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Recent studies on natural products as anti-HIV agents.

The discovery of medicinal agents capable of specifically inhibiting human immunodeficiency virus (HIV) is urgently needed due to its globally widespread infection. Most of clinically useful anti-HIV agents are nucleosides but their use is limited due to their severe toxicity and emerging drug resistance. More than 50% of world marketed drugs have their origin of the nature. Natural products, of which structural diversity is so broad, are good sources for the effective discovery of anti-HIV agents with decreased toxicity. Over the past decade, substantial progress has been made in research on the natural products for the anti-HIV agents. New natural products that have potent anti-HIV activities with novel structures were reviewed in this article. These compounds, isolated mainly from medicinal plants, in this review have been classified as secondary metabolites such as terpenes, phenolics, and naturally scarce peptides and sugars. Especially, terpenes and phenol substances have gained much interest due to their significant anti-HIV activities along with their structural diversity. Recent studies also showed that several polysaccharides are effective inhibitors of HIV replication. Most of chemotherapeutic targets reviewed in this article are found to be HIV reverse transcriptase (RT).

Alkaloids↗

Stimulation of collagen synthesis in fibroblast cultures by a triterpene extracted from Centella asiatica.

The drug "Titrated Extract from Centella asiatica" (TECA), used for its stimulating properties on the healing of wounds, is a mixture of 3 terpenes extracted from a tropical plant: asiatic acid (30%, w/w), madecassic acid (30%, w/w) and asiaticoside (40%, w/w). The effects of TECA and its individual components were checked on human foreskin fibroblast monolayer cultures. TECA increased the collagen synthesis in a dose-dependent fashion whereas a simultaneous decrease in the specific activity of neosynthesized collagen was observed. Asiatic acid was found to be the only component responsible for collagen synthesis stimulation. TECA and all three terpenes increased the intracellular free proline pool. This effect was independent of the stimulation of collagen synthesis.

Cell Count↗

Comparison of self-assessment and expert assessment of occupational exposure to chemicals.

OBJECTIVES: Occupational assessments of chemical exposure are often inadequate because of difficulties in obtaining sufficient numbers of measurements by trained professionals (experts). The objective of this study was to determine whether workers can provide unbiased data via self-assessments of exposure facilitated by the use of simple passive monitors for personal sampling. METHODS: Untrained workers obtained personal measurements of their exposures to gaseous contaminants (terpenes in sawmills and styrene in reinforced plastics factories) with passive monitors and written instructions. To study the validity of the self-assessments, an occupational hygienist performed exposure measurements on the same occupational groups after the workers had obtained two or more measurements independently. The potential bias of the self-assessments was evaluated by comparing the self-assessments with the expert assessments in mixed-effects statistical models. RESULTS: A total of 153 terpene (97 self and 56 expert) and 216 styrene (159 self and 57 expert) measurements were obtained from four sawmills and six reinforced plastics factories, respectively. No significant differences in the geometric mean exposures were observed between the self-assessments and the expert assessments in 3 of 4 sawmills and 5 of 6 reinforced plastics factories (P > 0.10). The potential bias of the self-assessments of exposure ranged from less than 0.1% to 102% and was less than 17% in 9 of the 10 groups investigated. CONCLUSIONS: The results indicate that untrained, unsupervised workers are able to collect consistently unbiased exposure data by employing currently available passive monitors.

Air Pollution, Indoor↗

[Urinary concentration and antibacterial effect of short and long acting tetracycyline].

Forty-one hospitalized patients, of whom the majority had asymptomatic bacteriuria, were randomly divided into 4 treatment groups. They were given either tetracycline HCl 1 g, or tetracycline HCl + terpenes 100 mg, or minocycline 200 mg, or cotrimoxazole (sulfamethoxazole 1.6 g + trimethoprim 320 mg), daily for one week. A large number of bacteria were resistant to tetracyclines. The urine was sterilized in 14 out of 24 patients receiving tetracyclines and in all of the 17 patients receiving cotrimoxazole. The mean urinary concentration of tetracycline was 20 times higher than that of minocycline. The 24-h urinary excretion of tetracyclines was slightly higher in patients receiving the combination of terpenes and tetracycline, but these differences in urinary excretion did not appear to have any influence on the antibacterial effect of tetracyclines.

