Exploring Anticancer Properties of Medicinal Plants against Breast Cancer by Downregulating Human Epidermal Growth Factor Receptor 2.

Plants have a history of being employed in managing breast cancer. However, no scientific evidence supports the idea that these plants can effectively reduce the level of HER2 expression. In this study, extracts from 10 medicinal plants were evaluated for their anticancer properties against HER2-positive breast cancer cells through various methods, including the SRB assay, comet assay, annexin V-FITC dual staining, and immunoblotting. All extracts exerted antiproliferative activity against HER2-positive breast cancer cells. Furthermore, Terminalia chebula (T. chebula), Berberis aristata (B. aristata), and Mucuna pruriens (M. pruriens) reduced HER2 expression in tested cell lines. In addition, an increased Bax/Bcl-2 ratio was observed after the treatment. A comparative proteomics study showed modulation in the proteome profile of breast cancer cells after treatment with T. chebula, B. aristata, Punica granatum, M. pruriens, and Acorus calamus. Metabolic profiling of lead plants revealed the existence of multiple anticancer compounds. Our study demonstrates the considerable potential of the mentioned plants as innovative therapies for HER2-positive breast cancer.

Plant-Based Approaches for Rheumatoid Arthritis Regulation: Mechanistic Insights on Pathogenesis, Molecular Pathways, and Delivery Systems.

Rheumatoid arthritis (RA) is classified as a chronic inflammatory autoimmune disorder, associated with a varied range of immunological changes, synovial hyperplasia, cartilage destructions, as well as bone erosion. The infiltration of immune-modulatory cells and excessive release of proinflammatory chemokines, cytokines, and growth factors into the inflamed regions are key molecules involved in the progression of RA. Even though many conventional drugs are suggested by a medical practitioner such as DMARDs, NSAIDs, glucocorticoids, etc., to treat RA, but have allied with various side effects. Thus, alternative therapeutics in the form of herbal therapy or phytomedicine has been increasingly explored for this inflammatory disorder of joints. Herbal interventions contribute substantial therapeutic benefits including accessibility, less or no toxicity and affordability. But the major challenge with these natural actives is the need of a tailored approach for treating inflamed tissues by delivering these bioactive agentsat an appropriate dose within the treatment regimen for an extended periodof time. Drug incorporated with wide range of delivery systems such as liposomes, nanoparticles, polymeric micelles, and other nano-vehicles have been developed to achieve this goal. Thus, inclinations of modern treatment are persuaded on the way to herbal therapy or phytomedicines in combination with novel carriers is an alternative approach with less adverse effects. The present review further summarizes the significanceof use of phytocompounds, their target molecules/pathways and, toxicity and challenges associated with phytomolecule-based nanoformulations.

Adhatoda vasica and Tinospora cordifolia extracts ameliorate clinical and molecular markers in mild COVID-19 patients: a randomized open-label three-armed study.

BACKGROUND: SARS-CoV-2 infections caused mild-to-moderate illness. However, a sizable portion of infected people experience a rapid progression of hyper-inflammatory and hypoxic respiratory illness that necessitates an effective and safer remedy to combat COVID-19. METHODS: A total of 150 COVID-19-positive patients with no to mild symptoms, between the age groups 19-65 years were enrolled in this randomized, open-labeled three-armed clinical trial. Among them, 136 patients completed the study with RT-PCR negative reports. The patients received herbal drugs orally (Group A (Adhatoda vasica; AV; 500 mg; n = 50); Group B (Tinospora cordifolia; TC; 500 mg; n = 43), and Group C (AV + TC; 250 mg each; n = 43)) for 14 days. Clinical symptoms, vital parameters, and viral clearance were taken as primary outcomes, and biochemical, hematological parameters, cytokines, and biomarkers were evaluated at three time points as secondary outcomes. RESULTS: We found that the mean viral clearance time was 13.92 days (95% confidence interval [CI] 12.85-14.99) in Group A, 13.44 days (95% confidence interval [CI] 12.14-14.74) in Group B, and 11.86 days (95% confidence interval [CI] 10.62-13.11) days in Group C. Over a period of 14 days, the mean temperature in Groups A, and B significantly decreased linearly. In Group A, during the trial period, eosinophils, and PT/INR increased significantly, while monocytes, SGOT, globulin, serum ferritin, and HIF-1α, a marker of hypoxia reduced significantly. On the other hand, in Group B hsCRP decreased at mid-treatment. Eosinophil levels increased in Group C during the treatment, while MCP-3 levels were significantly reduced. CONCLUSIONS: All the patients of the three-armed interventions recovered from COVID-19 and none of them reported any adverse effects from the drugs. Group C patients (AV + TC) resulted in a quicker viral clearance as compared to the other two groups. We provide the first clinical report of AV herbal extract acting as a modifier of HIF-1α in COVID-19 patients along with a reduction in levels of ferritin, VEGF, and PT/INR as the markers of hypoxia, inflammation, and thrombosis highlighting the potential use in progression stages, whereas the TC group showed immunomodulatory effects. Trial registration Clinical Trials Database -India (ICMR-NIMS), CTRI/2020/09/028043. Registered 24th September 2020, https://www.ctri.nic.in/Clinicaltrials/pdf_generate.php?trialid=47443&EncHid=&modid=&compid=%27,%2747443det%27.

