Screening of Indian plants for biological activity: Part XVI

Alcoholic extracts of 288 of plant materials from 199 plant species have been tested for various biological activities including chemotherapeutic and pharmacological screening. Biological activities, ranging from moderate to good degree, have been observed in 61 plants extracts. Follow up studies have been carried out in these extracts and some of them have shown moderate degree of activities at this Institute. However, none of the extracts was found to be good enough for further development. Results of the present studies, along with chemical investigations on different species of similar genera which were screened earlier, are also discussed.     

Death by proteases in plants: whodunit

Several studies have shown that protease inhibitors can suppress programmed cell death in various plant species and plant tissues. This is especially true of caspase inhibitors that can block programmed cell death and its marker DNA laddering. There are up to six different caspase-like activities that can be measured in plant extracts, the most prominent being caspase1-like and caspase3-like. These activities can be located in vacuoles and also in the nucleus or the cytoplasm. This represents a striking apparent similarity with animal programmed cell death. Because there are no caspase orthologue in plant genomes, a major challenge is to identify these proteases. Recently two proteases with caspase-like activities have been recognized as belonging to two different protease families that are not closely related to animal caspases. Various other protease families have been implicated and this suggests that complex protease networks have been recruited for the plant cell demise.     

What happened to plant caspases?

The extent of conservation in the programmed cell death pathways that are activated in species belonging to different kingdoms is not clear. Caspases are key components of animal apoptosis; caspase activities are detected in both animal and plant cells. Yet, while animals have caspase genes, plants do not have orthologous sequences in their genomes. It is 10 years since the first caspase activity was reported in plants, and there are now at least eight caspase activities that have been measured in plant extracts using caspase substrates. Various caspase inhibitors can block many forms of plant programmed cell death, suggesting that caspase-like activities are required for completion of the process. Since plant metacaspases do not have caspase activities, a major challenge is to identify the plant proteases that are responsible for the caspase-like activities and to understand how they relate, if at all, to animal caspases. The protease vacuolar processing enzyme, a legumain, is responsible for the cleavage of caspase-1 synthetic substrate in plant extracts. Saspase, a serine protease, cleaves caspase-8 and some caspase-6 synthetic substrates. Possible scenarios that could explain why plants have caspase activities without caspases are discussed.     

Bioactivity of marine organisms: Part VI--Screening of some marine flora from Indian coasts.

Alcoholic extracts of 50 botanically identified species of marine flora have been screened for a wide range of biological activities. Of these, 2 extracts exhibited anti-amoebic and antiviral activity each, 3 of them had anti-implantation activity; 9 had hypoglycaemic activity while hypotensive activity was associated with 11 extracts; 14 extracts were found to be diuretic and 1 of them had anti-inflammatory activity. Further, 10 of these extracts exhibited 2 types of activities while a combination of 3 and 4 types of activities was observed in one extract each. Follow-up studies have been carried out in some plants with confirmed activity. The active principles and results of these studies are reported.     

Bacopa monniera, a reputed nootropic plant: an overview

Bacopa monniera (BM), a traditional Ayurvedic medicine, used for centuries as a memory enhancing, anti-inflammatory, analgesic, antipyretic, sedative and antiepileptic agent. The plant, plant extract and isolated bacosides (the major active principles) have been extensively investigated in several laboratories for their neuropharmacological effects and a number of reports are available confirming their nootropic action. In addition, researchers have evaluated the anti-inflammatory, cardiotonic and other pharmacological effects of BM preparations/extracts. Therefore, in view of the important activities performed by this plant, investigation must be continued in the recently observed actions described in this paper. Moreover, other clinical studies have to be encouraged, also to evidence any side effects and possible interactions between this herbal medicine and synthetic drugs.     

石菖蒲提取物对四种储粮害虫的生物活性研究

研究了石菖蒲4种溶剂无水乙醇、丙酮、乙酸乙酯、石油醚提取物对玉米象、谷蠹、长角扁谷盗和锯谷盗4种储粮害虫的驱避作用和触杀作用。结果表明：石菖蒲4种溶剂提取物对4种试虫均有明显的驱避作用和触杀作用,处理60 h的平均驱避等级均达到Ⅲ级以上,对4种试虫的触杀死亡率均达到41.11%以上。     

Optimal methods for evaluating antimicrobial activities from plant extracts

The search for antimicrobial agents from plants has been a growing interest in the last few decades. However, results generated from many of these studies cannot be directly compared due to the absence of standardization in particular antimicrobial methods employed. The need for established methods with consistent results for the evaluation of antimicrobial activities from plant extracts has been proposed by many researchers. Nevertheless, there are still many studies reported in the literature describing different methodologies. The aim of this study was to find optimal methods to give consistent quantitative antimicrobial results for studying plant extracts. Three different agar-based assays (pour plate disc diffusion (PPDD), streak plate disc diffusion (SPDD) and well-in agar (WA)) and one broth-based (turbidometric (TB)) assay were used in this study. Extracts from two plant species (Duabanga grandiflora and Acalypha wilkesiana) were tested on two bacterial species, namely Escherichia coli and Staphylococcus aureus. Amongst the agar-based assays, PPDD produced the most reproducible results. TB was able to show the inhibitory effects of the test samples on the growth kinetic of the bacteria including plant extracts with low polarity. We propose that both agar- (i.e PPDD) and broth-based assays should be employed when assessing the antimicrobial activity of plant crude extracts.     

