A new labdane diterpenoid, in vitro and in silico cytotoxicity, and protease inhibitory effects of phytochemicals from Juniperus polycarposK. Koch leaves.

Cytotoxicity-guided purification of Juniperus polycarpos K. Koch leaves (Cupressaceae) led to the isolation of a new labdane diterpenoid, 3-(acetyloxy)-acetylisocupressic acid (1), together with isocupressic acid (2), 3,4-dimethoxycinnamoyl alcohol (3) and deoxypodophyllotoxin (4). The chemical structures of 1-4 were established by detailed 1D and 2D NMR, HRFAB-MS and LRESI-MS, as well as by comparing the spectral data with those reported in the literature. Compound 1 was ineffective against HepG2 cells and protease enzyme, while 2 showed potent cytotoxicity against HepG2 cells (IC50 of 3.73 µg/mL) compared to cisplatin (IC50 of 12.65 µg/mL). Computational analyses with CDK1 protein (a prominent protein in the cell cycle of HepG2 cells) revealed the binding affinity of 2 (-31.86 kcal/mol) was better than 1 (-19.70 kcal/mol) because the acetoxy groups did not allow binding deeply to the ATP binding site. Compounds 2 and 4 moderately inhibited the protease activity (IC50 = 52.7 and 63.0 µg/mL, respectively). Further in vitro and in vivo studies on the plant are strongly recommended.

Effects of Salvia mirzayanii extract administration on hyperglycemia improvement in diabetic rats: The role of GLUT4, PEPCK and G6Pase genes.

Diabetes is a dangerous metabolic disorder by increasing incidence in human societies worldwide. Recently, much attention has been focused on the development of hypoglycemic agents, particularly the derivatives of herbal drugs, in the treatment of diabetes. This research aimed to study the anti-diabetic effect of Salvia mirzayanii in the diabetic rat models. First, the plant material was extracted from the leaves, and orally administered to the rats. After treating the animals with the aqueous extract of S. mirzayanii at a dose of 600 mg/kg, animal body weight for 12 weeks, fasting blood glucose, oral glucose tolerance test (OGTT), and body weight changes were examined. To analyze the anti-diabetic function of S. mirzayanii, we measured the expression of glucose transporter-4 (GLUT4), phosphoenolpyruvate carboxykinase (PEPCK), and glucose 6-phosphatase (G6Pase) genes in healthy and streptozotocin (STZ)-diabetic rats. The expression levels of the genes of interest in muscle and liver tissues were determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry (IHC). There were no significant differences in fasting blood glucose and OGTT between normal control (NC) group and the diabetic control (DC) group treated with S. mirzayanii. In contrast, there was a significant difference with the untreated DC (P < 0.05). The treatment of diabetic rats with S. mirzayanii significantly increased the expression of GLUT4 in the muscle and decreased the expression levels of PEPCK and G6Pase in the liver compared to the DC group (P < 0.05). These findings clearly show that S. mirzayanii can improve hyperglycemia by increasing the GLUT4 expression, and inhibiting the gluconeogenesis pathway in the liver. In general, the obtained results provided a new insight into the efficacy of S. mirzayanii aqueous extract as an anti-diabetic herbal medicine.

Antiproliferative Effects of Ferulic, Coumaric, and Caffeic Acids in HepG2 Cells by hTERT Downregulation.

Objective: Phenolic acids are well-known phytochemicals that are detected in a wide variety of medicinal plants, and their antiproliferative effects on cancer cells are known, but their mechanisms are poorly revealed. In most of cancer cells, telomerase reverse transcriptase (hTERT) is a dominant factor of telomere length regulation. The hTERT expression promotes invasiveness in tumor cells and is a hallmark of cancer. Therefore, in this study, the probable inhibitory effects of caffeic (Caf), coumaric (Cum), and ferulic acids (Fer) are investigated on the hTERT expression pattern in HepG2 cells. Methods: The MTT, apoptosis assays, and real-time PCR analysis were applied to evaluate viability, cytotoxicity, and hTERT gene expression level, respectively. Results: All of the studied phenolic acids showed cytotoxic effects on HepG2 cells in a timely manner and presented a time-dependent inhibitory effect on the growth of HepG2 cells. They reduced percentage of viable cells and induced apoptosis. Also, these phenolic acids had significant inhibitory effects on hTERT gene expression. Conclusion: These findings suggest that cell viability along with hTERT gene expression in HepG2 cells could be reduced by Cum, Caf, and Fer. As different cancer cells are resistant to conventional chemotherapeutics, this type of results proposes the telomerase as a proper target of cancer therapy development by natural products.

