In Vitro and In Vivo Survey of Ethyl Acetate Extract of Acorus calamus (Sweet Flag) Rhizome on Toxoplasma gondii.

BACKGROUND: Toxoplasmosis is a zoonosis disease that can cause a variety range of manifestations in human specially fetus duration and immunodeficiency conditions. Due to toxicity and side effects of current treatment, we evaluated in vivo and in vitro effects of ethyl acetate extract of Acorus calamus rhizomes (rootstocks) on Toxoplasma gondii. METHODS: The plant, Acorus calamus, was collected from Sari, North of Iran in spring season. Ethyl acetate extract was provided from plant rhizomes using Soxhlet apparatus. The total phenolic and flavonoid contents of the extract were measured by the Folin-Ciocalteu method. The mortality effect of different concentrations (1-256 µg/ml) of the extract on Toxoplasma tachyzoites was assessed by flowcytometry and propidium iodide staining. For the therapeutic effect assessment, the tachyzoites were inoculated intraperitoneally to mice, and then these mice were orally and intraperitoneally administered different concentrations (32, 64, 128, and 256 mg/kg) of the extract. Also, an infected group received PBS including DMSO 1% as negative control, and an infected group administered sulfadiazine as positive control. For toxicity evaluation of this extract, a group only received dose 256 mg/kg. RESULTS: The plant extract was rich of phenolic compounds (41.27 ± 0.21 mg/g), whereas it contained fewer amounts of flavonoids (4.79 ± 0.01 mg/g). Results of in vitro experiments showed that there is an inverse relationship between the concentrations and the mortality of the parasites (IC50 = 200.01 ± 7.74 µg/ml). The highest percentage (62%) of dead tachyzoites was seen at maximum concentration of the extract. A significant longevity (8.9 days) was belonged to mice orally administered extract dose (256 mg/kg/day). CONCLUSION: The ethyl acetate extract of A. calamus rhizomes had significant anti-Toxoplasma activities either in vitro or in vivo. It may be connected to high amount of phenolic compounds. We suggest that the effects of different fractions and the admin types of the extract will be evaluated on the parasite.

Trachyspermum ammi aromatic water: A traditional drink with considerable anti-Candida activity.

BACKGROUND AND PURPOSE: Aromatic waters (AWs) are therapeutic distillates, which harbor both essential oil and water-soluble components of a plant. Due to the dispersion of the light amount of essence through the AWs, they have their specific pleasant smell, taste, and medicinal properties. In Iranian traditional medicine, Trachyspermum ammi AW is used to treat gastrointestinal disorders. The present study was conducted to determine the chemical composition of the essential oil extracted from T. ammi AW and its antifungal activities against Candida species. MATERIALS AND METHODS: The composition of the essential oil extracted from T. ammi AW was analyzed by gas chromatography-mass spectrometry. In addition, the evaluation of the antifungal activity of AW against Candida species was performed using broth microdilution methods as recommended by the Clinical Laboratory Standard Institute. Moreover, the biofilm formation inhibition, antioxidant properties, and experimental activity of AW were determined in an animal model. RESULTS: According to the results, thymol (78.08%) was the major compound of EO, followed by carvacrol (8.20%) and carvotanacetone (6.50%). Furthermore, T. ammi AW exhibited antifungal activities against the examined fungi and inhibited the biofilm formation of C. albicans at a concentration of up to 0.25 V/V. Histopathological analyses revealed that Candida colonization declined in the mice following the administration ofT. ammi AW in a therapeutic trial. CONCLUSION: It seems that the presence of phenolic monoterpenes in AW has resulted in antifungal effects. Pleasant odor and antioxidant properties are extra bonuses to the antimicrobial effects of this plant. Based on the findings, AW might have the potential to be used in the management of alimentary candidiasis or oral hygienic products.

Kinetics of α-glucosidase inhibition by different fractions of three species of Labiatae extracts: a new diabetes treatment model.

