Pharmacological mechanisms of black cohosh in Sprague-Dawley rats

Background

Studies indicate that extracts and purified components from black cohosh inhibit the growth of human breast cancer cells, but the molecular targets and signaling pathways have not yet been defined.

Purpose

This study examines the pharmacological mechanisms and toxicological effects in the short term of the herb black cohosh on female Sprague–Dawley rats.

Materials and methods

To assess effects on gene activity and lipid content, we treated female Sprague–Dawley rats with an extract of black cohosh enriched in triterpene glycosides (27%) at 35.7 or 0 mg/kg. Four animals for each group were sacrificed at 1, 6 and 24 h after treatment; liver tissue and serum samples were obtained for gene expression and lipid analysis.

Results

Microarray analysis of rat liver tissue indicated that black cohosh markedly downregulated mitochondrial oxidative phosphorylation genes. Phospholipid biosynthesis and remodeling, PI3-Kinase and sphingosine signaling were upregulated, driven largely by an upregulation of several isoforms of phospholipase C. Hierarchical clustering indicated that black cohosh clustered with antiproliferative compounds, specifically tubulin binding vinca alkaloids and DNA alkylators. In support of this, black cohosh repressed the expression of cyclin D1 and ID3, and inhibited the proliferation of HepG2, p53 positive, liver cancer cells. Black cohosh reduced the level of free fatty acids at 6 and 24 h and triglycerides at 6 h in the serum, but increased the free fatty acid and triglyceride content of the treated livers at 24 h.

Conclusion

Our results suggest that black cohosh warrants further study for breast cancer prevention and therapy.     

Transdermal behaviors comparisons among Evodia rutaecarpa extracts with different purity of evodiamine and rutaecarpine and the effect of topical formulation in vivo

Aim

Evodiamine (EVO) and rutaecapine (RUT), the major active components from Evodia rutaecarpa extract (EE), are recognized as a depended analgesic agent. This study was designed to investigate the effect of purity and chemical enhancers on the transdermal behavior of EVO and RUT, and the pharmacological effect of their topical cream in vivo.

Material and methods

Transdermal delivery across a full thickness pig abdominal skin was detected in vitro by Franz-type diffusion cell, with HPLC for quantification of the permeation of EVO and RUT. The activity of topical cream in vivo was evaluated by a mice pain model induced by formalin and hot plate.

Results

Transdermal characters of EVO and RUT showed a low transdermal rate, long lag time and low cumulative amount. The transdermal rate and cumulative amount could be promoted by lipophilic enhancers, whereas lag time was shortened by hydrophilic surfactant, but these permeation parameters were not markedly influenced by purity of EE (p > 0.05). The effect in vivo was confirmed by analgesic models in topical cream of EE, which produced a significant (p < 0.05) inhibitory effects on pain response in dose-dependent manner.

Conclusion

The purity of EVO and RUT from EE has no significant effect on their permeation through porcine skin, but oleic acid or nerolidol can markedly elevate the transdermal rate of EVO and RUT. High purity of EE is the best choice for topical preparation to increase the drug loading. The effect of EE in vivo is verified by formalin model and hot plate test.     

Cellular mechanisms underlying Hyperin-induced relaxation of rat basilar artery

Background and aim

Hyperin, a flavonol compound extracted from the Chinese herb Abelmoschus manihot L. Medic, is reported to exert protective actions in cerebral ischemic injury. The specific aim of the present study was to study the relaxation of Hyperin in rat isolated basilar artery and identify the underlying cellular mechanisms.

Methods

Rat isolated basilar artery segments were cannulated and perfused while being superfused with PSS solution. Vessel images were recorded by video microscopy and diameters measured. Membrane potential was recorded using glass microelectrodes to evaluate the basilar artery smooth muscle cell hyperpolarization.

Results

Perfusion of Hyperin (1 ~ 100 μM) elicited a concentration-dependent relaxation of basilar artery segments preconstricted with 0.1 μM U46619. The response was significantly inhibited by the removal of the endothelium. Hyperin also elicited marked and concentration-dependent hyperpolarization of smooth muscle cells. 30 μM nitro-L-arginine (an inhibitor of nitric oxide synthase) and indomethacin (an inhibitor of cyclooxygenase), partially inhibited Hyperin-induced relaxation and hyperpolarization leaving an attenuated, but significant, endothelium-dependent relaxation and hyperpolarization. This remaining effect was almost completely blocked by 1 mM tetraethylammonium (an inhibitor of Ca²⁺-activated K⁺ channels), or by 100 μM DL-propargylglycine, an inhibitor of cystathionine-γ-lyase (a synthase of the endogenous H₂S).

