Main Bioactive Metabolites Research Articles

Exosomal secretion may be a self-protective mechanism of its source cells under environmental stress: A study on human bronchial epithelial cells treated with hydroquinone.

Accumulating evidence reveals that exosome plays an important role in cell-to-cell communication in both physiological and pathological processes by transferring bioactive molecules. However, the role of exosomal secretion in the adaption of its source cells to the stimuli of environmental chemicals remains elusive. In this study, we revealed that the exposure of hydroquinone (HQ; the main bioactive metabolite of benzene) to human bronchial epithelial cells (16HBE) resulted in decreased ability of cell proliferation and migration, and simultaneously DNA damage and micronuclei formation. Interestingly, when exosomal secretion of HQ treated 16HBE cells was inhibited with the inhibitor GW4869, cellular proliferation and migration were further significantly reduced; concurrently, their DNA damage and micronuclei formation were both further significantly aggravated. Herein, we conclude that exosomal secretion of 16HBE cells may be an important self-protective function against the toxic effects induced by HQ.

Pharmaceuticals in environment: the effect of ivermectin on ribwort plantain (Plantago lanceolata L.).

The anthelmintic drug ivermectin (IVM), used frequently especially in veterinary medicine, enters the environment mainly via excrements in pastures and could negatively affect non-target organisms including plants. The present study was designed to follow up on our previous investigations into IVM metabolism and its effects in the common meadow plant ribwort plantain (Plantago lanceolata L.) during long-term exposure of both cell suspensions and whole plant regenerants. IVM uptake, distribution, and biotransformation pathways were studied using UHPLC-MS analysis. In addition, the IVM effect on antioxidant enzymes activities, proline concentration, the content of all polyphenols, and the level of the main bioactive secondary metabolites was also tested with the goal of learning more about IVM-induced stress in the plant organism. Our results showed that the ribwort plantain was able to uptake IVM and transform it via demethylation and hydroxylation. Seven and six metabolites respectively were detected in cell suspensions and in the roots of regenerants. However, only the parent drug IVM was detected in the leaves of the regenerants. IVM accumulated in the roots and leaves of plants might negatively affect ecosystems due to its toxicity to herbivorous invertebrates. As IVM exposition increased the activity of catalase, the concentration of proline and polyphenols, as well as decreased the activity of ascorbate peroxidase and the concentration of the bioactive compounds acteoside and aucubin, long-term exposition of the ribwort plantain to IVM caused abiotic stress and might decrease the medicinal value of this herb.

Ganoderma triterpenoids attenuate tumour angiogenesis in lung cancer tumour-bearing nude mice

Context Ganoderma lucidum (Leyss. ex Fr.) Karst. (Polyporaceae) triterpenoids (GLTs), the main components and bioactive metabolites of G. lucidum, have antitumour activity. Objective We investigated the effects of GLTs in lung cancer tumour-bearing nude mice and their potential mechanism. Materials and methods Forty BALB/c nude mice were randomly divided into four groups: saline control, GLT (1 g/kg/day), gefitinib (GEF, 15 mg/kg/day), and GLT (1 g/kg/day) + GEF (15 mg/kg/day) for 14 days. Cell viability was conducted using the Cell Counting Kit-8 assay. The tumour volume, inhibition rate, histopathological, microvessel density (MVD), mRNAs, and proteins were determined. Results GLTs inhibited the cell viability of A549 cells with an IC50 value of 14.38 ± 0.29 mg/L, while the IC50 value of GEF was 10.26 ± 0.47 μmol/L. The tumour inhibition rate in the GLT + GEF group (51.54%) was significantly decreased relative to the saline control… group (p < 0.05). The MVD in the GLT + GEF group (2.9 ± 0.7) was significantly decreased than that in the saline control group (12.8 ± 1.4, p < 0.05). The angiostatin, endostatin, and Bax protein expression in the GLT, GEF, and GLT + GEF groups were significantly increased compared to those in the saline control group, while the VEGFR2 and Bcl-2 protein expression were decreased. Discussion and conclusions Our study provided evidence that GLT and GEF combination therapy may be a promising candidate for the treatment of lung cancer and as an experimental basis for clinical treatment.