CONTEXT: Glucosidases are a group of enzymes playing crucial roles in digestion of carbohydrates. Glucosidase inhibitors can reduce carbohydrate digestion rate and have the potential to prevent development of type 2 diabetes. The Labiatae is one of the largest plant families grown globally and many studies that have isolated new pharmaceutical compounds. In folk medicine, some of Labiatae plants such as Zataria multiflora Boiss, Salvia mirzayanii Rech. F. & Esfand, and Otostegia persica Boiss are consumed for the treatment of diabetes. OBJECTIVES: This study investigates the inhibitory effects of different fractions of three mentioned species extracts on α-glucosidase. MATERIALS AND METHODS: Ethanol extracts of these plants leaves were fractionated using petroleum ether, chloroform, ethyl acetate, and n-butanol solutions. The duration of this study was 12 months. To measure enzyme inhibition, 5 µL of the enzyme, 20 µL of substrate and samples were used and for evaluation mode of inhibition, constant amounts of α-glucosidase were incubated with rising concentrations of substrate (PNPG). RESULTS: The results revealed that the ethyl acetate fraction of Zataria multiflora (IC50 = 0.35 ± 0.01 mg/mL) and petroleum ether fraction of Salvia mirzayanii (IC50 = 0.4 ± 0.11 mg/mL) were the most potent inhibitors of α-glucosidase in comparison with the other samples and acarbose as the standard (IC50 = 7 ± 0.19 mg/mL). All of the samples exhibited noncompetitive-uncompetitive inhibition. DISCUSSION AND CONCLUSION: It can be inferred from this study that α-glucosidase inhibitory potential of the studied extracts may be a marker of antidiabetic potential of these extracts.

Chemical Composition and the Effect of Walnut Hydrosol on Glycemic Control of Patients With Type 1 Diabetes.

BACKGROUND: Walnut hydrosol (WH) is used extensively by Iranian people with diabetes in order to control blood sugar (BS). There are few data regarding the effect of walnut on controlling diabetes. OBJECTIVES: A pilot study to determine the efficacy and safety of WH in patients with type 1 diabetes. MATERIALS AND METHODS: Eight patients with diabetes mellitus (DM) type 1 were enrolled in the study. They did not use any medicine except insulin. They were advised to drink 250 mL WH after meals twice a day for four weeks. Their BS level was measured and their insulin dose was changed according to their BS. After four weeks, they discontinued WH use and their BS level was checked for two weeks. Descriptive statistics was used to analyze the data. Also, the essential oil of the sample was extracted using a liquid extractor and then analysis of the constituents was performed. RESULTS: The average daily BS level and insulin dose decreased in seven subjects. Two subjects developed generalized pruritic erythematous skin rash. One patient presented hypoglycemic coma. She had no other coma risk factor. Seven compounds were identified in the walnut essential oil and the rate of monoterpenoid and sesquiterpenes hydrocarbons were 53.45% and 5.95%, respectively. The main constituents of the oil were carvacrol (33.21%), thymol (16%) and homoveratrole (15.83%). CONCLUSIONS: WH may control the glycemic level in people with diabetes, but it may be associated with minor and major side effects. Further in vitro studies, using these seven compounds, are recommended to determine the efficacy and complications of WH in people with diabetes.

An in vivo and in vitro investigation on hepatoprotective effects of Pimpinella anisum seed essential oil and extracts against carbon tetrachloride-induced toxicity.

OBJECTIVES: Protective effects of different extracts and essential oil from Pimpinella anisum L. seeds were examined against carbon tetrachloride (CCl4)-induced toxicity. The parameters such as serum transaminases, lactate dehydrogenase activity, hepatic glutathione content, liver lipid peroxidation and histopathological changes of liver were assessed as toxicity markers. In the in vitro model of this study, markers such as cell viability, cellular reduced and oxidized glutathione and lipid peroxidation in HepG2 cells were evaluated. MATERIALS AND METHODS: Human liver cancer cell line HepG2 and male Sprague-Dawley rats were treated with extracts and essential oil, and markers of hepatotoxicity were investigated. RESULTS: The data revealed that the n-hexane extract, effectively attenuated CCl4-induced toxicity in both in vitro and in vivo models in current investigation. CONCLUSION: As the oxidative stress markers were ameliorated, it might be concluded that anise seed possesses protective effects probably due to its antioxidant constituents.

Antimicrobial and cytotoxic activity of Ferula gummosa plant essential oil compared to NaOCl and CHX: a preliminary in vitro study.