Conclusion

These findings show that Hyperin produces significant hyperpolarization in rat basilar artery smooth muscle cells and relaxation through both endothelium-dependent and endothelium-independent mechanisms. The underlying mechanisms appeared to be multi-factorial involving nitric oxide, prostacyclin, and endothelium-derived hyperpolarizing factor (EDHF). Our data further suggest that endogenous H₂S is a component of the EDHF-mediated hyperpolarization and relaxation to Hyperin.     

Antimicrobial activity of bioactive compounds and leaf extracts in Jatropha tanjorensis

Objectives

Jatropha tanjorensis was investigated scientifically to generate evidence for the efficacies reported in traditional systems and the results are given here.

Methods

Different concentrations of the solvent extracts of leaves and four isolated compounds were tested against human pathogenic microorganisms such as gram-positive bacteria of Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermis, gram-negative bacteria of Aeromonas hydrophila, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris, Salmonella paratyphi, Salmonella paratyphi A, Vibrio alcaligenes, Vibrio cholerae and fungi of Aspergillus fumigatus, Candida albicans, Microsporum gypseum and Trichophyton rubrum by agar-well diffusion and disk diffusion methods.

Results

In agar-well diffusion method, maximum activity was recorded in a concentration-dependent manner. The extracts recorded activity against bacteria such as 17–26, 15–25 and 13–24 mm to methanol extract and 15–24, 14–23 and 12–22 mm to chloroform extract at 50, 25 and 12.5 mg/ml respectively and fungi such as 9–15 mm to A. fumigatus and 5–16 mm to T. rubrum. Maximum activity was 30–46, 27–43 and 17–40 mm to friedelin and 23–46, 28–44 and 18–41 mm to R (+) 4-hydroxy-2-pyrrolidinone against bacteria and 12–37, 8–34 and 31–33 mm to friedelin and 12–40, 11–35 and 10–33 mm to R (+) 4-hydroxy-2-pyrrolidinone against fungi at 10, 5 and 2.5 mg respectively.

Conclusions

The present study concludes that friedelin, β-amyrin, stigmasterol and R (+) 4-hydroxy-2-pyrrolidinone present in the methanol extract could be responsible for the broad spectrum of antimicrobial activity and provide scientific evidence.     

Cytotoxic, cytoprotective and antioxidant effects of isolated phenolic compounds from fresh ginger

Twenty-nine phenolic compounds were isolated from the root bark of fresh (Yunnan) ginger and their structures fully characterized. Selected compounds were divided into structural categories and twelve compounds subjected to in-vitro assays including DPPH radical scavenging, xanthine-oxidase inhibition, monoamine oxidase inhibition, rat-brain homogenate lipid peroxidation, and rat pheochromocytoma PC12 cell and primary liver cell viability to determine their antioxidant and cytoprotective properties. Isolated compounds were also tested against nine human tumor cell lines to characterize anticancer potency. Several diarylheptanoids and epoxidic diarylheptanoids were effective DPPH radical scavengers and moderately effective at inhibiting xanthine oxidase. An enone–dione analog of 6-shogaol (compound 2) was isolated and identified to be most effective at protecting PC12 cells from H₂O₂-induced damage. Almost all tested compounds inhibited lipid peroxidation. Three compounds, 6-shogaol, 10-gingerol and an enone-diarylheptanoid analog of curcumin (compound 6) were identified to be cytotoxic in cell lines tested, with KB and HL60 cells most susceptible to 6-shogaol and the curcumin analog with IC₅₀ < 10 μM. QSAR analysis revealed cytotoxicity was related to compound lipophilicity and chemical reactivity. In conclusion, we observed distinct compounds in fresh ginger to have biological activities relevant in diseases associated with reactive oxygen species.     

A review of phytotherapy of acne vulgaris: Perspective of new pharmacological treatments

Aim

This review focuses on plants currently used and those with a high potency for the future development of anti-acne products.

