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Tryptophan decarboxylase (TDC) and strictosidine synthase (STR) genes from Catharanthus roseus have been successfully over-expressed in the rol gene integrated cell suspensions of V. minor. Thirty seconds SAAT (sonication-assisted Agrobacterium transformation) treatment of plant cell suspension with LBA1119 having construct (hpt-Tdc2-Str-gus) generated three stable TDC + STR over-expressing cell lines--PVG1, PVG2, and PVG3. The transgenes were confirmed by β-glucuronidase GUS histochemical assay and PCR amplification of rol genes/GUS gene. All the three cell suspension lines were found to be slow growing. In comparison to the control cell suspensions (GI = 241.0 ± 5.8), PVG3 cell line registered a growth index (GI) of 208.0 ± 10.0 followed by PVG1 (GI = 140.0 ± 14.2) and PVG2 (GI = 85.0 ± 9.6). The PVG3 cell line was also up-scaled in the 5-l stirred tank bioreactor with GI of 745.6 ± 35.3 under optimized parameters. Only PVG3 line registered a twofold increase in total alkaloid content (2.1 ± 0.1% dry wt.) and showed vincamine presence (0.003 ± 0.001% dry wt.) which was further enhanced at the bioreactor level (2.7 ± 0.3 and 0.005 ± 0.001% dry wt., respectively). Real-time (RT) qPCR analysis of PVG3 showed more than sevenfold to eightfold increase in TDC and STR expression [relative quantity value (RQ) = 7.6 ± 0.8 (TDC); RQ = 8.5 ± 0.9 (STR)].

Hydroxylase/acetyltransferase elicitors and cyclooxygenase inhibitor along with various precursors from primary shikimate and secoiridoid pools have been fortified to vincamine less hairy root clone of Vinca minor to determine the regulatory factors associated with vincamine biosynthesis. Growth kinetic studies revealed that acetyltransferase elicitor acetic anhydride and terpenoid precursor loganin significantly reduce the growth either supplemented alone or in combination (GI = 140.6 ± 18.5 to 246.7 ± 24.3), while shikimate and tryptophan trigger biomass accumulation (GI = 440.2 ± 31.5 to 540.5 ± 40.3). Loganin also downregulates total alkaloid biosynthesis. Maximum flux towards vincamine production (0.017 ± 0.001 % dry wt.) was obtained when 20-day-old hairy roots were fortified with secologanin (10 mg/l) along with tryptophan (100 mg/l), naproxen (8.4 mg/l), hydrogen peroxide (20 μg/l), and acetic anhydride (32.4 mg/l). This was supported by RT PCR (qPCR) analysis where 2- and 3-fold increase in tryptophan decarboxylase (TDC; RQ = 2.0 ± 0.09) and strictosidine synthase (STR; RQ = 3.3 ± 0.36) activity, respectively, was recorded. The analysis of variance (ANOVA) for growth kinetics, total alkaloid content, and gene expression studies favored highly significant data (P

A vincamine positive hairy root clone of Vinca minor with integration of all the three- rolA, rolB and rolC genes and its subsequently raised cell suspensions were treated with culture filtrate of four endophytic fungi namely Chaetomium globosum; Aspergillus niveoglaucus; Paecilomyces lilacinus and Trichoderma harzianum. Addition of 10 % v/v T. harzianum culture filtrate boosted maximum biomass accumulation and highest total alkaloid content (TAC) both in the hairy roots and in the cell suspensions. Treated (10 % v/v T. harzianum) hairy roots registered growth index (GI) of 640 ± 28.3 and TAC of 3.6 ± 0.1 dry wt% in comparison to non-treated roots (GI = 420 ± 14.2; TAC = 3.1 ± 0.3 dry wt%). Treated cell suspensions showed GI of 600 ± 19.1 and TAC of 1.67 ± 0.03 dry wt% in comparison to non-treated cell suspensions (GI = 250 ± 12.6; TAC = 1.15 ± 0.02 dry wt%). The hairy roots and the cell suspensions were successfully up-scaled in the 5 l stirred tank bioreactor with respective GI of 850.0 and 654.0 under optimized conditions. On Real time (qPCR) analysis, treated hairy roots showed fourfold to sixfold enhanced tryptophan decarboxylase (TDC) transcript level [relative quantity value (RQ) = 4.64 ± 0.30 (shake flask); RQ = 5.95 ± 0.31 (bioreactor)] while treated cell suspensions showed only two fold increase in TDC transcript [RQ = 2.1 ± 0.26 (shake flask); RQ = 2.5 ± 0.21 (bioreactor)]. Similarly, fivefold to sixfold [RQ = 5.6 ± 0.20 (shake flask); RQ = 6.7 ± 0.49 (bioreactor)] and threefold to fourfold [RQ = 3.5 ± 0.18 (shake flask); RQ = 3.8 ± 0.68 (bioreactor)] increased transcript of strictosidine synthase (STR) in treated hairy roots and cell suspensions, respectively was observed. Treated shake flask roots showed 0.002 dry wt% vincamine that was further enhanced in the bioreactor grown treated roots (0.015 dry wt%). No vincamine was detected in the shake flask culture or in the bioreactor grown cell suspensions.

Phyllanthus amarus is a medicinal herb used in traditional Indian medicine for liver disorders. Several researches also show that it acts primarily in the liver, but the molecules were unidentified for liver protective activity. This study was to determine whether the lignans isolated from P. amarus attenuates the D-galactosamine (GalN) / Lipopolysaccharide (LPS)- induced acute hepatitis in mice. Standardize mixture of lignans (slPA) isolated from leaves of P. amarus using automated chromatographic technique was used for experiments. Experimental mice were orally pre-treated with slPA (10, 30 and 100 mg/kg) for 7 days before intra-peritoneal injection of GalN/LPS. Acute hepatitis in mice was confirmed by significant increase of pro-inflammatory cytokines, and hepatotoxic markers. Pre-treatment of slPA exhibit significant liver protection in dose dependant mannaer. In-silico molecular docking studies also suggests that lignans are preferentially more active due to strong binding affinity against pro-inflammatory cytokines; IL-1β, IL-6, and TNF-α. The electronic parameters of lignans for bioavailability, drug likeness and toxicity were within the acceptable limit. In-vivo and in-silico results suggest that pretreatment of slPA exhibit potent hepatoprotection against GalN/LPS-induced hepatitis in mice and the liver protective effects may be due to the inhibition of inflammatory mediators.

