Secretory avocado idioblast oil cells: evidence of their defensive role against a non-adapted insect herbivore

We tested the hypothesis that avocado idioblast oil cells play a defensive role against herbivorous insects. Toxicities of the intact avocado idioblast oil cells and the extracted idioblast oil were compared for three insect herbivores. Spodoptera exigua (Hübner) larvae are generalists that do not feed on avocados. By contrast, Sabulodes aegrotata (Guenée) and Pseudoplusia includens (Walker) larvae are generalist herbivores that readily feed on avocados. All bioassays were performed at a naturally occurring concentration of idioblast oil cells (2% w/w). Choice experiments showed that S. exigua larvae avoided diet treated with avocado idioblast oil cells and consume more control than treated diet. In contrast, idioblast oil cells had no significant antifeedant effects on the adapted S. aegrotata and P. includens larvae. Subsequent experiments designed to assess resistance mechanisms separated pre-ingestive (behavioral) and post-ingestive (physiological) effects of the avocado idioblast oil cells, and the extracted idioblast oil, on the two adapted herbivores. Post-ingestive adaptation was the mechanism that allows feeding. Because the impact of the avocado idioblast oil cells was greatest on the performance of non-adapted S. exigua, additional experiments determined that larvae fed diet containing the oil cells had higher mortality and reduced larval growth compared to controls. Developmental times were significantly prolonged for the survivors. Thus, increased mortality, reduced developmental rates, and antifeedant activity in the non-adapted insect indicate that defense against non-adapted herbivores may be an important function of idioblast cells in avocados.     

Effect of the essential oil fromAchillea fragrantissima onEscherichia coli cells

Escherichia coli cells treated with the essential oil from the plantAchillea fragrantissima released five polypeptides as well as K⁺ ions into the incubation medium. The oil also inhibited the respiration ofE. coli cells and reduced their ATP content. Electron micrographs showed that oil-treated cells were permeable to uranyl acetate. The effect of the essential oil on the cell membrane is discussed.     

Diversity of Essential Oil-Secretory Cells and Oil Composition in Flowers and Buds of Magnolia sirindhorniae and Its Biological Activities

Magnolia sirindhorniae Noot. & Chalermglin produces fragrant flowers. The volatile oil secretary cells, quantity and quality as well as antioxidant and antimicrobial activities of the oils extracted from buds and flowers, have been investigated. The distribution of essential oil secretory cell in bud and flower revealed that the density and size of the oil cells were significantly higher in flowers compared to buds. In different floral parts, carpel has a higher oil cell density followed by gynophore and tepal. The histochemical analysis revealed the essential oil is synthesized in oil secretory cells. The volatile oil yield was 0.25 % in the buds and 0.50 % in flowers. GC/FID and GC/MS analysis identified 33 compounds contributing 83.2–83.5 % of the total essential oil composition. Linalool is the main constituent contributing 58.9 % and 51.0 % in the buds and flowers oils, respectively. The essential oil extracted from the flowers showed higher antimicrobial efficacy against Klebsiella pneumoniae and Staphylococcus aureus. Similarly, the essential oil isolated from the flowers depicts higher free radical scavenging, and antioxidant activity compared to buds’ oil.     

杭白芷根中分泌道发生方式、分布及其挥发油积累过程研究

为探明杭白芷(Angelica dahurica var.formosana)根中分泌道发生方式、分布及其挥发油转运积累特征,该研究利用光镜及透射电子显微镜技术观察分泌道发生过程及挥发油转运特征,结合组织化学定位确定挥发油的主要积累部位。结果表明:杭白芷根中分泌道由中柱鞘细胞最先发生,次生结构中分泌道主要分布在韧皮部和皮层中;挥发油的合成不仅与分泌细胞中质体及细胞质有关,而且还与周围细胞关系密切;分泌细胞内高尔基体和内质网丰富,可能先通过形成小泡参与转运,再经由细胞壁向腔道内转移;相邻分泌细胞靠近角隅处的细胞壁分泌活动活跃,腔道内积累大量电子致密物质;成熟分泌道中分泌细胞及其腔道内积累大量油滴,因此挥发油主要积累场所为分泌细胞及其腔道。该研究明确了杭白芷根中分泌道的发生方式、分布及其挥发油积累部位,揭示了分泌道发育过程中挥发油的转运积累特征,为进一步阐明分泌组织生长发育与有效成分积累关系提供了理论依据。     

