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Antiviral activities of extracts and selected pure constituents of Ocimum basilicum.

1. Ocimum basilicum (OB), also known as sweet basil, is a well known medicinal herb in traditional Chinese medicine preparations. In the present study, extracts and purified components of OB were used to identify possible antiviral activities against DNA viruses (herpes viruses (HSV), adenoviruses (ADV) and hepatitis B virus) and RNA viruses (coxsackievirus B1 (CVB1) and enterovirus 71 (EV71)). 2. The results show that crude aqueous and ethanolic extracts of OB and selected purified components, namely apigenin, linalool and ursolic acid, exhibit a broad spectrum of antiviral activity. Of these compounds, ursolic acid showed the strongest activity against HSV-1 (EC50 = 6.6 mg/L; selectivity index (SI) = 15.2), ADV-8 (EC50 = 4.2 mg/L; SI = 23.8), CVB1 (EC50 = 0.4 mg/L; SI = 251.3) and EV71 (EC50 = 0.5 mg/L; SI = 201), whereas apigenin showed the highest activity against HSV-2 (EC50 = 9.7 mg/L; SI = 6.2), ADV-3 (EC50 = 11.1 mg/L; SI = 5.4), hepatitis B surface antigen (EC50 = 7.1 mg/L; SI = 2.3) and hepatitis B e antigen (EC50 = 12.8 mg/L; SI = 1.3) and linalool showed strongest activity against AVD-II (EC50 = 16.9 mg/L; SI = 10.5). 3. No activity was noted for carvone, cineole, beta-caryophyllene, farnesol, fenchone, geraniol, beta-myrcene and alpha-thujone. 4. The action of ursolic acid against CVB1 and EV71 was found to occur during the infection process and the replication phase. 5. With SI values greater than 200, the potential use of ursolic acid for treating infection with CVB1 and EV71 merits further investigation.

Acyclic Monoterpenes↗

Flavor volatiles and physical properties of vacuum-microwave- and air-dried sweet basil (Ocimum basilicum L.).

Basil (Ocimum basilicum L.) was dried using conventional hot air or the recently developed vacuum-microwave dryers. The effect of the drying method on the relative abundance of major flavor volatiles, rehydration rate, color, and structural integrity of the plant was evaluated. Dynamic headspace analysis of volatiles present in fresh or dried basil revealed that linalool and methylchavicol (estragole) were the two major headspace volatile compounds of the plant sample. Vacuum-microwave dehydrated basil yielded approximately 2.5 times the linalool and 1.5 times the methylchavicol of the air-dried samples. Furthermore, the vacuum-microwave-treated samples yielded more volatiles than fresh basil, due to chemical reactions during drying. Air-dried samples of basil had darker and fewer green hues than those prepared by vacuum microwave. Vacuum-microwave-dried samples had a higher rehydration rate, whereas the potential of the plant material to rehydrate was hindered in air-dried samples. This is likely attributed to the dramatic and pronounced structural collapse of the air-dried cells as revealed by the scanning electron microscope.

Food Handling↗

Biosorption of cesium-137 and strontium-90 by mucilaginous seeds of Ocimum basilicum.

Mucilaginous seeds of Ocimum basilicum were used in uptake studies with cesium-137 and strontium-90. Results showed that uptake was dependent on the structural integrity of the mucilage fibrils. Water imbibed seeds showed higher adsorption of both 137Cs and 90Sr in comparison to seeds pretreated with NaOH, HCl and Na-periodate solution. The uptake was pH dependent and while some divalent metal ions had no or little detrimental effect, the alkali metal ions Li+, Na+ and K+ decreased the uptake. The maximum adsorption capacity was 160 mg cesium g(-1) and 247 mg strontium g(-1) seed dry weight.

Cesium Radioisotopes↗

Light reflected from colored mulches affects aroma and phenol content of sweet basil (Ocimum basilicum L.) leaves.