Elicitors play an important role in challenging the plant defense system through plant-environment interaction and thus altering the secondary metabolite production. Culture filtrates of four endophytic fungi, namely, Chaetomium globosum, Aspergillus niveoglaucus, Paecilomyces lilacinus, and Trichoderma harzianum were tested on embryogenic cell suspensions of latex-less Papaver somniferum in dose-dependent kinetics. Besides this, abiotic elicitors salicylic acid, hydrogen peroxide, and carbon dioxide were also applied for improved sanguinarine production. Maximum biomass accumulation (growth index (GI) = 293.50 ± 14.82) and sanguinarine production (0.090 ± 0.008 % dry wt.) were registered by addition of 3.3 % v/v T. harzanium culture filtrate. Interestingly, it was further enhanced (GI = 323.40 ± 25.30; 0.105 ± 0.008 % dry wt.) when T. harzanium culture filtrate was employed along with 50 μM shikimate. This was also supported by real-time (RT) (qPCR), where 8-9-fold increase in cheilanthifoline synthase (CFS), stylopine synthase (STS), tetrahydroprotoberberine cis-N-methyltransferase (TNMT), and protopine 6-hydroxylase (P6H) transcripts was observed. Among abiotic elicitors, while hydrogen peroxide and carbon dioxide registered low level of sanguinarine accumulation, maximum sanguinarine content was detected by 250 μM salicylic acid (0.058 ± 0.003 % dry wt.; GI = 172.75 ± 13.40). RT (qPCR) also confirms the downregulation of sanguinarine pathway on CO2 supplementation. Various parameters ranging from agitation speed (70 rpm), impeller type (marine), media volume (2 l), inoculum weight (100 g), and culture duration (9 days) were optimized during upscaling in 5-l stirred tank bioreactor to obtain maximum sanguinarine production (GI = 434.00; 0.119 ± 0.070 % dry wt.). Addition of 3.3 % v/v T. harzanium culture filtrate and 50-μM shikimate was done on the 6th day of bioreactor run.

In North India, poultice of young unfolded leaves ofArgyreia speciosaLinn. (Convolvulaceae) is used for healing wounds. In order to find scientific evidence for the traditional utilization of leaves ofA. speciosain wound healing, this investigation was carried out. A linear incision wound of about 3 cm in length and 2 mm in depth and circular excision wound of 177 mm2full thickness were made on the dorsal region of separate groups (n=5) of anesthetized Swiss albino mice. A simple ointment, developed by including ethanol, ethanol-water, and water extracts (10% each, separately) ofA. speciosa, was applied topically to mice once daily for 14 days after wounding. To evaluate the effect of each extract, wound contraction, epithelization period, wound breaking strength, and hydroxyproline content were determined. The water extract ofA. speciosashowed accelerated wound healing activity as evidenced by fast wound contraction (96.30±0.52%;P<0.01), rapid epithelization period (11.40±0.60days;P<0.001), greater wound breaking strength (376.56±21.16 g;P<0.001), and higher hydroxyproline content (16.49±1.12 mg/g;P<0.05) of granulation tissue. The present report supports the traditional use ofArgyreia speciosaleaves for wound healing and signify its relevant therapeutic potential.

Breast cancer is the most prevalent cancer in women affecting about 12% of world's female population. It is a multifactorial disease, mostly invasive in nature. Diosgenin and related compounds are potent antiproliferative agents. Carbamate derivatives have been synthesized at C26 of furostene ring after opening spiroketal bond (F-ring) of diosgenin. Compound 10 possessed significant antiproliferative activity against human breast cancer cells by arresting the population at G1 phase of cell division cycle and induced apoptosis. Induction of apoptosis was observed through the caspase signalling cascade by activating caspase-3. Moreover, carbamate 10 exhibited moderate antiinflammatory activity by decreasing the expression of cytokines, TNF-α and IL-6 in LPS-induced inflammation in primary macrophage cells. Furthermore, compound 10 significantly reduced Ehrlich ascites carcinoma significantly in mice. It was well tolerated and safe in acute oral toxicity in Swiss albino mice. The concomitant anticancer and antiinflammatory properties of carbamate 10 are important and thus, can further be optimized for a better anti-breast cancer candidate.

In the present study, we report that neolignan1 (Diethyl-4,4ʹ-dihydroxy-8,3ʹ-neolign-7,7ʹ-dien-9,9ʹ-dionate) relaxes the superior mesenteric artery in a concentration dependent manner (pD2 value 5.392 ± 0.04; n = 8 for endothelium intact and 5.204 ± 0.03; n = 8 for endothelium denuded mesenteric rings, respectively). The relaxation response of neolignan1 was found to be endothelium independent and sensitive to 1H-[1,2,4] oxadiazolo [4,3-a]quinoxalin-1-on (ODQ; 1 μM) and tetraethyl ammonium (TEA; 1 mM). In-silico studies showed good LibDock score (92.66) of neolignan1 with BKCa channel and are in well corroboration with ex-vivo study. Further, neolignan1 significantly decreased the systolic blood pressure, diastolic blood pressure and mean arterial pressure in the Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME; 50 mg/kg) treated Wistar rats at the dose of 30 and 100 mg/kg given once orally for 15 days. In addition, neolignan1 is well tolerated up to 100 mg/kg when given as a repeated dose, once orally for 28 days in Swiss albino mice. Neolignan1 was well absorbed from oral route, reached peak at 4 h and eliminated below detection level by 12 h after administration. Our present study concludes that neolignan1 produced relaxation in superior mesenteric artery by opening of BKCa channel and produced significant antihypertensive activity in L-NAME treated Wistar rats and was well tolerated by the experimental animal.

Sweet basil (Ocimum basilicum) is well known for its diverse pharmacological properties and has been widely used in traditional medicine for the treatment of various ailments. Although a variety of secondary metabolites with potent biological activities are identified, our understanding of the biosynthetic pathways that produce them has remained largely incomplete. We studied transcriptional changes in sweet basil after methyl jasmonate (MeJA) treatment, which is considered an elicitor of secondary metabolites, and identified 388 candidate MeJA-responsive unique transcripts. Transcript analysis suggests that in addition to controlling its own biosynthesis and stress responses, MeJA up-regulates transcripts of the various secondary metabolic pathways, including terpenoids and phenylpropanoids/flavonoids. Furthermore, combined transcript and metabolite analysis revealed MeJA-induced biosynthesis of the medicinally important ursane-type and oleanane-type pentacyclic triterpenes. Two MeJA-responsive oxidosqualene cyclases (ObAS1 and ObAS2) that encode for 761- and 765-amino acid proteins, respectively, were identified and characterized. Functional expressions of ObAS1 and ObAS2 in Saccharomyces cerevisiae led to the production of β-amyrin and α-amyrin, the direct precursors of oleanane-type and ursane-type pentacyclic triterpenes, respectively. ObAS1 was identified as a β-amyrin synthase, whereas ObAS2 was a mixed amyrin synthase that produced both α-amyrin and β-amyrin but had a product preference for α-amyrin. Moreover, transcript and metabolite analysis shed light on the spatiotemporal regulation of pentacyclic triterpene biosynthesis in sweet basil. Taken together, these results will be helpful in elucidating the secondary metabolic pathways of sweet basil and developing metabolic engineering strategies for enhanced production of pentacyclic triterpenes.