Reassessing the molecular structures of some previously isolated abietane diterpenoids with a naphthalene moiety and the structure-activity relationship (SAR) of quinone diterpenoids.

Crystals of previously described para-naphthoquinone abietane diterpenoids 12,16-dideoxy-aegyptinone B and 12-deoxy-salvipisone were obtained from Zhumeria majdae Rech.f. & Wendelbo. However, single-crystal X-ray diffraction analysis followed by reinterpretation of their NMR data revealed that their structures require revision, and they should be revised to the two ortho-naphthoquinones, zhumerianone C and aethiopinone, respectively. Interestingly, a further search through literature revealed that there were more of such cases, in which differentiation between the ortho-/para-orientation had not been carried out correctly in the structure elucidation of naphthalene containing abietane diterpenoids. Therefore, in the current study, we pointed out some 1D and 2D NMR generalizations that would help the unambiguous deduction of the ortho-/para-orientation of naphthalene containing abietanes and revised the structure of some previously described compounds accordingly. Based on these generalizations, structures of sibiriquinones A and B, sahandinone, and sahandone were revised to the known structures 1,2-didehydromiltirone, miltirone, saprorthoquinone, and sahandone B, respectivelyand tebesinone B, arucadiol, and sahandol II were revised to three undescribed structures. It was also proposed that structures of palmitoyl arucadiol and compounds with the salvifolane skeleton need revision. Furthermore, these structure revisions shed light on the structure-activity relationship of the quinone diterpenoids, approving that the ortho-quinone is the critical structural component for cytotoxicity in these compounds.

Inhibition of α-Amylase and α-Glucosidase of Anthocyanin Isolated from Berberis integerrima Bunge Fruits: A Model of Antidiabetic Compounds.

Anthocyanins are components of the flavonoid group with different properties, such as antidiabetic properties. This study aimed to isolate anthocyanin from Berberis integerrima Bunge fruits and evaluate α-amylase and α-glucosidase inhibition by this mentioned anthocyanin. The anthocyanin of Berberis integerrima fruit was isolated using column chromatography, and the antidiabetic properties of the anthocyanin were determined by the levels of α-amylase and α-glucosidase inhibition. Km and Vmax were also evaluated using the GraphPad Prism 7. The results of this study showed that the anthocyanin content of the fruit extract was 14.36 ± 0.33 mg/g, and following purification, this amount increased to 34.51 ± 0.42 mg/g. The highest of α-glucosidase inhibition was observed in the purified anthocyanin with IC50 = 0.71 ± 0.085 mg/ml, compared to acarbose as the baseline with IC50 = 8.8 ± 0.14 mg/ml, p < 0.0001. Purified anthocyanin of the mentioned fruit with IC50 = 1.14 ± 0.003 mg/ml had the greatest α-amylase inhibition, which was similar to acarbose as the standard with IC50 = 1 ± 0.085 mg/ml, p < 0.05. The inhibition of α-glucosidase and α-amylase by purified anthocyanin showed uncompetitive inhibition, and the enzyme inhibition by unpurified anthocyanin showed mixed inhibition. The obtained findings showed that Berberis integerrima fruit can be mentioned as a source of anthocyanin with antidiabetic properties.

Improvement of Phenolic Compound Extraction by Using Ion Exchange Chromatography and Evaluation of Biological Activities of Polyphenol-enriched Fraction of Rosa canina Fruits.

Background: Rosa canina has been traditionally known as a medicinal plant. Different applications of fruits (Rose hip) comprise the food, perfume, and cosmetic industries. Objectives: This study aimed to prepare an enriched polyphenolic fraction from Rosa canina in addition to its biological activities. Methods: Poly phenolic enriched fraction was prepared using Amberlite XAD-7 for removing unwanted components. Phenols, flavonoids, and anthocyanins content analyses showed that they increased significantly compared to the extract. HPLC analysis showed that this fraction is a rich source of ascorbic acid. Results: The results of the DPPH, ferric-reducing antioxidant power (FRAP), ABTS, and nitric oxide assay confirmed that the antioxidant activities of the fraction had been increased compared to the extract. The oxygen radical absorbance capacity (ORAC) assay and cellular antioxidant activity of the fraction also confirmed its potential antioxidant activity. This fraction showed xanthine oxidase inhibitory activity at 100 µg/mL concentration. Comet assay analysis revealed that this fraction at 25 - 100 µg/mL concentrations inhibited H2O2 genotoxicity in human lymphocytes. Conclutions: This study suggests that the fruit of Rosa canina could be considered as a potential antioxidant, a xanthine oxidase inhibitor, and an antigenotoxic source, and the application of Amberlite XAD-7 improves extraction efficiencies through enrichment of phenolic compounds in this plant.