Liquiritigenin, isoliquiritigenin rich extract of glycyrrhiza glabra roots attenuates inflammation in macrophages and collagen-induced arthritis in rats.

Liquiritigenin (LTG) and its bioprecursor isoliquiritigenin(ISL), the main bioactives from roots of Glycyrrhiza genus are progressively documented as a potential pharmacological agent for the management of chronic diseases. The aim of this study was to evaluate the pharmacological potential of liquiritigenin, isoliquiritigenin rich extract of Glycyrrhiza glabra roots (IVT-21) against the production of pro-inflammatory cytokines from activated macrophages as well as further validated the efficacy in collagen-induced arthritis model in rats. We also performed the safety profile of IVT-21 using standard in-vitro and in-vivo assays. Results of this study revealed that the treatment of IVT-21 and its major bioactives (LTG, ISL) was able to reduce the production of pro-inflammatory cytokines (TNF-α, IL-6) in LPS-activated primary peritoneal macrophages in a dose-dependent manner compared with vehicle-alone treated cells without any cytotoxic effect on macrophages. In-vivo efficacy profile against collagen-induced arthritis in Rats revealed that oral administration of IVT-21 significantly reduced the arthritis index, arthritis score, inflammatory mediators level in serum. IVT-21 oral treatment is also able to reduce the NFкB-p65 expression as evidence of immunohistochemistry in knee joint tissue and mRNA level of pro-inflammatory cytokines in paw tissue in a dose-dependent manner when compared with vehicle treated rats. Acute oral toxicity profile of IVT-21 demonstrated that it is safe up to 2000 mg/kg body weight in experimental mice. This result suggests the suitability of IVT-21 for further study in the management of arthritis and related complications.

Screening and evaluation of approved drugs as inhibitors of main protease of SARS-CoV-2.

The COVID-19 pandemic caused by SARS-CoV-2 has emerged as a global catastrophe. The virus requires main protease for processing the viral polyproteins PP1A and PP1AB translated from the viral RNA. In search of a quick, safe and successful therapeutic agent; we screened various clinically approved drugs for the in-vitro inhibitory effect on 3CLPro which may be able to halt virus replication. The methods used includes protease activity assay, fluorescence quenching, surface plasmon resonance (SPR), Thermofluor® Assay, Size exclusion chromatography and in-silico docking studies. We found that Teicoplanin as most effective drug with IC50 ~ 1.5 µM. Additionally, through fluorescence quenching Stern-Volmer quenching constant (KSV) for Teicoplanin was estimated as 2.5 × 105 L·mol-1, which suggests a relatively high affinity between Teicoplanin and 3CLPro protease. The SPR shows good interaction between Teicoplanin and 3CLPro with KD ~ 1.6 µM. Our results provide critical insights into the mechanism of action of Teicoplanin as a potential therapeutic against COVID-19. We found that Teicoplanin is about 10-20 fold more potent in inhibiting protease activity than other drugs in use, such as lopinavir, hydroxychloroquine, chloroquine, azithromycin, atazanavir etc. Therefore, Teicoplanin emerged as the best inhibitor among all drug molecules we screened against 3CLPro of SARS-CoV-2.

Evaluation of medicinal herbs as a potential therapeutic option against SARS-CoV-2 targeting its main protease.