Development of a fermentation method using immobilized cells under unsterile conditions. 1. Protection of immobilized cells against anti-microbial substances

A method of protecting immobilized cells against inhibitory substances in the fermentation medium was investigated with the aim of developing a process for fermentation under unsterile conditions. It was found that yeast cells could be protected against the inhibitory effects of p-hydroxybenzoic acid esters by co-immobilizing the cells with vegetable oils. In such a system, the cells grow only in the water phase of the gel beads where most components of the fermentation medium are retained. On the other hand, the p-hydroxybenzoate that diffuses into the gel beads is retained mainly in the oil phase of the beads. Consequently, the p-hydroxybenzoate concentration in the water phase remains too low to inhibit the metabolic activities of the immobilized cells. The effectiveness of a vegetable oil in protecting the immobilized cells against an inhibitory substance depends on the partition coefficient of the substance between the oil and water, the concentration of the oil and the initial cell concentration.     

Effect of dispersing oil phase on the biodegradability of a solid alkane dissolved in non-biodegradable oil

Acinetobacter sp. CR was grown on a model oil, which consisted of an inert oil matrix of pristane with n-heneicosane dissolved in it as the sole carbon source, in a stirred-tank bioreactor. This bacterium takes up substrates from the oil phase by direct contact with the oil phase. A previously established mathematical model was applied to reveal the effect of agitation conditions on the growth and n-alkane degradation kinetics of the bacterium. Higher impeller speed resulted in both lower microbial growth and lower n-alkane degradation rate of the bacterium, although it increased the specific surface area of the oil, which was measured by a previously developed device. This result was due to the decreased number of cells adhering to the oil surface, i.e., intense agitation inhibited the adhesion of cells to the oil surface. The addition of a surfactant below a critical micelle concentration (CMC) inhibited the degradation of n-heneicosane dissolved in pristane, although the biodegradability of the substrate recovered gradually with the increase in the dose of surfactant over CMC. The results suggest that efforts to increase the specific surface area of the oil phase have the undesirable result of inhibiting oil degradation when the dominant microbial degraders take up substrates in oil by direct contact with the oil. Electronic Publication     

Functional and ultrastructural changes in Pseudomonas aeruginosa and Staphylococcus aureus cells induced by Cinnamomum verum essential oil

Aims: To study cellular damage induced by Cinnamomum verum essential oil in Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213. Methods and Results: The effect of cinnamon bark essential oil on these two strains was evaluated by plate counts, potassium leakage, flow cytometry and transmission electron microscopy (TEM). Exposure to this oil induced alterations in the bacterial membrane of Ps. aeruginosa, which led to the collapse of membrane potential, as demonstrated by bis‐oxonol staining, and loss of membrane‐selective permeability, as indicated by efflux of K⁺ and propidium iodide accumulation. Thus, respiratory activity was inhibited, leading to cell death. In Staph. aureus, cells treated with the oil entered a viable but noncultivable (VNC) state. The oil initially caused a considerable decrease in the metabolic activity and in the replication capacity of these bacterial cells. The loss of membrane integrity appeared later, as indicated by bis‐oxonol and Propidium iodide (PI) staining. Data provided by TEM showed various structural effects in response to cinnamon essential oil. In Ps. aeruginosa cells, coagulated cytoplasmic material was observed, and intracellular material was seen in the surrounding environment, while oil‐treated Staph. aureus showed fibres extending from the cell surface. Conclusions: Cinnamon essential oil damages the cellular membrane of Ps. aeruginosa, which leads to cell death. There is evidence of VNC Staph. aureus after exposure to the oil. Significance and Impact of the Study: Cinnamon essential oil shows effective antimicrobial activity and health benefits and is therefore considered a potential food additive. To use this oil as a natural food preservative, especially in combination with other preservation methods, a thorough understanding of the mechanism through which this oil exerts its antibacterial action is required.     