Assessment of the Cytotoxic Activity of Triphala: A Semisolid Traditional Formulation on HepG2 Cancer Cell Line.

Cancer chemotherapies may result in resistance, and therefore, contemporary treatments including natural products may find an increasing consideration. As per Persian medicine (PM), many natural products have been used for malignant and chronic diseases. Triphala, with a combination of Terminalia chebula Retz., Terminalia bellirica Retz., Phyllanthus emblica L., and honey, is a multi-ingredient traditional formulation attributed to anticancer activities in PM. This study is aimed at evaluating the cytotoxic activity of this preparation on HepG2, the human liver cancer cell line. Hydroalcoholic extracts were prepared from the formulation and its components. Compared with the control and Cisplatin, the extracts were tested using MTT assay at different concentrations. All concentrations of the preparation, as well as Cisplatin, were effective significantly against HepG2 cells. All extract preparations at multiple concentrations were significantly effective as evidenced by MTT assay when compared to the control group. The IC50 level for Triphala extract was 77.63 ± 4.3 µg/ml. Based on the results, Triphala and its components have cytotoxic activity on the HepG2 cancer cell line and they can reduce the survival rate significantly.

Preparation and Evaluation of Possible Antioxidant Activities of Rose Traditional Tablet"(Qurs-e-Vard)" A Selected Traditional Persian Medicine (TPM) Formulation via Various Procedures.

BACKGROUND: Free radicals can lead to liver dysfunction. Quality control of traditional formulations ensures their safe, pure, and pharmaceutical efficacy. "Qurs-e-Vard", containing petals of Rosa damascena Mill., fruits of Rhus coriaria L. and roots of Glycyrrhiza glabra L. has been suggested as a hepatoprotective preparation in Traditional Persian Medicine (TPM). OBJECTIVE: This study was directed at the evaluation of the phytochemical characterization, standardization, and in vitro antioxidant activity determination of a solid formulation and its components. METHODS: Some qualitative and quantitative controls were performed like ash value, heavy metals investigation, and microbial contamination. The phytochemical assays were used for obtaining total phenolic and flavonoid contents with spectrophotometric methods. 2, 2-Diphenyl-1-c (DPPH) and Nitric Oxide (NO) assays were run for determining Radical scavenging activities of the formulation and its components. Ferric reducing antioxidant power assay (FRAP) was determined as well. RESULTS: Total phenolic contents of hydroalcoholic and aqueous extracts of the polyherbal formulation measured respectively, (376±0.93) and (297.6±0.96) mg of gallic acid/g of dry matter. Total flavonoid contents of the formulation were also measured (36.27±0.98) for hydroalcoholic extract and (17.79±0.86) mg of quercetin/g of dry matter for aqueous extract. The IC50 of hydroalcoholic and aqueous extract was obtained (88.14±1.15) and (140.78±2.98) µg/ml, respectively. NO scavenging percentages (200µg/ml) of hydroalcoholic and aqueous extracts were measured (59.11±2.15) and (65.08±2.35). FRAP values of hydroalcoholic and aqueous extracts were achieved (255.24±3.45) and (134.57±3.45) µg/ml as well. CONCLUSION: Our findings indicated that this polyherbal formulation and its components have justifiable antioxidant effects.

Valorization of Waste Water of Rosa damascena Oil Distillation Process by Ion Exchange Chromatography.

