In this study, we included patients who have used different medical noninvasive treatments and were unsuccessful in losing weight from the obesity clinic in Al Zahra Hospital, Isfahan, from 2014 to 2015. Morbidly obese patients were interviewed using some open-ended questions, and then, directional content analysis of data was done. RESULTS: Analysis of data showed five main categories including (1) physical health, (2) psychological health, (3) social relationships, (4) environment, and (5) "about the causes of obesity" with some subcategories for each category. CONCLUSION: This study is the first step of designing a quality of life questionnaire while we focused on spiritual and cultural states of Iranian people.

Furostanol Saponins from the Bulbs of Welsh Onion, Allium fistulosum L.

An extensive phytochemical analysis of the polar extracts from bulbs of Welsh onion Allium fistulosum L. led to the isolation of nine saponins, four of them, named fistulosaponins G (1a/1b), H (2), I (3a/3b), and J (4), have never been reported previously. Fistulosaponins G and I were isolated as a couple of isomers in equilibrium. On the basis of 2D NMR and mass spectrometry data, the structure of the novel compounds were elucidated as (25R)-26-[(ß-D-glucopyranosyl)oxy]-3ß,22ß-dihydroxyfurost-5-en-1ß-yl O-α-L-rhamnopyranosyl-(1 → 4)-O-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranoside (1a) with its 22α epimer (1b), (25R)-26-[(ß-D-glucopyranosyl)oxy]-3ß-hydroxyfurost-5,20-dien-1ß-yl O-α-L-rhamnopyranosyl-(1 → 4)-O-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranoside (2), (25R)-26-[(ß-D-glucopyranosyl)oxy]-3ß,22ß-dihydroxyfurost-5-en-1ß-yl O-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranoside (3a) with its 22α epimer (3b), and (25R)-26-[(ß-D-glucopyranosyl)oxy]-3ß-hydroxyfurost-5,20-dien-1ß-yl O-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranoside (4). This is the first report of furostanol saponins in A. fistulosum bulbs. In addition, data on the antibacterial tests of the isolated saponins against Escherichia coli and Enterococcus faecalis are reported.

Jatrophanes from Euphorbia squamosa as potent inhibitors of Candida albicans multidrug transporters.

A series of structurally related jatrophane diterpenoids (1-6), including the new euphosquamosins A-C (4-6), was purified from the Iranian spurge Euphorbia squamosa and evaluated for its capacity to inhibit drug efflux by multidrug transporters of Candida albicans. Three of these compounds showed an interesting profile of activity. In particular, deacetylserrulatin B (2) and euphosquamosin C (6) strongly inhibited the drug-efflux activity of the primary ABC-transporter CaCdr1p, an effect that translated, in a yeast strain overexpressing this transporter, into an increased sensitivity to fluconazole. These compounds were transported by CaCdr1p, as shown by the observation of an 11-14-fold cross-resistance of yeast growth, and could also inhibit the secondary MFS-transporter CaMdr1p. In contrast, euphosquamosin A (4) was selective for CaCdr1p, possibly as a result of a different binding mode. Taken together, these observations suggest jatrophane diterpenes to be a new class of potent inhibitors of multidrug transporters critical for drug resistance in pathogenic yeasts.

Comparing different sterol containing solid lipid nanoparticles for targeted delivery of quercetin in hepatocellular carcinoma.

