Effect of ethanol extract from Chrysanthemum cinerariifolium leaves on Ki-67 proliferation and dysplasia severity in a rat model of oral squamous cell carcinoma.

Background: Oral squamous cell carcinoma (OSCC) is a malignant tumor that can rapidly infiltrate the oral epithelial tissue and cause high mortality worldwide because the available therapies are less effective. Chrysanthemum cinerariifolium leaf contains secondary metabolites as anti-inflammatory, antioxidant, anticancer, and antimutagenic. Aims: The study aimed to analyze the ethanolic extract of C. cinerariifolium leaf in reducing proliferation (Ki-67) and the degree of dysplasia in OSCC rats. Methods: This study used male Sprague Dawley induced by 7,12-dimethylbenz(a)anthracene (DMBA) 0.5% and divided into five treatment groups, namely positive control/C+ (sick), negative control/C- (healthy), and treatment group induced with DMBA and given extract C. cinerariifolium leaf with successive doses of T1, T2, and T3 (50, 100, and 200 mg/kg bw). The oral epithelium was stained with hematoxylin and eosin and immunohistochemically stained with a Ki-67 monoclonal antibody. The statistical analysis utilizes the one-way analysis of variance test. Results: The results showed that T1 at a dose of 200 mg/kg bw could significantly reduce Ki-67 expression and the degree of oral epithelial dysplasia (OED; p < 0.05) close to healthy controls. Conclusion: The conclusion shows that C. cinerariifolium leaf extract can be a therapy against OSCC by decreasing cell proliferation and the degree of OED.

The use of Dalmatian pyrethrum daisy and an excipient in the treatment of seborrheic dermatitis.

The active ingredients of the Pyretrin-D trichological cosmetic series, namely benzyl benzoate, Dalmatian pyrethrum daisy, Cistus incanus, tea tree oil and geranium oil, almond acid and arginine were tested in respect to the treatment of seborrheic dermatitis. The paper describes the application of Dalmatian pyrethrum daisy and the excipient. Methods and devices used to confirm the effectiveness of the tested formulations included the TrichoScope Polarizer Dino-Lite (MEDL4HM) and the scanning electron microscope (SEM).

Improvement of fertility parameters with Tribulus Terrestris and Anacyclus Pyrethrum treatment in male rats.

OBJECTIVE: Anacyclus Pyrethrum (AP) and Tribulus Terrestris (TT) have been reported as male infertility treatment in several studies; however, in Iranian traditional medicine these two plants are prescribed simultaneously. In this study, we aimed to determine the effects of AP and TT extracts both separately and simultaneously on the male Wistar rat fertility parameters. MATERIALS AND METHODS: 32 male Wistar rats were divided into 4 groups: Control, TT, AP, and AT treated groups. Treatment continued for 25 days and rats were weighed daily. Their testes were dissected for histological studies. Sperm analysis including sperm count, viability and motility were performed. Serum was obtained to evaluate testosterone, LH and FSH levels. Histological studies were conducted to study Leydig, and Sertoli cells, spermatogonia and spermatid cell numbers, and to measure seminiferous diameter and epithelium thickness. RESULTS: Sperm count increased in all the treatment groups. Sperm viability and motility in AT and AP groups were elevated. TT and AT groups showed signifi cantly increased testosterone level compared to control group (P=004, P=0.000, respectively) and TT, AP and AT treatment groups showed increased LH level (P=0.002, P=0.03 and P=0.000, respectively) compared to control, while only AT group showed increased FSH (p=0.006) compared to control. Histological studies showed signifi cant increase of spermatogonia, Leydig and Sertoli cell numbers and epithelial thickness in AT group compared to other groups. All the treatment groups had higher number of Leydig, spermatogonia and spermatid cells. CONCLUSION: TT and AP improved sexual parameters; however, their simultaneous administration had higher improving effects on studied parameters.