OBJECTIVES: The usage of medicinal plants as natural antimicrobial agents has grown in many fields including dental medicine. The aim of this in vitro study was three-fold: (i) to determine the chemical compositions of the Ferula gummosa essential oil (FGEO), (ii) to compare the antimicrobial efficacy of the oil with sodium hypochlorite (NaOCl) and chlorhexidine (CHX), (iii) to assess the toxic behavior of FGEO in different concentrations compared to 5% NaOCl and 0.2% CHX. MATERIALS AND METHODS: Gas chromatography/mass spectrometry (GC/MS) was used to determine the chemical compositions of the oil. The disk diffusion method and a broth micro-dilution susceptibility assay were exploited to assess the antimicrobial efficacy against Enterococcus faecalis, Staphylococcus aureus, Streptococcus mitis, and Candida albicans. The cytocompatibility of the FGEO was assessed on L929 fibroblasts, and compared to that of NaOCl and CHX. RESULTS: Twenty-seven constituents were recognized in FGEO. The major component of the oil was ß-pinene (51.83%). All three irrigants significantly inhibited the growth of all examined microorganisms compared to the negative control group. FGEO at 50 µg/mL was effective in lower concentration against Enterococcus faecalis than 5% NaOCl and 0.2% CHX, and was also more potent than 0.2% CHX against Candida albicans and Staphylococcus aureus. FGEO was a cytocompatible solution, and had significantly lower toxicity compared to 5% NaOCl and 0.2% CHX. CONCLUSIONS: FGEO showed a promising biological potency as a root canal disinfectant. More investigations are required on the effectiveness of this oil on intracanal bacterial biofilms.

Evaluation of alpha- amylase inhibition by Urtica dioica and Juglans regia extracts.

OBJECTIVES: One strategy for the treatment of diabetes is inhibition of pancreatic α- amylase. Plants contains different chemical constituents with potential for inhibition of α-amylase and hence maybe used as therapeutic. MATERIALS AND METHODS: Urtica dioica and Juglans regia Linn were tested for α-amylase inhibition. Different concentrations of leaf aqueous extracts were incubated with enzyme substrate solution and the activity of enzyme was measured. For determination of the type of inhibition, Dixon plot was depicted. Acarbose was used as the standard inhibitor. RESULTS: Both plant extracts showed time and concentration dependent inhibition of α-amylase. 60% inhibition was seen with 2 mg/ml of U. dioica and 0.4 mg/ml of J. regia aqueous extract. Dixon plots revealed the type of α-amylase inhibition by these two extracts as competitive inhibition. CONCLUSION: Determination of the type of α-amylase inhibition by these plant extracts could provide by successful use of plant chemicals as drug targets.

Types of headache and those remedies in traditional persian medicine.

The history of headache, as a common neurological complication, goes back to almost 9000 years ago. Many ancient civilizations present references to headaches and the coherent treatment strategies. Accordingly, several documents comprising headache complications embodying precise medical information stem from Traditional Persian Medicine (TPM) that can provide useful opportunities for more comprehensive treatment. We conducted a survey on headache through original important pharmacopeias and other important medical manuscripts of TPM which were written during 9(th) to 19(th) centuries and have derived all headache categories and herbal remedies. An extensive search of scientific data banks, such as Medline and Scopus, has also been exercised to find results relating to the anti-inflammatory, anti-nociceptive, and analgesic effects of denoted medicinal herbs. The concept of headache and treatments in TPM covers over 20 various types of headache and more than 160 different medicinal plants administered for oral, topical, and nasal application according to 1000 years of the subject documents. Nearly, 60% of remarked medicinal herbs have related anti-inflammatory or analgesic effects and some current headache types have similarities and conformities to those of traditional types. Beside historical approaches, there are many possible and available strategies that can lead to development of new and effective headache treatment from medicinal plants so that this study can provide beneficial information on clinical remedies based on centuries of experience in the field of headache which can stand as a new candidate for further investigations.

Silymarin and milk thistle extract may prevent the progression of diabetic nephropathy in streptozotocin-induced diabetic rats.