Methods

All relevant literature databases were searched up to 25 March 2011. The search terms were plant, herb, herbal therapy, phytotherapy, and acne, acne vulgaris and anti-acne. All of the human, animal, and in vitro studies, and reviews were included. Anti-bacterial, anti-inflammatory, anti-oxidant, and anti-androgen effects were the key outcomes.

Results

Studies on cell lines revealed that flavonoid, alkaloid, essential oil, phenol and phenolic compound, tannin, xanthone and xanthone derivative, and the bisnaphthquione derivative are effective in treatment of acne. Animal studies showed that diterpene acid, phenylpropanoid glycosides, acteoside and flavonoids have anti-inflammatory activity. Eleven human studies revealed that Camellia sinensis has 5α-reductase inhibitory and anti-inflammatory activities. Also anti-bacterial effect was shown by oleoresin of Commiphora mukul.

Conclusion

In addition to the standardization of these herbs, screening herbs as anti-acne agents may help to find new sources of therapy for acne.     

Two new amide alkaloids with anti-leukaemia activities from Aconitum taipeicum

Two new amide alkaloids, named 3-isopropyl-tetrahydropyrrolo [1,2-a] pyrimidine-2,4(1H,3H)-dione (1) and 1-acetyl-2,3,6-triisopropyl-tetrahydropyrimidin-4(1H)-one (2), were isolated from the roots of Aconitum taipeicum. Their chemical structures were characterized on the basis of MS, 1D- and 2D NMR. The anti-leukaemia activities of the compounds were also tested. The results indicated that the compounds exhibited more significant cell growth inhibitory activities against HL-60 cells than adriamycin, with the IC₅₀ of 1.1 ± 0.03 μg/mL and 1.6 ± 0.07 μg/mL respectively. In addition, two compounds showed anti-tumor activities against K562 cells as well.     

Kujigamberol, a new dinorlabdane diterpenoid isolated from 85million years old Kuji amber using a biotechnological assay

A new compound, 15,20-dinor-5,7,9-labdatriene-18-ol (1), named kujigamberol, was isolated from amber, fossilized tree resin from the Kuji area in Japan, has been dated as being 85 million years old (late Cretaceous). Kujigamberol was identified using the hypersensitive mutant yeast (zds1? erg3? pdr1? pdr3?) with respect to Ca²⁺-signal transduction. The structure was elucidated on the basis of spectroscopic analysis including 1D NMR, 2D NMR and HR-EI-MS. It was different from known diterpenoids with a similar activity isolated from Baltic amber (agathic acid 15-monomethyl ester (2), dehydroabietic acid (3) and pimaric acid (4)). Kujigamberol showed glycogen synthase kinase-3β (GSK-3β) inhibition activity involving the growth restored activity against the mutant yeast and was cytotoxic to HL60 cells (IC₅₀ = 19.6 μM).     

Cytotoxic prenylated flavonoids from Morus alba

A phytochemical fractionation of the methanol extract of the Morus alba leaves led to the isolation of eleven flavonoids (1–11). The structure of the new 3′-geranyl-3-prenyl-2′,4′,5,7-tetrahydroxyflavone (1) was elucidated by means of spectroscopic methods. The cytotoxicity of the isolated compounds against human cervical carcinoma HeLa, human breast carcinoma MCF-7, and human hepatocarcinoma Hep3B cells was evaluated. Of note, morusin (9) was the most potent with an IC₅₀ value of 0.64 μM against HeLa cells.     

Antiproliferative steroidal glycosides from Digitalis ciliata

Two new compounds, a furostanol glycoside (1) and a pregnane glycoside (4), along with eight known compounds, belonging to the classes of spirostane (2,3), pregnane (5–7) and cardenolide (8–10) glycosides, were isolated from the seeds of Digitalis ciliata. Their structures were elucidated by 1D and 2D-NMR experiments as well as ESI-MS analysis. For the first time pregnane glycosides of the diginigenin series have been isolated from D. ciliata. The cytotoxic effects of compounds 1–10 on cell viability of several cancer cell lines, namely human breast cancer (MCF-7), human glioblastoma (T98G), human lung adenocarcinoma (A549), human colon carcinoma (HT-29), and human prostate cancer (PC-3) cell lines were evaluated. Compounds 1, 4, 7 and 8 showed antiproliferative effects against MCF-7, HT-29 and A549 cancer cells with IC₅₀ values ranging from 8.3 to 20 μM. The effects of compounds 1–10 on cell proliferation were evaluated on these three cancer cell lines by cell cycle analysis of DNA content using flow cytometry. Compounds 7, 8 and 10 induced significant changes in G₂/M cell cycle phase of all analyzed cells. The obtained results indicate that compounds 7, 8 and 10 are cytostatic compounds effective in reducing cell proliferation by inducing accumulation of the cells in the G₂/M phase of the cell cycle.     