Water steam distillation is a classical method of rose oil production from the flowers of Rosa damascena Mill. This process produces considerable amount of waste water. In this study, ion-exchange column chromatography (Amberlite was the stationary phase) was used to prepare polyphenol-enriched fraction of waste water with improved biological activity. Phenol, flavonoid, and anthocyanin contents were examined before and after using column. Antioxidant activities, DNA protection ability, xanthine oxidase inhibition, and cytotoxicity of this fraction were also determined. The use of Amberlite increased phenol, flavonoid, and anthocyanin contents in fraction compared to the sample before fractionation. The IC50 values of various antioxidant assays comprises 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), ferric-reducing antioxidant power assay (FRAP), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) which were 226.66 ± 1.25, 126.03 ± 0.11, and 241.43 ± 0.33 for waste water, and these values for fraction were 63.21 ± 0.90, 34.6 ± 0.17, and 50.59 ± 0.75 µg/ml, respectively. The Trolox equivalent values of fraction in oxygen radical absorbance capacity (ORAC) assay were 0.34 ± 0.04, and the EC50 values in cellular antioxidant activity were 91.24 ± 0.32 µg/ml. The xanthine oxidase inhibition capacity of fraction (100 µg/ml) was 96.4 ± 0.02% µg/ml. The comet assay analysis showed that this fraction (25-100 µg/ml) protects human lymphocytes against H2O2-induced DNA damages significantly. The IC50 values of cytotoxicity assay were 248.145 ± 35.56 and 227.14 ± 16.51 µg/ml after 24 and 48 h, respectively. There has been great attention to the valorization of waste materials. Recovered fraction could be considered as a proper antioxidant, DNA damage-protection agent, and xanthine oxidase inhibitor. Using a nontoxic solid phase such as Amberlite is a fruitful way to concentrate bioactive ingredients which can be used in pharmaceutical and nutraceutical industry.

DNA protection, antioxidant and xanthine oxidase inhibition activities of polyphenol-enriched fraction of Berberis integerrima Bunge fruits.

OBJECTIVES: The aim of this study was to prepare fraction and determine the biological activities of the polyphenol-enriched fraction of Berberis integerrima Bunge fruits. MATERIALS AND METHODS: In this assay fraction was extracted by column chromatography, using Amberlite column as the stationary phase. Phenol and flavonoids in the extract and fraction were analyzed by high performance liquid chromatography (HPLC). DNA protection ability, antioxidant and xanthine oxidase inhibition capacities of this fraction were also examined. RESULTS: Phenol and flavonoid content measurement and HPLC analyses of this fraction confirmed that phenol and flavonoids were increased in fraction in comparison to extract (before using Amberlite column). In antioxidant measurement assay, the trolox equivalent values were 1.05± 0.04 and 0.8±0.11 in oxygen radical absorbance capacity (ORAC) and the EC50 values for cellular antioxidant activity were 55.51±0.21 and 95.67±0.13 µg/ml for quercetin and the fraction, respectively. The xanthine oxidase inhibition percentages were 97.6±0.003 and 90.2 6±0.003 in 100 µg/ml concentration of fraction and vitamin C respectively. Comet assay analysis showed that this fraction protects human lymphocytes against H2O2-induced DNA damages at 12.5 to 100 µg/ml concentrations. CONCLUSION: This study suggests that Amberlite column as the stationary phase help to improve phenolic compound in separating fractions. The results showed that B. integerrima fruits are rich in phenolic compounds and they are potent antioxidants with protective effects on oxidative damages. They might be used as functional ingredients in food and supplements.

Insight into DNA protection ability of medicinal herbs and potential mechanisms in hydrogen peroxide damages model

DNA damage is one of the most important consequences of oxidative stress in the cells.If DNA repair is unable to modify these inducible DNA damages,genomic instability may lead to mutation,cancer,aging and many other diseases.Single cell gel electrophoresis or comet assay is a common and versatile method to quantify these types of DNA damages.DNA damages induced by hydrogen peroxide (H2O2) are one of the proper models for measurement of protective ability of different compounds.So the main aim of this review is to provide an overview about protection ability of medicinal plants and their potential mechanism against H2O2 induced DNA damages.In this review,relevant researches on the effect of medicinal plants on DNA damages induced by H2O2 and possible molecular mechanisms are discussed.It seems that,medicinal plants are considered as therapeutic key factors to protect DNA from consequences caused by oxidative stress.Sufficientin vitro evidences introduce them as DNA protective agents through different mechanisms including antioxidant activity and some other cellular mechanisms.Moreover,in order to correlate the antigenotoxicity effects with their potential antioxidant property,most of medicinal plants were evaluated in term of antioxidant activity using standard methods.This review highlights the preventive effects of herbal medicine against oxidative DNA damages as well as provides rational possibility to engage them in animal studies and future clinical investigations.