Melissa officinalis ssp. altissima extracts: A therapeutic approach targeting psoriasis in mice.

Melissa officinalis L., commonly known as lemon balm, is one of the most well known edible and medicinal plants of the Lamiaceae family. It is quoted in almost all known medical treatises, from Antiquity up to modern era, such as Corpus Hippocraticum, Dioscorides' De Materia Medica and later on in medieval medical manuscripts and Pharmacopoeias. Actually, it is widely used as herbal medicine for the relief of mild symptoms of mental stress, to aid sleep and for symptomatic treatment of mild gastrointestinal complaints including bloating and flatulence. In Greece, the empirical physicians "vikoyiatroi" recommended the decoction of dry flowers and leaves to treat tracheobronchitis, hysteria, epilepsy, heart arrhythmias, as hypnotic and against skin disorders. The present study was conducted to investigate the potential beneficial effects in psoriasis in mice of M. officinalis ssp. altissima and to carry out the chemical analysis in order to reveal its main bioactive secondary metabolites. Non polar and polar extracts of M. officinalis ssp. altissima aerial parts were prepared by using dichloromethane and methanol, successively; in addition a decoction was made upon oral information by local users in Crete, where the plant was collected. All three preparations were chemically analyzed in order to isolate their main constituents. Chemical structures of all isolated compounds were determined by 1D, 2D-NMR and UV-Vis spectroscopy. Furthermore, the antioxidant potential of extracts and decoction was evaluated through DPPH radical scavenging capability. The in vivo in mice anti-psoriatic efficacy of all preparations was estimated through clinical and histopathological assessment and measurements of TEWL and hydration. The dichloromethane extract yielded ursolic acid, 2α-hydroxy-ursolic acid, pomolic acid, 3β-stearyloxy-urs-12-ene, oleanolic acid, noropacursane and campesterol; the methanol extract afforded rosmarinic acid and methyl rosmarinate, while from the decoction caffeic acid, 3-(3,4-dihydroxyphenyl)lactic acid and rosmarinic acid were isolated. The psoriasis evaluation, based on PASI score, photodocumentation and histopathological estimation showed that the decoction primarily and the dichloromethane extract secondly could significantly contribute to psoriasis treatment. The decoction seems able to reestablish skin physiology by decreasing dryness and enhancing skin barrier function. Moreover, the decoction showed the best antioxidant activity, while the dichloromethane extract the weakest. The triterpene derivatives of the dichloromethane extract are likely to be responsible for its anti-psoriatic activity. The abundant polyphenolic load of the decoction contributes to its high antioxidant activity and the most effective results against psoriasis. The anti-psoriatic activity of the decoction confirmed the traditional use of this plant as antioxidant, wound healing and skin barrier repair agent.

Reversal effect of ginsenoside Rh2 on oxaliplatin-resistant colon cancer cells and its mechanism.

Chemotherapy is an important treatment modality for colon cancer, however, drug resistance is the main factor leading to treatment failure. Ginsenoside Rh2 (G-Rh2), the main bioactive metabolite of ginseng, is known to possess the ability to potently induce cell apoptosis, inhibit cell proliferation and reverse multidrug resistance in a variety of cancer cells. The present study examined the effect of G-Rh2 on oxaliplatin (L-OHP)-resistant colon cancer cells and its potential mechanism. L-OHP-resistant colon cancer cells (LoVo/L-OHP) and LoVo cells were used in the present study. The effect of G-Rh2 on LoVo/L-OHP and LoVo cell proliferation was measured using a 3-(4,5 dimethylthiazol-z-yl)-3,5-diphenyltetrazolium bromide assay. The effects of G-Rh2 on LoVo/L-OHP and LoVo cell apoptosis were detected by flow cytometry. The mRNA and protein expression of apoptosis-related genes Bax, Bcl-2 and caspase-3, drug resistance-related genes P-glycoprotein (P-gp) and Smad4, were determined in LoVo/L-OHP and LoVo cells treated with G-Rh2 by reverse transcription-quantitative polymerase chain reaction and western blot analyses. G-Rh2 treatment significantly inhibited the proliferation and induced the apoptosis of LoVo/L-OHP and LoVo cells. In addition, G-Rh2 treatment resulted in a significant increase in pro-apoptotic factors, Bax and caspase-3, and decrease in anti-apoptotic factor Bcl-2 in the LoVo/L-OHP and LoVo cells. Furthermore, G-Rh2 treatment significantly decreased the levels of P-gp and increased the levels of Smad4 in the LoVo/L-OHP and LoVo cells. It was found that L-OHP had no significant effects on LoVo/L-OHP cell proliferation or apoptosis, whereas G-Rh2 + L-OHP treatment significantly inhibited LoVo/L-OHP cell proliferation and induced apoptosis. L-OHP had no significant effects on the expression of P-gp, Smad4, Bcl-2, Bax or caspase-3 in LoVo/L-OHP cells. Treatment with G-Rh2 + L-OHP significantly reduced the expression of P-gp and Bcl-2, and enhanced the expression levels of Smad4, Bax and caspase-3. These findings demonstrated that G-Rh2 reversed the drug resistance of LoVo/L-OHP cells to L-OHP, and this may be mediated by inhibiting cell proliferation and promoting apoptosis and regulating the expression of drug resistance genes. These results suggest that G-Rh2 may function as a potent anticancer drug for drug resistance in colon cancer treatment.