Papaver somniferum produces therapeutically useful benzylisoquinoline alkaloids (BIAs) like papaverine, thebaine, codeine, and morphine that accumulate in its capsular latex. Morphine is a potent analgesic but is also abused as a narcotic, which has increased the demand for non-narcotic thebaine that can be converted into various analgesics. To curtail the narcotic menace, many distinct genotypes of the plant have been developed that are deficient in morphine and/or latex. Sujata is one such latex-less low alkaloid-producing variety developed from the alkaloid-rich gum harvest variety Sampada. Its utility for gene prospecting and studying differential gene regulation responsible for its low alkaloid, nutritive seed oil, and latex-less phenotype has been exploited in this study. BIA profiling of Sujata and Sampada capsules at the early and late stages indicated that except for thebaine, Sujata had a depressed alkaloid phenotype as compared to Sampada. Comparative transcript-based analysis of the two genotypes was carried out in the early stage capsule (higher thebaine) using subtractive hybridization and microarray. Interrogation of a P. somniferum array yielded many differentially expressing transcripts. Their homology-based annotation classified them into categories--latex related, oil/lipid related, alkaloid related, cell wall related, and others. These leads will be useful to characterize the highly sought after Sujata phenotype.

Withania somnifera (Ashwagandha), globally known as Indian Ginseng, is an Ayurvedic “Rasayana” herb which possesses medicinal and pharmacodynamic properties. It is used as one of the ingredients in the Indian herbal tea. Its use as functional food with immunostimulating and rejuvenating properties is very popular and available over the counter in the Asian market. Here we report its enrichment to achieve glycone and aglycone fractions with respect to its three key withanolides and immune-stimulant potential. Modulations of two cytokines, tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10) were also estimated. Pharmacokinetic experiments using LC–MS were performed in mice after a single oral administration of crude extract and enriched aglycone fraction. The proposed scheme is most effective for the enrichment of 12-deoxy-withastramonolide as compared to other two withanolides i.e. withaferin-A and withanolide-A. Acid and base treatment not only degrades the key marker compounds but also alters their respective HPTLC finger print. The slow bioavailability and elimination of withaferin A from the crude methanol extract administration were evidenced higher T1/2 (h) and AUC(0–t) (h μg/L) as compared to WSAg. This study established the immune enhancing properties of the enriched fractions of W. somnifera and also confirming that the effect is cell mediated and humoral. The enriched aglycone fraction could find use as an immune-stimulatory agent in humans.

Ethnopharmacological relevance: Many of the effective therapeutic strategies have been derived from ethnopharmacologically used natural products. Pluchea lanceolata is an herb employed in Indian folk medicine for malaria like fever but it lacks proper pharmacological intervention. Aim of the study: To evaluate antimalarial and safety profile of Pluchea lanceolata: an in-vitro, in-vivo for its ethnopharmacological validation. Materials and methods: Methanol, butanol, ethyl acetate, chloroform, hexane extracts and its isolate, taraxasterol acetate (TxAc) were obtained from air dried aerial part of Pluchea lanceolata. These were tested in-vitro against chloroquine-sensitive strain of Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase activity. The most potent hexane extract and TxAc were further validated for in-vivo antimalarial and safety evaluation. TxAc, a pentacyclic-triterpene isolated from the most active fraction was further evaluated with special emphasis on inflammatory mediators involved in malaria pathogenesis. Murine malaria was induced by intra-peritoneal injection of Plasmodium berghei infected red blood cells to the male Swiss inbred mice. Mice were orally treated following Peters 4-Day suppression test. In-vivo antimalarial efficacy was examined by evaluating the parasitaemia, percent survival, mean survival time, blood glucose, haemoglobin and pro-inflammatory mediators involved in malaria pathogenesis. Results: Hexane extract and TxAc showed promising antimalarial activity in-vitro and in-vivo condition. TxAc attributed in inhibition of the pro-inflammatory cytokines as well as afford to significant increase in the blood glucose and haemoglobin level when compared with vehicle treated infected mice. We have not observed the synergistic action of combinations of chloroquine and TxAc from our experimental results. In-vitro and in-vivo safety evaluation study revealed that hexane extract is non toxic at higher concentration. Conclusion: Present study further validates the ancient Indian traditional knowledge and use of Pluchea lanceolata as an antimalarial agent. Study confirms the suitability of Pluchea lanceolata as a candidate for further studies to obtain a prototype for antimalarial medicine.

Catharanthus roseus is the sole source of two most important monoterpene indole alkaloid (MIA) anti- cancer agents: vinblastine and vincristine. MIAs possess a terpene and an indole moiety derived from terpenoid and shikimate pathways, respectively. Geranyl diphosphate (GPP), the entry point to the formation of terpene moiety, is a product of the condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by GPP synthase (GPPS). Here, we report three genes encoding proteins with sequence similarity to large subunit (CrGPPS.LSU) and small subunit (CrGPPS.SSU) of heteromeric GPPSs, and a homomeric GPPSs. CrGPPS.LSU is a bifunctional enzyme producing both GPP and geranyl geranyl diphosphate (GGPP), CrGPPS.SSU is inactive, whereas CrGPPS is a homomeric enzyme forming GPP. Co-expression of both subunits in Escherichia coli resulted in heteromeric enzyme with enhanced activity producing only GPP. While CrGPPS.LSU and CrGPPS showed higher expression in older and younger leaves, respectively, CrGPPS.SSU showed an increasing trend and decreased gradually. Methyl jasmonate (MeJA) treatment of leaves sig- nificantly induced the expression of only CrGPPS.SSU. GFP localization indicated that CrGPPS.SSU is plastidial whereas CrGPPS is mitochondrial. Transient overexpression of AmGPPS.SSU in C. roseus leaves resulted in increased vindoline, immediate monomeric precursor of vinblastine and vincristine. Although C. roseus has both heteromeric and homomeric GPPS enzymes, our results implicate the involvement of only heteromeric GPPS with CrGPPS.SSU regulating the GPP flux for MIA biosynthesis.

Objective To validate the traditional use of Cissampelos pareira as an antidiabetic agent in streptozotocin–nicotinamide induced diabetic male mice. Methods Antidiabetic effect of aqueous extract of C. pareira leaves (CPAE) was evaluated at 250 mg/kg and 500 mg/kg body weight, p.o. doses in male albino mice over the period of 14 days. Random blood glucose level and body weight were observed periodically. Liver glycogen level, organ coefficient and other biochemical parameters were also determined after completion of the study. Result Significant ( p p p Conclusions The present study concluded that the aqueous extract of C. pareira at 500 mg/kg body weight was capable in reducing diabetic attritions so it might be a valuable candidate for diabetes treatment.

Crude extracts of Punica granatum fruit peel, its isolates, and semisynthetic analogs were investigated for their Cathepsin D protease reticence. Hexane extract (HE), chloroform extract (CE), and ethyl acetate extract (EAE) exhibited significant inhibition of Cathepsin D activity. The bioactivity-guided isolation of hexane- and chloroform-soluble fractions yielded piperine and ursolic acid as bioactive constituents. Upon semisynthetic modifications of ursolic acid, two of the analogs (7 and 8) exhibited higher inhibition of Cathepsin D activity: IC50 at 12.87 and 9.40 μM, respectively. P. granatum fruit peel extracts may act as breast cancer chemopreventive agent.

