Morphological, phytochemical, and transcriptome analyses provide insights into the biosynthesis of monoterpenes in Monarda citriodora.

MAIN CONCLUSION: Morphological, phytochemical, and transcriptome analyses revealed candidate genes involved in the biosynthesis of volatile monoterpenes and development of glandular trichomes in Monarda citriodora. Monarda citriodora Cerv. ex Lag. is a valuable aromatic plant due to the presence of monoterpenes as major constituents in its essential oil (EO). Thus, it is of sheer importance to gain knowledge about the site of the biosynthesis of these terpenoid compounds in M. citriodora, as well as the genes involved in their biosynthesis. In this study, we studied different types of trichomes and their relative densities in three different developmental stages of leaves, early stage of leaf development (L1), mid-stage of leaf development (L2), and later stage of leaf development (L3) and the histochemistry of trichomes for the presence of lipid and terpenoid compounds. Further, the phytochemical analysis of this plant through GC-MS indicated a higher content of monoterpenes (thymol, thymoquinone, γ-terpinene, p-cymene, and carvacrol) in the L1 stage with a substantial decrease in the L3 stage of leaf development. This considerable decrease in the content of monoterpenes was attributed to the decrease in the trichome density from L1 to L3. Further, we developed a de novo transcriptome assembly by carrying out RNA sequencing of different plant parts of M. citriodora. The transcriptome data revealed several putative unigenes involved in the biosynthesis of specialized terpenoid compounds, as well as regulatory genes involved in glandular trichome development. The data generated in the present study build a strong foundation for further improvement of M. citriodora, in terms of quantity and quality of its essential oil, through genetic engineering.

HPLC-PDA Method for Quantification of Bioactive Compounds in Crude Extract and Fractions of Aucklandia costus Falc. and Cytotoxicity Studies against Cancer Cells.

Aucklandia costus Falc. (Synonym: Saussurea costus (Falc.) Lipsch.) is a perennial herb of the family Asteraceae. The dried rhizome is an essential herb in the traditional systems of medicine in India, China and Tibet. The important pharmacological activities reported for Aucklandia costus are anticancer, hepatoprotective, antiulcer, antimicrobial, antiparasitic, antioxidant, anti-inflammatory and anti-fatigue activities. The objective of this study was the isolation and quantification of four marker compounds in the crude extract and different fractions of A. costus and the evaluation of the anticancer activity of the crude extract and its different fractions. The four marker compounds isolated from A. costus include dehydrocostus lactone, costunolide, syringin and 5-hydroxymethyl-2-furaldehyde. These four compounds were used as standard compounds for quantification. The chromatographic data showed good resolution and excellent linearity (r2 ˃ 0.993). The validation parameters, such as inter- and intraday precision (RSD < 1.96%) and analyte recovery (97.52-110.20%; RSD < 2.00%),revealed the high sensitivity and reliability of the developed HPLC method. The compounds dehydrocostus lactone and costunolide were concentrated in the hexane fraction (222.08 and 65.07 µg/mg, respectively) and chloroform fraction (99.02 and 30.21 µg/mg, respectively), while the n-butanol fraction is a rich source of syringin (37.91 µg/mg) and 5-hydroxymethyl-2-furaldehyde (7.94 µg/mg). Further, the SRB assay was performed for the evaluation of anticancer activity using lung, colon, breast and prostate cancer cell lines. The hexane and chloroform fractions show excellent IC50 values of 3.37 ± 0.14 and 7.527 ± 0.18 µg/mL, respectively, against the prostate cancer cell line (PC-3).

Medicinal plants traditionally used in health care practices by inhabitants of Paddar region of Jammu and Kashmir, India.

