In vitro fungitoxicity of the essential oil of Syzygium aromaticum

The in vitro antifungal activity of clove oil was studied against four test fungi namely Alternaria alternata, Fusarium chlamydosporum, Helminthosporum oryzae and Rhizoctonia bataticola by the agar well diffusion method. These test fungi were found to be highly sensitive to clove oil at a concentration of 100 μl/well. The inhibition zone diameter was found to be in the range of 55–65 mm. The toxicity of clove oil on the germination and growth of A. alternata was further examined in liquid medium. Concentration- and time-dependent toxicity was recorded from 0.05 to 20% (v/v) concentration. The minimum fungistatic concentration was found to be 0.05%. Above this concentration, lysis of conidia and inhibition of mycelial growth were detected. Microscopic analysis showed 20–40% lysis of conidia after 72 h of incubation at 5% concentration. However at higher clove oil concentration (10%), up to 20% of conidia were lysed within 24 h of incubation. Similar concentration- and time-dependent toxicity was observed at different concentrations and time intervals. The findings indicated that clove oil possesses fungicidal activity against phytopathogenic fungi. Further study is required to determine whether it could have value in the management of plant infectious diseases. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cytotoxicity of clove (Syzygium aromaticum) oil and its major components to human skin cells

The essential oil extracted from clove (Syzygium aromaticum) is used as a topical application to relieve pain and promote healing in herbal medicine and also finds use in the fragrance and flavouring industries. Clove oil has two major components, eugenol and beta-caryophyllene, which constitute 78% and 13% of the oil, respectively. Clove oil and these components are generally recognized as 'safe', but the in-vitro study here demonstrates cytotoxic properties of both the oil and eugenol, towards human fibroblasts and endothelial cells. Clove oil was found to be highly cytotoxic at concentrations as low as 0.03% (v/v) with up to 73% of this effect attributable to eugenol. beta-caryophyllene did not exhibit any cytotoxic activity, indicating that other cytotoxic components may also exist within the parent oil.     

Antioxidant effects of clove bud (Syzygium aromaticum) extract used with different extenders on ram spermatozoa during cryopreservation

Clove bud (Syzygium aromaticum) extract was added at concentrations of 0, 35, 75, and 115 μg/ml to ovine semen extenders in order to investigate the antioxidant activities of clove bud extract and its effects on semen quality parameters after cryopreservation of ram spermatozoa. The basic extender was composed of Tris, egg yolk, and glycerol. Two other extenders were prepared by substitution of egg yolk with either LDL or egg yolk + SDS. The DPPH inhibition test was employed to assess the antioxidant activity of clove bud extract. Results showed that, compared to vitamin E, clove bud extract had a higher antioxidant activity. Better sperm motility and movement characteristics (P < 0.05) were observed in the semen diluted with medium containing egg yolk + SDS than in that containing egg yolk and LDL. Progressive motility and movement characteristics of the sperm were significantly improved (P < 0.05) by adding 35 and/or 75 μg/ml of clove bud extract to semen extenders. Sperm viability and plasma membrane integrity were also higher (P < 0.05) in the semen exposed to medium containing egg yolk + SDS and 75 μg of clove buds extract after cryopreservation processes. Higher levels of clove bud extract, however, had adverse effects on all the sperm quality parameters and significantly reduced (P < 0.05) the motility, movement parameters, viability, and plasma membrane integrity of ovine sperm. It was concluded that the clove bud extract had an antioxidant potential that makes it useful for addition to semen extenders and that the best results are obtained with a maximum clove bud extract of 75 μg/ml. Moreover, the combination of egg yolk and a detergent was found to improve sperm quality after the cooling and freeze–thawing processes.     