Microorganisms play an important role in the development of periodontal diseases in human that lead to serious complications. Since ages, medicinal plants, singly or in combinations are used for the treatment of various dental disorders. However, their scientific validation and characterization is scarce. Therefore, the goal of this study was to develop knowledge based poly-herbal formulation as a remedy for oral hygiene. After validations of antimicrobial potential, extracts of 11 plants were combined in different ratios, and finally one poly-herbal formulation was selected for further characterization. Antimicrobial activity was determined through disc and well diffusion assays. Further level of tumour necrosis factor (TNF-α) for assessing anti-inflammatory activity and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for safety assessment was determined. High performance thin layer chromatography (HPTLC) fingerprint profile was developed using thin layer chromatography (TLC) scanner and densitometer. The poly-herbal formulation, PHF 12-05 showed antimicrobial activity against both pathogenic bacteria and fungi with zone of inhibition ranging from 7 to 13 mm. PHF 12-05 was found to possess anti-inflammatory activity by significantly (P

The present study describes anticancer effect of gallic acid based indanone derivative (1). Indanone 1 exhibited in vivo anticancer activity against Erhlich ascites carcinoma in Swiss albino mice by inhibiting tumor growth by 54.3% at 50 mg/kg b.wt. It showed antitubulin effect by inhibiting tubulin polymerase enzyme. In cell cycle analysis, it inhibited G2/M phase and induced apoptosis. It significantly suppressed VEGF-R1, VEGF-R2 and HIF-α in human breast cancer MCF-7 cells, thus exhibiting antiangiogenic activity. In acute oral toxicity, indanone 1 was well tolerated and was found to be non-toxic up to 1000 mg/kg b.wt. in Swiss albino mice. Pharmacokinetic studies in rabbits revealed rate of absorption, half life, volume of distribution with high plasma and blood clearance after i.v. administration. Indanone 1, is a safe and moderately active anticancer agent.

Background Flacourtia indica is especially popular among the various communities of many African countries where it is being used traditionally for the treatment of malaria. In our previous report, we have identified some phenolic glycosides from the aerial parts of F. indica as promising antiplasmodial agents under in vitro conditions. Purpose Antimalarial bioprospection of F. indica derived phenolic glycoside in Swiss mice (in vivo) with special emphasis on its mode of action. Methods Chloroquine sensitive strain of Plasmodium falciparum was routinely cultured and used for the in vitro studies. The in vivo antimalarial potential of phenolic glycoside was evaluated against P. berghei in Swiss mice through an array of parameters viz., hematological, biochemical, chemo-suppression and mean survival time. Results 2-(6-benzoyl-β- d -glucopyranosyloxy)-7-(1α, 2α, 6α-trihydroxy-3-oxocyclohex-4-enoyl)-5-hydroxybenzyl alcohol (CPG), a phenolic glycoside isolated from the aerial parts of F. indica was found to exhibit promising antiplasmodial activity by arresting the P. falciparum growth at the trophozoite stage. Spectroscopic investigations reveal that CPG possesses a strong binding affinity with free heme moieties. In addition, these interactions lead to the inhibition of heme polymerization in malaria parasite, augmenting oxidative stress, and delaying the rapid growth of parasite. Under in-vivo condition, CPG exhibited significant antimalarial activity against P. berghei at 50 and 75 mg/kg body weight through chemo-suppression of parasitemia and ameliorating the parasite induced inflammatory and oxidative (hepatic) imbalance in the experimental mice. Conclusion CPG was found to be a potential antimalarial constituent of F. indica with an explored mechanism of action, which also offers the editing choices for developing CPG based antimalarial chemotypes.

Not much is known about the mechanism of endophyte-mediated induction of secondary metabolite production in Catharanthus roseus. In the present study two fungal endophytes, Curvularia sp. CATDLF5 and Choanephora infundibulifera CATDLF6 were isolated from the leaves of the plant that were found to enhance vindoline content by 229–403%. The isolated endophytes did not affect the primary metabolism of the plant as the maximum quantum efficiency of PSII, net CO2 assimilation, plant biomass and starch content of endophyte-inoculated plants was similar to endophyte-free control plants. Expression of terpenoid indole alkaloid (TIA) pathway genes, geraniol 10-hydroxylase (G10H), tryptophan decarboxylase (TDC), strictosidine synthase (STR), 16-hydoxytabersonine-O-methyltransferase (16OMT), desacetoxyvindoline-4-hydroxylase (D4H), deacetylvindoline-4-O-acetyltransferase (DAT) were upregulated in endophyte-inoculated plants. Endophyte inoculation upregulated the expression of the gene for transcriptional activator octadecanoid-responsive Catharanthus AP2-domain protein (ORCA3) and downregulated the expression of Cys2/His2-type zinc finger protein family transcriptional repressors (ZCTs). The gene for the vacuolar class III peroxidase (PRX1), responsible for coupling vindoline and catharanthine, was upregulated in endophyte-inoculated plants. These endophytes may enhance vindoline production by modulating the expression of key structural and regulatory genes of vindoline biosynthesis without affecting the primary metabolism of the host plant.

Cinnamate-4-hydroxylase (C4H) converts trans-cinnamic acid (CA) to p-coumaric acid (COA) in the phenylpropanoid/lignin biosynthesis pathway. Earlier we reported increased expression of AaCYP71AV1 (an important gene of artemisinin biosynthesis pathway) caused by CA treatment in Artemisia annua. Hence, AaC4H gene was identified, cloned, characterized and silenced in A. annua with the assumption that the elevated internal CA due to knock down may increase the artemisinin yield. Accumulation of trans-cinnamic acid in the plant due to AaC4H knockdown was accompanied with the reduction of p-coumaric acid, total phenolics, anthocyanin, cinnamate-4-hydroxylase (C4H) and phenylalanine ammonia lyase (PAL) activities but increase in salicylic acid (SA) and artemisinin. Interestingly, feeding trans-cinnamic acid to the RNAi line increased the level of artemisinin along with benzoic (BA) and SA with no effect on the downstream metabolites p-coumaric acid, coniferylaldehyde and sinapaldehyde, whereas p-coumaric acid feeding increased the content of downstream coniferylaldehyde and sinapaldehyde with no effect on BA, SA, trans-cinnamic acid or artemisinin. SA is reported earlier to be inducing the artemisinin yield. This report demonstrates the link between the phenylpropanoid/lignin pathway with artemisinin pathway through SA, triggered by accumulation of trans-cinnamic acid because of the blockage at C4H.