Adult↗

Application of topological indexes for evaluation of the TLC separation of selected essential oil components.

The selected essential oil components (menthol, (+)bomeol, geraniol, linalool, carvone, camphor, (IR)-(-)fenchone) were separated by adsorption thin-layer chromatography using benzene as mobile phase. Investigated terpenes were characterized by selected topological indexes based on the adjacency and distance matrix. From among all the topological indexes counted only Si(o) index (the sum of the distance between the oxygen atom and all the remaining atoms in graph) as well as cluster Randić indexes (4chi(c), and 4chi(v)c) allow for estimation of the chromatograms obtained. With the fact mentioned above, there exists the possibility of prediction of relative situation on a chromatogram of the terpenes investigated. For the rest of topological indexes that possibility was not stated.

Administration, Topical↗

OH-initiated oxidation of monoterpenes: reaction of alpha-pinene.

The aim of this paper is to study the reaction products of alpha-pinene, beta-pinene, limonene, 3-carene and sabinene with OH radicals by FT-IR spectroscopy and by HPLC-MS-MS, to evaluate the secondary aerosol formation. All gas phase reaction products were quantified using reference compounds. As source of OH radicals were used H2O2 and CH3ONO. The experiments were performed at low terpene concentration (0.9-2.1 ppm) and at high terpene concentration (4.1-13.2 ppm), using H2O2 and CH3ONO as sources of OH radicals.

Aerosols↗

[Analysis of volatile constituents from Anemarrhena asphodeloides by GC-MS].

OBJECTIVE: To study the chemical constituents of the volatile oil from the rhizome of Anemarrhena asphodeloides. METHOD: The volatile oil was steam distillation. Chemical constituents were separated and analyzed by GC-MS. The relative content of each component was determined by area nomalization. RESULT: 24 volatile compounds were isolated and identified for the first time, representing 70.83% of the total oil. CONCLUSION: The main constituents of this oil were aldehydes (31.15%), terpene and their oxide (20.66%), alkyls (8.35%), Furan heterocyclic compounds (6.41%), non terpene alcohol (4.26%). There are 12 compounds with contents over 3%. Among them, borneol has the highest content (9.35%).

Aldehydes↗

Combined effect of cyclic monoterpenes and ethanol on percutaneous absorption of diclofenac sodium.

The combined effect of cyclic monoterpenes and ethanol on the percutaneous absorption of diclofenac sodium (DFS) from gel ointments was investigated in vivo in rats. The enhancing activity of terpenes was significantly affected by the concentration of ethanol formulated in the gel ointments. At a lower concentration of ethanol (20%), 1,8-cineole was observed to be the most effective. On the other hand, d-limonene showed strong activity when the large amount of ethanol was formulated (40%). A synergistic effect between terpenes and ethanol on the percutaneous absorption of DFS was significantly observed in cases of 1,8-cineole and l-menthol using an analysis of variance (ANOVA). When the diclofenac (DF) free form was formulated in gel ointment, the percutaneous absorption was significantly reduced. The reduction of the percutaneous absorption was closely related to the decrease in pH of the gel ointment owing to the free form of DF which was formulated.

Analysis of Variance↗

Diversity and unity in the nuclear hormone receptors: a terpenoid receptor superfamily.

The remarkable structural unity among the different members of the nuclear hormone receptor superfamily stands in striking contrast to the diversity of the chemical structures of their ligands. Of the three currently known classes of ligands, steroids, retinoids, and thyroid hormones, the first two share a common biosynthetic pathway. Both are terpenes, which are derived by assembly of isoprene units. This biosynthetic link suggests that the receptors for three other classes of terpenoid hormones, the insect juvenile hormones and the plant hormones gibberellic acid and abscissic acid, may also be members of the superfamily. A number of putative nuclear hormone receptors that do not have known ligands have been isolated. At least some of the ligands for these orphan members of the receptor superfamily may be found on the list of biologically active terpenes. Finally, the terpenoid connection raises interesting issues for the evolution of the receptor superfamily.