The role of seasonality on the inhibitory effect of Brazilian green propolis on the oxidative metabolism of neutrophils

The reactive oxygen species (ROS) produced by neutrophils are involved in the pathogenesis of several diseases, for which the intake of antioxidants could benefit patients either as a prophylactic or therapeutic treatment. Propolis is among the known antioxidants, and its chemical composition may vary under the influence of seasonality, which may interfere in its biological properties. This work evaluates the role of seasonality on the production of some important compounds of propolis samples produced monthly from November 2001 through October 2002 as well as the effect of these samples on the oxidative metabolism of stimulated neutrophils, by using both luminol and lucigenin to produce chemiluminescence (CLlum and CLluc, respectively). The cytotoxicity of the most active extracts to neutrophils was also investigated. The inhibitory effect of the propolis samples varied significantly during the studied period for both assays (3.4 ± 1.1 to 16.0 ± 1.1 μg/mL for CLlum and 6.2 ± 2.0 to 30.0 ± 5.0 μg/mL for CLluc), which was also observed in the quantitative profile of the main analyzed compounds (aromadendrin-4′-methyl ether, artepillin C, and baccharin). This effect started to become more prominent during the fall and, among all the studied extracts, the one obtained in May displayed the highest inhibitory effect on CL production (3.4 ± 1.1 μg/mL for CLlum and 6.2 ± 2.0 μg/mL for CLluc). The HPLC qualitative profiles of the extracts of propolis samples were quite similar, but there was a huge variation in terms of quantitative profile. It seems that aromadendrin-4′-methyl ether and baccharin play an essential role in the antioxidant activity, while artepillin C is not very important for this effect. The extracts presenting the highest antioxidant activity were produced in May, June, and August, and they did not display cytotoxicity at 25 μg/mL; quercetin, used as control, was not toxic to neutrophils at 8.5 μg/mL.     

Trichalasins C and D from the plant endophytic fungus Trichoderma gamsii

Two new cytochalasans, trichalasins C (1) and D (2) together with known cytochalasans aspochalasins D (3), M (4) and P (5) were isolated from one endophytic fungus Trichoderma gamsii inhabiting in traditional medicinal plant Panax notoginseng (BurK.) F.H.Chen. The structures for the new compounds 1 and 2 were determined by NMR and HRESIMS, and their relative configurations were established by analysis of coupling constants and NOESY correlations. Compound 3 displaying inhibitory activity with EC₅₀ value 5.72 μM, whereas the EC₅₀ values for compounds 1, 2 and 4, 5 are more than 40 μM.     

The antifungal constituents from the seeds of Itoa orientalis

Three new phenolic constituents, itolide A (1), itolide B (2), itoside P (3), and 1D-3-deoxy-3-hydroxymethyl-myo-inositol (4), which is described herein for the first time as a natural product, were isolated along with four other known compounds (5 to 8) from the methanol extract of the seeds of Itoa orientalis Hemsl by the activity-guided fractionation. Their structures were determined by spectroscopic means. Compounds 1 to 8 exhibited antifungal activities against Sclerotium rolfsii with IC₅₀ values ranging from 60.12 to 240.00 μM and against Rhizoctonia solani with IC₅₀ values ranging from 45.34 to 233.14 μM, respectively, and compounds 1, 2, 5 exhibited cytotoxic activity against Tn5B1-4 insect cell line with EC₅₀ values of 203.68, 93.41 and 40.37 μM, respectively.     

A new abietane diterpene from Glyptostrobus pensilis

A new abietane diterpene, glypensin A (1) and four known compounds, 12-acetoxy-ent-labda-8(17), 13E-dien-15-oic acid (2), quercetin 3-O-α-L-arabinofuranoside (3), quercetin 3-O-β-D-galactopyranoside (4), β-sitosterol (5) were isolated from the branches and leaves of Glyptostrobus pensilis (Staut.) Koch. Their structures were determined by MS, 1D- and 2D-NMR means. Compound 1 showed cytotoxicity on human chronic myeloid leukemia cell line K562 (IC₅₀ = 21.2 μM).     