BACKGROUND Flacourtia indica (Burm. f.) Merr. is a medicinal plant indigenous to India and is broadly used worldwide for the treatment of a variety of health ailments. The present study was experimented on hyperlipidemic Charles Foster rats with the aim to explore the possible mechanism responsible for the antidyslipidemic activity of the hydromethanolic extract from F. indica leaves (FIL). METHODS Hyperlipidemia was induced by a single intraperitoneal dose of Triton WR-1339 in Charles Foster rats. The plasma lipid levels were estimated in control and treated groups. The antioxidant potential of F. indica was assessed in both enzymatic and non-enzymatic systems. An acute toxicity study of high-performance liquid chromatography (HPLC)-fingerprinted extract was carried out in Swiss albino mice. RESULTS The F. indica extract at a dose of 150 mg/kg significantly lowers the plasma level of total cholesterol (17%), triglycerides (13%), and phospholipids (16%) by increasing post-heparin lipolytic activity (19%) and lecithin-cholesterol-acyltransferase activity (20%) in Triton-induced hyperlipidemic rats. In addition, the F. indica extract showed significant in vitro antioxidant and anti-adipogenic activity. HPLC analysis indicates the presence of flavanones and flavones in the extract, and the extract was found to be non-toxic up to a dose of 2000 mg/kg body weight in the acute oral toxicity study. CONCLUSIONS These finding suggest that F. indica holds significant potential in preventing clinical deterioration induced by dyslipidemia along with oxidative stress.

Vinca minor is the sole source of vincamine, an alkaloid known to be used in a variety of cerebral disorders. Three stable variant shoot lines (V10, V20 and V30) with tolerance thresholds of 10, 20 and 30 mg/l 5-methyltryptophan (5-MT; analogue of tryptophan), respectively, were selected. These lines showed twofold to threefold increase in tryptophan content and 1.5- to 2-fold increment in the total alkaloids in comparison to the wild line shoots. A maximum of 16-fold enhancement in vincamine production was recorded in V30 line followed by eightfold in V20 line. Inter simple sequence repeat (ISSR)-PCR amplification of all the three lines showed total of 65 bands; out of which 60 were monomorphic (92.3 %) and 5 were polymorphic (7.7 %). Tryptophan being a limiting factor in the indole alkaloid pathway plays a crucial role in modulating the flux towards vincamine production and its over-production positively resulted into enhanced vincamine production.

Ocimum sanctum is well known for its wide range of therapeutic activities. In this article, a methodology to define the safety of the herb with its chromatographic fingerprint has been established. We report the fingerprint method for three standardized extracts of O. sanctum and the correlation of the chromatographic peaks with their in vivo pharmacological safety profile. HPLC fingerprints with the statistical similarity correlation were validated and used in acute and sub-acute toxicity studies in Charles Foster rats. Under optimized chromatographic conditions, numbers of constituents of O. sanctum were successfully separated by the reverse-phase HPLC methods. Various groups have been marked in the chemical fingerprint. The effects on body weight and biochemical indices were mainly related to the chromatographic peaks of group II and V. On the basis of characteristic UV spectra, it was observed that the peaks of group II were related with phenolic moieties. Further, the peak number 11 of group II was characterized as rosmarinic acid—a naturally occurring polyphenolic compound with antioxidant and anti-inflammatory properties by reference comparison. All three extracts in both acute and sub-acute toxicity experiments were well tolerated, and no adverse changes were observed in mortality, morbidity, gross pathology, body weight, and biochemical parameters. This accurate and reproducible method can be used for the chemical identification in standardized extract of O. sanctum leaves, batch-to-batch quality assurance, adulteration, and their safety evaluation.

Mango ginger (Curcuma amada) is a spice of high usage in pickles, sauce, culinary formulations and traditional/folk systems of medicine for therapeutic actions in Asian countries. After establishing scheme for isolation and characterisation of labdane diterpene dialdehyde [labda-8(17), 12-diene-15, 16-dial]-an anti-tubercular agent, we developed a new validated HPLC-PDA method for its quantification in C. amada. Chromatography was performed with reverse phase column in isocratic condition at a flow rate of 1.0 mL/min using mobile phase of acidified water and acetonitrile. Specificity of determination was achieved with UV (190–400 nm) and mass spectrum. Good linearity was obtained with correlation coefficients >0.99. In the analytical range the precision and accuracy (RSD%) values were ⩽2%. Present validated method allowed both the identification and determination of labda-8(17), 12-diene-15, 16-dial, in C. amada rhizome. The method was applied to screen the labdane diterpene dialdehyde in the samples of different geographic locations.

The lipophilic character of phytol derivatives has been studied using reverse-phase planar chromatographic procedures. Methanol-water and acetonitrile-water binary mixtures as mobile phases with C18 and cyano as stationary phases were used in order to determine the lipophilicity parameters and C0. The classical values were compared with the factor scores obtained by principal component analysis based also on the TLC retention data. Moderate to high correlation between the values and slopes (specific hydrophobic surface area) was observed, which reflects the involvement of secondary interactions in the TLC retention behavior. The phytol derivatives considered in this study were screened for their antituberculosis activity against the H37Rv strain. Chromatographically obtained lipophilicity parameters were correlated with the calculated log P values and minimum inhibitory concentration (MIC) values. Principal component analysis established the dominant pattern in the chromatographic indices. Quantitative structure-retention relationship and quantitative structure-activity (antitubercular) relationship investigations were performed on the lipophilicities and molecular descriptors of phytol derivatives using partial least squares. The predicted biological activities support the fact that the chromatographic processes of the investigated phytol derivatives influence the partitioning over biomembrane and are controlled mainly by the lipophilicity.

Fifteen hairy root lines of Catharanthus roseus were compared for growth, alkaloid production and shoot bud regeneration potential in relation to their morphology and pattern of rol gene(s) integration. These lines could be classified into 10 distinct morphotypes on the basis of their thickness and branching behaviour. Twelve root lines showed the presence of both rolB and rolC and two indicated the integration of only rolC. Genes rolA, B and C were only detected in N5 line. The root lines did not differ significantly for their total alkaloids content (1.10–1.30% dry wt.) but showed variations in relative concentration of ajmalicine (0.007–0.08% dry wt.), serpentine (0.01–0.08% dry wt.) and catharanthine (0.01–0.04% dry wt.). Line PG2 with highest biomass accumulation (GI = 326.73) and line PI3 with highest catharanthine content were up-scaled in 5-l bioreactor wherein a 9–11 folds increment in biomass accumulation was obtained with two folds enhancement in ajmalicine (0.029 % dry wt.) in PI3 lin...