Quantification of bioactive lignans in sesame seeds using HPTLC densitometry: Comparative evaluation by HPLC-PDA

Sesamin and sesamolin constitute the main bioactive secondary metabolites of sesame seeds (Sesamum indicum L., Pedaliaceae). In the present work, a rapid HPTLC-based methodology was developed in compliance with the requirements of the European Pharmacopoeia for the quantification of these two lignans in sesame seeds. A comparative study was simultaneously performed with HPLC-PDA for assessing the sesamin and sesamolin content of diverse samples. Both methods were validated and the results were subsequently subjected to statistical analysis in order to compare their performance as well as to investigate possible correlations. The methods were shown to be adequately correlated in terms of performance, as revealed by Pearson’s rank correlation coefficients (>0.99 for sesamin and >0.98 for sesamolin) and Bland-Altman analysis (relative method bias 0.06–0.21, SD of bias 0.05–0.07). HPTLC densitometry could thereby serve as a valid and reliable tool for the rapid determination of the major lignans in sesame seed samples.

Chlamyphilone, a Novel Pochonia chlamydosporia Metabolite with Insecticidal Activity.

Metabolites from a collection of selected fungal isolates have been screened for insecticidal activity against the aphid Acyrthosiphon pisum. Crude organic extracts of culture filtrates from six fungal isolates (Paecilomyces lilacinus, Pochonia chlamydosporia, Penicillium griseofulvum, Beauveria bassiana, Metarhizium anisopliae and Talaromyces pinophilus) caused mortality of aphids within 72 h after treatment. In this work, bioassay-guided fractionation has been used to characterize the main bioactive metabolites accumulated in fungal extracts. Leucinostatins A, B and D represent the bioactive compounds produced by P. lilacinus. From P. griseofulvum and B. bassiana extracts, griseofulvin and beauvericin have been isolated, respectively; 3-O-Methylfunicone and a mixture of destruxins have been found in the active fractions of T. pinophilum and M. anisopliae, respectively. A novel azaphilone compound, we named chlamyphilone, with significant insecticidal activity, has been isolated from the culture filtrate of P. chlamydosporia. Its structure has been determined using extensive spectroscopic methods and chemical derivatization.

A rapid, sensitive, and selective liquid chromatography–mass spectrometry method for simultaneous quantification of astragaloside IV and cycloastragenol in mouse plasma and its application to a pharmacokinetic study

Astragaloside IV is the primary active ingredient of Astragalus membranaceus (Fisch.) Bunge with multiple pharmacological effects and a promising therapeutic agent for a number of diseases such as diabetic nephropathy and heart disease. To study the pharmacokinetics of astragaloside IV in mice, a rapid, sensitive, and selective liquid chromatography–mass spectrometry method was developed for the simultaneous quantification of astragaloside IV and its main bioactive metabolite, cycloastragenol. Samples were prepared using only 20 μL of mouse plasma and digoxin as an internal standard. After protein precipitation, the supernatant was treated using a phospholipid removal plate to remove ion-suppressive phospholipids. The analytes and internal standard in the filtered solutions were separated on a C18 column and detected under positive ion and selected ion monitoring mode within just 3 min. The assay exhibited good linearity in the concentration range 1–200 ng/mL. The intra- and interday precisions expressed as relative standard deviations were all below 8.6%, and the accuracy expressed as a relative error was ≤8.8%. The method was successfully applied to a pharmacokinetic study of healthy mice upon administration of 30 mg/kg/d astragaloside IV.