Basil (Ocimum basilicum L.) is an herb the leaves of which are used to add a distinct aroma and flavor to food. It was hypothesized that the size and chemical composition of sun-grown basil leaves could be influenced by the color of light reflected from the soil surface and by the action of the reflected light through the natural growth regulatory system within the growing plants. Leaf morphology, aroma compounds, and soluble phenolics were compared in basil that had been grown over six colors of polyethylene row covers. Altering the ratios of blue, red, and far-red light reflected to growing plants influenced both leaf morphology and chemistry. Leaves developing over red surfaces had greater area, moisture percentage (succulence), and fresh weight than those developing over black surfaces. Basil grown over yellow and green surfaces produced significantly higher concentrations of aroma compounds than did basil grown over white and blue covers. Leaves grown over yellow and green mulches also contained significantly higher concentrations of phenolics than those grown over the other colors. Clearly, the wavelengths (color) of light reflected to growing basil plants affected leaf size, aroma, and concentrations of soluble phenolics, some of which are antioxidants.

Color↗

Hypolipidaemic activity of aqueous Ocimum basilicum extract in acute hyperlipidaemia induced by triton WR-1339 in rats and its antioxidant property.

Hyperlipidaemia, atherosclerosis and related diseases are becoming a major health problem in developing countries. Ocimum basilicum is one of the medicinal plants widely used in Morocco to reduce plasma cholesterol and to reduce the risk of atherosclerosis-related diseases. However, mechanisms underlying the reported hypolipidaemic effect of this plant have not been investigated. This study evaluates the lipid lowering effect of aqueous Ocimum basilicum extract in Triton WR-1339-induced hyperlipidaemic rats. Hyperlipidaemia was developed in animals by intraperitoneal injection of Triton (200 mg/kg). After injection of Triton the animals were divided into three treatment groups: hyperlipidaemic, hyperlipidaemic plus herb extract and hyperlipidaemic plus fenofibrate treated rats. At 7 h after the Triton injection, levels of plasma cholesterol, triglycerides and LDL-cholesterol in rats treated also with the Ocimum basilicum extract (0.5 g/100 g body weight) were, respectively, 50%, 83% and 79% lower than Triton-treated rats and HDL-cholesterol was 129% higher than in rats given Triton alone. At 24 h following Ocimum basilicum administration, total cholesterol, triglycerides and LDL-cholesterol levels decreased by 56%, 63% and 68%, respectively, in comparison with the Triton treated group and HDL-cholesterol was not increased significantly. The hypolipidaemic effect exerted by Ocimum basilicum extract was markedly stronger than the effect induced by fenofibrate treatments. Further it was demonstrated that Ocimum basilicum aqueous extract displayed a very high antioxidant power. These results indicate that Ocimum basilicum extract may contain hypolipidaemic and antioxidant substances and its use as a therapeutic tool in hyperlipidaemic subjects may be of benefit and encourage further investigation in this field.

Animals↗

Methyleugenol in Ocimum basilicum L. Cv. genovese gigante.

Ocimum basilicum cv. Genovese Gigante is the basil cultivar used the most in the production of a typical Italian sauce called pesto. The aromatic composition of plants at different growth stages was determined. Plants from different areas of northwestern Italy were analyzed at 4 and 6 weeks after sowing and showed methyleugenol and eugenol as the main components. The content of these compounds was correlated with plant height rather than plant age. Particularly, methyleugenol was predominant in plants up to 10 cm in height, whereas eugenol was prevalent in taller plants. These results are important in the evaluation of risk to human health posed by dietary ingestion of methyleugenol contained in pesto.

Carcinogens↗

UV-B is required for normal development of oil glands in Ocimum basilicum L. (sweet basil).

Plants of Ocimum basilicum L. grown under glass were exposed to short treatments with supplementary UV-B. The effect of UV-B on volatile essential oil content was analysed and compared with morphological effects on the peltate and capitate glandular trichomes. In the absence of UV-B, both peltate and capitate glands were incompletely developed in both mature and developing leaves, the oil sacs being wrinkled and only partially filled. UV-B was found to have two main effects on the glandular trichomes. During the first 4 d of treatment, both peltate and capitate glands filled and their morphology reflected their 'normal' mature development as reported in the literature. During the following days there was a large increase in the number of broken oil sacs among the peltate glands as the mature glands broke open, releasing volatiles. Neither the number of glands nor the qualitative or quantitative composition of the volatiles was affected by UV-B. There seems to be a requirement for UV-B for the filling of the glandular trichomes of basil.