The COVID-19 disease caused by the SARS-CoV-2 has emerged as a worldwide pandemic and caused huge damage to the lives and economy of more than hundred countries. As on May 10, 2020, more than 4,153,300 people stand infected from the virus due to an unprecedented rate of transmission and 282,700 have lost their lives because of the disease. In this context, medicinal plants may provide a way to treat the disease by targeting specific essential proteins of the virus. We screened about 51 medicinal plants and found that Tea (Camellia sinensis) and Haritaki (Terminalia chebula) has potential against SARS-COV-2 3CLpro , with an IC50 for Green Tea as 8.9 ± 0.5 µg/ml and Haritaki 8.8 ± 0.5 µg/ml. The in-silico studies suggested that Tea component Thearubigins binds to the cysteine 145 of protease active site and could be a pharmacoactive molecule. We predict that the inhibition in protease activity may be able to halt the SARS-CoV-2 replication cycle and therefore, we propose Green Tea, Black Tea, and Haritaki plant extracts as potential therapeutic candidates for SARS-CoV-2 infection. Further investigation on role of bioactive constituents of extracts is needed to establish the molecular basis of inhibition and towards expedited drug discovery.

Mindful awareness for female dental students through yoga, motivational video, and a combination of two on stress reduction.

OBJECTIVE: Assessment of the potential effect of yoga, motivational videos, and a combination of two on stress reduction in female dental students. METHODS: The current study design is a nonrandomized trial carried out among female dental students (n = 120). The selected subjects were allocated into three groups, namely, group A: yoga group assigned under a professional yoga instructor, group B: motivational video group, and group C: a combination of two (A and B) involving yoga sessions and motivational videos; following which formative examinations were carried out at 6 weeks which were referred to as stressor 1 and those carried out at 12 weeks were referred to as stressor 2. Using Spielberger's State-Trait Anxiety Inventory for Adults (STAI-A) recordings were measured at baseline, stressor 1, and stressor 2. RESULTS: There was a significant decrease in trait anxiety scores at both stressors in all the groups with a significant reduction in depression score from stressor 1 to 2 in group 1 and group 3 subjects. The significant decrease occurred in STAI-A scores in group 1 and group 3 subjects at both stressors (P < 0.001) with no change observed in group 2 subjects. CONCLUSION: The present study is one of a kind and positively correlates yoga and motivational videos with stress reduction. Both of these interventions proved to be beneficiary for physical as well as mental health of study subjects.

Simultaneous quantification of five bioactive phenylethanoid, iridoid, and flavonol glycosides in Duranta erecta L.: Ultra performance liquid chromatography method validation and uncertainty measurement.

Duranta erecta L.is a valuable herb used in traditional system of medicine in India and China for the treatment of malaria and abscesses. The presence of phenylethanoid (e.g., acteoside as major bioactive) in addition to iridoid and flavonoids glycosides are mainly responsible for reported pharmacological properties. Although D. erecta is widely used in many Asian countries, the quality assurance of raw herb and its derived products are still in question, due to the lack of validated protocol. Current Indian guidelines for quality assurance of phytopharmaceutical product demands accurate determination of minimum four chemicals. Moreover, there is no chromatographic method to simultaneously analyse acteoside, isoacteoside, durantoside I, quercetin, and methylapigenin-7-O-ß-D-glucopyranuronate in D. erecta. The present study establishes a systematic approach for the quality evaluation of D. erecta with efficient and validated UPLC-PDA method that simultaneously analyses the five bioactive marker chemicals in D. erecta. A fast, sensitive, and reliable ultraperformance liquid chromatography-photodiode array (UPLC-PDA) method was developed for the quantification five marker compounds-acteoside, isoacteoside, durantoside-I, quercetin, and methylapigenin-7-O-ß-D-glucopyranuronate in D. erecta. The optimized UPLC conditions on Reverse phase column Phenomenex Luna® (C18, 2.5µm, 2.0×100mm)) and binary gradient elution resulted in best separation for all five targeted phytoconstituents. Validation characteristics viz. linearity (r2 >0.999), accuracy (%RSD<2.0), precision (%RSD, 1.62-2.59), recovery (99-101%), and sensitivity (LOD- 0.27-0.40µg mL-1; LOQ- 0.90-1.35µgmL-1) were satisfying the ICH criteria. Uncertainty in the measurement was also estimated using method validation data and other sources to fulfil the technical requirement of ISO 17025:2017. It is the first validated method that provides the simultaneous and accurate analysis of five bioactive phytoconstituents of D. erecta in short time with defined traceability and accuracy profile. Further, it delivers a holistic quality analysis of raw medicinal herb and its preparation.