An Overview on Indications and Chemical Composition of Aromatic Waters (Hydrosols) as Functional Beverages in Persian Nutrition Culture and Folk Medicine for Hyperlipidemia and Cardiovascular Conditions.

Hydrosol beverages in Persian nutrition culture and ethnomedicine are the side products of essential oil industry that are used as delicious drinks or safe remedies. To investigate indications and chemical composition of hydrosol beverages for hyperlipidemia and cardiovascular conditions, Fars province was selected as the field of study. Ethnomedical data were gathered by questionnaires. The constituents of hydrosols were extracted with liquid/liquid extraction and analyzed by gas chromatography-mass spectrometry. Statistical analysis were used to cluster their constituents and find the relevance of their composition. A literature survey was also performed on plants used to prepare them. Thymol was the major or second major component of these beverages, except for wormwood and olive leaf hydrosols. Based on clustering methods, although some similarities could be found, composition of barberry, will fumitory, dill, and aloe hydrosols have more differences than others. These studies may help in developing some functional beverages or new therapeutics.

Evaluation of Antioxidant Potentials and α-Amylase Inhibition of Different Fractions of Labiatae Plants Extracts: As a Model of Antidiabetic Compounds Properties.

In an attempt to identify herbal drugs which may become useful in the prevention of diabetes, antioxidant potentials and α-amylase inhibition by the ethanol extracts of two plants belonging to Lamiaceae family, Otostegia persica and Zataria multiflora, and their different fractions were studied. Also, inhibition of α-amylase by Salvia mirzayanii and its fractions was evaluated. All of the samples exhibited antioxidant activities, among which ethyl acetate fraction of Zataria multiflora (17.21 ± 0.17 mg GAE/g) was found to contain the highest amounts of phenols and the ethyl acetate fraction of Zataria multiflora (218 ± 2.76 mg QUE/g) had the most values of flavonoids. Ethyl acetate fraction of Zataria multiflora (IC50 = 3.05 ± 0.51 µg/ml) was shown to have the most reducing power and the ethyl acetate fraction of Zataria multiflora (IC50 = 32.17 ± 1.82 µg/ml) exhibited the highest DPPH radical scavenging. The ethyl acetate fraction of Otostegia persica (99.39 ± 0.94%) showed the highest α-amylase inhibitory activity which was similar to acarbose used as a standard. Mode of α-amylase inhibition of the most samples was uncompetitive except for ZMC, OPP, OPC, and SMP which presented competitive inhibition. The present findings showed that studied samples may have some compounds with antioxidant and antidiabetic effects.

Efficacy of cryotherapy plus topical Juniperus excelsa M. Bieb cream versus cryotherapy plus placebo in the treatment of Old World cutaneous leishmaniasis: A triple-blind randomized controlled clinical trial.

BACKGROUND: Cutaneous leishmaniasis is one of the highly prevalent endemic diseases in the Middle East and North Africa. Many treatment modalities have been recommended for this condition but success rates remain limited. Herbal remedies have also been used for treatment but evidence-based clinical trials with these products are sparse. In-vitro and in-vivo studies have shown the anti-leishmanial and curative effects of extract of fruits and leaves of Juniperus excelsa (J. excelsa). The aim of this study was to determine the efficacy of topical J. excelsa M. Bieb extract as an adjuvant to cryotherapy for the treatment of human CL. MATERIALS AND METHODS: This study was designed as a two-arm triple-blind randomized placebo-controlled clinical trial using a parallel design. Seventy-two patients with clinical diagnosis of CL confirmed by leishmania smears were allocated to receive either a topical formulation of leaf of J. excelsa extract (group A) or placebo (group B) for 3 months. Both groups received cryotherapy as baseline standard treatment. Patients were evaluated before and weekly after the intervention was initiated until complete cure. RESULTS: Overall, 82% of patients in group A, experienced complete cure and 9% of them had partial cure. On the other hand, 34% in group B reported complete cure, while 14% of them had partial cure at the end of treatment protocol with a significant difference between the two groups (P< 0.001). The mean duration to healing of the lesions in patients who received J. excelsa extract was statistically significantly shorter than the placebo group (p = 0.04). No significant side effect was seen in the J. excelsa extract group except for mild to moderate local irritation after a few weeks in a few numbers of patients. CONCLUSION: The results of this study showed that topical J. excelsa extract can be used as an adjuvant treatment modality in addition to cryotherapy for accelerating the time to cure in addition to increasing the complete cure rate in CL. TRIAL REGISTRATION: ClinicalTrials.gov IRCT2015082523753N1.