Improvement of fertility parameters with tribulus terrestris and anacyclus pyrethrum treatment in male rats

ABSTRACT Objective Anacyclus Pyrethrum (AP) and Tribulus Terrestris (TT) have been reported as male infertility treatment in several studies; however, in Iranian traditional medicine these two plants are prescribed simultaneously. In this study, we aimed to determine the effects of AP and TT extracts both separately and simultaneously on the male Wistar rat fertility parameters. Materials and Methods 32 male Wistar rats were divided into 4 groups: Control, TT, AP, and AT treated groups. Treatment continued for 25 days and rats were weighed daily. Their testes were dissected for histological studies. Sperm analysis including sperm count, viability and motility were performed. Serum was obtained to evaluate testosterone, LH and FSH levels. Histological studies were conducted to study Leydig, and Sertoli cells, spermatogonia and spermatid cell numbers, and to measure seminiferous diameter and epithelium thickness. Results Sperm count increased in all the treatment groups. Sperm viability and motility in AT and AP groups were elevated. TT and AT groups showed significantly increased testosterone level compared to control group (P=004, P=0.000, respectively) and TT, AP and AT treatment groups showed increased LH level (P=0.002, P=0.03 and P=0.000, respectively) compared to control, while only AT group showed increased FSH (p=0.006) compared to control. Histological studies showed significant increase of spermatogonia, Leydig and Sertoli cell numbers and epithelial thickness in AT group compared to other groups. All the treatment groups had higher number of Leydig, spermatogonia and spermatid cells. Conclusion TT and AP improved sexual parameters; however, their simultaneous administration had higher improving effects on studied parameters.

Pyrethric acid of natural pyrethrin insecticide: complete pathway elucidation and reconstitution in Nicotiana benthamiana.

In the natural pesticides known as pyrethrins, which are esters produced in flowers of Tanacetum cinerariifolium (Asteraceae), the monoterpenoid acyl moiety is pyrethric acid or chrysanthemic acid. We show here that pyrethric acid is produced from chrysanthemol in six steps catalyzed by four enzymes, the first five steps occurring in the trichomes covering the ovaries and the last one occurring inside the ovary tissues. Three steps involve the successive oxidation of carbon 10 (C10) to a carboxylic group by TcCHH, a cytochrome P450 oxidoreductase. Two other steps involve the successive oxidation of the hydroxylated carbon 1 to give a carboxylic group by TcADH2 and TcALDH1, the same enzymes that catalyze these reactions in the formation of chrysanthemic acid. The ultimate result of the actions of these three enzymes is the formation of 10-carboxychrysanthemic acid in the trichomes. Finally, the carboxyl group at C10 is methylated by TcCCMT, a member of the SABATH methyltransferase family, to give pyrethric acid. This reaction occurs mostly in the ovaries. Expression in N. benthamiana plants of all four genes encoding aforementioned enzymes, together with TcCDS, a gene that encodes an enzyme that catalyzes the formation of chrysanthemol, led to the production of pyrethric acid.

Synthesis of citrophilus mealybug sex pheromone using chrysanthemol extracted from Pyrethrum ( Tanacetum cinerariifolium).

A commercial pyrethrum extract was used as a source of chrysanthemol for the synthesis of the citrophilus mealybug ( Pseudococcus calceolariae) sex pheromone. The chrysanthemic acid esters (pyrethrins I) were isolated and subsequently reduced to obtain chrysanthemol, which was used for ester pheromone synthesis. Field tests showed that the pheromone synthesized using plant-derived chrysanthemol was as attractive to male P. calceolariae as the pheromone obtained using a commercial isomeric chrysanthemol mixture.

Pyrethrum-extract induced autophagy in insect cells: A new target?

Pyrethrum extract (PY) is a natural insecticide that is extensively used across the world, and its insecticidal activity is attributed to the presence of six active esters known as pyrethrins. PY targets the nervous systems of insects by delaying the closure of voltage-gated sodium ion channels in the nerve cells. However, limited information is available regarding the toxicity and detailed mechanisms of PY activity. This study is aimed at understanding the toxicity effect and the underlying mechanisms of PY in cellular level, which have not yet been investigated on the non-nervous system of insects. Results of the MTT assay showed that the viability of Sf9 cells was inhibited by PY in a time- and concentration-dependent manner, and observation under a microscope revealed accumulation of intracellular vacuoles. Monodansylcadaverine staining analysis and transmission electron microscope images revealed typical autophagic morphological changes in PY-treated Sf9 cells. Autophagy-related proteins such as LC3, p62, and beclin-1 were detected using by Western blotting. Protein expression levels of LC3-II and beclin-1 were upregulated while that of p62 was markedly downregulated in a dose-dependent manner upon the PY treatment in Sf9 cells. In conclusion, these results indicate that PY could induce autophagy in the non-nervous system of insects which may contribute to its insecticidal mechanism.