In vitro anticancer activity of extracted oil from Parrotiopsis jacquemontiana (Decne) Rehder

BackgroundParrotiopsis jacquemontiana, commonly referred to as “Beranj” in the local community, is widely used traditionally and has numerous health benefits. However, no studies have been conducted to investigate its anticancer potential, particularly its extracted oil.PurposeThe present study was put forth to appraise the anticancer potential of Parrotiopsis jacquemontiana extracted oil against liver (hcclm3 and hepg2) and breast cancer (mda-mb 231 and mcf-7) cell lines relative to normal cell lines (lo2 and mcf-10a) via MTT assay.MethodsFlow cytometry indicated the apoptotic effect whereas invasion and migration capabilities of oil against cancer cells were determined by Matrigel invasion chamber and wound-scratch assays.ResultsThe results of oil revealed a time and dose-dependent increase in cell proliferation inhibition, conferring to least IC50 shown against hcclm3 (144.9 ± 0.75 μg/ml) and mda-mb 231 (145.7 ± 0.32 μg/ml) cell line at 72 h, whereas no cytotoxic effect on normal cells was observed. In addition, the oil significantly (p < 0.001) suppressed the migration and invasion of hcclm3 and mda-mb 231 cells, showing noteworthy anti-metastatic potential. Furthermore, cell death was confirmed by Annexin‒V/PI staining where the maximum apoptotic percentage was calculated for oil (200 μg/ml) alongside mda-mb 231 conferring to 15.36 ± 1.22, 26.7 ± 1.2, and 36.43 ± 1.65 at 24, 48, and 72 h whereas 12.33 ± 1.05, 19.36 ± 1.62, and 29.3 ± 0.79 was recorded alongside hcclm3 at similar time intervals, respectively.ConclusionIn conclusion, the extracted oil exhibited strong anti-proliferative, anti-metastatic, and apoptotic effects and therefore may have potential applications in cancer treatment, however, further studies of oil regarding the action mechanisms and compounds involved in anticancer therapy are necessary.     

响应面法优化超临界CO2萃取茉莉花蕾精油及抗氧化活性的研究

为研究超临界CO2萃取茉莉花蕾获得精油的最佳工艺及抗氧化活性,实现资源的综合利用。实验考察了夹带剂用量、原料颗粒大小、萃取压力、萃取温度和动态萃取时间对茉莉精油产率的影响,进一步以响应面实验设计优化挥发油的提取工艺参数,以GC-MS分析挥发油组成及相对含量,并测定精油清除DPPH自由基及还原铁离子能力。结果表明,最佳工艺参数为:夹带剂乙醇用量为0.2mL/g,花粉颗粒为40目,萃取压力20MPa,萃取温度46℃,动态萃取时间1.6h。在此条件下茉莉精油的产率为13.67%,远高于同时蒸馏萃取法(2.87%)和超声辅助提取(2.45%)。超临界萃取精油的主要香气成分与茉莉鲜花基本一致,并且具有较好的清除自由基和还原铁离子能力。实验证明超临界萃取技术提取茉莉花蕾得到茉莉精油品质高,可为其资源综合利用提供参考。     

In vitro antibacterial effect of the essential oil of Thymus longiflorus Boiss

The essential oil of Thymus longiflorus Boiss was tested for its in vitro antibacterial activity. The results showed antibacterial effects against Gram-positive and Gram-negative bacteria, especially against Pseudomonas fluorescens and Mycobacterium phlei.     

In vitro antioxidant effects and cytotoxicity of polysaccharides extracted from Laminaria japonica

A water-soluble crude polysaccharide (WPS) was obtained from Laminaria japonica by hot water extraction. Three major polysaccharide fractions (WPS-1, WPS-2 and WPS-3) were purified from WPS by anion-exchange chromatography. Monosaccharide components analysis indicated that galactose was the predominant monosaccharide in WPS and WPS-3, accounting for 56.25% and 54.11%, respectively. And fucose was the predominant monosaccharide in WPS-1 and WPS-2, accounting for 46.91% and 45.1%, respectively. Antioxidant activity tests revealed that WPS-2 showed significant function of scavenging hydroxyl free radical and WPS-1 exhibited the highest inhibitory effects on superoxide radical. Cytotoxicity of all polysaccharide fractions was evaluated by MTT assay and Hoechst 33258 staining. Results showed that WPS-1 and WPS-2 significantly inhibited the growth of A375 cells and low anti-proliferative effects of WPS-2 on vascular smooth muscle cells (VSMCs) were observed. These results suggested that the polysaccharide fraction of WPS-2 might be explored as a potential safe antioxidant and antitumor agent.     