Chemical Composition and Antimicrobial Activities of the Essential Oil From Salvia mirzayanii Leaves.

Resistance of many pathogens to available drugs is a global challenge and is leading to growing interest in natural alternative products. In this study, chemical composition and in vitro antibacterial and antifungal activities of the essential oil from Salvia mirzayanii were investigated. The chemical constituents of essential oil from S mirzayanii were analyzed by gas chromatography-mass spectrometry. The antimicrobial activity was determined by broth microdilution. The main identified compounds were 1,8-cineole (41.2 ± 1.3%), linalool acetate (11.0 ± 0.5%), and α-terpinyl acetate (6.0 ± 0.4%) (mL of essential oil/g of plant material). The MIC95 were 0.03 to 0.5 µL/mL and 16 to 128 µL/mL for gram-positive and gram-negative bacteria, respectively. These results indicated that Salvia mirzayanii essential oil significantly inhibited the growth of standard and clinically isolated tested yeasts by MIC50 0.03 to 1 µL/mL. Potent antibacterial and antifungal activities of Salvia mirzayanii essential oil may be considered in future study, particularly against antibiotic-resistant cases.

A Survey on Chemical Constituents and Indications of Aromatic Waters Soft Drinks (Hydrosols) Used in Persian Nutrition Culture and Folk Medicine for Neurological Disorders and Mental Health.

In Persian nutrition culture, drinking aromatic waters (hydrosols, distillate) has a long history as functional beverages or therapeutic remedies. The co-distilled water with essential oils, which contains partial amounts of more water-soluble volatile compounds are diluted and used as beverages. Since the solubility of volatile components is different in water, the overall composition, and thus the biological activities of aromatic waters seem to be different from the essential oils they were co-distilled with. Despite the essential oils, chemical constituents of many aromatic waters have not been evaluated scientifically. This research investigated hydrosols used for mental and neurological health maintenance in Persian nutrition culture and their chemical constituents. Constitutions of these hydrosols were extracted by liquid/liquid extraction method and identified by gas chromatography-mass spectrometry. Furthermore, cluster analysis was used to evaluate the relevance of these hydrosols chemical constituents. About 93 compounds were identified from 20 aromatic waters. the major or second major constituents were thymol (azarol howthorn, frankincense, lemon balm, valerian, shadab), phenethyl alcohol (damask rose, dog-rose, starflower), carvacrol (basil, creeping buttercup, lemon balm); eugenol (shadab, dog-rose, starflower, basil), camphor (yarrow and wormwood), carvone (oriental plane), caryophyllene (cuminum), cinnamaldehyde (Chinese cinnamon), p-cymen-7-ol (musk willow), limonene (lemon verbena), linalool and α-terpineol (bitter orange), menthol (date palm) and methyl 5-vinylnicotinate (olive). Although, these hydrosols prepared from plants belong to different genus and families, but cluster analysis showed obvious similarities between their chemical constituents. Results of this investigation showed in many cases that the constituents of aromatic waters are different from the pure essential oil.

Natural herbicide activity of Satureja hortensis L. essential oil nanoemulsion on the seed germination and morphophysiological features of two important weed species.

The aim of the present study was to obtain an oil/water (O/W) nanoemulsion (NE) containing garden savory (Satureja hortensis) essential oil (EO) and evaluating its herbicidal activity against Amaranthus retroflexus and Chenopodium album. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were employed to determine the chemical composition of the EO. Carvacrol (55.6%) and γ-terpinene (31.9%) were the major EO components. Low energy method was applied, allowing achievement of EO nanodroplets. The NE also presented low polydispersity, and the mean droplet was below 130nm even after storage for 30d. Laboratory tests showed that the NE at different concentrations (100, 200, 400, 800, and 1000µL.L-1) significantly (P≤0.05) reduced the germination indices and the seedling's growth in dose-response. The inhibitory effect was the greatest at 800µL.L-1 NE. Overall, root length was more inhibited as compared to shoot length. Post-emergence application of NE at different concentrations (1000, 2000, 3000, 4000 and 5000µL.L-1 of EO) on 2-4 true leaves' stage of the weeds caused significant (P≤0.05) decrease in the growth factors in dose-dependent manner. Complete lethality was observed by 4000µL.L-1 NE sprayed on the weeds. Spraying of NE significantly (P≤0.05) reduced chlorophyll content in the tested weeds. Increasing in relative electrolyte leakage (REL) 1 and 5d after treatment represented significant cell membrane disruption and increased cell membrane permeability. Transmission electron microscope (TEM) pictures confirmed NE droplet size and demonstrated membrane destruction. The study approved that the NE of S. hortensis EO has herbicidal properties as it has high phytotoxic effect, and interferes with the germination, growth and physiological processes of the weeds. The production of NE from S. hortensis EO is a low energy method that offers a promising practical natural herbicide for weed control in organic agricultural systems.