A new coumestan with immunosuppressive activities from Flemingia philippinensis

A new coumestan, 1,3,9-trihydroxy-8-methoxycoumestan, named flemicoumestan A 1 together with nine isoflavone-related compounds were isolated from the ethyl acetate extract of the roots of Flemingia philippinensis. Their structures were elucidated and confirmed by spectroscopic methods and literature data. Compounds 3 and 4 showed strong lymphocyte proliferation inhibitory activity with an IC₅₀ value of 1.04-2.76 μM, and the low cytotoxicity with the CC₅₀ value of 71.01 and 56.36 μM, respectively.     

Soil aggregation may be a relevant indicator of nutrient cation availability

Key message

Aggregation was studied in two acidic forest soils (NE France) to investigate the potential link between available Ca and Mg content and soil aggregate size distribution and properties. Clay content influenced the aggregation status while clay mineralogy influenced aggregate stability and dynamics. Aggregation status and reactivity of soil components contributed to the difference of exchangeable Ca and Mg content in topsoil between the two sites.

Context

Though nutrient fluxes are important to define forest soil chemical fertility, the quantification of nutrient reservoirs in the soils and their availability to tree uptake is essential. A thorough understanding of nutrient availability requires an investigation of nutrient location and distribution in the soil solid phase.

Aims

The general aim was to investigate the potential link between available Ca and Mg content and soil aggregate size distribution and their properties (chemical, physical, mineralogical).

Methods

Soil aggregates were separated according to three size classes (200–2000 μm; 50–200 μm; <?50 μm) in two forest soils of the Lorraine plateau (France), both classified as Luvisols ruptic. The physical, chemical, and mineralogical properties were measured for each aggregate class.

Results

We showed that the relative abundance of an intermediate aggregate class [200–50 μm] was relevant to explain the difference of exchangeable Ca and Mg between sites. These aggregates were the poorest in organic and reactive mineral components and were unstable, which may mitigate the retention of Ca and Mg by ion-exchange.

Conclusion

This study highlights the role of aggregation and reactivity of soil components as relevant determinants of cation availability to tree uptake in soils.

     

Patterns of within-stem variations in wood specific gravity and water content for five temperate tree species

? Key message

Intensive measurements of basic specific gravity and relative water content of lumens show that within-stem variations strongly depend on species and cannot be summarised through the typical patterns reported in the literature; breast height measurements are not always representative of the whole stem.

? Context

Knowledge of the distribution of wood properties within the tree is essential for understanding tree physiology as well as for biomass estimations and for assessing the quality of wood products.

? Aims

The radial and vertical variations of basic specific gravity (BSG) and relative water content of lumens (RWC _L ) were studied for five species: Quercus petraea/robur, Fagus sylvatica, Acer pseudoplatanus, Abies alba and Pseudotsuga menziesii. The observations were compared with typical patterns of variations reported in the literature.

? Methods

Wood discs were sampled regularly along tree stems and X-rayed in their fresh and oven-dry states.

? Results

At breast height, BSG was found to clearly increase radially (pith to bark) for two species and to decrease for one species. For F. sylvatica and A. alba, the radial variations of BSG were rather U-shaped, with in particular inner wood areas showing respectively lower and higher BSG than the corresponding mature wood. RWC _L increased generally from inner to outer area but wet sapwood was clearly distinguishable only for the coniferous species. Vertical variations of BSG and RWC _L were strongly dependant on the species with usually non-linear patterns.

? Conclusion

The observed variations of BSG were only partially in agreement with the reported typical radial patterns. Despite the vertical variations, the mean BSG of a cross-section at breast height appeared to be a good estimator of the mean BSG of the whole stem (although the difference was statistically significant for coniferous species), whereas breast height measurement of RWC _L was not representative of the whole stem.

     

Self-thinning in four pine species: an evaluation of potential climate impacts

Key message

Self-thinning lines are species- and climate-specific, and they should be used when assessing the capacity of different forest stands to increase biomass/carbon storage.