The leaf explants of Stevia rebaudiana were exposed to a range of EMS (chemical mutagen) and gamma radiation (physical mutagen) doses. Among all the treatments, doses of 0.4 % v/v EMS and 0.95 KR gamma radiation were found to be most effective treatment for selection of variants via direct shoot bud induction. The field-established plants designated as EMS (E), gamma (G), and control (C) plants were subjected to photosynthetic parameter studies where E and G plants showed lower chlorophyll content, transpiration rate, CO2 exchange rate, and stomatal conductance. The implanted field soil at the time of harvesting of plants showed reduction in organic carbon (OC), electrical conductivity (EC), phosphorus (P)/potassium (K), and total Kjeldahl nitrogen (TKN). The ISSR profiling of E, G, and C scored total of 107 bands out of which 63 % were polymorphic bands, and plant E was found to be more distant from the C plant phylogenetically. On phytochemical analysis, the G plant registered twofold enhanced rebaudioside A with lower stevioside (3.2 ± 0.22 % dry wt.) content, whereas E plant showed more than 1.5- and 2.0-fold increase in stevioside and rebaudioside A as compared to control plants. Both the E (3.1 ± 0.15 % dry wt.) and G (2.3 ± 0.21 % dry wt.) plants registered lower steviol. The enhanced steviol glycoside profile was supported by the RT-PCR analysis of UGT74G1 and UGT76G1 that corresponds to stevioside and rebaudioside A biosynthesis, respectively. While the E plant showed 5–6-fold increase in the UGT74G1 (RQ = 5.51 ± 0.5) and UGT76G1 (RQ = 6.61 ± 0.5) gene expression, the G plant showed 5-fold increase in UGT76G1 (RQ = 5.29 ± 0.2) gene expression.

Micelle-mediated extraction and cloud-point preconcentration method was developed for the separation and determination of andrographolide (ANDRO) and neo-andrographolide (NEO) from Andrographis paniculata by planar and high performance liquid chromatography with photodiode array detection. Comparative efficiency and feasibility of employing non-ionic surfactants, namely, Triton X-100, Triton X-114, and Genapol X-80 as an alternative and effective solvent have been demonstrated. Ultrasonic assisted extraction, when compared with common organic solvents, at 5% surfactant (Triton X-100) concentration has yielded higher extraction efficiency of ANDRO and NEO than common organic solvents. The preconcentration factor was approximately 7.0 and the cloud-point extraction recoveries for the two labdane diterpenoids were 90.28 and 98.80%, respectively. The HPLC method consisting monolithic column and isocratic elution with water: acetonitrile (75:25, v/v) has shown advantages over C18 column, in terms of noninterfe...

1-Chloro-2-formyl indenes and tetralenes have been synthesized using Vilsmeier-Haack-Arnold reaction onto indanones and tetralones. Most of these analogues exhibited antitubercular activity against Mycobacterium tuberculosis H37Rv strain with MICs ranging from 30 to 500 μg/mL. Analogue 13 was further modified to some derivatives. The most active analogue 23 showing MIC at 30 μg/mL was further evaluated for acute oral toxicity in Swiss albino mice and was found to be safe up to 300 mg/kg dose.

Summary An online-hyphenated high-performance liquid chromatography-photodiode array-mass spectrometry (HPLC-PDA-MS) analytical method was developed for the simultaneous determination of six lignans of therapeutic importance in four Phyllanthus spp. (P. amarus, P. maderaspatensis, P. urinaria, and P. virgatus). HPLC with monolithic reverse phase silica column (4.6 × 100 mm) and simple isocratic elution of methanol-water mixed with dioxane facilitated the separation of lignans of diverse nature such as diarylbutyrolactone, tetrahydrofuran, isomeric aryltetralin, and diarylbutane type for quantitative analysis. Targeted lignans viz. heliobuphthalmin lactone (1), virgatusin (2), hypophyllanthin (3), phyllanthin (4), nirtetralin (5), and niranthin (6) were confirmed unambiguously in four Phyllanthus species by their abundant molecular adduct ions, retention time, UV, and mass spectra as compared with those of reference compounds. Advantages and limitations of both detection techniques for qualitative (fingerp...

A method was developed based on multiple approaches wherein DNA and chemical analysis was carried out toward differentiation of important species of Sida complex that is being used for commercial preparation. Isolated DNA samples were successfully performed through PCR amplification using ISSR markers and degree of genetic diversity among the different species of Sida is compared with that of chemical diversity. For genetic fingerprint investigation, selected 10 ISSR primers generating reproducible banding patterns were used. Among the total of 63 amplicons, 62 were recorded as polymorphic, genetic similarity index deduced from ISSR profiles ranged from 12 to 51%. Based on similarity index, S. acuta and S. rhombifolia found to be most similar (51%). High number of species-specific bands played pivotal role to delineate species at genetic level. Investigation based on HPTLC fingerprints analysis revealed 23 bands representing to characteristic chemicals and similarity index ranged from 73 to 91%. Prominent distinguishable bands were observed only in S. acuta, while S. cordifolia and S. rhombifolia shared most bands making them difficult to identify on chemical fingerprint basis. This report summarizes the genotypic and chemotypic diversity and the use of profiles for authentication of species of Sida complex.

Thirty‐two distinct accessions of Papaver somniferum were screened for morphinan alkaloid content in the straw. The combined content of major morphinan alkaloids (morphine+codeine+thebaine) was found to vary in the range 0.2260–0.0683%. Two genotypes each, were selected as prototypes for low [I‐48 (0.0683%) and I‐344 (0.0878%)] and high [Pps‐1 (0.2260%) and N‐3 (0.2074%)] morphinan alkaloid content for studying DNA polymorphism. RAPD analysis of these four genotypes using 80 primers could not detect the polymorphism. However, AFLP analysis of these genotypes with 12 EcoRI/MseI primer pairs could distinctly group the high‐ and low‐morphinan alkaloid genotypes separately. Furthermore, 50 AFLP fragments, specific to high‐straw morphinan alkaloid genotypes (Pps‐1 and N‐3) and 28 DNA fragments specific to low‐straw morphinan alkaloid genotypes (I‐48 and I‐344) could be identified. This investigation is the first report on the polymorphism identified in the genotypes differing in their straw morphinan ...

Methods based on HPTLC and RP-HPLC with UV detection for rapid quantitative determination of two major plant growth promoters in Callicarpa macrophylla, calliterpenone (1) and calliterpenone monoacetate (2) are described. The recoveries of the two compounds were between 97.5–100.8% by HPTLC method and 99.3–100.9% by HPLC assay. The relative standard deviations of the two compounds ranged between 1.26–1.68 (Intra-day) and 1.06–1.68 (Inter-day) for HPTLC and 0.02–0.92 (Intra-day) and 0.03–0.92 (Inter-day) for HPLC. The methods were used for routine analysis of two compounds in the leaves of the plant.

An LC method is developed for the quantitation of rhoifolin in Uraria picta, a plant of high usage frequency in all Asian traditional systems of medicine. An isocratic RP-LC method using C18 column, UV detection 265 nm and specificity with PDA and MS is speeding up, reliable and comprehensive analysis of rhoifolin in U. picta. Good linearity was obtained in the working range (0.02–0.10 mg mL−1), with correlation coefficients >0.99. LOD and LOQ were 2.33 and 7.69 ng, respectively. The method was validated following international guidelines. The described method can be utilized for assays and stability tests of U. picta extracts as well as Ayurvedic drugs based on Prishniparni.