In vitro antioxidant activity of acetylated and benzoylated derivatives of polysaccharide extracted from Ulva pertusa (Chlorophyta)

The antioxidant activity of natural ulvan and its derivatives (acetylated and benzoylated ulvans) in vitro was determined, including scavenging activity against superoxide and hydroxyl radicals, reducing power, and chelating ability. Obvious differences in antioxidant activity between natural ulvan and its derivatives were observed, moreover, the antioxidant activity of acetylated and benzoylated ulvans was stronger than that of natural ulvan.     

Anti-Giardia activity of Syzygium aromaticum essential oil and eugenol: effects on growth, viability, adherence and ultrastructure

The present work evaluates the anti-Giardia activity of Syzygium aromaticum and its major compound eugenol. The effects were evaluated on parasite growth, adherence, viability and ultrastructure. S. aromaticum essential oil (IC₅₀ = 134 μg/ml) and eugenol (IC₅₀ = 101 μg/ml) inhibited the growth of G. lamblia. The essential oil inhibited trophozoites adherence since the first hour of incubation and was able to kill almost 50% of the parasites population in a time dependent manner. The eugenol inhibited G. lamblia trophozoites adherence since the third hour and not induce cell lyses. The main morphological alterations were modifications on the cell shape, presence of precipitates in the cytoplasm, autophagic vesicles, internalization of flagella and ventral disc, membrane blebs, and intracellular and nuclear clearing. Taken together, our findings lead us to propose that eugenol was responsible for the anti-giardial activity of the S. aromaticum essential oil and both have potential for use as therapeutic agents against giardiasis.     

In vitro antioxidant activities of sulfated polysaccharide fractions extracted from Corallina officinalis

Sulfated polysaccharides (F1, F2) from seaweed Corallina officinalis were isolated through anion-exchange column chromatography. Their chemical characteristics were determined by GC, HPLC, FT-IR and UV spectra. F1 and F2 contained only two monosaccharides, namely galactose and xylose. The antioxidant activities of F1, F2 and the de-sulfated polysaccharides (DF-1, DF-2) in vitro were investigated, including hydroxyl radicals scavenging effect, superoxide radical scavenging capacity, DPPH radical activity and reducing power. As expected, antioxidant assay showed that the two sulfated polysaccharide fractions (F1, F2) possessed considerable antioxidant properties and had more excellent abilities than de-sulfated polysaccharides (DF-1, DF-2).     

薄荷油超临界CO2萃取条件的优化和筛选

以样品中的薄荷脑含量为指标,通过单因素和正交实验对影响薄荷(Mentha haplocalyx Briq.)油超临界CO2萃取的因素进行研究,筛选出薄荷油超临界CO2萃取的最佳条件.研究结果表明,影响样品中薄荷脑萃取率的因素从大到小依次为萃取压力、萃取温度、萃取时间、CO2流量.样品中薄荷脑含量最高的超临界CO2萃取条件为萃取压力10 MPa、萃取温度50℃、CO2流量30 L·h-1且萃取时间1.5 h.     

In vitro evaluation of antibacterial,anticollagenase, and antioxidant activities of hop components (Humulus lupulus) addressing acne vulgaris

Seven naturally derived components from hop plant (Humulus lupulus L.) extracts were tested for evaluation of biological activities affecting acne vulgaris. Five strains, Propionibacterium acnes, Staphylococcus epidermidis, Staphylococcus aureus, Kocuria rhizophila and, Staphylococcus pyogenes, were selected as the main acne-causing bacteria. Hop extracts xanthohumol and the lupulones showed strong inhibitory activities against all of the strains. Although hydrogenated derivatives did not show the same level of activity, naturally occurring xanthohumol, humulones, and lupulones all showed moderate to strong anticollagenase inhibitory activities. Antioxidant capacity was also evaluated with seven different methods based on different reactive oxygen species. Xanthohumol showed the highest activity in total oxygen radical absorbance capacity as well as singlet oxygen absorbance capacity.     