Quercetin (QT) is a potential chemotherapeutic drug with low solubility that seriously limits its clinical use. The aim of this study was enhancing cellular penetration of QT by sterol containing solid lipid nanoparticles (SLNs) which make bilayers fluent for targeting hepatocellular carcinoma cells. Three variables including sterol type (cholesterol, stigmasterol and stigmastanol), drug and sterol content were studied in a surface response D-optimal design for preparation of QT-SLNs by emulsification solvent evaporation method. The studied responses included particle size, zeta potential, drug loading capacity and 24 h release efficiency (RE24%). Scanning electron and atomic force microscopy were used to study the morphology of QT-SLNs and their thermal behavior was studied by DSC analysis. Cytotoxicity of QT-SLNs was determined by MTT assay on HepG-2 cells and cellular uptake by fluorescence microscopy method. Optimized QT-SLNs obtained from cholesterol and QT with the ratio of 2:1 that showed particle size of 78.0 ± 7.0 nm, zeta potential of -22.7 ± 1.3 mV, drug loading efficiency of 99.9 ± 0.5% and RE24 of 56.3 ± 3.4%. IC50 of QT in cholesterol SLNs was about six and two times less than free QT and phytosterol SLNs, respectively, and caused more accumulation of QT in HepG2 cells. Blank phytosterol SLNs were toxic on cells.

Psychometric characteristics of the persian (farsi) version of attachment style questionnaire.

BACKGROUND: Attachment relationship provides a secure base for the infants from which to explore the environment and a safe haven to return to in times of danger. Attachment style shapes the behavior of individuals in adulthood. There are many different measures of attachment and a lot of controversy about what they measure and how they relate to each other. Hence, we tried to evaluate the psychometric properties of one of such questionnaires on a sample of the Iranian population. METHODS: "Attachment style questionnaire" designed by Van Oudenhoven measures four dimensions: secure, preoccupied, fearful and dismissing. Psychometric properties of the questionnaire were evaluated in a cross sectional study on 730 adults in Isfahan, Iran. Statistical analysis of data was performed by the explanatory factor analysis with the principal component method, Cronbach's alpha, Pearson correlation coefficients, and the multiple analysis of variance (MANCOVA). RESULTS: The Cronbach's alpha for all items was 0.704. As a whole, the internal consistency was good. There was a high inter-scale correlation between preoccupied and fearful, also the secure style correlated negatively with fearful and preoccupied. The stability coefficient of the attachment scales were 0.625, 0.685, 0.777 and 0.605 for secure, fearful, preoccupied and dismissing styles respectively (P<0.001). Regarding construct validity, factor analysis showed that some items require iterations to fit the Iranian population. CONCLUSION: This study showed that the Persian version of ASQ has a reasonable reliability and validity in general and the questionnaire is appropriate for use among the Iranian population in future studies.

New amide and diterpene alkaloids with anticholinesterase activity from Delphinium cyphoplectrum roots.