Anacyclus Pyrethrum Extract Exerts Anticancer Activities on the Human Colorectal Cancer Cell Line (HCT) by Targeting Apoptosis, Metastasis and Cell Cycle Arrest.

INTRODUCTION: Colorectal cancer is the second leading cause of cancer mortality in the USA. There are a number of medicinal plants triggering apoptosis response in cancer cells, thus have a therapeutic potential. On the other hand, due to traditional uses and availability of Anacyclus pyrethrum extract, we decided to evaluate the efficacy of this medicinal herb on human colorectal cancer cell line (HCT). MATERIALS AND METHODS: In the present study, the cytotoxic effects of Anacyclus pyrethrum extract were assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, and trypan blue viability dye. Then, flow cytometry assay was exploited to measure cell death and apoptosis stage. The scratch test was exploited to assess the effect of Anacyclus pyrethrum on the migration of cancer cells. The expression levels of Caspase 3, Bcl-2, MMP1, and Vimentin genes were quantified by real-time PCR. Finally, cell cycle was analyzed by flow cytometry. RESULTS: MTT assay showed that Anacyclus pyrethrum extract significantly inhibited the cell growth. According to the flow cytometry assay result, the herbal extract was able to induce apoptosis in colorectal cancer cells. Our findings also demonstrated that the plant extract substantially increases the caspase 3 mRNA expression, while decreases Bcl-2, MMP1, and Vimentin. Cell cycle arrest occurred in G1 stage, due to the results of flow cytometry. CONCLUSION: These results indicate that Anacyclus pyrethrum extract can successfully induce apoptosis in HCT cells. Therefore, it could be used as a novel therapeutic candidate for colorectal cancer treatment.

Antiplasmodial and antitrypanosomal activity of pyrethrins and pyrethroids.

In a screen of 1800 plant and fungal extracts for antiplasmodial, antitrypanosomal, and leishmanicidal activity, the n-hexane extract of Chrysanthemum cinerariifolium (Trevir.) Vis. flowers showed strong activity against Plasmodium falciparum. We isolated the five pyrethrins [i.e., pyrethrin II (1), jasmolin II (2), cinerin II (3), pyrethrin I (4), and jasmolin I (5)] from this extract. These were tested together with 15 synthetic pyrethroids for their activity against P. falciparum and Trypanosoma brucei rhodesiense and for cytotoxicity in rat myoblast L6 cells. The natural pyrethrins showed antiplasmodial activity with IC(50)s between 4 and 12 µM, and antitrypanosomal activity with IC(50)s from 7 to 31 µM. The pyrethroids exhibited weaker antiplasmodial and antitrypanosomal activity than the pyrethrins. Both pyrethrins and pyrethroids showed moderate cytotoxicity against L6 cells. Pyrethrin II (1) was the most selective antiplasmodial compound, with a selectivity index of 24.

Effect of a mixed herb-enriched diet on the innate immune response and disease resistance of Paralichthys olivaceus against Philasterides dicentrarchi infection.

We investigated the effect of a mixed herb-enriched diet obtained from pomegranate Punica granatum, Dalmatian chrysanthemum Chrysanthemum cinerariaefolium, and mastic-leaved prickly-ash Zanthoxylum schinifolium on innate immune mechanisms (e.g., phagocytosis activity, respiratory burst activity, alternative complement activity, lysozyme activity, and disease resistance) of olive flounder Paralichthys olivaceus against the scuticociliate Philasterides dicentrarchi. All experimental groups were challenged with P. dicentrarchi (1 x 10(5) ciliates/mL) through intraperitoneal administration of the pathogen (50 microL) on day 1. On day 7, the infected groups were fed 0, 5, 50, and 100 mg/kg of the enriched diets. The innate immune parameters, cumulative mortality, and relative percent survival (RPS) were assessed at weeks 1, 2, 3, and 4. Administration of 50 or 100 mg/kg of the herbal-enriched diet enhanced immunity throughout the experimental period. However, at the 5-mg/kg dosage, the enriched diet did not enhance the innate immune estimates at any time. At doses of 50 and 100 mg/kg, administration of the diet preceding the challenge with P. dicentrarchi decreased the percentage cumulative mortality in the experimental groups and thereby increased RPS values. This study reports that administration of 50 or 100 mg/kg mixed herbal-enriched diet could positively influence the innate immune response to P. dicentrarchi and enhance the health status of olive flounder with respect to this microbe.