ETHNOPHARMACOLOGICAL RELEVANCE: The present study is the first quantitative assessment of ethno-medicinal plants of Paddar region of Jammu and Kashmir. AIM OF THE STUDY: The study aimed to document the relative importance of medicinal plants used in human ailments by the villagers in the Paddar region of Jammu and Kashmir, India. MATERIAL AND METHODS: Data were collected from 132 informants (72 male and 60 female) through semi structured interviews and group discussions. Use report (UR) and Informant consensus factor (ICF) were employed to quantitatively examine the data. RESULTS: The inhabitants of Paddar reported the use of 98 plants species of 55 families to treat 63 ailments. Rosaceae (10 spp.) was the most frequently used family in the study area. Herbs were dominantly (66 spp., 50%) utilized in herbal preparation and leaves the mostly used plant parts (25%). The highest informant consensus factor (ICF = 0.96) was obtained for parasitic problems. Important new uses for species stated by informants to treat human diseases were; Viburnum grandiflorum Wall. ex DC., Sium latijugum C.B.Clarke, Corylus jacquemontii Decne., Capsella bursa-pastoris (L.) Medik., Cannabis sativa L., Taraxacum campylodes G.E.Haglund, Euphorbia helioscopia L., Juglans regia L., Cotoneaster acuminatus Lindl., Ficus palmata Forssk., Plantago lanceolata L., and Eleusine coracana (L.) Gaertn. CONCLUSIONS: The current study contributes towards the preservation of indigenous plants' based knowledge. Although the therapeutic value of most of the preferred medicinal plants has already been validated, some medicinal plants lack proper scientific validation. We recommend further phytochemical investigations and pharmacological validations of Viburnum grandiflorum, C. jacquemontii, F. palmata, Viola pilosa, Cotoneaster acuminatus, Eleucine coracana, Sium latijugum, Aquilegia pubiflora, Euphorbia helioscopia, Plantago lanceolata and Pinus gerardiana.

Biotransformation of Geraniol to Geranic Acid Using Fungus Mucor irregularis IIIMF4011.

Geraniol is an important component in essential oils of aromatic plants such as lemongrass, rosa grass, and many others. It can be converted to different high-value products by using microbes/enzymes. The present study aims at the isolation and screening of microbes showing efficient production of geranic acid (a high-value product) from geraniol (a low-value monoterpene). Mucor irregularis IIIMF4011, isolated from the soil sample of Cymbopogon citratus (Lemongrass), showed biotransformation of geraniol to geranic acid. After optimization of reaction parameters, 97-100% conversion of geraniol to geranic acid was obtained after 72 h of incubation at 28 °C. Furthermore, the biotransformation reaction was also carried out in a 3 L fermentor (working volume 1.5 L), and 98.89% conversion was observed. Therefore, an efficient process of geranic acid production using M. irregularis IIIMF4011 was developed.

Medicinal plants traditionally used in the health care practices by the indigenous communities of the Trans-Himalayan region of Ladakh, India.