Ultra performance liquid chromatography coupled with principal component and cluster analysis of Swertia chirayita for adulteration check.

This article describes the study to standardize phytochemically and distinguish Swertia chirayita from that of possible substitution/adulteration using ultra performance liquid chromatography (UPLC) with photodiode array detector (PDA) and chemometric tools viz. principal component analysis (PCA) and hierarchical clustering analysis (HCA). Five ecotypes of Swertia chirayita and five possible substitutions, e.g.,Swertia bimaculata (SB), Swertia chordata (SCH), Swertia ciliata (SCL), Swertia paniculata (SP), and Halenia elliptica (HE) collected from different Indian Himalayan region. Samples evaluated for 04 marker compounds- swertiamarin (SM), mangiferin (MF), gentiopicroside (GP), and sweroside (SW). Reverse phase column (Waters Acquity BEH C18, 50 mm × 2.1 mm , 1.7 µm) provided high resolution for all target analytes with binary gradient elution. The detector response was linear (concentration 2.5-125 µg/mL, R2 > 0.999). The limit of detection (LOD) and quantification (LOQ) of targeted compounds was in the range of 1.40-2.06 and 4.57-6.27 µg/mL respectively. The combined relative standard deviation (%RSD) for intra-day and inter-day precision values were less than 2%. The recoveries study comply the method suitability. Chromatogram similarity analysis based on congruence coefficient was higher than 0.925 for the chirayita ecotypes while much lower than 0.629 for possible substitutes. HCA showed that the samples could be clustered (all 5 clusters in two-level) reasonably into different ecotypes and substitutes. HCA together with loading plots has indicated different chemical properties of all five groups. PCA results showed that the discrimination of chirayita ecotypes is because of the presence of SW while SM may have more influence on the targeted substitutes to discriminate from chirayita ecotypes. Therefore, UPLC fingerprint in association with chemometric tools provides a reliable and accurate quality assessment and detection of possible adulteration.

Enhanced expression of ginsenoside biosynthetic genes and in vitro ginsenoside production in elicited Panax sikkimensis (Ban) cell suspensions.

Dual metabolite, i.e., ginsenoside and anthocyanin, co-accumulating cell suspensions of Panax sikkimensis were subjected to elicitation with culture filtrates of Serratia marcescens (SD 21), Bacillus subtilis (FL11), Trichoderma atroviridae (TA), and T. harzianum (TH) at 1.25% and 2.5% v/v for 1- and 3-week duration. The fungal-derived elicitors (TA and TH) did not significantly affect biomass accumulation; however, bacterial elicitors (SD 21 and FL11), especially SD 21, led to comparable loss in biomass growth. In terms of ginsenoside content, differential responses were observed. A maximum of 3.2-fold increase (222.2 mg/L) in total ginsenoside content was observed with the use of 2.5% v/v TH culture filtrate for 1 week. Similar ginsenoside accumulation was observed with the use of 1-week treatment with 2.5% v/v SD 21 culture filtrate (189.3 mg/L) with a 10-fold increase in intracellular Rg2 biosynthesis (31 mg/L). Real-time PCR analysis of key ginsenoside biosynthesis genes, i.e., FPS, SQS, DDS, PPDS, and PPTS, revealed prominent upregulation of particularly PPTS expression (20-23-fold), accounting for the observed enhancement in protopanaxatriol ginsenosides. However, none of the elicitors led to successful enhancement in in vitro anthocyanin accumulation as compared to control values.

Elicitors' influenced differential ginsenoside production and exudation into medium with concurrent Rg3/Rh2 panaxadiol induction in Panax quinquefolius cell suspensions.

Cobalt nitrate, nickel sulphate, hydrogen peroxide, sodium nitroprusside, and culture filtrates of Pseudomonas monteili, Bacillus circularans, Trichoderma atroviridae, and Trichoderma harzianum were tested to elicit ginsenoside production in a cell suspension line of Panax quinquefolius. Abiotic elicitors preferentially increased panaxadiols whereas biotic elicitors upregulated the panaxatriol synthesis. Cobalt nitrate (50 µM) increased total ginsenosides content by twofold (54.3 mg/L) within 5 days. It also induced the Rc synthesis that was absent in the control cultures. Elicitation with P. monteili (2.5 % v/v, 5 days) also supported 2.4-fold enhancement in saponin yield. Elicitation by T. atroviridae or hydrogen peroxide induced the synthesis of Rg3 and Rh2 that are absent in ginseng roots. The highest ginsenosides productivity (3.2-fold of control) was noticed in cells exposed to 1.25 % v/v dose of T. atroviridae for 5 days. Treating cells with T. harzianum for 15 days afforded maximum synthesis and leaching (8.1 mg/L) of ginsenoside Rh1.