BACKGROUND: The cholinergic hypothesis posits a robust correlation between the onset of Alzheimer's disease and a pronounced deficit in acetylcholine, a pivotal neurotransmitter crucial for the central cholinergic nervous system's function, pivotal for memory and learning. Diterpene alkaloids exhibit intricate and distinctive chemical structures that facilitate their passage through the blood-brain barrier. Moreover, their potent pharmacological attributes render them promising candidates for addressing central nervous system disorders. OBJECTIVES: This investigation aims to scrutinize the alkaloidal composition of Delphinium cyphoplectrum (Ranunculaceae) roots, further exploring their anticholinesterase inhibitory activity and mode of inhibition. METHOD: Innovative chromatography techniques were repetitively employed to purify the alkaloids. Acetylcholinesterase (AChE) inhibition assays were conducted using Ellman's tests. The mode of inhibition was meticulously characterized through Michaelis-Menten, and Lineweaver-Burk plots. Conducting molecular docking studies, we employed the AUTO DOCK 4.2 software package. RESULTS: Eight alkaloids were identified including five C19-diterpene alkaloids (6,14,16,18-tetramethoxy-1,7,8-trihydroxy-4-methylaconitane (1), 6,16,18-trimethoxy-1,7,8,14-tetrahydroxy-4-methylaconitane (2), 6,8,16,18-tetramethoxy-1,7,14-trihydroxy-4-methylaconitane (3), 6,14,16-trimethoxy-1,7,8,18-tetrahydroxy-4-methylaconitane (4), and 14-O-acetyl-8,16-dimethoxy-1,6,7,18-tetrahydroxy-4-methylaconitane (5)), an epoxy C18-diterpene alkaloid (6,8,16-trimethoxy-1,7,14-trihydroxy-3,4-epoxyaconitane (6)), a known (pyrrolidin-2-one (7) and an undescribed amide alkaloid (1-(2'-hydroxylethylamine)-3,5,5,-trimethyl-1,5-dihydro-2H-pyrrol-2-one (8). All diterpene alkaloids underwent assessment for acetylcholinesterase (AChE) inhibition assay and displayed noteworthy AChE activity, surpassing that of the reference drug (with IC50 values of 13.7, 21.8, 23.4, 28.2, 40.4, and 23.9 for compounds 1-6, respectively, in comparison to 98.4 for Rivastigmine). Analysis of Michaelis-Menten and Lineweaver-Burk plots represents an uncompetitive mode of inhibition for compound 1 on AChE. Notably, computational docking simulations indicated that all diterpene alkaloids were accommodated within the same enzymatic cleft as the reference ligand, and displaying superior free binding energy values (from - 10.32 to -8.59 Kcal.mol-1) in contrast to Rivastigmine (-6.31 Kcal.mol-1). CONCLUSION: The phytochemical analysis conducted on the roots of Delphinium cyphoplectrum yielded the identification of eight alkaloidal compounds including one C18-diterpene, five C19-diterpene, one pyrrolidine and one amide alkaloids. AChE inhibition assay and molecular simulations unveiled remarkable significant potency attributed to the C19-diterpene alkaloids by the order of 1 > 2 > 3,6 > 4 > 5. Presence of hydroxyl group on C-1, C-7, C-8, C-14, and C-18 increased the effect. The best in vitro activity was recorded for compound 1 able to bind to Asp72 in the narrow region of PAS, while interacting by pi-sigma with Phe330 at the hydrophobic region of the gorge involving the acyl and choline binding site. This observation underscores the substantial promise of this category of natural products in the realm of drug discovery for Alzheimer's Disease, offering a compelling avenue for further research and therapeutic development.

Phytochemical study of Seriphidium khorassanicum (syn. Artemisia khorassanica) aerial parts: sesquiterpene lactones with anti-protozoal activity.

Two new eudesmane-type sesquiterpene lactones, 1ß,3α,8α-trihydroxy-11ß,13-dihydroeudesma-4(15)-en-12,6α-olide (1) and 1ß,4α,8α-trihydroxy-11ß,13-dihydroeudesma-12,6α-olide (2), and an unprecedented elemane-type sesquiterpene lactone, 1ß,2ß,8α-trihydroxy-11ß,13-dihydroelema-12,6α-olide (3) along with a known eudesmanolide artapshin (4) were isolated from Seriphidium khorassanicum. Structures were elucidated by NMR, HR-ESI-MS, and ECD spectral data analysis. The anti-protozoal activity was evaluated against Leishmania major promastigotes and amastigote-infected macrophages. They showed dose- and time-dependent activity against L. major amastigotes with IC50 values in the range of 4.9 to 25.3 µM being favourably far below their toxicity against normal murine macrophages with CC50 values ranging from 432.5 to 620.7 µM after 48 h of treatment. Compound 3 exhibited the strongest activity and the highest selectivity index (SI) with IC50 of 4.9 ± 0.6 µM and SI of 88.2 comparable with the standard drug, meglumine antimoniate (Glucantime), with IC50 and SI values of 15.5 ± 2.1 µM and 40.0, respectively.

Vavilosides A1/A2-B1/B2, new furostane glycosides from the bulbs of Allium vavilovii with cytotoxic activity.