Cell Surface Extensions↗

Chemomodulatory efficacy of basil leaf (Ocimum basilicum) on drug metabolizing and antioxidant enzymes, and on carcinogen-induced skin and forestomach papillomagenesis.

Basil or sweet basil (Ocimum basilicum) is cultivated throughout India and is known for its medicinal value. The effects of doses of 200 and 400 mg/kg body weight of hydroalcoholic extract (80% ethanol, 20% water) of the fresh leaves of Ocimum basilicum on xenobiotic metabolizing Phase I and Phase II enzymes, antioxidant enzymes, Glutathione content, Lactate dehydrogenase and lipid peroxidation in the liver of 8-9 weeks old Swiss albino mice were examined. Furthermore, the anticarcinogenic potential of basil leaf extract was studied, using the model of Benzo(a)pyrene-induced forestomach and 7,12 dimethyl benz(a)anthracene (DMBA)-initiated skin papillomagenesis. The hepatic glutathione S-transferase and DT-diaphorase specific activities were elevated above basal level by basil leaf treatment (from p < 0.005 to p < 0.001). Basil leaf extract was very effective in elevating antioxidant enzyme response by increasing significantly the hepatic glutathione reductase (GR) (p < 0.005), superoxide dismutase (SOD) (p < 0.05), and catalase activities (p < 0.005). Reduced glutathione (GSH), the major intracellular antioxidant, showed a significant elevation in the liver (p < 0.005) and also in all the extrahepatic organs (from p < 0.05 to p < 0.005). In the forestomach, kidney and lung, glutathione S-transferase and DT-diaphorase levels were augmented significantly, varying from p < 0.01 to p < 0.001. There were significant decreases in lipid peroxidation and lactate dehydrogenase activity. Chemopreventive response was evident from the reduced tumor burden (the average number of papillomas/mouse, p < 0.005 to p < 0.001), as well as from the reduced percentage of tumor bearing-animals. Basil leaf, as deduced from the results, augmented mainly the Phase II enzyme activity that is associated with detoxification of xenobiotics, while inhibiting the Phase I enzyme activity. There was an induction in antioxidant level that correlates with the significant reduction of lipid peroxidation and lactate dehydrogenase formation. Moreover, Basil leaf extract was highly effective in inhibiting carcinogen-induced tumor incidence in both the tumor models at peri-initiational level.

9,10-Dimethyl-1,2-benzanthracene↗

[Study on the constituents of volatile oil from Ocimum basilicum].

OBJECTIVE: To study the constituents of volatile oil from Ocimum basilicum. METHOD: GC-MS technique was used. RESULT: Forty-five compounds was elucidated through consulting literature and searching database by computer. CONCLUSION: The main constituent in volatile oil from O. basilicum are 1, 7-dimethyl-1, 6-octadien-3-ol, which accounts for 29.87%.

Gas Chromatography-Mass Spectrometry↗

In vitro rosmarinic acid accumulation in sweet basil (Ocimum basilicum L.).

Leaf-derived suspension cultures of sweet basil, Ocimum basilicum L. accumulated rosmarinic acid up to 10 mg g(-1) dry wt, a value up to 11 times higher than in callus cultures or in leaves of donor plants. Immobilized cells accumulated less than 15 microg rosmarinic acid g(-1).

Cell Culture Techniques↗

Phenolics composition and antioxidant activity of sweet basil (Ocimum basilicum L.).

The antioxidant activity of a methanolic extract of Ocimum basilicum L. (sweet basil) was examined using different in vitro assay model systems. The crude extract was fractionated on a Sephadex LH-20 column, and six fractions were identified. The DPPH scavenging assay system and the oxidation of the soy phosphotidylcholin liposome model system were used to evaluate the antioxidant activity of each fraction. Fraction IV showed the strongest activity followed by fractions V and VI. Phenolic compounds responsible for the antioxidative activity of the fractions were characterized by atmospheric pressure chemical ionization liquid chromatography-mass spectrometry. The major antioxidant compound in fraction IV was confirmed as rosmarinic acid by (1)H NMR and characteristic fragmentations in the mass spectrum. Moreover, the native of antioxidant activity of rosmarinic acid in the liposome system was examined. The results showed that one rosmarinic acid can capture 1.52 radicals, and furthermore, the existence of a synergistic effect between alpha-tocopherol and rosmarinic acid was revealed.