ETHNOPHARMACOLOGICAL RELEVANCE: Ladakh, "the land of high-rising passes," is a cold arid desert located in the India's northernmost part of the Trans-Himalayan region. Traditional knowledge of medicinal plants in this fragile ecosystem is an important part of the primary healthcare system, particularly in remote areas where modern medical facilities are not fully operational. There is a need to update the traditional information on medicinal plants from time to time to understand any addition of plants or uses to assist in developing new drugs through pharmacological and phytochemical studies. AIM OF THE STUDY: The study was conducted to document the traditional knowledge and current therapeutic practices of the indigenous communities of Ladakh. Besides, the study strives to evaluate previous studies from Ladakh to identify plants that have not been previously reported for medicinal use. MATERIAL AND METHODS: Ethnomedicinal information was collected from 350 local informants through open and semi-structured questionnaires and field surveys in 35 villages of 8 regions of Ladakh. The primary data were analyzed for use reports (UR) and the informant consensus factor (ICF). The reported diseases were classified according to the International Classification of Primary Care-2 (ICPC-2). New reported medicinal plants and unreported medicinal uses in this study were identified and compared to 36 prior studies conducted in Ladakh. RESULTS: In the study, 176 medicinal plants belonging to 45 families and 131 genera were reported to treat 116 diseases of 16 (ICPC-2) groups with a total of 4841 use reports. Based on the highest use reports (UR), the most utilized species were Thymus linearis Benth. (134), followed by Aconitum heterophyllum Wall. (127) The highest informant consensus factor (ICF) was reported for Pregnancy, Childbearing, and Family Planning (ICF = 1) followed by respiratory (ICF = 0.91), and digestive (ICF = 0.90) disorders. Based on the comparative literature review, a total of 518 plant species (502 previously reported plus 16 newly cited) are being used in the Ladakh region. A total of 133 use reports for 16 newly cited plants using 30 diseases were recorded. The highest degree of overlapping of medicinal plants and uses for medicines was recorded in regions with comparable ethnicity, and shorter geographical distances. Based on the IUCN Red List, recorded medicinal plants include 7 critically endangered, 9 endangered, and 13 vulnerable species. CONCLUSION: Ladakh's indigenous populations use a diverse range of medicinal plants to treat a variety of illnesses. The introduction of species and medicinal uses not previously cited in the primary health care system demonstrates that shared knowledge of traditional medicine among Ladakhi is still rich. The medicinal value of preferred medicinal plants has already been validated, but some medicinal plants lack scientific validation. We recommend further scientific studies on Aconitum violaceum Jacquem. ex Stapf,Anaphalis nepalensis var. monocephala (DC.) Hand.-Mazz., Allardia nivea Hook. f. & Thomson ex C.B. Clarke, Atriplex hortensis L., Eriophyton tibeticum (Vatke) Ryding, Iris lactea Pall. and Rheum webbianum Royle.

A Comprehensive Review on Phytochemistry and Pharmacology of Rosa Species (Rosaceae).

The Rosa L. genus is a significant plant family in the Rosaceae group, consisting of around 200 species, primarily shrubs. In India, it has 37 species, most located in the Western Himalayan region of Jammu and Kashmir and Himachal Pradesh. Roses are highly regarded for their beauty and growth and are popular worldwide for their nutritional, therapeutic, ornamental, and cosmetic value. The rose hips are utilized in creating various food and drink items, such as jams, jellies, teas, and alcoholic beverages. The Rosa species has various pharmacological activities, including anti-inflammatory, antidiabetic, hepatoprotective, antimicrobial, anti-proliferative/anticancer, anti-arthritic, neurological, and anti-obesity activity. This review aims to provide an in- -depth overview of the phytochemistry and pharmacology of the Rosa species in India, focusing on Rosa indica L., which has significant potential for future chemical and biological research.

Biotransformation of eugenol by an endophytic fungus Daldinia sp. IIIMF4010 isolated from Rosmarinus officinalis.

Natural value-added compounds produced from biological sources have attained immense significance in medicinal, food, flavourings, and agrochemical industries. Further, biotransformation is a powerful tool used to produce value-added compounds cost-effectively and selectively. In the present study, biotransformation of eugenol using an endophytic fungus Daldinia sp. IIIMF4010 isolated from the fresh leaves of the plant Rosmarinus officinalis leads to the production of two known value-added compounds. The biotransformation reaction of eugenol (50 mM) resulted in the production of eugenol-ß-D-glucopyranoside (6.2%) and vanillin (21.8%). These biotransformed products were further characterized by liquid chromatography-mass spectroscopy (LC-MS) and nuclear magnetic resonance (NMR).

Development and analysis of de novo transcriptome assemblies of multiple genotypes of Cymbopogon spp. reveal candidate genes involved in the biosynthesis of aromatic monoterpenes.