Chemical and biological comparisons on supercritical extracts of Tanacetum cinerariifolium (Trevir) Sch. Bip. with three related species of chrysanthemums of Sardinia (Italy).

In this manuscript, the authors compare the chemical composition and the biological effects of extracts of some Sardinian plant species: Glebionis coronaria (L.) Spach [=Chrysanthemum coronarium L.], locally known as 'caragantzu', Glebionis segetum (L.) Fourr. [=Chrysanthemum segetum L.], known as 'caragantzu masedu', and Sardinian endemic species Plagius flosculosus (L.) Alavi and Heywood [=Chrysanthemum flosculosus L.], known as 'caragantzu burdu'. In addition, the authors compare the pyrethrins contained in these species with an extract of Tanacetum cinerariifolium (Trevir.) Sch. Bip. [=Chrysanthemum cinerariifolium (Trevir.) Vis.], a commercial species rich in pyrethrins. The volatile fractions from chrysanthemum flowers were obtained by supercritical fluid extraction (SFE) with CO(2) at 90 bar and 50 degrees C and by hydrodistillation. Pyrethrins were extracted, together with other high molecular mass compounds, by SFE at high pressure, 300 bar and 40 degrees C. The composition of the volatile oils is determined by GC-MS analysis and the amount of pyrethrins by HPLC analysis. Moreover, the antibacterial and antimycotic activities of volatile fractions were investigated in order to compare to their traditional uses.

[Chemical constituents from Pyrethrum tatsienense].

OBJECTIVE: To isolate and identify the chemical constituents from the whole plant of Pyrethrum tatsienense, which has been used as traditonal herb medicine in the treatment of some diseases. METHODS: The compounds were isolated by column chromatography and their structures were elucidated through spectroscopic analysis (NMR). RESULTS: Six compounds were isolated and identified as: linoeic acid, pinoresinol, (+)-pinoresinol-4'-O-beta-D- glucopyranoside, betulabuside A, dihydrosyringin. CONCLUSION: All these compounds were obtained from Pyrethrum tatsienense for the first time.

Triterpenoids from pyrethrum tatsienense.

Two novel and eight known triterpenoids were isolated from alcoholic extract of the whole plant of Pyrethrum tatsienense. The structures of new compounds were elucidated as olean-12-en-3beta,11alpha,16beta-triol-3-O-palmitate (1), ursa-12-en-3beta,11beta,16beta-triol-3-O-palmitate (2) by spectroscopic methods including intensive 2D NMR techniques (H1-H1 COSY, gHMQC, gHMBC) and FAB-MS, and the known compounds were identified on the basis of comparing their NMR data with those of corresponding compounds in the literature.

Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world.

Botanical insecticides have long been touted as attractive alternatives to synthetic chemical insecticides for pest management because botanicals reputedly pose little threat to the environment or to human health. The body of scientific literature documenting bioactivity of plant derivatives to arthropod pests continues to expand, yet only a handful of botanicals are currently used in agriculture in the industrialized world, and there are few prospects for commercial development of new botanical products. Pyrethrum and neem are well established commercially, pesticides based on plant essential oils have recently entered the marketplace, and the use of rotenone appears to be waning. A number of plant substances have been considered for use as insect antifeedants or repellents, but apart from some natural mosquito repellents, little commercial success has ensued for plant substances that modify arthropod behavior. Several factors appear to limit the success of botanicals, most notably regulatory barriers and the availability of competing products (newer synthetics, fermentation products, microbials) that are cost-effective and relatively safe compared with their predecessors. In the context of agricultural pest management, botanical insecticides are best suited for use in organic food production in industrialized countries but can play a much greater role in the production and postharvest protection of food in developing countries.

Supercritical carbon dioxide processing of pyrethrum oleoresin and pale.