A phytochemical analysis of the bulbs of Allium vavilovii M. Pop. & Vved. was attained for the first time extensively, affording to the isolation of four new furostanol saponins, named vavilosides A1/A2-B1/B2 (1a/b-2a/2b), as two couple of isomers in equilibrium, together with ascalonicoside A1/A2 (3a/3b) and 22-O-methyl ascalonicoside A1/A2 (4a/4b), previously isolated from shallot, Allium ascalonicum. High concentrations of kaempferol, kaempferide, and kaempferol 4(I)-glucoside were also isolated. The chemical structures of the new compounds, established through a combination of extensive nuclear magnetic resonance, mass spectrometry and chemical analyses, were identified as (25R)-furost-5(6)-en-1ß,3ß,22α,26-tetraol 1-O-α-L-rhamnopyranosyl-(1→2)-O-ß-D-galactopyranosyl 26-O-α-L-rhamnopyranoside (vaviloside A1), (25R)-furost-5(6)-en-1ß,3ß,22ß,26-tetraol 1-O-α-L-rhamnopyranosyl-(1→2)-O-ß-D-galactopyranosyl 26-O-α-L-rhamnopyranoside (vaviloside A2), (25R)-furost-5(6)-en-1ß,3ß,22α,26-tetraol 1-O-α-L-rhamnopyranosyl-(1→2)-O-ß-D-xylopyranosyl 26-O-α-L-rhamnopyranoside (vaviloside B1), (25R)-furost-5(6)-en-1ß,3ß,22ß,26-tetraol 1-O-α-L-rhamnopyranosyl-(1→2)-O-ß-d-xylopyranosyl 26-O-α-L-rhamnopyranoside (vaviloside B2). The isolated saponins showed cytotoxic activity on J-774, murine monocyte/macrophage, and WEHI-164, murine fibrosarcoma, cell lines with the following rank: vaviloside B1/B2>ascalonicoside A1/A2>vaviloside A1/A2.

The psychometric properties of the persian migraine-specific quality of life questionnaire version 2.1 in episodic and chronic migraines.

BACKGROUND: Migraine-specific quality of life (MSQ) is a valid and reliable questionnaire. Linguistic validation of Persian MSQ questionnaire, analysis of psychometric properties between chronic and episodic migraine patients, and capability of MSQ to differentiate between chronic and episodic migraines were the aims of this study. METHOD: Participants were selected from four different neurology clinics that were diagnosed as chronic or episodic migraine patients. Baseline data included information from MSQ v. 2.1, MIGSEV, SF-36, and symptoms questionnaire. At the third week from the baseline, participants filled out MSQ and MIGSEV. Internal consistency (Cronbach alpha) and test-retest reproducibility (intraclass correlation coefficients) were used to assess reliability. Convergent and discriminant validities were also assessed. RESULTS: A total of 106 participants were enrolled. Internal consistencies of MSQ among all patients, chronic and episodic migraines, were 0.92, 0.91, and 0.92, respectively. Test-retest correlation of MSQ dimensions between visits 1 and 2 varied from 0.41 to 0.50. Convergent, item discriminant, and discriminant validities were approved. In all visits MSQ scores were lower in chronic migraine than episodic migraine; however, the difference was not statistically significant. CONCLUSION: Persian translation of MSQ is consistent with original version of MSQ in terms of psychometric properties in both chronic and episodic migraine patients.

Dentists' knowledge, attitude, and practice regarding oral cancer in Iran.

Dentists are usually the first group who can examine patients for oral cancer and the early diagnosis highly depends on their knowledge. In this study, we aim to survey the Iranian dentists' knowledge, attitude, and behaviors regarding oral cancer. A valid and reliable self-administered questionnaire was designed and sent to 150 dentists. Responses to the questionnaires were analyzed using descriptive and analytical statistics. Just one fifth of the dentists reported to perform oral cancer examination for all of their patients in age of 40 and above and about 34 % of them were knowledgeable. They mostly claimed that the lack of knowledge is the main barriers to the provision of routine oral cancer examinations. The opinion of dentists about the effectiveness of continuing education courses supports the development of these courses on oral cancer. Besides, more emphasis should be placed on oral cancer prevention in dental schools.

The effects of Astragalus polysaccharides, tragacanthin, and bassorin on methotrexate-resistant acute lymphoblastic leukemia.