Animals↗

Vincent van Gogh and the thujone connection.

During his last two years Vincent van Gogh experienced fits with hallucinations that have been attributed to a congenital psychosis. But the artist admitted to episodes of heavy drinking that were amply confirmed by colleagues and there is good evidence to indicate that addiction to absinthe exacerbated his illness. Absinthe was distilled from an alcoholic steep of herbs. Wormwood (Artemisia absinthium) was the most significant constituent because it contributed thujone. This terpene can cause excitation, convulsions that mimic epilepsy, and even permanent brain damage. Statements in van Gogh's letters and from his friends indicate that he had an affinity for substances with a chemical connection to thujone; the documented examples are camphor and pinene. Perhaps he developed an abnormal craving for terpenes, a sort of pica, that would explain his attempts to eat paints and so on, which were previously regarded as unrelated absurdities.

Alcoholic Beverages↗

Increased mannose 6-phosphate/insulin-like growth factor II receptor and transforming growth factor beta 1 levels during monoterpene-induced regression of mammary tumors.

The monoterpenes represent a potentially new class of breast cancer therapeutic agents. We have shown that d-limonene induces the regression of advanced rat mammary adenocarcinomas. These regressing tumors have an increased cellular concentration of both the mannose-6-phosphate/insulin-like growth factor II receptors and transforming growth factor beta 1. The terpene-induced regression of mammary tumors may result in part from the mitoinhibitory and differentiation properties of active transforming growth factor beta 1. Furthermore, the activation of transforming growth factor beta 1 in these tumors is likely to be facilitated by the increased concentration of the mannose-6-phosphate/insulin-like growth factor II receptors in the mammary tumor cells. Tumors not responding to terpene therapy lacked a rise in the mannose-6-phosphate/insulin-like growth factor II receptor level which may relate to the fact that this gene is hemizygous due to maternal imprinting.

9,10-Dimethyl-1,2-benzanthracene↗

Effect of d-limonene, alpha-pinene and cineole on in vitro transdermal human skin penetration of chlorpromazine and haloperidol.

The aim of this research is to carry out a comparative study of the ability to cross human skin of two neuroleptic drugs: chlorpromazine (CAS 50-53-3) and haloperidol (CAS 52-86-8), in the absence and in the presence of three terpenes (cineole, d-limonene and alpha-pinene) with the purpose of considering the possibility of improving their transdermal penetration profile. Franz diffusion cells were used, in conjunction with human skin as permeation membrane. The permeation parameters calculated were permeability constant (Kp), flux (J) and lag time (Tl) in the presence and in the absence of enhancers. None of the three enhancers assayed improved the penetration profile of chlorpromazine, and d-limonene even reduced the transdermal permeability (enhancement index, EI = 0.67) since its coefficient of relative activity was reduced, (Xr = 0.73). Cineole and d-limonene increased the permeation profile of haloperidol, giving EI values of 1.95 and 4.21, respectively, and leading to a fourfold increase in the flux value for both enhancers. alpha-Pinene did not modify the permeation profile of haloperidol. None of the three terpenes assayed had a significant effect on the lag time of chlorpromazine or haloperidol. In these experimental conditions the concentration values predicted at steady state of chlorpromazine formulated without enhancers are within the therapeutic range. In contrast, therapeutic levels of haloperidol cannot be predicted in the absence of enhancers such as d-limonene or cineole.

Antipsychotic Agents↗

Interaction of air pollution and hyperbaric oxygen on virus replication.

A study was undertaken to weigh the effects of air pollution and oxygen at high pressure on the susceptibility of mice to Coxsackievirus B1 infection. Animals exposed to air pollutants or oxygen at high pressure were found to contain higher amounts of recoverable virus than control animals. Animals exposed to both air pollutants and oxygen at high pressure were found to contain the greatest levels of recoverable virus. This same group of animals was also found to have hearts which were smaller than any other group. Animals maintained in an ambient atmosphere had higher levels of recoverable virus and smaller hearts than animals exposed to terpene vapors and hyperbaric oxygen. The results of this study suggest that terpene vapors may nullify the activity of oxygen at high pressure.