In vitro evaluation of glutathione peroxidase (GPx)-like activity and antioxidant properties of some Ebselen analogues

Four analogues of Ebselen were synthesized and their glutathione peroxidase activity and antioxidant property evaluated and compared to Ebselen. Among the studied compounds, only diselenide [3] exhibited both glutathione peroxidase activity and radical-scavenging capability. Compounds [3] and [4] showed a strong inhibitory effect (53% and 43%, respectively) on the lipid peroxidation of linoleic acid compared to Ebselen and selenide derivatives ([1] and [2]) which were less active (28%, 26% and 18% inhibition, respectively). A concentration-dependent inhibitory effect was also found in the model of the formation of ABTS*+ radical cation: 65% and 89% inhibition for compound [3] at 10(-4) M and 5 x 10(-5) M, respectively, and 68% and 90% for compound [4], compared to 14% and 52% inhibition for Ebselen and the diselenides [1] and [2] (29%, 46% and 45%, 68%, respectively). By EPR spin trapping technique, the following inhibitory profile of the Ebselen analogues was observed towards the formation of thiyl radicals: Ebselen = [3]>[1]>[2]>[4]. Studies with compound [3] are in progress on oxidative stress cell models.     

In vitro evaluation of antioxidant and anti-inflammatory properties of genistein-modified hemodialysis membranes

Genistein-modified poly(amide):poly(vinyl pyrrolidone) (PA:PVP/G) hemodialysis membranes have been fabricated by coagulation via solvent (dimethyl sulfoxide, DMSO)/nonsolvent (water) exchange. The antioxidant and anti-inflammatory properties of the unmodified PA:PVP membranes were evaluated in vitro using human blood. It was found that these unmodified PA:PVP membranes were noncytotoxic to peripheral blood mononuclear cells (PBMC) but raised intracellular reactive oxygen species (ROS) levels. Pure genistein (in DMSO solution) was not only nontoxic to PBMC, but also suppressed the ROS levels in a manner dependent on genistein dosage. A similar dose-dependent suppression of ROS was found in genistein-modified PA (i.e., PA/G) membranes. However, the PVP addition had little or no effect in the suppression of ROS levels for the ternary PA:PVP/G system; the membrane ROS suppression was largely controlled by the genistein dosage. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin (IL-6) in whole blood were measured by ex vivo stimulation with lipopolysaccharide (LPS). The unmodified PA:PVP membranes drastically increased the level of TNF-α; however, the concentration of IL-1β and IL-6 remained almost the same. The PA/G membranes reduced the concentration of IL-1β and TNF-α even at very low genistein loadings, but it required a higher genistein loading to realize a similar effect in the case of IL-6. Of particular importance is that the genistein-modified blend membranes (PA:PVP/G) showed greater suppression of the concentrations of all three cytokines (TNF-α, IL-1β, and IL-6) in comparison with those of the PA/G membranes, signifying the role of PVP in the enhanced anti-inflammatory properties of these genistein-modified membranes. Ultraviolet-visible (UV-vis) spectroscopy was employed to quantify any genistein leaching during the in vitro testing.     

In vitro evaluation of Datura innoxia (thorn-apple) for potential antibacterial activity

Various parts of Datura innoxia were examined for potential antibacterial activity by preparing their crude aqueous and organic extracts against Gram-negative bacteria (Escherichia coli and Salmonella typhi) and Gram-positive bacteria (Bacillus cereus, Bacillus subtilis and Staphylococcus aureus). The results of agar well diffusion assay indicated that the pattern of inhibition depends largely upon the plant part, solvent used for extraction and the organism tested. Extracts prepared from leaves were shown to have better efficacy than stem and root extracts. Organic extracts provided potent antibacterial activity as compared to aqueous extracts. Among all the extracts, methanolic extract was found most active against almost all the bacterial species tested. Gram-positive bacteria were found most sensitive as compared to Gram-negative bacteria. Staphylococcus aureus was signifi cantly inhibited by almost all the extracts even at very low MIC followed by other Gram-positives. For Escherichia coli (a Gram-negative bacterium), the end point was not reached for ethyl acetate extract while it was very high for other extracts. The study promises an interesting future for designing a potentially active antibacterial agent from Datura innoxia.     

In vitro and ex vivo anti-diabetic and anti-hyperglycemic properties of Zataria multiflora essential oil

Molecular Biology Reports - The underlying mechanism involved in the onset of many diseases such as diabetes is oxidative stress. Zataria multiflora has a very high antioxidant power that can be...     