Background and purpose: One strategy to overcome methotrexate (MTX) resistance in acute lymphoblastic leukemia is suppressing MDR1 expression. It has been proved Astragalus polysaccharides (APS) exert their anticancer effect by reversing drug resistance. Due to the structural similarity of tragacanthin and bassorin with APS, we aimed to investigate the effects of the aforementioned polysaccharides on the expression of the MDR1 gene in the MTX-treated CCRF-CEM cells. Experimental approach: Cytotoxicity of APS, bassorin, and tragacanthin on CCRF-CEM, CCRF-CEM/MTX (cells treated with MTX at IC50), and CCRF-CEM/R cells (CCRF-CEM cells resistant to MTX) was evaluated by MTT assay. The effect of all three compounds on MDR1 expression was evaluated using RT-PCR. Findings/Results: All the concentrations of tragacanthin, bassorin, and APS (except at 0.8-100 µg/mL in CCRF-CEM) decreased the viability of all the cells compared to the negative control group; and against the positive control (MTX-treated cells), only bassorin at 20-100 µg/mL in CCRF-CEM/R and tragacanthin at 50 and 100 µg/mL in CCRF-CEM/MTX and at 2-100 µg/mL in CCRF-CEM/R decreased cell viability. Tragacanthin diminished MDR1 expression in CCRF-CEM/MTX and CCRF-CEM/R cells, which MTX had already induced. Conclusion and implication: According to the results of this study, tragacanthin was a potent cytotoxic agent against CCRF-CEM cells and enhanced the chemosensitivity of CCRF-CEM/MTX and CCRF-CEM/R cells to MTX by down-regulation of MDR1 gene expression. Therefore, it could be a promising compound against cancer. Other possible mechanisms of action of tragacanthin should be evaluated and further in vitro and in vivo investigations are required.

Diterpenoids from Euphorbia gedrosiaca as Potential Anti-Proliferative Agents against Breast Cancer Cells.

Isolated diterpenes from various species of Euphorbia are important compounds for drug discovery with a broad spectrum of structures and biological effects. In this study, Euphorbia gedrosiaca, one of the endemic species of Iran, was analyzed in terms of the presence and structural determination of diterpenoid compounds. They were extracted with dichloromethane/acetone (2:1) from aerial parts of this plant and purified by chromatographic methods such as MPLC and HPLC. Four premyrsinane compounds and one myrsinane diterpene were isolated from Euphorbia gedrosiaca. They were characterized by extensive 1D and 2D NMR and HRMS analyses. Additionally, their activities were evaluated against two breast cancer cell lines, MDA-MB-231 and MCF-7, by MTT proliferation assay. They exhibited cytotoxic effects in a dose-dependent manner with promising results, which can help to find possible therapeutic application of diterpenoids in breast cancer treatment.

Neuropharmacological Potential of Diterpenoid Alkaloids.

This study provides a narrative review of diterpenoid alkaloids (DAs), a family of extremely important natural products found predominantly in some species of Aconitum and Delphinium (Ranunculaceae). DAs have long been a focus of research attention due to their numerous intricate structures and diverse biological activities, especially in the central nervous system (CNS). These alkaloids originate through the amination reaction of tetra or pentacyclic diterpenoids, which are classified into three categories and 46 types based on the number of carbon atoms in the backbone structure and structural differences. The main chemical characteristics of DAs are their heterocyclic systems containing ß-aminoethanol, methylamine, or ethylamine functionality. Although the role of tertiary nitrogen in ring A and the polycyclic complex structure are of great importance in drug-receptor affinity, in silico studies have emphasized the role of certain sidechains in C13, C14, and C8. DAs showed antiepileptic effects in preclinical studies mostly through Na+ channels. Aconitine (1) and 3-acetyl aconitine (2) can desensitize Na+ channels after persistent activation. Lappaconitine (3), N-deacetyllapaconitine (4), 6-benzoylheteratisine (5), and 1-benzoylnapelline (6) deactivate these channels. Methyllycaconitine (16), mainly found in Delphinium species, possesses an extreme affinity for the binding sites of α7 nicotinic acetylcholine receptors (nAChR) and contributes to a wide range of neurologic functions and the release of neurotransmitters. Several DAs such as bulleyaconitine A (17), (3), and mesaconitine (8) from Aconitum species have a drastic analgesic effect. Among them, compound 17 has been used in China for decades. Their effect is explained by increasing the release of dynorphin A, activating the inhibitory noradrenergic neurons in the ß-adrenergic system, and preventing the transmission of pain messages by inactivating the Na+ channels that have been stressed. Acetylcholinesterase inhibitory, neuroprotective, antidepressant, and anxiolytic activities are other CNS effects that have been investigated for certain DAs. However, despite various CNS effects, recent advances in developing new drugs from DAs were insignificant due to their neurotoxicity.