Chemical Composition and Cytotoxic Evaluation of the Essential Oil of Phyllogonium viride (Phyllogoniaceae,Bryophyta)

The present study aimed to determine the chemical composition and biological activity of the essential oil obtained from Phyllogonium viride Brid. (Phyllogoniaceae, Bryophyta), whose samples were collected in southern Brazil. For the first time, the cytotoxic activity of the essential oil of P. viride in breast and colorectal tumor cells (MCF-7 and HCT-116) was evaluated, as well as the cytotoxic potential of this oil in non-tumoral cells of human immortalized keratinocytes (HaCaT) via MTT assay. The compounds majorly found in P. viride essential oil were β-bazzanene (20.30 %), β-caryophyllene (17.06 %), β-chamigrene (14.02), and germacrene B (11.72 %). Treatment with P. viride essential oil in the different tested cell lines did not induce any toxicity in most of the tested concentrations. These data contribute to generating new scientific information about this understudied plant species. Furthermore, the chemical characterization of the compounds present in the essential oil of P. viride can lead to greater elucidation of its biotechnological potential.     

A study of the ultrastructure of the shoot apex and leaf cells in two liverworts,with special reference to the oil bodies

Summary In this investigation attention has been paid to the general ultrastructure of the shoot apical and leaf cells in the liverwortsBazzania trilobata andLophozia ventricosa but especially to the different developmental stages of their oil bodies. These species have been chosen because their oil bodies differ from each other in size and shape.The appearance of the different organelles, nucleus, chloroplasts, mitochondria, ER, and Golgi bodies, are in their main features the same as those of higher plants described in the literature. The dark cytoplasm seen in the leaf cells ofLophozia in the vicinity of the oil bodies but without any surrounding membrane when fixed in double fixative 2, seems to be specific to this species. On the other hand, granular dense bodies were visible in the cells of the shoot apex ofBazzania, which shrank in size as the development of the oil bodies proceeded and were lacking in the mature leaf cells.In both species investigated, the oil bodies have the same component parts: (1) an outer membrane enveloping the whole body, (2) inside this, a granular stroma layer of varying thickness enveloping (3) specific globules of varying size and number, each of which is surrounded by (4) a thin inner membrane (Fig. 28).The oil bodies develop in at least two ways and usually in one way for each species. InBazzania they seem to develop from vacuole-like formations in the shoot apex or in the leaf primordia into which substances have segregated. InLophozia they seem to originate by aggregation and fusion of lipid bodies.     

Feinstrukturelle Untersuchungen zur Entwicklung der Ölzellen bei dem LebermoosRiella

Summary Liverworts are characterized by the possession of typical cell elements, the oil bodies. In the submerse, thalloid liverwortRiella helicophylla (Sphaerocarpales) oil bodies are existing in idioblastic cells only, called oil body cells. Each oil body cell contains only one oil body. The oil body originates from small vacuoles. Their membranes are extremely high in contrast and asymmetric. The lipophilic substances are probably produced inside the oil bodies. At the end of the development of an oil body cell lipophilic and hydrophilic material will be separated from each other inside the oil body. The result is an oil body, consisting of one large spherical oil globule surrounded by a thin layer of hydrophilic matrix.

     

Electronic paramagnetic resonance investigation of the activity of Origanum vulgare L. essential oil on the Listeria monocytogenes membrane

Aims: To evaluate the effect of oregano essential oil on Listeria monocytogenes cytoplasmic membrane. Methods and Results: Nitroxide free‐radical Electron Paramagnetic Resonance was applied on L. monocytogenes after 30 min exposure to oregano essential oil concentrations ranging from 0 to 1·25%. The impact of essential oil on the number of viable cells was evaluated by plate count. Growth dynamics of survivors in BHI and TSB were evaluated by turbidometry. After exposure to essential oil concentrations up to 0·50%, the membrane fluidity was changed and its order increased. When L. monocytogenes was exposed to higher concentrations, membrane order parameters slightly returned to the values of untreated cells. However, when the cells were exposed to EO in the presence of sodium azide, which impairs energy metabolism, the membrane fluidity was progressively enhanced, even at the lowest EO concentration (0·25%). Microbiological analyses confirmed a progressive reduction of viable count, at increasing essential oil concentrations. Both in BHI and TSB, the Lag phase length increased in treated cells with respect to controls, suggesting a cell damage recovery. Conclusions: The combined approach including microbiological and EPR analyses provided relevant information on membrane modification and cell response to essential oils. Significance and Impact of the Study: EPR approach was demonstrated to be an effective and helpful tool to comprehend the modifications exerted by essential oil on the bacterial membrane.     