Context

The capacity of forests to store carbon can help to mitigate the effects of atmospheric CO₂ rise and climate change. The self-thinning relationship (average size measure ～ stand density) has been used to identify the potential capacity of biomass storage at a given density and to evaluate the effect of stand management on stored carbon. Here, a study that shows how the self-thinning line varies with species and climate is presented.

Aims

Our main objective is thus testing whether species identity and climate affect the self-thinning line and therefore the potential amount of carbon stored in living biomass.

Methods

The Ecological and Forest Inventory of Catalonia was used to calculate the self-thinning lines of four common coniferous species in Catalonia, NE Iberian Peninsula (Pinus halepensis, Pinus nigra, Pinus sylvestris and Pinus uncinata). Quadratic mean diameter at breast height was chosen as the average size measure. The self-thinning lines were used to predict the potential diameter at a given density and study the effect of environmental variability.

Results

Species-specific self-thinning lines were obtained. The self-thinning exponent was consistent with the predicted values of ?3/2 and ?4/3 for mass-based scaling for all species except P. sylvestris. Species identity and climatic variability within species affected self-thinning line parameters.

Conclusion

Self-thinning lines are species-specific and are affected by climatic conditions. These relationships can be used to refine predictions of the capacity of different forest stands to increase biomass/carbon storage.

     

Short-day photoperiods affect expression of genes related to dormancy and freezing tolerance in Norway spruce seedlings

Key Message

Gene expression analysis showed that prolonged short day (SD) treatment deepened dormancy and stimulated development of freezing tolerance of Picea abies seedlings. Prolonged SD treatment also caused later appearance of visible buds in autumn, reduced risks for reflushing, and promoted earlier spring bud break.

Context

Short day (SD) treatment of seedlings is a common practice in boreal forest tree nurseries to regulate shoot growth and prepare the seedlings for autumn planting or frozen storage.

Aims

The aim of this study was to examine responses of Norway spruce (Picea abies (L.) Karst.) to a range of SD treatments of different length and evaluate gene expression related to dormancy induction and development of freezing tolerance.

Methods

The seedlings were SD treated for 11 h a day during 7, 14, 21, or 28 days. Molecular tests were performed, and the expression profiles of dormancy and freezing tolerance-related genes were analyzed as well as determination of shoot growth, bud set, bud size, reflushing, dry matter content, and timing of spring bud break.

Results

The 7-day SD treatment was as effective as longer SD treatments in terminating apical shoot growth. However, short (7 days) SD treatment resulted in later activation of dormancy-related genes and of genes related to freezing tolerance compared to the longer treatments which had an impact on seedling phenology.

Conclusion

Gene expression analysis indicated an effective stimulus of dormancy-related genes when the SD treatment is prolonged for at least 1–2 weeks after shoot elongation has terminated and that seedlings thereafter are exposed to ambient outdoor climate conditions.

     

Anticandidal activity of curcumin and methyl cinnamaldehyde

Cinnamaldehyde, its derivatives and curcumin are reported to have strong antifungal activity. In this work we report and compare anticandidal activity of curcumin (CUR) and α-methyl cinnamaldehyde (MCD) against 38 strains of Candida (3; standard, fluconazole sensitive, 24; clinical, fluconazole sensitive, 11; clinical, fluconazole resistant). The minimum inhibitory concentrations (MIC₉₀) of CUR ranged from 250 to 650 μg/ml for sensitive strains and from 250 to 500 μg/ml for resistant strains. MIC₉₀ of MCD varied between 100 and 250 μg/ml and 100–200 μg/ml for sensitive and resistant strains, respectively. Higher activity of MCD as compared to CUR was further reinforced by spot assays and growth curve studies. At their respective MIC₉₀ values, in the presence of glucose, average inhibition of H⁺-efflux caused by CUR and MCD against standard, clinical and resistant isolates was 24%, 31%, 32% and 54%, 52%, 54%, respectively. Inhibition of H⁺-extrusion leads to intracellular acidification and cell death, average pHi for control, CUR and MCD exposed cells was 6.68, 6.39 and 6.20, respectively. Scanning electron micrographs of treated cells show more extensive damage in case of MCD. Haemolytic activity of CUR and MCD at their highest MIC was 11.45% and 13.00%, respectively as against 20% shown by fluconazole at typical MIC of 30 μg/ml. In conclusion, this study shows significant anticandidal activity of CUR and MCD against both azole-resistant and sensitive clinical isolates, MCD is found to be more effective.