Possible refining of crude hexane extract (CHE) from pyrethrum flowers and further refining of Pyrethrum Board of Kenya (PBK) pale product is investigated with both liquid and supercritical carbon dioxide. The experiments were carried out in a small pilot plant with a 200 mL extractor and three cyclonic separators in series. To understand the dynamics of pyrethrin extraction, CHE was extracted in a single step; pyrethrin concentration was found to be improved from 0.16 to 0.50 g/g. The effects of temperature and pressure on the quality of the extract were studied at 29 degrees C and 80 bar and at 40 degrees C and 100 bar. Liquid CO(2) processing (29 degrees C, 80 bar) yielded slightly better product quality. A comparison study of CHE and PBK pale processing with supercritical CO(2) (40 degrees C, 100 bar) showed that the final products were similar in terms of pyrethrin content. Extraction of both PBK pale and CHE in two steps with different operating conditions improved their purity.

Constituents of Compositae plants III. Anti-tumor promoting effects and cytotoxic activity against human cancer cell lines of triterpene diols and triols from edible chrysanthemum flowers.

Fifteen pentacyclic triterpene diols and triols, consisting of: six taraxastanes, faradiol (1), heliantriol B0 (2), heliantriol C (3), 22alpha-methoxyfaradiol (4), arnidiol (5), and faradiol alpha-epoxide (6); five oleananes, maniladiol (7), erythrodiol (8), longispinogenin (9), coflodiol (10), and heliantriol A(1) (11); two ursanes, brein (12) and uvaol (13); and two lupanes, calenduladiol (14) and heliantriol B2 (15), isolated from the non-saponifiable lipid fraction of the edible flower extract of chrysanthemum (Chrysanthemum morifolium) were evaluated for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate, in Raji cells as a primary screening test for anti-tumor-promoters. All of the compounds tested showed inhibitory effects against EBV-EA activation with potencies either comparable with or stronger than that of glycyrrhetic acid, a known natural anti-tumor-promoter. Evaluation of the cytotoxic activity of six compounds, 1-3 and 5-7, against human cancer cell lines revealed that compound 5 possesses a wide range of cytotoxicity, with GI50 values (concentration that yields 50% growth) of mostly less than 6 microM.

Method development and validation of a high-performance liquid chromatographic method for pyrethrum extract.

A robust analytical methodology is developed for the quantitative determination of all six insecticidal components of pyrethrum extract: jasmolin I, cinerin I, pyrethrin I, jasmolin II, cinerin II, and pyrethrin II. This method, based on the separation technique normal-phase high-performance liquid chromatography, offers selectivity, accuracy, precision, linearity, range, ruggedness, and robustness as well as efficiency and ease.

Purification of chrysancorin, a novel antifungal protein with mitogenic activity from garland chrysanthemum seeds.

A novel antifungal protein, designated chrysancorin, was isolated from seeds of Chrysanthemum coronarium var. spatiosum with a procedure involving ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue resin, ion exchange chromatography on SP-Sepharose and FPLC-gel filtration on Superdex 75. The N-terminus of chrysancorin displays sequence similarity to the genomic sequence of chromosome 1 from Arabidopsis thaliana BAC T19E23. Chrysancorin exhibits a molecular mass of 13.4 kDa in gel filtration and SDS-polyacrylamide gel electrophoresis. It stimulates the proliferation of mouse splenocytes and inhibits the activity of human immunodeficiency virus-1 reverse transcriptase. The protein possesses antifungal activity against Botrytis cinerea, Mycosphaerella arachidicola and Physalospora piricola, but not against Rhizoctonia solani, Fusarium oxysporum and Coprinus comatus. However, we could not detect antibacterial activity against a variety of bacteria.

Stereoselective total synthesis of chrysanthemol.

Chrysanthemol (1), a trans-eudesmane type sesquiterpene from Chrysanthemum indicum L., possesses certain anti-inflammatory activity. Its total synthesis was approached from two alternative routes and finally accomplished in ten steps from R-(+)-carvone via alpha-eudesmol (10) as the key intermediate. The overall yield is 2.4% and the spectral data of the synthetic target compound were identical with that of natural chrysanthemol (1). Seven intermediary compounds were tested for inhibitory effects on the carragenan-induced swelling of mouse paw but demonstrated no obvious activities.