沙棘果油对过氧化氢诱导氧化损伤的保护作用

沙棘(Hippophae rhamnoides)是一种具有药用价值的植物,沙棘果油具有抗氧化、抗炎症及抗肿瘤等多种药理作用。为了探讨沙棘果油对H₂O₂造成氧化性损伤的细胞生长的影响及其抗氧化性,该研究选择H₂O₂对RAW264.7细胞氧化损伤模型,通过DPPH(1,1-二苯基-2-三硝基苯肼)自由基清除实验检测沙棘果油体外抗氧化能力,用[3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐]MTT法和流式细胞仪检测超氧化物阴离子荧光探针(DHE)信号,分别检测不同浓度沙棘果油对H₂O₂损伤细胞的存活率和超氧化物阴离子水平。结果表明:(1)在DPPH自由基清除实验中,当沙棘果油浓度小于4.9%时,沙棘果油的抗氧化能力大于维生素C。(2)通过MTT法发现,浓度为3.125%的沙棘果油对H₂O₂损伤细胞的存活率显著升高(P<0.01)。(3)从DHE检测发现,在同一检测时间点,随着沙棘果油浓度增加,DHE阳性细胞比例显著下降(P<0.01); 在不同检测时间点,随着沙棘果油浓度增加,DHE阳性细胞比例显著升高(P<0.01)。沙棘果油对过氧化氢诱导的RAW264.7细胞氧化损伤模型有一定修复作用,可能与细胞内超氧化物阴离子水平受到抑制有关,它具有抗氧化性损伤的潜能。     

Use of microorganism-immobilized polyurethane foams to absorb and degrade oil on water surface

Highly oil-absorbent polyurethane foam (PUF) materials were obtained by polymerizing polyether polyol mixture and carbodiimide-modified d-methyl diisocyanate in a weight ratio of 10:2. The foam materials were prepared to contain inorganic nutrients (slow-release fertilizer; SRF) and oil-degrading yeast cells, Yarrowia lipolytica 180, to be applied for removal of oil films on surface waters through absorption and biodegradation after oil spills. PUFs absorbed 7–9 times their own weight of Arabian light crude oil and the oil absorbency appeared to improve as the ratio of surface area to foam weight increased. PUFs showed excellent floatability which was maintained for more than 6 months in sea water, and less than 5% of the absorbed oil was released when the foams were left on water for more than 10 days. For immobilization of yeast cells into PUFs, various immobilization techniques were tested to compare their oil degrading ability and the maintenance thereof. All immobilized cells showed oil degrading abilities as good as those of free cells immediately after the preparation of PUFs, however, the activity of chitin-immobilized cells remained at a high level for the longest period of preservation. The high efficiency of oil absorption and oil degradation by PUF-immobilized yeast cells suggested that PUF-immobilized cells have a high potential as a bioremediation technique for the treatment of oil films on surface waters. Received: 27 September 1999 / Received revision: 6 March 2000 / Accepted: 17 March 2000     

Protective effects of Cinnamomum verum,Cinnamomum cassia and cinnamaldehyde against 6-OHDA-induced apoptosis in PC12 cells