In vitro and in vivo evaluation of antioxidant and neuroprotective properties of antipsychotic D2AAK1

Neurochemical Research - The susceptibility of neurons to free radical toxicity partially underlies the pathomechanism of neurodegenerative diseases. On the other hand, excitotoxicity also...     

Acaricidal activity of essential oil of Syzygium aromaticum,hydrolate and eugenol formulated or free on larvae and engorged females of Rhipicephalus microplus

The cattle tick, Rhipicephalus microplus (Canestrini, 1888) (Ixodida: Ixodidae), is the most important ectoparasite in cattle‐breeding areas and is responsible for severe economic losses. Synthetic acaricides have been used to control this parasite. However, the need for safer products has stimulated the search for new acaricides, such as those to be obtained from medicinal plants. The essential oil of Syzygium aromaticum (clove) has many biological properties and shows great potential for use in veterinary applications. In the context of the need for new agents, this study investigated the in vitro properties of the hydrolate, essential oil and the main constituent of S. aromaticum, eugenol, in formulated and free applications against larvae and females of R. microplus. Eugenol and the essential oil caused 100% mortality in larvae at starting applications of 2.5 mg/mL and 5.0 mg/mL, respectively. The hydrolate showed no activity. Both eugenol and essential oil had good efficacy in adult immersion tests at 50 mg/mL and achieved 100% efficacy at a concentration of 100 mg/mL. The results of these tests reaffirm the important potential of clove essential oil and eugenol.     

In vitro pro- and antioxidant properties of estrogens

The pro- and antioxidant properties of estrogens are subject of debate. The apparent discrepancy is largely caused by the chemical heterogeneity in the estrogen family and by their concentration and the environment in which they are found. To gain some insight into this debate, we determined whether estradiol (E(2)), estrone (E(1)), the 2-, 4- and 16alpha-hydroxyestrogens and also the 2- and 4-methoxyestrogens are: (1) good electron-donors; (2) capable of O(2) consumption and DNA strand break induction; (3) capable of inhibiting lipid peroxidation in vitro. E(2), E(1) and 16alpha-hydroxyestrone (16alpha-OHE(1)) were not pro-oxidants and were rather weak antioxidants, while the 2- and 4-hydroxyestrogens demonstrated both properties inducing DNA strand breaks damage as well as inhibiting lipid peroxidation. The 4-hydroxyestrogens consumed O(2) and induced DNA strand breaks to a level approximately 2.5-fold higher than the 2-hydroxyestrogens, but these hydroxyestrogens exhibited similar antioxidant capacity, as measured by inhibition of lipid peroxidation. The 4-methoxyestrogens cannot induce oxidative damage to DNA but can inhibit lipid peroxidation, although being less potent than the 2-methoxyestrogens and the 2- and 4-hydroxyestrogens. The 2-methoxyestrogens were both potent electron donors and inhibitors of lipid peroxidation. Although 2-methoxyestrogens cannot generate superoxide in vitro, they may also be considered pro-oxidants in vivo.     

In vitro antileishmanial activity and cytotoxicity of essential oil from Lippia sidoides Cham

Leishmaniasis is a widespread tropical infection caused by different species of Leishmania protozoa. There is no vaccine available for Leishmania infections and conventional treatments are very toxic to the patients. Therefore, antileishmanial drugs are urgently needed. In this study we have analyzed the effects of essential oils from Lippia sidoides (LSEO) and its major compound thymol on the growth, viability and ultrastructure of Leishmania amazonensis. The essential oil and thymol showed significant activity against promastigote forms of L. amazonensis, with IC₅₀/48 h of 44.38 and 19.47 μg/mL respectively. However, thymol showed toxicity against peritoneal macrophages and low selectivity against the promastigotes when compared with the crude LSEO. On the other hand, no cytotoxic effect was observed in macrophages treated with the crude essential oil. Incubation of L. amazonensis-infected macrophages with LSEO showed a marked reduction in amastigote survival within the macrophages. Significant morphological alterations as accumulation of large lipid droplets in the cytoplasm, disrupted membrane and wrinkled cells were usually seen in treated parasites. The LSEO's activity against both promastigote and the amstigote forms of L. amazonensis, together with its low toxicity to mammalian cells, point to LSEO as a promising agent for the treatment of cutaneous leishmaniasis.