OBJECTIVES: To investigate the effect of silymarin and milk thistle extract on the progression of diabetic nephropathy (DN) in rats. METHODS: Diabetes was induced with a single intraperitoneal (IP) injection of streptozotocin (STZ) (60 mg/kg). Silymarin (100 mg/kg/d) or the extract (1.2 g/kg/d) was gavaged for 4 weeks. Blood glucose (BS), serum urea (S(u)), serum creatinine (S(cr)), and 24-h urine protein (Up) were measured and glomerular filtration rate (GFR) was calculated. Concentration of thiobarbituric acid reactive species (TBARS) and activities of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) were evaluated in the renal tissue. RESULTS: Data were expressed as mean +/- SEM. Silymarin or the extract had no significant effect on BS, S(cr), and GFR. Both milk thistle extract and silymarin, respectively, decreased S(u) (mg/dL) (87.1 +/- 7.78, p < 0.001; 84.5 +/- 7.15, p < 0.001), Up (mg) (5.22 +/- 1.56, p = 0.014; 5.67 +/- 0.86, p = 0.034), and tissue TBARS (nmol/mg protein) (0.67 +/- 0.04, p < 0.001; 0.63 +/- 0.07, p < 0.001) in diabetic rats, compared to diabetic control (DC) (S(u): 131.0 +/- 4.55, Up: 8.3 +/- 0.84, TBARS: 0.94 +/- 0.06). Both the extract and silymarin could increase the activity of CAT (IU/mg protein) (25.5 +/- 4.0, p = 0.005; 20 +/- 1.8, p = 0.16) and GPx (IU/mg protein) (0.86 +/- 0.05, p = 0.005; 0.74 +/- 0.04, p = 0.10), respectively, in diabetic rats compared to DC (CAT = 14.4 +/- 2.0, GPx = 0.57 +/- 0.02). CONCLUSION: Milk thistle extract, to a lesser extent silymarin, can attenuate DN in rats possibly by increasing kidney CAT and GPx activity and decreasing lipid peroxidation in renal tissue.

Anticonvulsant Activity of Teucrium polium Against Seizure Induced by PTZ and MES in Mice.

Teucrium polium (Labiatae) is a plant that widely grows in Iran. Some of species of Teucrium are used for a considerable range of actions in traditional medicine and T. polium has frequently been used as anticonvulsant. In this study, we investigated the protective effects of T. polium ethanolic aqueous extracts and related fractions on seizures induced by pentylenetetrazole (PTZ) and maximal electroshock stimulation (MES). Moreover, presence of alkaloids, terpenoids, tannins and flavonoid contents were evaluated. It was found that aqueous extract (ED50 = 22.4 mg/kg body weight) and related n-butanol fraction (ED50 = 12.6 mg/kg body weight) have antiseizure effects comparing to control groups. There was no difference between preventing of PTZ-induced death and MES-induced hindlimb tonic extension (HLTE) in ethanolic extract comparing to control groups. Our results showed that the amount of flavonoid quantity present in aqueous extract is higher than that of ethanolic extract. These data also showed that the quantity of the flavonoid in n-butanol fraction of aqueous extract is more than other fractions. In conclusion, it was realized that flavonoid rich extracts are more potent than other fractions in showing antiseizure effects.

Radical scavenging and reducing power of Salvia mirzayanii subfractions.

In this research, the radical scavenging activity and reducing power of the ethyl acetate fraction and subfractions of Salvia mirzayanii (SM) have been investigated. The plant material was initially extracted with ethanol. The fractionation was carried out using liquid-liquid extraction, then the ethyl acetate fraction, which showed the greatest antioxidant activity, was selected. This fraction was submitted to column chromatography on a Sephadex LH 20 column eluted with pure MeOH to obtain subfractions A-G. No significant differences exist between the IC(50) of Salvia mirzayanii ethyl acetate subfraction C (IC(50) = 37.9 +/- 0.85), F (IC(50) = 40.05 +/- 1.4) and quercetin (38.84 +/- 0.86), (P > 0.05), indicating that the radical scavenging capacity of these two subfractions and quercetin (antioxidant standard) were similar. The reducing power of the ethyl acetate fraction was less than that of all subfractions, except for subfraction A. The greatest amount of phenolic compounds was found in subfraction E (55.23 +/- 4.2) and the lowest in subfraction F (5.23 +/- 0.18). The greatest total flavonoid content was established in subfraction D (1.84 +/- 0.01) and the lowest was in subfraction A (0.108 +/- 0.007).

Antileishmanial, antiplasmodial and cytotoxic activities of 12,16-dideoxy aegyptinone B from Zhumeria majdae Rech.f. & Wendelbo.

The ethanol extract of Zhumeria majdae showed potent antileishmanial and antiplasmodial activity in vitro. Bioactivity guided fractionation of the extract led to the isolation of 12,16-dideoxy aegyptinone B. This compound exhibited potent in vitro antileishmanial activity with an IC(50) of 0.75 microg/mL and a strong antiplasmodial activity with IC(50) values of 1.3 and 1.4 microg/mL against chloroquine sensitive and resistant strains, respectively. This compound was further found to have mild cytotoxicity towards cancer cell lines.