Despite the high economic value of the monoterpene-rich essential oils from different genotypes of Cymbopogon, the knowledge about the genes and metabolic route(s) involved in the biosynthesis of aromatic monoterpenes in this genus is limited. In the present study, a comprehensive transcriptome analysis of four genotypes of Cymbopogon, displaying diverse quantitative and qualitative profiles of volatile monoterpenes in their essential oils has been carried out. The comparative analysis of the deduced protein sequences corresponding to the transcriptomes of the four genotypes revealed 4609 genotype-specific orthogroups, which might contribute in defining genotype-specific phenotypes. The transcriptome data mining led to the identification of unigenes involved in the isoprenogenesis. The homology searches, combined with the phylogenetic and expression analyses provided information about candidate genes concerning the biosynthesis of monoterpene aldehyde, monoterpene alcohol, and monoterpene esters. In addition, the present study suggests a potential role of geranial reductase like enzyme in the biosynthesis of monoterpene aldehyde in Cymbopogon spp. The detailed analysis of the candidate pathway genes suggested that multiple enzymatic routes might be involved in the biosynthesis of aromatic monoterpenes in the genus Cymbopogon. The present study provides deeper insights into the biosynthesis of monoterpenes, which will be useful for the genetic improvement of these aromatic grasses.

Identification of Lipoxygenase gene repertoire of Cannabis sativa and functional characterization of CsLOX13 gene.

Lipoxygenase (LOX) enzymes play a pivotal role in the biosynthesis of oxylipins. The phyto-oxilipins have been implicated in diverse aspects of plant biology, from regulating plant growth and development to providing tolerance against biotic and abiotic stresses. C. sativa is renowned for its bioactive secondary metabolites, namely cannabinoids. LOX route is assumed to be involved in the biosynthesis of hexanoic acid, which is one of the precursors of cannabinoids of C. sativa. For obvious reasons, the LOX gene family deserves thorough investigation in the C. sativa. Genome-wide analysis revealed the presence of 21 LOX genes in C. sativa, which can be further grouped into 13-LOX and 9-LOX depending upon their phylogeny as well as the enzyme activity. The promoter regions of the CsLOX genes were predicted to contain cis-acting elements involved in phytohormones responsiveness and stress response. The qRT-PCR-based expression analysis of 21 LOX genes revealed their differential expression in different plant parts (root, stem, young leaf, mature leaf, sugar leaf, and female flower). The majority of CsLOX genes displayed preferential expression in the female flower, which is the primary site for the biosynthesis of cannabinoids. The highest LOX activity and expression level of a jasmonate marker gene were reported in the female flowers among all the plant parts. Several CsLOX genes were found to be upregulated by MeJA treatment. Based on the transient expression in Nicotiana benthamiana and the development of stable Nicotiana tabacum transgenic lines, we demonstrate that CsLOX13 encodes functional lipoxygenase and play an important role in the biosynthesis of oxylipins.

Costunosides A-C: cytotoxic sesquiterpene lactones from the rhizomes of Aucklandia costus Falc.

Three new eudesmane type rare sesquiterpene lactone galactosides, costunosides A-C (1-3) were isolated from the rhizomes of Aucklandia costus along with ten known compounds (4-13). Costunosides A-C (1-3) are the first example of naturally eudesmane glycosides containing a ß-galactopyranoside moiety. The structure and relative configurations of these compounds were established by comprehensive analysis of MS and, in particular 1D/2D NMR spectroscopic data. The isolated compounds were tested against a panel of human cancer cell lines, where compounds 3, 6 and 7 have shown promising cytotoxic activity against PC-3, HCT-116 and A549 cell lines with IC50 values in the range of 3.4 µM to 9.3 µM, respectively. Costunosides A-C (1-3) were also screened for inhibition assay of acetyl-cholinesterase (AChE), and butyrylcholinesterase (BChE) and found inactive at a concentration of 10 µM.

Discovery of Anti-NRLP3 Inflammasome, Immunomodulatory Phytochemicals from the Extract of Habenaria intermediaD. Don: An Unexplored Plant Species.