Bioactive dicaffeoylquinic acid derivatives from the root extract of Calea urticifolia

Calea urticifolia (Mill.) DC., Asteraceae, is a native plant of the Yucatan Peninsula used in traditional medicine to treat inflammation and pain. The bioassay-guided purification of the ethanol root extract allowed the isolation of the main bioactive metabolites, which were identified as an inseparable mixture of thymol (1) and 3-methyl-4-isopropylphenol (2), together with 3,4-O-dicaffeoylquinic acid methyl ester (3), 3,4-O-dicaffeoyl-epi-quinic acid methyl ester (4), 3,5-O-dicaffeoyl-epi-quinic acid methyl ester (5) and 3,5-O-dicaffeoylquinic acid (6). The results showed that the analgesic activity detected in the root extract of C. urticifolia could be attributed mainly to the mixture of 1 and 2 and to the novel 3,4-O-dicaffeoyl-epi-quinic acid methyl ester (4). Alternatively, the similarity on the antiinflammatory and antioxidant activities of the dicaffeoylquinic acid derivatives 3–5 suggests that the former might be related to their ability as radical scavengers.

Specific rescue by ortho-hydroxy atorvastatin of cortical GABAergic neurons from previous oxygen/glucose deprivation: role of pCREB.

The statin atorvastatin (ATV) given as a post-treatment has been reported beneficial in stroke, although the mechanisms involved are not well understood so far. Here, we investigated invitro the effect of post-treatment with ATV and its main bioactive metabolite ortho-hydroxy ATV (o-ATV) on neuroprotection after oxygen and glucose deprivation (OGD), and the role of the pro-survival cAMP response element-binding protein (CREB). Post-OGD treatment of primary cultures of rat cortical neurons with o-ATV, but not ATV, provided neuroprotection to a specific subset of cortical neurons that were large and positive for glutamic acid decarboxylase (large-GAD(+) neurons, GABAergic). Significantly, only these GABAergic neurons showed an increase in phosphorylated CREB (pCREB) early after neuronal cultures were treated post-OGD with o-ATV. We found that o-ATV, but not ATV, increased the neuronal uptake of glutamate from the medium; this provides a rationale for the specific effect of o-ATV on pCREB in large-GABAergic neurons, which have a higher ratio of synaptic (pCREB-promoting) vs extrasynaptic (pCREB-reducing) N-methyl-D-aspartate (NMDA) receptors (NMDAR) than that of small-non-GABAergic neurons. When we pharmacologically increased pCREB levels post-OGD in non-GABAergic neurons, through the selective activation of synaptic NMDAR, we observed as well long-lasting neuronal survival. We propose that the statin metabolite o-ATV given post-OGD boosts the intrinsic pro-survival factor pCREB in large-GABAergic cortical neurons invitro, this contributing to protect them from OGD.

Effect of Fermentation on Compositional Changes of Cinnamomum osmophloeum Kaneh Leaves

Cinnamomum osmophloeum Kaneh is known as “indigenous cinnamon” with the chemical constituents of its leave’s essential oil are similar to the famous C. cassia inner bark oil. Its oil has long been used as a medicinal plant. Fermentation is one of the processes in tea production, which could change the compound’s composition. This research aims to study the compositional changes of C. osmophloeum leaves during fermentation compared to unfermented leaves and commercial tea leaves. The main bioactive secondary metabolites in C. osmophloeum leaves extract are two flavonol glycosides. Both of this glycosides changed into aglycone during fermentation. By using HPLC and LC-MS analysis the major components and their derivative were identified. The retention time of kaempferol aglycone was 35.21 minute and the concentration showed increased from 0.46 to 46.8 µg. mL-1 after fermentation. There are 3 major groups lactic acid bacteria isolated from fermented C. osmophloeum Kaneh leaves, Bacillus coagulans, Lactobacillus plantarum, and Pediococcus pentosaceus, which it plays the key role during the compositional changes of glycosides into aglycone.