Ceramic nanoparticles: Recompense, cellular uptake and toxicity concerns.

Over the past few years, nanoparticles and their role in drug delivery have been the centre of attraction as new drug delivery systems. Various forms of nanosystems have been designed, such as nanoclays, scaffolds and nanotubes, having numerous applications in areas such as drug loading, target cell uptake, bioassay and imaging. The present study discusses various types of nanoparticles, with special emphasis on ceramic nanocarriers. Ceramic materials have high mechanical strength, good body response and low or non-existing biodegradability. In this article, the various aspects concerning ceramic nanoparticles, such as their advantages over other systems, their cellular uptake and toxicity concerns are discussed in detail.

Effects of a standardized Ayurvedic formulation on diabetes control in newly diagnosed Type-2 diabetics; a randomized active controlled clinical study.

OBJECTIVES: The purpose of this study was to investigate the efficacy of a standardized polyherbal formulation consists of aqueous extracts from six herbs, in patients with Type-2 diabetes mellitus. DESIGN: Randomized, active control study. INTERVENTIONS: 93 patients, newly diagnosed with Type-2 diabetes mellitus were randomly allocated to group 1 (received polyherbal capsules 500 mg/day, up titrated weekly to a maximum of 3 g/day) and group 2 (received Metformin 500 mg/day, up titrated weekly to a maximum of 2 g/day). MAIN OUTCOME MEASURES: The primary endpoint was effect on the change from baseline in blood glucose (Fasting blood Glucose and Postprandial blood glucose), and glycosylated hemoglobin (HbA1c). The secondary outcome includes the effect on lipid levels, liver enzymes and renal function test. RESULTS: After 24 weeks, mean laboratory measured fasting and post prandial blood glucose showed a decrease of 25.52% and 24.22% in polyherbal formulation (PHF) treated group, compared to 31.46% and 24% decrease in Metformin treated group (estimated treatment difference -10.8; 95% CI -22.63 to 1.03 and -0.36; -12.1 to 11.38, respectively). Reduction in HbA1c was also similar for PHF and Metformin (estimated treatment difference 0.01; 95% CI -0.51 to 0.53). However, the decrease in the mean total cholesterol level was more pronounced in PHF treated group (estimated mean difference 61.3; 95% CI 55.32 to 67.28) than Metformin treated group (estimated mean difference 41.12; 95% CI 34.92 to 47.32). Also, there was statistical significance between the treatment groups in total cholesterol level at the end of six months treatment (estimated treatment difference 20.18; 95% CI 12.34 to 28.02). CONCLUSION: The study demonstrated that daily intake of this PHF decreased the glycemic level and improved lipid homeostasis, while maintaining the other serum biochemical levels to the normal, and therefore it may be useful for the patients with Type-2 diabetes. This trial is registered in the Clinical Trials Registry - India (CTRI) (CTRI/2014/03/004490).

RP-HPLC-DAD method for the identification of two potential antioxidant agents namely verminoside and 1-O-(E)-caffeoyl-ß-gentiobiose from Spathodea campanulata leaves.

A simple and reliable high-performance liquid chromatographic method was successfully developed for the study of fingerprint chromatograms of extract and fractions from the leaves of Spathodea campanulata (SC) using verminoside (1) and 1-O-(E)-caffeoyl-ß-gentiobiose (2) as marker compounds. Antioxidant activity of SC was determined by using free radical of 2,2-diphenyl-1-picryl-hydrazyl-hydrate as an experimental model. The docking study of selected target, tyrosinase and ligands (ascorbic acid, compounds 1 and 2) was performed through Autodock Vina v0.8. Fingerprints of methanol, chloroform, ethylacetate, n-butanol and water extracts could resolve 13, 11, 22, 16 and 5 peaks, respectively. Extract, fractions and compounds 1 and 2 previously isolated from SC displayed remarkable antioxidant activity with radical-scavenging activity ranging from 2.5 to 6.7 µg/mL. In silico study identified compounds 1 and 2 as potential inhibitors of tyrosinase correlating with the observed antioxidant activity in vitro.