Air Pollutants↗

[Observations on organic components of thermal mud: morphohistochemical and biochemical studies on lipid components of mud of the Terme dei Papi (Laghetto del Bagnaccio, Viterbo). Chemical bases of the interpretation of biological and therapeutic actions of thermal mud].

In previous findings the lipidic fractions extracted acc. to Folch from the mature muds of the majority of the Italian thermal springs hot baths was studied, with the aim to identify the organic substrates of their therapeutical activity. The organic components of the "mature" peloids are produced by the metabolism of the microphytozooplankton growing spontaneously in the clay-substrate, in contact with the hot water. The Popes thermal springs (Bagnaccio's Lake) are characterised by an unique environmental situation, because the muds are naturally matured in the hot thermal water, but not in artificial baths. The morphohistochemical aspects of thermal algae growing in the Bagnaccio's lake have been studied by means of Computerised Optic Probe Video-Microscopy, using a not contact zoom objective 70-400x. Peloid types, both the "white" and the "black" contains yellow pigments, fragments of hyphae, monocellular algae, Diatomeae, Cyanophyceae and few other species. The biochemical aspects of the muds extracts are characterised by the presence of Phospholipids (PC, PE, PS, SP), a series of Hydrocarbons ranging from C30 to C38, Phytosterols (beta-sitosterol, Campesterol, Stigmasterol and traces of Cholesterol), Free Fatty Acids (Palmitic, Palmitoleic, Myristic, Stearic, Oleic and Linoleic, heptadecenoic and heptadecanoic) and Terpenes (beta-amirrhyne, 24-methylene-cyclo-arthanole). In our opinion, the therapeutic effects of the mature muds are related to its organic components, with special regards to Phospholipids, Phytosterols and Terpenes. The richness of these components in the Popes Thermal springs seems to be great interest in the dermatological and cosmetic applications, other then the traditional use.

Animals↗

Direct analysis of oligomeric tackifying resins in rubber compounds by automatic thermal desorption gas chromatography/mass spectrometry

Two analytical methods, automatic thermal desorption gas chromatography/mass spectrometry (ATD-GC/MS) and pyrolysis gas chromatography/mass spectrometry (Py-GC/MS), were applied as direct methods for the analysis of oligomeric tackifying resins in a vulcanized rubber. The ATD-GC/MS method, based on discontinuous volatile extraction, was found to be an effective means for direct analysis of the oligomeric tackifying resins contained in a vulcanized rubber. The oligomeric tackifying resins, such as t-octylphenolformaldehyde (TOPF) resin, rosin-modified terpene resin, and cashew resin, could be directly analyzed in vulcanized rubber by ATD-GC/MS. Much simpler total ion chromatograms were obtained by ATD-GC/MS than by flash pyrolysis with a Curie-point pyrolyzer, permitting much easier interpretation. Ions at m/z 206, 135, and 107 were fingerprints in the characteristic mass spectra obtained by ATD-GC/MS for TOPF resin in the vulcanized rubber. 1H-Indene, styrene, and isolongifolene were observed as their characteristic mass spectra in the pyrolyzate of the rosin-modified terpene resin. From the cashew resin, phenol, 3-methylphenol, and 4-(1,1,3, 3-tetramethylbutyl)phenol were obtained as the characteristic pyrolyzates by discontinuous thermal extraction via ATD-GC/MS. Copyright 1999 John Wiley & Sons, Ltd.

Journal Article↗

Mechanism(s) of in vitro percutaneous absorption enhancement of tamoxifen by enhancers.