Antileishmanial Activities of Greek Juniper (Juniperus excelsa M.Bieb.) Against Leishmania major Promastigotes.

Petroleum ether, chloroform, ethyl acetate, and n-butanol fractions of Greek juniper (Juniperus excelsa M.Bieb. from the family Cupressaceae) were evaluated for antileishmanial activities against Leishmania major promastigotes compared to meglumine antimoniate (Glucantime). In vitro toxicity assay was performed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and microplate ELISA reader. Extracts were prepared in ethanol/dimethyl sulfoxide (80/20) at 10 to 0.62 mg/mL. The standard was prepared in phosphate-buffered saline at 500 to 15.62 mg/mL. Both leaf and fruit extracts and related fractions showed strong inhibitory effects against promastigotes, significantly different from that of the standard. The leaf extract and the respective petroleum ether fraction showed maximum effectiveness compared to other fractions and also fruit extract and fractions (IC90 = 1.89 ± 0.03 and 0.90 ± 0.03 mg/mL, respectively). Regarding the potent activities of nonpolar fractions of Greek juniper leaf extract, these fractions can be suggested for further investigation.

Trachyspermum ammi 10 % topical cream versus placebo on neuropathic pain, a randomized, double-blind, placebo-controlled trial.

A four-week, double-blind, randomized, placebo-controlled trial was conducted to assay the effectiveness of Ajwain 10 % (Trachyspermum ammi Sprague) topical cream on neuropathic pain. Intervention encompassed Ajwain 10 % and placebo creams. Ninety-two patients who specifically mentioned daily and nocturnal burning feet were randomly assigned to receive one of those interventions. Presence and decline in patients' numbness, tingling and allodynia were also evaluated. Major outcome measure was alteration in feet burning intensity (final week versus baseline week) regarding to a visual analog scale on a 0-10 cm scale (0 being "no pain", 10 being "worst pain"). Significant reduction in feet burning scores as well as numbness, tingling and allodynia were found in Ajwain group compared to placebo. This trial examining a cream of Ajwain essential oil versus placebo revealed the significance difference between two groups. This medicament can be a good candidate for the alleviation of feet burning, a neuropathic complication.

Development and Assessment of a Gas Chromatographic Based Method for the Quantification of Thymol from Cream Based Formulation.

INTRODUCTION: Herbal medicines have been used for different illnesses. However, standardization of these medicaments should be done before introducing for treatment purposes. Ajwain an essential oil, is traditionally used for neuropathic pain. OBJECTIVE: To develop and assess a gas chromatographic-based method for the quantification of thymol in Ajwain essential oil, current work was performed. METHODOLOGY: Both pure thymol and Ajwain creams were prepared and subjected to hydrodistillation method under temperature-controlled procedure to re-extract the applied essential oil and pure thymol. Previously, Ajwain seeds essential oil composition was analyzed and identified using GC/MS. After re-extraction, GC/FID was applied quantitatively to determine the thymol content in the Ajwain and thymol creams. The parameters represented in International Conference on Harmonization (ICH) guidelines were considered for the determination. RESULTS: Thymol content in a 50 g laminated tube of Ajwain cream was calculated as 2.34 g ± 0.02. Regarding the total thymol content of a 50 g laminated tube of Thymol cream (2.43 g), recovery percent for Ajwain cream was calculated as 96.29 %. CONCLUSION: Using hydrodistillation for an essential oil- containing cream sample via Clevenger proved to be a simple and convenient method to work up and extract active volatile components of such semisolid formulation. However, the extraction yield was profoundly related to the condenser temperature. The current employed determination method is introduced as a rapid and reliable method and thus, can be suggested for the quality control assessment of phytopharmaceutical semisolid preparations containing thymol and similar volatile constituents.