Antinociceptive and anti-inflammatory activities of Satureja hortensis seed essential oil, hydroalcoholic and polyphenolic extracts in animal models.

OBJECTIVE: The aim of this study was to evaluate the anti-inflammatory and analgesic effects of Satureja hortensis seed extracts and to analyze their essential oil constituents. MATERIALS AND METHODS: Hydroalcoholic and polyphenolic extracts and essential oil of S. hortensis seeds were prepared using standard methods. Analgesic activity was assessed in male mice (25-35 g) using standard methods (acetic acid and formalin tests). For the evaluation of anti-inflammatory activity, the carrageenan-induced rat paw edema test was used. The mice were pretreated with 50, 100 or 200 mg/kg, i.p., hydroalcoholic or polyphenolic extracts or 100 or 200 µl/kg, p.o. RESULTS: Hydrodistillation of the seeds of S. hortensis afforded a pale yellowish oil in a yield of 0.05% (v/w). Pretreatment of mice with hydroalcoholic or polyphenolic extracts or essential oil significantly (p < 0.001) reduced acetic acid-induced abdominal twitches. Hydroalcoholic extracts also significantly reduced pain responses in early and late phases of the formalin test whereas the polyphenolic extract and essential oil were only effective in the late phase of the formalin test. All three fractions were found to reduce paw edema in the carrageenan test. CONCLUSION: These results clearly demonstrate the analgesic and anti-inflammatory activity of S. hortensis seeds and since the hydroalcoholic extract relieved pain in the first phase of the formalin test, it seems that at least part of its analgesic activity may be mediated centrally. The results of this study substantiated the traditional use of S. hortensis plant seeds in painful and inflammatory ailments.

Evaluation of DNA damage of hydro-alcoholic and aqueous extract of Echium amoenum and Nardostachys jatamansi.

BACKGROUND: Today most of herbal medicines are marketing without any standard safety profiles. Although common assumption is that these products are nontoxic but this assumption may be incorrect and dangerous, so toxicological studies should be done for herbal drugs. According to the frequent use of Echium amoenum as immunostimulant and useful in conditions including pain, cough, sore throat and arthritis, and Nardostachys jatamansi as tranquilizer and sleep inducer and evidences of some toxicities, we assessed the probable effect of their extracts on DNA of hepG2 cells using the comet assay. MATERIALS AND METHODS: Different concentrations of above extracts of the plants are incubated with hepG2 cells for 24 h. A mixture of cell suspension and agarose gel were put on slides, then slides were embedded in a lysing solution and were put in electrophoresis buffer (pH = 13). Then the electrophoresis procedure took place in an alkaline solution and after neutralization stage, colorization was done by ethidium bromide and comets were observed using a fluorescence microscope. At least 100 cells of each sample were evaluated and three parameters including comet length, percent of DNA in tail, and tail moment were assessed. RESULTS: Both Aqueous and hydro-alcoholic extract of E. amoenum were genotoxic in the concentrations of 25 mg/ml and aqueous and hydro-alcoholic extract of N. jatamansi were genotoxic in the concentrations 5 and 10 mg/ml, respectively. CONCLUSIONS: Although E. amoenum and N. jatamansi are highly used in medicine, these herbs have genotoxic effects in determined concentrations and they should be used cautiously.

Phytochemical Study of Euphorbia turcomanica Boiss.