The effects of enhancers (5% terpenes; i.e., eugenol, limonene, and menthone) in combination with 50% propylene glycol in water (50% PG) on the in vitro percutaneous absorption of tamoxifen through the porcine epidermis, on biophysical changes in the stratum corneum (SC) lipids, on macroscopic barrier properties, and on binding of the drug to the SC were investigated. These enhancers in combination with 50% PG significantly increased (p<0.05) the permeability coefficient of tamoxifen in comparison with that of the control (50% PG in water). Fourier transform infrared spectroscopy (FT-IR) was employed to investigate the biophysical changes in the SC lipids. The FT-IR results showed that treatment of the SC with 5% terpenes/50% PG did not shift the asymmetric and symmetric C-H stretching absorbances peak positions to higher wavenumbers but resulted in a decrease in the peak heights and areas in comparison with the untreated SC. Treatment with menthone and limonene in combination with 50% PG significantly increased (p<0.05) the partition coefficient of tamoxifen in comparison with treatment with 50% PG alone. Also, exposure of the SC to 5% terpenes in combination with 50% PG significantly increased (p < 0.05) the in vitro transepidermal water loss (TEWL) in comparison with 50% PG alone. Thus, an enhancement by menthone, eugenol, and limonene in the permeability of the SC to tamoxifen is due to lipid extraction and macroscopic barrier perturbation. Moreover, the effective diffusion coefficient of tamoxifen through the epidermis was enhanced following the treatment with either 5% eugenol/50%PG or 5% limonene/50%PG compared with 50%PG alone.

Animals↗

Volatile flavour of wine: correlation between instrumental analysis and sensory perception.

The aroma of wine consists of 600 to 800 aroma compounds from which especially those, typical for the variety, are already present in the grapes. There are significant varietal differences between the aromagrams ('fingerprint patterns'). Thus the amount of some flavour compounds ('key substances') shows typical dependence on the variety. Especially monoterpene compounds play an important role in the differentiation of wine varieties. The German white wines can be differentiated into three groups only by quantitative determination of 12 monoterpenes ('terpene profile'). These groups are: 'Riesling type', 'Muscat type' and 'Silvaner-Weissburgunder type'. Such 'terpene profiles' are also useful for the separation of real Riesling wines from others called Riesling (e.g. Welschriesling, Kap Riesling, Emerald Riesling) but not produced from grapes of the variety Riesling. Including further components and by means of statistical methods (discriminant analysis) even the different varieties within the mentioned groups for instance the 'Riesling'-group (e.g. Riesling, Kerner, Ehrenfelser, Bacchus, Müller-Thurgau) can be separated from each other. An analytical characterization of the neutral ('Silvaner-type') grape varieties Silvaner, Ruländer (Pinot gris), Weissburgunder (Pinot blanc) is also possible with about 20 compounds (e.g. monoterpenes, alcohols). Computing at the same time free and glycosidically bound aroma components (monoterpenes, alcohols, norisoprenes) in discriminant analysis the characterization of the neutral grape varieties can be considerably improved. To identify compounds causing 'off-flavours' sniffing technique is the method of choice. The off-flavour is pinpointed during gas chromatographic separation of the complex aroma mixture by effluent sniffing. Once allocated, the chemical nature of the off-flavours is elucidated by spectroscopic methods. Substances contributing to the green pepper taint, the strawberry note, moussiness, corkiness, etc. in wine could be found in this way.

Humans↗

Determination of active components of Ginkgo biloba in human urine by capillary high-performance liquid chromatography/mass spectrometry with on-line column-switching purification.

Ginkgo biloba is one of the most popular herbal nutritional supplements, with terpene lactones and flavonoids being the two major active components. An on-line purification high-performance liquid chromatography/mass spectrometry (HPLC/MS) method was successfully developed for the quantitative determination of flavonoids and terpene lactones excreted in human urine after ingesting the herbal supplement. Satisfactory separation was obtained using a C18 capillary column made in-house with sample clean-up and pre-concentration achieved using a C18 pre-column with column switching. High selectivity and limits of detection of 1-18 ng/mL were achieved using a selected ion monitoring (SIM) scan in negative ion mode; the on-line solid-phase extraction (SPE) recovery of the active components in Ginkgo biloba determined in this study was greater than 75%.

Capillary Electrochromatography↗