Cinnamon (Cinnamomum verum and C. cassia) is a medicinal plant, widely-used as a culinary spice. It possesses various therapeutic effects and can slow down the progression of neurological disorders impressively. In this article, the effects of hydro-alcohol extract and essential oil of C. verum and C. cassia and its main bioactive component cinnamaldehyde, has been examined on 6-OHDA-exposed PC12 cells as an in vitro model of Parkinson's disease. The cytotoxicity and cell apoptosis has been induced by 6-OHDA in PC12 cells. The protective effect was determined by measuring cell viability, the amount of reactive oxygen species (ROS), and apoptosis. Cell viability and apoptosis were assessed using resazurin assay, flow cytometry of propidium iodide (PI) stained cells, and western blot analysis. 6-OHDA resulted in the death and apoptosis of cells while, pretreatment with the extract and essential oil of C. verum and C. cassia at 20 µg/ml and cinnamaldehyde at 5 and 10 µM for 24 h could significantly increase the viability (p?<?0.001), and decrease ROS content (p?<?0.05). Pretreatment with the extracts increased survivin and decreased cyt-c whereas, pretreatment with the essential oil decreased cyt-c, increased survivin, and reduced P-p44/42/p44/42 levels to a level near that of the related control. The extract and essential oil of C. verum and C. cassia can be effective against 6-OHDA cytotoxicity. It is suggested that, the synergistic effects of cinnamaldehyde and other components of extract and essential oil promote cinnamon’s medicinal properties.

     

Antiproliferative Effect and Cell Cycle Alterations Induced by Salvia officinalis Essential Oil and Its Three Main Components in Human Colon Cancer Cell Lines

Colon cancer is one of the most common human malignancies, and chemotherapy cannot yet prevent recurrence in all patients. Essential oils are phytocomplexes with antiproliferative properties. In this study, we elucidated the antiproliferative properties and the effect on cell cycle progression of Sicilian Salvia officinalis essential oil and its three main compounds, α‐thujone, 1,8‐cineole (eucalyptol) and camphor, on three human colon cancer cell lines. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography. Cell proliferation was evaluated by MTT assay, and the cell cycle distribution was determined by flow cytometry. Thirty‐four compounds were identified in the tested essential oil. Growth inhibition was observed after 72 h, with an impact on cell cycle progression and no effect on the viability of normal colonic epithelial cells. The study shows that S. officinalis essential oil and its three main components have an in vitro antiproliferative effect on colon cancer cells.     

Comparison of solid‐phase cytometry and the plate count method for the evaluation of the survival of bacteria in pharmaceutical oils

Aim: To compare the survival of four bacterial strains (Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Pseudomonas aeruginosa) in pharmaceutical oils, including jojoba oil/tea tree oil, carbol oil, jojoba oil and sesame oil. Methods and Results: Oils were spiked with the test bacteria in a concentration of 10⁴ CFU ml^?1. Bacteria were extracted from oils with phosphate‐buffered saline containing 0·5% Tween 20. Aliquots of the pooled water layers were analysed by solid‐phase cytometry and plate counting. Plate counts dropped to zero for all test strains exposed for 24 h to three of the four oils. In contrast, significant numbers of viable cells were still detected by SPC, except in the jojoba oil/tea tree oil mixture and partly in sesame oil. Conclusions: Exposure of bacteria for 24 h to the two oils containing an antimicrobial led to a loss of their culturability but not necessarily of their viability. The antibacterial activity of the jojoba oil/tea tree oil mixture supersedes that of carbol oil. Significance and Impact of the Study: These in vitro data suggest that the jojoba oil/tea tree oil mixture more than carbol oil inhibits bacterial proliferation when used for intermittent self‐catherization.     

The anterior-like cells in Dictyostelium are required for the elevation of the spores during culmination

 Shortly after initiation of Dictyostelium fruiting body formation, prespore cells begin to differentiate into non-motile spores. Although these cells lose their ability to move, they are, nevertheless, elevated to the tip of the stalk. Removal of the amoeboid anterior-like cells, located above the differentiating spores in the developing fruiting body, prevents further spore elevation although the stalk continues to elongate. Furthermore, replacement of the anterior-like cells with anterior-like cells from another fruiting body largely restores the ability to lift the spores to the top of the stalk. However, if amoeboid prestalk cells are used to replace the anterior-like cells, there is no restoration of spore elevation. Finally, when a droplet of mineral oil replaces differentiating spores, it is treated as are the spores: the mineral oil is elevated in the presence of anterior-like cells and becomes arrested on the stalk in the absence of anterior-like cells. Because a similar droplet of mineral oil is totally ignored by slug tissue, it appears that there is a dramatic transformation in the treatment of non-motile matter at this point in Dictyostelium development. Received: 26 January 1998 / Accepted: 27 May 1998     