The attraction to the Euphorbia genus, with its remarkable diversity in species, is due to its variety of chemical compositions. Euphorbia turcomanica is one of the species of the spurge family growing wildly in Iran. This research aims to investigate the presence of secondary metabolites, specially terpenoid compounds, in terms of structural determination. Samples of E. tucomanica were finely powdered and macerated with acetone/dichloromethane 2:1. Repeated column chromatography using silica gel, MPLC, and HPLC methods followed by the analysis of data obtained from spectroscopic means was carried out to purify and identify the terpenoid constituents. The chemical structures of nine known terpenoids were determined for the first time from E. turcomanica during this study. Loliolide (1), a monoterpene, and eight steroids and triterpenes, including simiarenol (2), isomultiflorenol (3), cycloart-25-ene-3ß,24-diol (4), ß-sitosterol (5), cycloart-23-ene-3ß,25-diol (6), 3α, 11α-dihydroxyurs-12-ene (7), 3ß, 24ß, 25-trihydroxycycloartane (8), and 7α-hydroxystigmasterol (9) were isolated and identified. E. tucomanica, with a rich terpenoid profile, can be one of the valuable and economic sources providing compounds for drug development.

Gas Chromatography-Mass Spectrometry Analysis of Anbarnesa Smoke and Its Antiviral Activity.

Background: Anbarnesa is the female donkey dung typically collected after the labor and in early springtime. Materials and Methods: The chemical composition of the smoke collected from Anbarnesa was evaluated by gas chromatography-mass spectrometry (GC/MS), and its antiviral activity was analyzed based on 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Results: As a result, twenty-two constituents representing 97.1% of the Anbarnesa smoke could be identified. Hexadecanoic acid (29.4%), cis-9-octadecenoic acid (17.7%), and octadecanoic acid (10.8%) were the smoke's main constituents, respectively. Antiviral activity was evaluated using MTT assay. The CC50 value of the compound on Hep2 and Verro cells was 2271.2 µg/mL and 5077.5 µg/mL, respectively. Furthermore, the 50% inhibitory concentration value on adenovirus and herpes simplex type-1 was 802.55 µg/mL and >5077.5, respectively. Conclusions: it was revealed that Anbarnesa was nontoxic in 1/64, 1/128, and 1/256 dilutions, while the toxicity was detected in 1/32 dilution after 72 h. In addition, in 1/8 and 1/16 dilutions, cell toxicity was identified in the first hour.

New Cytotoxic Premyrsinane-Type Diterpenes from Euphorbia aleppica Against Breast Cancer Cells.

Euphorbia is used in traditional medicine to remove warts, possibly due to its cytotoxic or antiviral effects. This study investigated its phytochemistry and bioactive compounds. Euphorbia aleppica from the Euphorbiaceae family was collected from Kuhdasht, Lorestan, Iran. Plant material was dried and ground. Extraction was performed by maceration using a dichloromethane-acetone solvent. After removing fatty contents, fractionation was done by open column chromatography. Based on the initial H-NMR spectra, fractions containing diterpenoid compounds were identified. The Sephadex column and HPLC performed isolation. The HPLC was done with a regular YMC silica column using a hexane: Ethyl acetate (70: 30) solvent. The selected sub-fractions were identified by one and two-dimensional corelative NMR spectra. Accurate mass spectra confirmed the molecular formula of the obtained structures. Cytotoxicity was assessed using a standard MTT assay against breast cancer cells. The NMR and mass analysis identified compound 1 as a newly described and compound 2 as a pre-defined compound as 3, 7, 15ß-triacetyl-5α-tigliate-13(17)-α-epoxy-14-oxopremyrsinane and 3, 7, 14, 15, 17-pentaacetyl-5-tigliate-13(17)-epoxypremyrsinane, respectively. Compound 1 showed moderate cytotoxicity, and compound 2 exhibited a potent cytotoxic effect dose-dependently against MCF-7 and MDA-MB 231 breast cancer cells, probably because of 14-O-acetyl and 17-O-acetylated hemiacetal groups.