Biosynthesis of the psychotropic plant diterpene salvinorin A: Discovery and characterization of the Salvia divinorum clerodienyl diphosphate synthase.

Salvia divinorum commonly known as diviner's sage, is an ethnomedicinal plant of the mint family (Lamiaceae). Salvia divinorum is rich in clerodane-type diterpenoids, which accumulate predominantly in leaf glandular trichomes. The main bioactive metabolite, salvinorin A, is the first non-nitrogenous natural compound known to function as an opioid-receptor agonist, and is undergoing clinical trials for potential use in treating neuropsychiatric diseases and drug addictions. We report here the discovery and functional characterization of two S.divinorum diterpene synthases (diTPSs), the ent-copalyl diphosphate (ent-CPP) synthase SdCPS1, and the clerodienyl diphosphate (CLPP) synthase SdCPS2. Mining of leaf- and trichome-specific transcriptomes revealed five diTPSs, two of which are class II diTPSs (SdCPS1-2) and three are class I enzymes (SdKSL1-3). Of the class II diTPSs, transient expression in Nicotiana benthamiana identified SdCPS1 as an ent-CPP synthase, which is prevalent in roots and, together with SdKSL1, exhibits a possible dual role in general and specialized metabolism. In vivo co-expression and invitro assays combined with nuclear magnetic resonance (NMR) analysis identified SdCPS2 as a CLPP synthase. A role of SdCPS2 in catalyzing the committed step in salvinorin A biosynthesis is supported by its biochemical function, trichome-specific expression and absence of additional class II diTPSs in S.divinorum. Structure-guided mutagenesis revealed four catalytic residues that enabled the re-programming of SdCPS2 activity to afford four distinct products, thus advancing our understanding of how neo-functionalization events have shaped the array of different class II diTPS functions in plants, and may promote synthetic biology platforms for a broader spectrum of diterpenoid bioproducts.

Simultaneous determination of six active metabolites in Astragalus mongholicus (Fisch.) Bge. under salt stress by ultra-pressure liquid chromatography with tandem mass spectrometry.

Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao (A. mongholicus, family Leguminosae) is one of the most important traditional Chinese herbs because it contains lots of bioactive metabolites, which have beneficial and pharmacological effects on health. Simultaneously, it has been proved to be a salt-tolerant plant—one of the potential species to control the soil salinization. Therefore, a sensitive and specific ultra-pressure liquid chromatography coupled with tandem mass spectrometric (UPLC-MS/MS) method was developed and validated for the simultaneous determination of six main bioactive metabolites, astragaloside IV, cycloastragenol, calycosin-7-O-β-d-glucoside, calycosin, ononin and formononetin in different organs of A. mongholicus. The detection was accomplished by multiple-reaction monitoring (MRM) scanning via electrospray ionization source operating in the positive ionization mode. Calibration curves offered linear ranges of two orders of magnitude with R2 > 0.99. The method was fully validated for the linearity, intra-day and inter day precisions, accuracy, recovery, matrix effect and stability. Then this method was successfully applied to detect the content of major bioactive metabolites in different plant organs of A. mongholicus under salt stress. Significant variations in the content of six bioactive metabolites were observed after been processed by different levels of salinity in different part of plant. The results support for further exploration of the salt-tolerant mechanisms in A. mongholicus and its possibility as the species that control the soil salinization. Meanwhile, we established a UPLC-MS/MS assay of the trace components in seedling of A. mongholicus in this study.

Delineation of Platelet Activation Pathway of Scutellarein Revealed Its Intracellular Target as Protein Kinase C.