A dual purpose cell line of an Indian congener of ginseng--Panax sikkimensis with distinct ginsenoside and anthocyanin production profiles.

The age-dependent production kinetics of ginsenosides and an anthocyanin pigment in a cell suspension line of Panax sikkimensis was followed in vitro. Highest total saponin content [7.37 mg/g dry weight (DW)] and biomass accumulation (% biomass increase = 209.67) in this line occurred after 3 and 5 weeks of culture, respectively. Accumulation of individual protopanaxatriol (Re, Rg1, and Rg2) and protopanaxadiol (Rb1, Rb2, and Rc) ginsenosides showed a variable pattern of accumulation independent of cell biomass buildup during the 7-week culture cycle. However, total content of triol ginsenosides was always significantly more than the diol group of ginsenosides, being 183.2-, 63.5-, and 72.1-folds at third, fourth, and fifth week stage of cell growth. Interestingly, in addition to these ginsenosides, the cell line also co-accumulated an anthocyanin pigment in vitro. The pigment content increased gradually from 8.66 to 14.29 mg/g DW after first to fifth week followed by a marginal fall to 12.79 and 10.95 mg/g DW during next 2 weeks. Therefore, in terms of total recovery of saponins (77.4 mg/l) and anthocyanin (199.16 mg/l), harvesting of cells after 3 and 5 weeks of growth was most profitable, respectively. The possible utility of this dual purpose cell line in nutraceutical industry is discussed.

Protective mechanism of lignans from Phyllanthus amarus against galactosamine/ lipopolysaccharide-induced hepatitis: an in-vivo and in-silico studies.

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-1ß, 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.

Acute and sub-chronic toxicity of HPLC fingerprinted extract of Conyza sumatrensis (Retz.) E.H. Walker in rodents.

ETHNOPHARMACOLOGICAL RELEVANCE: Conyza sumatrensis (CS) is an extensively used medicinal herb in the tropics for varied ailments related to pain, inflammation and malaria. Though in constant folkloric use, scientific validations are proving valuable. AIM OF THE STUDY: Evaluate the safety profile of methanol extract from CS in mice and rats through acute and sub-chronic toxicity studies respectively. MATERIAL AND METHODS: Acute toxicity study involved the single oral administration of CS at 1000, 2000 and 3000mg/kg in mice, while the sub chronic toxicity was carried upon in rats at doses 250, 500 and 1000mg/kg/day for 28 days. Besides body weight, general behaviour and mortality, serum biochemical parameters and liver histology were assessed after 7 and 28 days for acute and sub-chronic study respectively. The parameters were again checked on days 14 and 56 in order to assess the recovery from damage, if any. HPLC fingerprinting of the aqueous and methanol extract was performed through C18 column using water: acetonitrile as mobile phase with observations at 240nm. RESULTS: In the acute toxicity test, single oral dose of 1000, 2000 and 3000mg/kg of CS did not result in any behavioural changes or mortality, indicating non toxicity. In sub-chronic toxicity studies in rats, no mortality was observed at 250, 500 and 1000mg/kg/day when administered orally for a period of 28 days. Except Serum Glutamate Pyruvate Transaminase (SGPT) level in acute study and Alkaline Phosphatase (ALP), SGPT and Serum Glutamate Oxaloacetate Transaminase (SGOT) level in sub-chronic study, all the observational, haematological and biochemical parameters studied showed non-significant changes. Histological examination of liver did not reveal any treatment-related effects in any of the studies. Moreover, haematological and biochemical changes orchestrated by CS got normalised after 14 and 56 days post-treatment in acute and sub-chronic toxicity respectively. The HPLC fingerprint could resolve 11 and 28 peaks from aqueous and methanol extracts respectively. CONCLUSION: The experiments indicate the methanol extract to be safe even at high and repeated doses in pre-clinical studies even though the constituents are more than in aqueous extract.

Characterization of volatile components of Zingiber roseum essential oil using capillary GC on modified cyclodextrins.