The present study describes the isolation, identification, and quantification of biomarker compounds in plant extracts of Habenaria intermedia D. Don (Orchidaceae). The isolation of the compounds was carried out from H. intermedia D. Don by repeated column chromatography of petroleum ether and ethanol fractions of extract of tubers. These compounds were characterized by 1H and 13C NMR and mass spectral data. A new quantitative method was established by using high-performance liquid chromatography (HPLC)-PDA. As a result, seven compounds were isolated and characterized. This is the first report of isolation of these compounds from this plant species H. intermedia D.Don. Out of seven isolated compounds, five were used for the quantitative study. A reliable and suitable HPLC method was developed for the well-resolved chromatogram of compounds. The proposed method was applied successfully to the detection and quantification of compounds. This study also represents the immunomodulatory and anti-inflammasome biological studies of isolated natural products. Loroglossol (HBR-4) has been reported to possess immunomodulatory activity. The immunostimulating assay indicated that HBR-4 could significantly promote the cell proliferation, especially via IL-2, TNF-α, and IFN-γ secretion from spleen cells. These results suggested the potential utilization of HBR-4 as an attractive functional health supplement candidate for hypoimmunity population. Additionally, cyclophosphamide-induced immunosuppression was counteracted by treatment with HBR-4, revealing significant increase in hemagglutinating antibody responses and hemolytic antibody responses. The current work revealed the potential anti-inflammasome and immunomodulatory activities of H. intermedia D. Don compounds and validates the usage of this prominent Rasayna plant.

Trilliumosides K and L, two novel steroidal saponins from rhizomes of Trillium govanianum, as potent anti-cancer agents targeting apoptosis in the A-549 cancer cell line.

Two novel steroidal saponins, trilliumosides K (1) and L (2), were isolated from the rhizomes of Trillium govanianum led by bioactivity-guided phytochemical investigation along with seven known compounds: govanoside D (3), protodioscin (4), borassoside E (5), 20-hydroxyecdysone (6), 5,20-hydroxyecdysone (7), govanic acid (8), and diosgenin (9). The structure of novel compounds 1-2 was established using analysis of spectroscopic data including 1D and 2D nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HR-ESI-MS) data. All isolated compounds were evaluated for in vitro cytotoxic activity against a panel of human cancer cell lines. Compound 1 showed significant cytotoxic activity against the A-549 (Lung) and SW-620 (Colon) cancer cell lines with IC50 values of 1.83 and 1.85 µM, respectively whereas the IC50 value of Compound 2 against the A-549 cell line was found to be 1.79 µM. Among the previously known compounds 3, 5, and 9, the cytotoxic IC50 values were found to be in the range of 5-10 µM. Comprehensive anti-cancer investigation revealed that Compound 2 inhibited in vitro migration and colony-forming capability in the A-549 cell line. Additionally, the mechanistic analysis of Compound 2 on the A-549 cell line indicated distinctive alterations in nuclear morphology, increased reactive oxygen species (ROS) production, and decreased levels of mitochondrial membrane potential (MMP). By upregulating the pro-apoptotic protein BAX and downregulating the anti-apoptotic protein BCL-2, the aforementioned actions eventually cause apoptosis, a crucial hallmark in cancer research, which activates Caspase-3. To the best of our knowledge, this study reports the first mechanistic anti-cancer evaluation of the compounds isolated from the rhizomes of T. govanianum with remarkable cytotoxic activity in the desired micromolar range.

Comparative morpho-anatomical standardization and chemical profiling of root drugs for distinction of fourteen species of family Apocynaceae.