Erigeron breviscapus has been widely used in traditional Chinese medicine (TCM) and its total flavonoid component is commonly used to treat ischemic stroke, coronary heart disease, diabetes and hypertension. Scutellarin is the major ingredient of E. breviscapus and scutellarein is one of the main bioactive metabolites of scutellarin in vivo, but the latter's pharmacological activities have not been fully characterized. Provided evidence that could inhibit platelet aggregation, the effect of scutellarein on rat washed platelets and its underlying mechanisms were evaluated in our research. Scutellarein inhibited platelet adhesion and aggregation induced by multiple G protein coupled receptor agonists such as thrombin, U46619 and ADP, in a concentration-dependent manner. Furthermore, the mild effect of scutellarein on intracellular Ca(2+) mobilization and cyclic AMP (cAMP) level was observed. On the other hand, the role of scutellarein as potential protein kinase C (PKC) inhibitor was confirmed by PKC activity analysis and molecular docking. The phorbol myristate acetate-induced platelets aggregation assay with or without ADP implied that the scutellarein takes PKC(s) as its primary target(s), and acts on it in a reversible way. Finally, scutellarein as a promising agent exhibited a high inhibition effect on ADP-induced platelet aggregation among its analogues. This study clarifies the PKC-related signaling pathway involved in antiplatelet action of scutellarein, and may be beneficial for the treatment of cardiovascular diseases.

Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS.

As the main bioactive metabolites of Ganoderma lucidum, triterpenoids have various pharmacological effects. In this paper, the nutritional requirements and culture conditions of a submerged culture of G. lucidum were optimized using the response surface methodology; maximum mycelia biomass and intracellular triterpenoid production reached 1.87 g/100 ml and 93.21 mg/100 ml, respectively, for a culture consisting of wort 4.10% (0.041 g/ml) and yeast extract 1.89% (0.0189 g/ml), pH 5.40. For the first time, we established that wort, which is cheap and abundant, can replace the more commonly used glucose as the sole source of carbohydrate. Using high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS), 10 major ganoderic acids were tentatively identified based on the predominant fragmentation pathways with the elimination of H2O and CO2, as well as cleavage of the D-ring.

Bacillus subtilis subsp. subtilis CBMDC3f with antimicrobial activity against Gram-positive foodborne pathogenic bacteria: UV-MALDI-TOF MS analysis of its bioactive compounds.

In this work a new Bacillus sp. strain, isolated from honey, was characterized phylogenetically. Its antibacterial activity against three relevant foodborne pathogenic bacteria was studied; the main bioactive metabolites were analyzed using ultraviolet matrix assisted laser desorption-ionization mass spectrometry (UV-MALDI MS). Bacillus CBMDC3f was phylogenetically characterized as Bacillus subtilis subsp. subtilis after rRNA analysis of the 16S subunit and the gyrA gene (access codes Genbank JX120508 and JX120516, respectively). Its antibacterial potential was evaluated against Listeria monocytogenes (9 strains), B. cereus (3 strains) and Staphylococcus aureus ATCC29213. Its cell suspension and cell-free supernatant (CFS) exerted significant anti-Listeria and anti-S. aureus activities, while the lipopeptides fraction (LF) also showed anti-B. cereus effect. The UV-MALDI-MS analysis revealed surfactin, iturin and fengycin in the CFS, whereas surfactin predominated in the LF. The CFS from CBMDC3f contained surfactin, iturin and fengycin with four, two and four homologues per family, respectively, whereas four surfactin, one iturin and one fengycin homologues were identified in the LF. For some surfactin homologues, their UV-MALDI-TOF/TOF (MS/MS; Laser Induced Decomposition method, LID) spectra were also obtained. Mass spectrometry analysis contributed with relevant information about the type of lipopeptides that Bacillus strains can synthesize. From our results, surfactin would be the main metabolite responsible for the antibacterial effect.

Tissue-specific metabolites profiling and quantitative analyses of flavonoids in the rhizome of Belamcanda chinensis by combining laser-microdissection with UHPLC-Q/TOF-MS and UHPLC–QqQ-MS