The essential oil from different parts of Zingiber roseum plants was extracted by hydrodistillation, and analyzed using enantio-GC, capillary-GC and GC-MS. Two chiral selectors, 6-tert-butyldimethylsilyl-2,3-diethyl-beta-cyclodextrin (TBDE-beta-CD), and 2,3,6-methyl-beta-cyclodextrin (PM-beta-CD) doped into 14% cyanopropylphenyl/86% dimethylpolysiloxane, and 35% diphenyl/65% dimethylpolysiloxane, respectively were compared in order to clarify the stereochemistry and enantioselectivity of terpenoids using chiral gas chromatography. The enantiomeric excess for (1R)-(+)-alpha-pinene, (1R)-(+)-beta-pinene, and (R)-(+)-limonene were characteristic for the rhizome. In TBDE-beta-cyclodextrin coated chiral columns, a significant increase in separation factor (alpha) for beta-pinene, limonene, linalool and alpha-terpineol enantiomers was observed when compared with methyl substituted beta-cyclodextrin. The increase in chain length of the alkyl substituents may be the possible cause for enantiomer separation in beta-cyclodextrin cavity. In addition, enantioreversal of alpha-pinene enantiomers in 6-tert-butyldimethylsilyl-2,3-diethyl-beta-cyclodextrin was noticed as a unique feature. The enantiomeric compositions of Z. roseum fruit and flower essential oils were similar, but, in contrast, the rhizome oil contained an entirely different composition. Therefore, these results aid in the authentication of the natural origin of Z. roseum essential oils.

Effects of wild-type and α-tocopherol-enriched transgenic Brassica juncea on the components of xenobiotic metabolism, antioxidant status, and oxidative stress in the liver of mice.

Alpha (α)-tocopherol is the most biologically active and preferentially retained form of vitamin E in the human body and is known for its antioxidant and gene regulatory functions. Its increased intake is implicated in protection against diseases that involve an oxidative stress component. We have evaluated the chemopreventive potential of a diet supplemented with natural α-tocopherol-enriched transgenic (TR) Brassica juncea seeds. The modulation of phase I and phase II xenobiotic metabolism and of antioxidative enzymes was compared in the livers of mice fed on a control diet or on a diet supplemented with 2, 4, and 6 % (w/w) of wild-type (WT) or TR seeds. A dose-dependent increase in the specific activities of these enzymes was observed in those animals fed on diet supplemented with TR seeds. In comparison, an increase in the specific activities of antioxidative enzymes was substantial only at higher doses of WT seeds. Consequently, oxidative stress measured in terms of lipid peroxidation and lactate dehydrogenase activity was found to be lower in the case of mice fed with the supplemented diet. However, the chemopreventive potential of TR seeds was more pronounced than that of WT seeds. This study demonstrates the feasibility of fortifying diets with natural α-tocopherol for chemopreventive benefits by means of transgenic manipulation of a commonly used oilseed crop.

Growth and asiaticoside production in multiple shoot cultures of a medicinal herb, Centella asiatica (L.) Urban, under the influence of nutrient manipulations.

Growth and in vitro asiaticoside accumulation in multiple shoot cultures of Centella asiatica (L.) Urban was studied as a function of nutrient manipulations in the culture media. Shoot cultures raised in liquid Murashige and Skoog medium supplemented with 2.5 mg/l kinetin attained a growth index (GI) of 6.06 along with the highest asiaticoside content of 3.8 mg/g dry weight on the 35th day of the culture cycle. The shoot growth and asiaticoside accumulation were found to be influenced by the relative proportions of NH(4)(+)-N:NO(3)(-)-N or Cu(2+) concentration in the medium. Asiaticoside content in shoots increased from 5.3 to 8.9 and 8.7 mg/g dry weight when total nitrogen concentration of 60 mM in the control medium was reduced to 50 and 40 mM with a corresponding change in NH(4) (+):NO(3)(-) ratio from 20:40 to 20:30 or 20:20, respectively. Total nitrogen level higher than 60 mM drastically reduced the asiaticoside concentration in these in vitro shoot cultures. Medium devoid of Cu(2+) significantly favored higher asiaticoside accumulation in the cultured tissue (7.05 mg/g dry weight) along with an improved biomass production (GI = 7.7) when compared with shoots reared on the control medium with 0.10 µM Cu(2+) (GI = 5.8; asiaticoside content = 4.4 mg/g dry weight). Carbohydrate enrichment of the medium by increasing the sucrose concentration from 3.0 to 5.0 or 7.0% was also beneficial for biomass and asiaticoside production with GI = 17.1 and 16.9 and asiaticoside content = 7.2 and 5.2 mg/g dry weight, respectively, in comparison to control cultures maintained on medium containing 3.0% sucrose. The procedure described here provides a viable production platform for generating clean and quality material from Centella with high bioactive content.