BACKGROUND: The root drugs of the family Apocynaceae are medicinally important and used in Indian Systems of Medicine (ISM). There is often a problem of misidentification and adulteration of genuine samples with other samples in the market trade. Keeping in view the adulteration problem of raw drug material, comparative macroscopic and microscopic (qualitative and quantitative) characterisation and chemical analysis (TLC and LC-MS profiling) of a total of 14 economically important root drugs of family Apocynaceae were done for practical and rapid identification. A total of 33 qualitative botanical characteristics of root samples were subjected to Principal Component Analysis (PCA) and Cluster analysis to identify taxonomically significant characteristics in the distinction of root drug samples at the species level. RESULTS: Comparative qualitative and quantitative data on morphological, macroscopic, and microscopic characters were generated for the studied 14 species. Despite the similarity in some root characters, a combined study involving the surface, anatomical, and powder features helped distinguish root samples at the species level. The relative relationship between selected species was represented as clustering or grouping in the dendrogram. PCA analysis determined significant characters leading to species grouping and identification. Results showed that clustering of xylem vessels in cross-section, pore size, and distribution in the cut root, the shape of starch grains, the thickness of cork zone were among the most notable characters in species distinction. Chemical profiling revealed unique fingerprints and content of chemical compounds, which were significant in identification of root drug samples. CONCLUSIONS: The comparative botanical standards and chemical profiles developed in the present study can be used as future reference standards for the quick, easy, and correct identification of root drug samples to be used in the herbal drug industry. Further, the identified significant microscopic characters have the potential for taxonomic studies in species delimitation.

A Coumarin-Donepezil Hybrid as a Blood-Brain Barrier Permeable Dual Cholinesterase Inhibitor: Isolation, Synthetic Modifications, and Biological Evaluation of Natural Coumarins.

Plants have immensely contributed to the drug discovery for neurodegenerative diseases. Herein, we undertook the phytochemical investigation of Nardostachys jatamansi (D.Don) DC. rhizomes followed by semisynthetic modifications to discover cholinesterase (ChE) and beta-site amyloid precursor protein cleaving enzyme 1 (BACE-1) inhibitors. The 8-acetyl-7-hydroxycoumarin isolated from the bioactive extract moderately inhibits acetylcholinesterase (AChE) and BACE-1 with IC50 values of 22.1 and 17.7 µM, respectively. The semisynthetic trifluoromethyl substituted coumarin chalcone display a 5-fold improvement in BACE-1 inhibition (IC50 3.3 µM). Another semisynthetic derivative, a coumarin-donepezil hybrid, exhibits dual inhibition of both ChEs with IC50 values of 1.22 and 3.09 µM, respectively. Molecular modeling and enzyme kinetics revealed that the coumarin-donepezil hybrid is a non-competitive inhibitor of AChE. It crosses the blood-brain barrier and also inhibits Aß self-aggregation. The results presented herein warrant a detailed investigation of the coumarin-donepezil hybrid in preclinical models of Alzheimer's disease.

Coronarin K and L: Two Novel Labdane Diterpenes From Roscoea purpurea: An Ayurvedic Crude Drug.

The main objective of cancer treatment with chemotherapy is to kill the cancerous cells without affecting the healthy normal cells. In the present study, bioactivity-guided purification of the n-chloroform soluble fraction from the methanol extract of Roscoea purpurea resulted in the identification of two new labdane diterpenes: coronarin K (1) and coronarin L (2), along with eight known compounds, coronarin A (3), bisdemethoxycurcumin (4), kaempferol 3-O-methyl ether (5), kaempferol (6), fenozan acid (7), 3-(3-methoxy,4-hydroxyphenyl)-2-propenoic acid ferulic acid (8), caffeic acid (9), and gallic acid (10). The structural identification of new compounds (1 and 2) were determined by detailed analysis of 1D (1H and 13C) and 2D NMR (COSY, HSQC, and HMBC) spectroscopic data. The relative configurations of 1 and 2 were determined with the help of NOESY correlations and comparison of optical rotations with known labdane diterpenes, with established stereochemistry, while structure of known compounds was established by direct comparison of their NMR data with those reported in the literature. This is the first report of isolation of this labdane diterpenes and phenolic classes of secondary metabolites in R. purpurea. In the preliminary screening, the methanol extract and its fractions were tested for the cytotoxic activity against a panel of four cancer cell lines (A549, HCT-116, Bxpc-3, and MCF-7); extract and its chloroform fraction were found to be active against the lung cancer cell line, A-549, with IC50 value <25 µg/ml. Owing to the notable cytotoxic activity of the chloroform fraction, the compounds (1-5) were screened for their cytotoxicity against all the cell lines by MTT assay. Coronarin K, 1 showed significant cytotoxic potential against lung cancer cell lines (A-549), with IC50 value of 13.49 µM, while other compounds did not show activity below 22 µM.