Antioxidants↗

Chemical characterization of basil (Ocimum basilicum L.) found in local accessions and used in traditional medicines in Iran.

Ocimum species are used in traditional Iranian medicine, as a culinary herb, and as a well-known source of flavoring principles. Horticultural characteristics, including quantitative and qualitative traits along with the chemical variation of phenolic acids, of 23 accessions of basil (Ocimum basilicum L.) from Iran were studied. Morphological studies of accessions showed a high level of variability in recorded traits. Quantification of phenolic acids was determined using high-performance liquid chromatography and showed drastic variations between accessions. Chemical studies revealed that rosmarinic acid is the predominant phenolic acid present in both flower and leaf tissues. Unusual basil accessions were identified that can serve as genetic sources of phenolic acids for crop improvement.

Agriculture↗

Purification and characterization of Ocimum basilicum L. polyphenol oxidase.

A partial characterization of polyphenol oxidase (PPO) activity in Ocimum basilicum L. is described. PPO in O. basilicum L. was extracted and purified through (NH4)2SO4 precipitation, dialysis, and a Sepharose 4B-l-tyrosine-p-aminobenzoic acid affinity column. The samples obtained from (NH4)2SO4 precipitation and dialysis were used for the characterization of PPO. At the end of purification by affinity chromatography, 11.5-fold purification was achived. The purified enzyme exhibited a clear single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular mass of the enzyme was estimated to be approximately 54 kDa. The contents of total phenolic and protein of O. basilicum L. extracts were determined. The total phenolic content of O. basilicum L. was determined spectrophotometrically according to the Folin-Ciocalteu procedure and was found to be 280 mg 100 g(-1) on a fresh weight basis. The protein content was determined according to the Bradford method. The enzyme showed activity to 4-methylcatechol, catechol, and pyrogallol substrates, but not to tyrosine. Therefore, of these three substrates, 4-methylcatecol was the best substrate due to the highest V(max)/K(m) value, followed by pyrogallol and catechol. The optimum pH was at 6, 8, and 9 for 4-methylcatechol, catechol, and pyrogallol, respectively. The enzyme had an optimum temperature of 20, 40, and 50 degrees C for 4-methylcatechol, catechol, and pyrogallol, respectively. It was found that optimum temperature and pH were dependent on the substrates studied. The enzyme activity with increasing temperature and inactivation time for 4-methylcatechol, catechol, and pyrogallol substrates decreased due to heat denaturation of the enzyme.

Catechol Oxidase↗

Anthocyanin inheritance and instability in purple basil (Ocimum basilicum L.).

The instability of the purple pigments (anthocyanins) in purple basil varieties (Ocimum basilicum L.) limits their use as ornamental plants and as a potential anthocyanin source. Several self-pollinated generations of all purple plants were unsuccessful in stabilizing anthocyanin expression. In this study we investigated the inheritance and stability patterns of leaf traits using the Purple Ruffles variety. The results from the complete diallele crosses indicated anthocyanin expression in vegetative tissue is controlled by two dominant genes and ruffled leaf texture is controlled by a single recessive gene. Genes controlling leaf margin and leaf base structures were tightly linked to leaf texture. Essential oil production and oil constituents in leaves did not change as a result of the reversion in color. Color stability in cuttings was affected by the environment and the location where cuttings were taken. An accumulation of secondary metabolites (apigenin, genistein, and kaempferol) in green-reverted sectors on purple leaves was detected using reverse-phase high-performance liquid chromatography (HPLC) analysis; this suggested a potential block in the anthocyanin pathway. We hypothesize the reversion mutation is occurring in an anthocyanin regulatory gene.