Cultivation ofCandida lipolytica 4-1 on hydrocarbons

The length of the carbon chain of the hydrocarbon substrate affects markedly the fatty acid composition in the cell lipids of the yeastCandida lipolytica 4-1. During cell growth onn-hexadecane dissolved in deparaffinated gas oil, direct incorporation of palmitic acid into lipids takes place. During growth onn-dodecane, on the other hand, an immediate synthesis and incorporation into oleic acid is observed. The cells contain only little lauric acid (maximum 11%). During fermentation of then-alkanes dissolved in deparaffinated gas oil which contains a mixture of isoalkanes, alkylated aromatic and cyclic hydrocarbons, free fatty acids accumulate in the oil phase. The fatty acid composition in the oil differs markedly according to the growth stage of cells. At the beginning of the logarithmic phase of growth, the fatty acid composition in the oil phase reflects the acid composition in the cell lipids, toward the end of cell growth, the cooxidation products of the isoalkanes accumulate. The aqueous phase contains lower fatty acids and cooxidation products of isoalkanes and of alkylated aromatic and alicyclic hydrocarbons. Part III. Oxidation and Utilization of Individual Pure Hydrocarbons—seeFolia Microbiol. 14,334 (1969).     

Oil-in-water emulsions characterization by laser granulometry and impact on γ-decalactone production in <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis>

Oil-in-water emulsions composed of methyl ricinoleate (MR) or castor oil (CO) as the organic phase, stabilized by Tween 80, are in the basis of the biotechnological production of γ-decalactone. Yarrowia lipolytica was used due to its ability to grow on hydrophobic substrates and to carry out the biotransformation. The characterization of oil droplets size distribution by laser granulometry was performed under different oil concentrations. The impact of the presence of cells on droplets size was also analyzed as well as the relevance of washing inoculum cells. Furthermore, the granulometric characterization of the emulsions was related with γ-decalactone production and it was observed that, in the presence of non-washed cells, the smaller droplets disappeared, using both oils, which increased γ-decalactone concentration. This suggests that the access of cells to the substrate occurs by their adhesion around larger oil droplets.     

Antimicrobial and anti‐inflammatory activity of five Taxandria fragrans oils in vitro

The antimicrobial activity of five samples of Taxandria fragrans essential oil was evaluated against a range of Gram‐positive (n= 26) and Gram‐negative bacteria (n= 39) and yeasts (n= 10). The majority of organisms were inhibited and/or killed at concentrations ranging from 0.06–4.0% v/v. Geometric means of MIC were lowest for oil Z (0.77% v/v), followed by oils X (0.86%), C (1.12%), A (1.23%) and B (1.24%). Despite differences in susceptibility data between oils, oils A and X did not differ when tested at 2% v/v in a time kill assay against Staphylococcus aureus. Cytotoxicity assays using peripheral blood mononuclear cells demonstrated that T. fragrans oil was cytotoxic at 0.004% v/v but not at 0.002%. Exposure to one or more of the oils at concentrations of ≤0.002% v/v resulted in a dose responsive reduction in the production of proinflammatory cytokines IL‐6 and TNF‐α, regulatory cytokine IL‐10, Th1 cytokine IFN‐γ and Th2 cytokines IL‐5 and IL‐13 by PHA stimulated mononuclear cells. Oil B inhibited the production of all cytokines except IL‐10, oil X inhibited TNF‐α, IL‐6 and IL‐10, oil A inhibited TNF‐α and IL‐6, oil C inhibited IL‐5 and IL‐6 and oil Z inhibited IL‐13 only. IL‐6 production was significantly inhibited by the most oils (A, B, C and X), followed by TNF‐α (oils A, B and X). In conclusion, T. fragrans oil showed both antimicrobial and anti‐inflammatory activity in vitro, however, the clinical relevance of this remains to be determined.