Corrigendum: Coronarin K and L: Two Novel Labdane Diterpenes From Roscoea purpurea: An Ayurvedic Crude Drug.

[This corrects the article DOI: 10.3389/fchem.2021.642073.].

GC-MS based metabolomic approach to understand nutraceutical potential of Cannabis seeds from two different environments.

Cannabis sativa L. is a valuable plant that has regained its importance for medicinal use. Gas chromatography-mass spectrometry based untargeted metabolomic study was conducted in seeds of two accessions from different environments. A total of 236 metabolites were observed, and 43 metabolites were found differentially significant (p ≤ 0.05) in both the accessions. Based on the qualitative and quantitative accumulation of the nutraceutically important amino acids, cannabinoids, alkaloids, and fatty acids, the high altitude temperate Himalayan accession (CAN2) was found to have an advantage over the low altitude subtropical accession (CAN1). Seed oil from CAN2 showed the exclusive presence of linoleic acid and α- linolenic acid. The reducing power and DNA nicking assay on the methanolic extracts suggested higher antioxidant and nutraceutical potential in CAN2 and corroborated with the metabolic content of phenols and flavonoids. The environmental effect on the antioxidant and nutraceutical value in seeds is further discussed.

Cocculus hirsutus-Derived Phytopharmaceutical Drug Has Potent Anti-dengue Activity.

Dengue is a serious public health concern worldwide, with ∼3 billion people at risk of contracting dengue virus (DENV) infections, with some suffering severe consequences of disease and leading to death. Currently, there is no broad use vaccine or drug available for the prevention or treatment of dengue, which leaves only anti-mosquito strategies to combat the dengue menace. The present study is an extension of our earlier study aimed at determining the in vitro and in vivo protective effects of a plant-derived phytopharmaceutical drug for the treatment of dengue. In our previous report, we had identified a methanolic extract of aerial parts of Cissampelos pareira to exhibit in vitro and in vivo anti-dengue activity against all the four DENV serotypes. The dried aerial parts of C. pareira supplied by local vendors were often found to be mixed with aerial parts of another plant of the same Menispermaceae family, Cocculus hirsutus, which shares common homology with C. pareira. In the current study, we have found C. hirsutus to have more potent anti-dengue activity as compared with C. pareira. The stem part of C. hirsutus was found to be more potent (∼25 times) than the aerial part (stem and leaf) irrespective of the extraction solvent used, viz., denatured spirit, hydro-alcohol (50:50), and aqueous. Moreover, the anti-dengue activity of stem extract in all the solvents was comparable. Hence, an aqueous extract of the stem of C. hirsutus (AQCH) was selected due to greater regulatory compliance. Five chemical markers, viz., Sinococuline, 20-Hydroxyecdysone, Makisterone-A, Magnoflorine, and Coniferyl alcohol, were identified in fingerprinting analysis. In a test of primary dengue infection in the AG129 mice model, AQCH extract at 25 mg/kg body weight exhibited protection when administered four and three times a day. The AQCH was also protective in the secondary DENV-infected AG129 mice model at 25 mg/kg/dose when administered four and three times a day. Additionally, the AQCH extract reduced serum viremia and small intestinal pathologies, viz., viral load, pro-inflammatory cytokines, and vascular leakage. Based on these findings, we have undertaken the potential preclinical development of C. hirsutus-based phytopharmaceutical, which could be studied further for its clinical development for treating dengue.

Synthesis and biological evaluation of novel bavachinin analogs as anticancer agents.