Anthocyanins↗

Aroma content of fresh basil (Ocimum basilicum L.) leaves is affected by light reflected from colored mulches.

Sweet basil (Ocimum basilicum L.) is an herb that is used to add a distinct aroma and flavor to food. Volatile compounds emitted from fully expanded fresh leaves grown in drip-irrigated plots that were covered with six colors of mulch were compared. The colors reflected a range of photosynthetic photon flux, far-red, red, and blue light from the soil surface to developing leaves. Our objective was to determine whether reflection from the different colors could influence concentrations of volatile compounds emitted from the fresh leaves. Volatile compounds were isolated by headspace sampling and quantified by gas chromatography. Twenty-six compounds were identified, of which the terpenoids linalool and 1,8-cineole comprised more than 50% of the total yield. Concentrations of volatile compounds from leaves that developed over green, blue, yellow, white, and red mulches followed the same patterns as they did for air-dried leaves of the same cultivar. However, the concentration of volatile compounds from fresh leaves was about 50-fold higher than those found in the previous study of air-dried leaves.

Acyclic Monoterpenes↗

Infraspecific taxonomy and essential oil chemotypes in sweet basil, Ocimum basilicum.

Essential oil compositions of fresh and freeze-dried leaves were determined for 16 accessions of Ocimum basilicum belonging to different varieties to see whether they could be used as infraspecific taxonomic characters. One accession of O. x citriodorum was also studied. Some 30 monoterpenoids, sesquiterpenoids and phenylpropanoids were identified, the major components (more than 20% of the total essential oil composition in one or more accessions) being geranial and neral in O. x citriodorum, and linalool, methyl chavicol, eugenol, methyl eugenol and geraniol in O. basilicum. Based on a combination of the latter compounds, five major essential oil profiles could be distinguished in the accessions studied for O. basilicum. These profiles were largely the same for fresh and freeze-dried material of the same plant, although in the dried leaves, methyl chavicol and eugenol concentrations had generally declined in comparison to those of linalool. There appeared to be little correlation between essential oil patterns and varietal classification within O. basilicum. In view of the chemical heterogeneity of O. basilicum and its use as an essential oil-producing crop, culinary herb, medicinal plant and insect-controlling agent, in all of which chemicals play an important role, the infraspecific classification of this taxon should take chemical characters into consideration. A system for the classification of essential oil chemotypes in O. basilicum is proposed.

Gas Chromatography-Mass Spectrometry↗

Somatic embryogenesis and plant regeneration from leaf callus of Ocimum basilicum L.

A effective protocol for complete plant regeneration via somatic embryogenesis has been developed for Ocimum basilicum L. Callus was initiated from leaf explant of young plant on supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) 1.0 mg l(-1), 3% sucrose and 0.9% agar. The calli showed differentiation of globular structure embryos when transferred to MS medium containing 2,4-D 0.5 mg l(-1) and BAP 1.0 mg l(-1). The maximum globular structure embryos were further enlarged and produced somatic embryos in MS basal medium supplemented with BAP 1.0 mg l(-1)+NAA 1.0 mg l(-1) + KN 0.5 mg l(-1). Continued formation of globular embryo and germination of embryos occurred in this medium. Complete plantlets were transferred onto specially made plastic cup containing soilrite followed by their transfer to the garden soil. Survival rate of the plantlets under ex vitro condition was 80%.

Cell Culture Techniques↗

Immobilization of invertase through its carbohydrate moiety on Ocimum basilicum seed.

Yeast invertase, a glycoprotein, was covalently coupled to Ocimum basilicum seeds either through its protein or carbohydrate moiety. Of the various methods investigated, binding of the enzyme through its carbohydrate moiety resulted in the retention of considerably higher amounts of enzyme activity. Immobilized invertase showed a shift in the pH optimum toward the alkaline side without appreciable change in temperature optimum. However, the immobilized preparation was more thermostable than the free enzyme. Invertase bound to the seeds could be used repeatedly for the hydrolysis of sucrose syrups in a batch process without appreciable loss in activity. The seeds could serve as an inexpensive, ready-to-use, natural pellicular polysaccharide support for immobilizing enzymes.

Carbohydrates↗