The rhizome of Belamcanda chinensis (L.) DC. is a traditionally used medicinal material in China. Due to increasing demand, B. chinensis has been cultivated widely, and thus the study on its rational utilization of medicinal part and guidelines for the optimal cultivation and harvest is an important issue. Considering flavonoids were the main bioactive secondary metabolites of B. chinensis, fluorescence microscopy, laser microdissection (LMD), ultra-high performance liquid chromatography-quadrupole/time-of-flight-mass spectrometry (UHPLC-Q/TOF-MS), and UHPLC coupled with triple quadrupole mass spectrometer (UHPLC–QqQ-MS) were applied to profile and determine flavonoids in various tissues in this study. Consequently, 43 peaks were detected by UHPLC-Q/TOF-MS, and 26 flavonoid compounds combined with seven triterpene compounds were identified or tentatively identified in the tissue extractions. The results indicated that the hydrophobic compounds, especially flavonoid or isoflavonoid aglycones and xanthone mainly accumulated in the cork, whereas the hydrophilic compounds, namely the flavonoid and isoflavonoid glycosides were usually found in the cortex or center (the part inside of endodermis). Samples of rhizomes from different growth ages and origins were simultaneously analyzed. It was shown that the bulb or lateral part of the rhizome generally possessed more total flavonoids than the vertical part or the primordium. The present study established a new practical method to evaluate the quality of the rhizome of B. chinensis and to explore the relationship between distribution patterns of secondary metabolites and growth years of plants, thus important information for cultivation and processing was provided.

Bioequivalence and Pharmacogenetics

The current drug development framework grants the innovator companies the opportunity to market a new drug exclusively for a defined time period under patent protection. After the patent protection period ends, other companies are permitted to produce their own generic version provided they meet bioequivalance. Accordingly, the test compound and the reference compound pharmacokinetic parameters are expected to be within 80% to 125% in terms of Cmax (maximum plasma concentration) and AUC (area under the curve). Generally, bioequivalence trials are carried out in healthy volunteers as a crossover, open, two-way, randomized design where each volunteer takes both test and reference medication in different periods. The remit of a bioequivalence trial is investigating the similarity of test and reference compounds without observing the pharmacodynamics or therapeutic concentration of the medication. If the trial medication has a bioactive metabolite, in this case, both the main molecule and bioactive metabolite should demonstrate bioequivalancy in terms of pharmacokinetic parameters. Both inter- and intra-individual pharmacokinetic variability can be impacted through pharmacogenetic differences and gene expression regulation at an individual and population level. Surprisingly, pharmacogenetics has been considered very little in bioequivalence studies. This expert review article aims to initiate a discussion on the ways in which these two fields of biomedicine offer synergy for innovation, and the ways forward to address the extant challenges at the unique intersection of pharmacogenetics and bioequivalance research and development. Keywords: Bioequivalence, bioavailability, clinical trials, gene expression, personalized medicine, pharmacogenetics, polymorphism.

Purification of antibiotics from the biocontrol agent Streptomyces anulatus S37 by centrifugal partition chromatography

A novel actinomycete strain, Streptomyces anulatus S37, has been isolated from the rhizosphere of healthy Moroccan Vitis vinifera on the basis on its ability to promote grapevine growth and to induce natural defences against various phytopathogens. In the present work, the main bioactive metabolites produced by S. anulatus S37 were isolated. A crude n-BuOH extract of the S37 fermentation broth was firstly partitioned in a biphasic solvent system composed of n-heptane, methanol, and water (5:1.5:3.5, v/v). The most active organic fraction (1.1g) as revealed by TLC-bioautography was subsequently separated by a two-step centrifugal partition chromatography procedure. The first separation was performed in the ascending mode at 6mL/min with the biphasic solvent system n-heptane, ethyl acetate, methanol and water (2:1:2:1, v/v), to finally recover 40mg of a pure compound identified as streptochlorin by NMR spectroscopy. In a second separation, the solvent system n-heptane, acetonitrile, and water (5:5:4, v/v) was used in the ascending mode at 3mL/min to purify 135mg of nigericin and 53mg of piericidin A1. Assays performed with the three compounds have confirmed their inhibitory impact on the growth of Botryris cinerea in dual confrontation and also on V. vinifera L. plantlets.

Bioenergetic and antioxidant effects of a fresh leaf extract from Cynara scolymus L.

Fresh leaf artichoke extracts contain high amounts of cynarin and related caffeoylquinic acids which possess high antioxidant activity. We have investigated mitochondrial stimulation and confirmed radical scavenging activity of cynarin, the lead compound of the special extract [15 – 30:1] from fresh leafs instead of dried material from Cynara cardunculus var. scolymus (L.) Benth., and its main bioactive metabolites caffeic acid, ferulic acid, dihydroferulic acid and their amide derivatives formed after administration of the fresh leaf artichoke extract to humans.