A library of 28 analogs of bavachinin including aliphatic and aromatic ethers, epoxide, chalcone, oxime, semicarbazide, oxime ether and triazole derivatives have been synthesized and evaluated for cytotoxicity against four different human cancer cell lines. Bio-evaluation studies exhibited better cytotoxic profile for many analogs compare to bavachinin. Best results were observed for a 1,2,3-triazole analog (17i) with IC50 values 7.72, 16.08, 7.13 and 11.67 µM against lung (A549), prostate (PC-3), colon (HCT-116) and breast (MCF-7) cancer cell lines respectively. This analog showed three and four fold improvement in cytotoxicity against HCT-116 and A549 cell lines than parent molecule (1). Structure activity relationship (SAR) study for all synthesized analogs was carried out. Further, mechanistic study of the lead molecule (17i) revealed that it inhibits colony formation and in vitro migration of human colon cancer cells (HCT-116). Also, it induced the morphological changes and mediated the apoptotic cell death of HCT-116 cells with perturbance in mitochondrial membrane potential (MMP) and PARP cleavage.

A cross-cultural analysis of Jammu, Kashmir and Ladakh (India) medicinal plant use.

ETHNOPHARMACOLOGICAL RELEVANCE: Jammu & Kashmir (J&K) is a predominantly Himalayan state in the north-western part of India. It has three geographically distinct divisions viz., Jammu, Kashmir and Ladakh, which are immensely rich in their biological and cultural diversity. Medicinal plants are an important element of indigenous medical system of the region. The main goal of the present article is to examine the use of ethnomedicinal plants in three divisions of J&K and to discuss cross-cultural consensus on the use of medicinal plants in these divisions. The article also discusses the gaps in the current state of knowledge on ethnomedicinal plants of the region and gives recommendations for the future studies. MATERIALS AND METHODS: Scientific literature on ethnomedicinal field studies conducted in J&K state of India available in the journals, edited books and other scientific databases viz., CAB international, DOAJ, Google Scholar, PubMed, Science direct, SciFinder, Scopus and Web of Science were searched. Only field based ethnomedicinal surveys from last four decades up to December 2013 reporting first hand information on the medicinal plants used to treat human health related ailments by indigenous communities of J&K were included in this study. Venn diagram was used to analyze the cross-cultural consensus on the use of ethnomedicinal plants in the three divisions of J&K. RESULTS: A total of 948 plant taxa (923 angiosperms, 12 gymnosperms and 13 pteridophytes) belonging to 129 families, 509 genera, 937 species and 11 varieties have so far been reported to have a traditional medicinal use by indigenous communities of J&K. Asteraceae (60 genera, 132 spp.) was the most frequently used family followed by Fabaceae (32 genera, 50 spp.) and Lamiaceae (27 genera, 55 spp.). 514, 415 and 397 medicinal plants were used in Jammu, Kashmir and Ladakh divisions, respectively. Sixty eight plant taxa were used in all the three divisions, whereas 95 plants were common between Ladakh and Jammu, 127 plants between Ladakh and Kashmir, and 216 plants between Jammu and Kashmir. Maximum numbers of plant taxa were used for treating dermatological problems (321), followed by cold, cough and throat related ailments (250), fever (224), joint and muscle related ailments (215), gastrointestinal disorders (210), urogenital ailments (199), respiratory ailments (151), body pain (135) and gynecological disorders (127). CONCLUSIONS: This is the first study from the J&K state, which has examined the medicinal plant use in three divisions of J&K and discussed the promising medicinal plant species with cross-cultural consensus. The analysis of the data suggested that while large numbers of plants are used medicinally in each division, there is a low interregional consensus and high variation between medicinal plants used in these divisions, which is due to both cultural divergence as well as biological distinctness. The issues related to current status of knowledge on medicinal plants used by indigenous communities of J&K have been discussed and some recommendations have been made for future studies on medicinal plants in J&K region.