Chemical characterization and acute and sub-acute toxicity study of Trikatu, a generic herbal formulation of Indian system of medicine, was carried out in Charles Foster (CF) rats for safety profiling. In acute toxicity experiment, Trikatu at 2,000 mg/kg body weight once orally was well tolerated by the experimental animals (both male and female) and no changes were observed in mortality, morbidity, gross pathology, gain in weight, vital organ weight, hematological (total white blood cells (WBC) and red blood cells (RBC) count), biochemical parameters such as serum creatinine, serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum lipid profile and tissue biochemical parameters such as reduced glutathione and malonaldehyde content as oxidative stress markers. In sub-acute experiment, Trikatu was administered at 5, 50 and 300 mg/kg body weight once daily for 28 days in female CF rats, and non-significant changes were found in most of the parameters studied such as acute experiment except significant increase in low density lipoprotein (LDL) cholesterol level at 50 and 300 mg/kg body weight, decrease in high density lipoprotein (HDL) cholesterol level at 300 mg/kg body weight, increase in SGPT activity at 50 mg/kg body weight and decrease in WBC count at 300 mg/kg body weight on 28(th) day post treatment.

A formulation of Ocimum sanctum, Phyllanthus emblica, and Glycyrrhiza glabra in a defined ratio termed CIM-Candy was prepared based on Ayurvedic methodology. To study the humoral immune response, rabbit red blood corpuscles (rRBCs) were used as an antigen. Mice were immunized on day 7 and 14 days the start of the experiment using an intraperitoneal injection of 200 μl of rRBCs (2×108 cells/ml). Humoral immune response, body weight variation, and hematology parameters were studied. CIM-Candy at doses of 10 and 100 mg/kg body weight enhanced the immune function by significantly increasing (p

This paper describes a sensitive, selective, specific, robust, and validated densitometric high-performance thin-layer chromatographic (HPTLC) method for the simultaneous determination of 3 key withanolides, namely, withaferin-A, 12-deoxywithastramonolide, and withanolide-A, in Ashwagandha (Withania somnifera) plant samples. The separation was performed on aluminum-backed silica gel 60F254 HPTLC plates using dichloromethanemethanolacetonediethyl ether (15 + 1 + 1 + 1, v/v/v/v) as the mobile phase. The withanolides were quantified by densitometry in the reflection/absorption mode at 230 nm. Precise and accurate quantification could be performed in the linear working concentration range of 66330 ng/band with good correlation (r2 0.997, 0.999, and 0.996, respectively). The method was validated for recovery, precision, accuracy, robustness, limit of detection, limit of quantitation, and specificity according to International Conference on Harmonization guidelines. Specificity of quantification was confirmed using retention factor (Rf) values, UV-Vis spectral correlation, and electrospray ionization mass spectra of marker compounds in sample tracks.

Three iridoid glycosides 6-O-(3''-O-benzoyl)-alpha-L-rhamnopyranosylcatalpol (1a), 6-O-(3''-O-trans-cinnamoyl)-alpha-L-rhamnopyranosylcatalpol (2a) and 6-O-(3''-O-cis-cinnamoyl)-alpha-L-rhamnopyranosylcatalpol (3a) were isolated from aerial parts of Gmelina arborea and structures were elucidated by spectral analysis. Additionally a known iridoid 6-O-(3'', 4''-O-dibenzoyl)-alpha-L-rhamnopyranosylcatalpol (4) was also isolated and identified.

Medicinal plants have been traditionally used for treating liver diseases since centuries. Several leads from plant sources have been found as potential hepatoprotective agents with diverse chemical structures. Although, a big list of hepatoprotective phytomolecules was reported in the scientific literature, only a few were potent against various types of liver damages. Of which, silymarin, andrographolide, neoandrographolide, curcumin, picroside, kutkoside, phyllanthin, hypophyllanthin, and glycyrrhizin have largely attracted the scientific community. This review focuses discussion on the chemistry, biological activity, mode of action, toxicity, and future prospects of these leads.

A sensitive, selective and robust qualitative and quantitative densitometric high-performance thin layer chromatographic method was developed and validated for the determination of iridoid glycoside in the aerial part of Gambhari ( Gmelina arborea ). Iridoid gycoside 6- O -(2″,3″-dibenzoyl)-α- l -rhamnopyranosylcatalpol (IG) was used as a chemical marker for the standardization of G. arborea plant extracts. The separation was performed on aluminum Kieselgel 60F 254 TLC plates using chloroform–methanol as mobile phase. The quantitation of IG was carried out using the densitometric reflection/absorption mode at 240 and 430 nm after post-chromatographic derivatization with vanillin–sulphuric acid reagent. A precise and accurate quantification can be performed in the linear working concentration range of 1000–5000 ng/spot with good correlation ( r 2 = 0.994). The method was validated for peak purities, precision, robustness, limit of detection (LOD) and quantitation (LOQ), etc., as per ICH guidelines. Specificity of quantitation was confirmed using retention factor ( R f ), UV–vis spectral correlation and ESI-MS spectra of marker compound (IG) in sample track.

Variability in composition of paclitaxel, the precursor 10-deacetylbaccatin III (10-DAB), and related taxoids of different polarity, namely 2-acetoxy brevifoliol, brevifoliol, 2-deacetoxy-decinnamoyl taxinine-J, and cephalomannine in Himalayan Yew (Taxus wallichiana) was studied using octadecylsilane and octasilane columns and an isocratic elution of CH3OH:H2O (0.05%AcOH)-(62:38) with PDA detection. The specific procedure for sample preparation was optimized with HPLC chromatographic protocol, validated following the ICH (International Conference on Harmonization) guidelines, and the precision and accuracy of the analysis confirmed for all six taxoids of interest. The present study established significant variation in the composition of taxoids in yew (T. Wallichiana) samples from two locations (Arunachal and Uttranchal regions) of Himalaya.

A quantitative method using silica gel 60F254 high performance thin layer chromatography plates, automated bandwise sample application, and automated visible mode densitometric method has been developed for the determination of 24β-ethylcholesta-5,22E,25-triene-3β-ol (ECTO) in the aerial part of Clerodendrum phlomidis. ECTO was used as a chemical marker for the standardization of C. phlomidis plant extracts. The separation was performed on silica gel 60F254 TLC plates using chloroform-methanol (98.5: 1.5, v/v) as mobile phase. The quantitation of ECTO was carried out using the densitometric reflection/absorption mode at 650 nm after post chromatographic derivatization with anisaldehyde reagent. A precise and accurate quantification can be performed in the linear working concentration range of 150–400 ng band−1 with good correlation (r 2 = 0.996). The method was validated for peak purities, precision, robustness, limit of detection (LOD) and quantitation (LOQ), etc. as per ICH guidelines.

A sensitive, selective, and robust high-performance TLC (HPTLC) method using chiral TLC plates for qualitative and quantitative analysis of phyllanthin (A), hypophyllanthin (B), niranthin (C), and nirtetralin (D), the active lignans of Phyllanthus species, was developed and validated. The effectiveness and role of various stationary phases viz TLC silica gel 60F(254), HPTLC silica gel 60F(254), and chiral TLC plates in the quantitation were evaluated. A precoated chiral TLC plate was found suitable for the simultaneous analysis of four pharmacologically active lignans. For achieving good separation, the optimized mobile phase of n-hexane/acetone/1,4-dioxane (9:1:0.5 by volume) was used (R(f) = 0.30, 0.36, 0.41, and 0.48 for compounds A, B, C, and D, respectively). A densitometric determination of the above compounds was carried out in reflection/absorption mode at 620 nm. Optimized chromatographic conditions provide well-separated compact bands for the tested lignans. The calibration curves were found linear in the concentration range of 100-500 ng/band. Recoveries of A-D were 99.98, 100.51, 99.22, and 98.74%, respectively. The method was validated according to ICH guidelines. The method reported here is reproducible and applied for the quantitative analysis of the above lignans in the leaves of four Phyllanthus species, i. e., P. amarus, P. maderaspatensis, P. urinaria, and P. virgatus.

A reverse phase high performance liquid chromatographic method is developed for the quantitation of glabridin in Glycyrrhiza glabra, using C18 column with acetonitrile-water containing 2% AcOH (70:30) as an eluent. Glabridin is detected by UV absorption at 280 nm after separation by the chromatographic system. Good linearity was obtained in the working range of the concentration (0.01–0.1 mg mL−1), with correlation coefficients 0.999. Limit of detection and limit of quantitation were 0.0195 and 0.065 mg mL−1. The method was validated under ICH guidelines. The described method can be utilized for routine analysis (assays and stability tests) of G. glabra extracts and Ayurvedic medicine based on Yashti-madhu.

A high-performance liquid chromatographic method for the determination of novel bioactive nitrile glycosides niaziridin and niazirin in the leaves, pods and bark of Moringa oleifera is reported. Niaziridin is a bioenhancer for drugs and nutrients. The analytical conditions for reversed-phase HPLC with UV detection were as follows: column, Chromolith RP-18 e, 4.6 × 100 mm 0.5 μm (Merck); column temperature, 25 °C; mobile phase, a 20:80 (% v/v) mixture of acetonitrile: Phosphate buffer – pH 3.8; flow rate, 0.7 ml/min; detection at 220 nm. Method precision (relative standard deviation) was 1.81% for niaziridin and 1.94% for niazirin. Niaziridin (0.015% and 0.039%) and niazirin (0.038% and 0.033%) are present in leaves and pods, respectively. Niaziridin and niazirin were not detected in the bark of M. oleifera. Relatively higher amount of niazirin was present in leaves in comparisons to the pods while niaziridin content was about three times higher in the pods than the leaves of the M. oleifera. The method is robust to evaluate niaziridin and niazirin in samples from M. oleifera as well as for quality assurance of pharmacologically active standardized extract.

The monoterpene indole alkaloids (MIAs) are generally derived from strictosidine, which is formed by condensation of the terpene moiety secologanin and the indole moiety tryptamine. There are conflicting reports on the limitation of either terpene or indole moiety in the production of MIAs in Catharanthus roseus cell cultures. Formation of geraniol by geraniol synthase (GES) is the first step in secologanin biosynthesis. In this study, feeding of C. roseus leaves with geraniol, but not tryptophan (precursor for tryptamine), increased the accumulation of the MIAs catharanthine and vindoline, indicating the limitation of geraniol in MIA biosynthesis. This was further validated by molecular and in planta characterization of C. roseus GES (CrGES). CrGES transcripts exhibited leaf and shoot specific expression and were induced by methyl jasmonate. Virus-induced gene silencing (VIGS) of CrGES significantly reduced the MIA content, which was restored to near-WT levels upon geraniol feeding. Moreover, over-expression of CrGES in C. roseus leaves increased MIA content. Further, CrGES exhibited correlation with MIA levels in leaves of different C. roseus cultivars and has significantly lower expression relative to other pathway genes. These results demonstrated that the transcriptional regulation of CrGES and thus, the in planta geraniol availability plays crucial role in MIA biosynthesis.

Gallic acid, one of the most abundant plant phenolic acids, has been modified to cathepsin D protease inhibitors. The strategy of modification was proposed basing on some previously reported structure and activity relationship (SAR) studies. The synthesized naphthophenone fatty acid amide derivatives have been evaluated for in vitro cathepsin D inhibition activity. Two of them have shown significant inhibition activity with IC50 values of 0.06 and 0.14 μM, respectively, as compared against pepstatin (0.0023 μM), the most potent inhibitor known so far. The study revealed that such attempts on gallic acid based pharmacophores might result in potent inhibitors of cathepsin D.

Background: Ocimum sanctum and Ocimum kilimandscharicum are cultivated in Indian subcontinent both for the religious and medicinal properties. Traditionally, the leaves have been reported for their enormous therapeutic potentials but the roots which are otherwise considered as a waste part have not been explored for their pharmacological activity. Methods: Total phenolic content, free radical scavenging and ferric reducing antioxidant potential of various extracts from Ocimum sanctum and Ocimum kilimandscharicum were assessed and compared. In vitro antioxidant activity was estimated spectrophotometrically and the most potent ethyl acetate extract was chemically characterized by developing the chemical fingerprint and quantifying the probable constituents with the help of HPLC/LC-MS techniques. Results: The ethyl acetate extract of both the species exhibited significant free radical scavenging potential and also reduced the ferric ions. It was observed that ethyl acetate extract have superior ferric reducing potential than other tested extracts, which were evidenced by high ferrous sulphate equivalent value of 77.05 ± 1.54 and 80.98 ± 0.80 at 100 µg/ml for O. sanctum and O. kilimandscharium respectively. The ferric reducing capacity of ethyl acetate extract for both the species was also evidenced by an elevated optical density of 1.64 ± 0.12 and 2.14 ± 0.08. Ocimum sanctum exhibited better antioxidant capacity (11.31 ± 0.20 AScE) as compared to Ocimum kilimandscharium (9.08 ± 0.27 AScE). The total phenolic and flavonoid content were estimated by spectrophotometric method and tentatively characterized by HPLC/LC-MS profiling which revealed the presence of rosmarinic acid, caffeic acid along with its derivatives such as caffeoyl-dihydroxyphenyllactoyl-tartaric acid. Conclusion: The ethyl acetate extract of both the species being rich in phenolic and flavonoid contents exhibited potent antioxidant activity. The presence of flavonoid in ethyl acetate extracts further co-relates the antioxidative properties of roots extracts. To the best of our knowledge and understanding this is the first report of comparative chemical profiling by RP-HPLC/LC-MS and antioxidant potential of roots of two Ocimum species. Peer Review Details Peer review method: Single-Blind (Peer-reviewers: 02) Peer-review policy Plagiarism software screening?: Yes Date of Original Submission: 27 July 2017 Date accepted: 23 August 2017 Peer reviewers approved by: Dr. Lili Hami Editor who approved publication: Dr. Phuc Van Pham