Antioxidant and Antimicrobial Activity in Some Indian Herbal Plants: Protective Effect against Free Radical Mediated DNA Damage

Indian herbal plant species Lantana indica, Adhatoda vasica, Pandanus furcatus, Tylophora indica and Centella asiatica, traditionally used in ethno medicines to treat common infections and various disorders, have been studied for their antimicrobial and antioxidant activity. The methanolic extracts of the plant leaves exhibited significant and dose-dependent antioxidant activities in DPPH radical scavenging, ferric ion reducing and phosphomolybdate assays. These leaf extracts showed antimicrobial activity against selected Gram +ve and Gram ?ve bacterial strains. A. vasica and L. indica extracts possessed maximum antioxidant and antimicrobial activity, respectively. The activities could be correlated to phenolics and flavonoid content of the leaf extracts which ranged from 30.25 to 91.98 mg GAE g^?1 dw leaf extract and 2.67 to 96.45 mg RE g^?1 dw leaf extract respectively. The aqueous extracts of plant leaves significantly protected the DNA damage against the oxidative damage caused by hydroxyl radicals.     

Challenges and complexity of functionality evaluation of flavan-3-ol derivatives

Flavan-3-ol derivatives are common plant-derived bioactive compounds. In particular, (–)-epigallocatechin-3-O-gallate shows various moderate biological activities without severe toxicity, and its health-promoting effects have been widely studied because it is a main ingredient in green tea and is commercially available at low cost. Although various biologically active flavan-3-ol derivatives are present as minor constituents in plants as well as in green tea, their biological activities have yet to be revealed, mainly due to their relative unavailability. Here, I outline the major factors contributing to the complexity of functionality studies of flavan-3-ol derivatives, including proanthocyanidins and oligomeric flavan-3-ols. I emphasize the importance of conducting structure-activity relationship studies using synthesized flavan-3-ol derivatives that are difficult to obtain from plant extracts in pure form to overcome this challenge. Further discovery of these minor constituents showing strong biological activities is expected to produce useful information for the development of functional health foods.     

Mechanisms underlying antigonadotropic effects of some traditional plant extracts in pituitary cell culture.

Previous works have demonstrated that stem bark extracts of Cola nitida (Sterculiaceae), Afrormosia laxiflora and Pterocarpus erinaceus (Fabaceae) provoked a blockade of female rat ovulation and estrous cycle by inhibiting pituitary LH release in vivo. In addition, these plant extracts exerted an inhibitory effect on LH release of rat pituitary cells. Therefore, these data could explain inhibitory effects of plant extracts on LH release in vivo. The present study was undertaken to examine the mechanisms by which these plants exert their antigonadotropic activities. So, we studied the biological activities of these plant extracts on pituitary cells in culture. Data show that C. nitida, A. laxiflora and P. erinaceus extracts only inhibit LH release and have no effect on FSH release. In fact, A. laxiflora, P. erinaceus and C. nitida extracts diminish LH release in culture medium without acting on rat pituitary cell content. Plant extracts form complexes with basic glycoproteins (but not with acid glycoproteins) and prevent them from entering the cells.     

Cotinus coggygria Scop.: An overview of its chemical constituents,pharmacological and toxicological potential

The Anacardiaceae Lindl. family comprises of many species which are used in nutrition and in traditional folk medicine for the treatment of several human diseases. Cotinus coggygria Scop. commonly known as “smoke tree”, is a commercial ornamental plant with high medicinal usages, belongs to the family Anacardiaceae. The present review provides a comprehensive report of empirical investigations on important pharmacological activities and phytochemical screening of essential oils and extracts. Relevant information was collected from scientific journals, books, and reports via library and electronic search using Medline, PubMed, Google Scholar, ScienceDirect, Web of Science, and Scopus. The plant has been extensively investigated in a broad range of studies to provide scientific evidence for folklore claims or to find new therapeutic uses. Numerous activities namely antioxidative, antibacterial, antifungal, antiviral, anticancer, antigenotoxic, hepatoprotective and anti-inflammatory have been demonstrated for all parts of these plants by in vivo and in vitro studies. Essential oils and extracts showed various pharmacological and biological properties which make them an effective remedy for various kinds of illnesses. Considering data from the literature, it could be demonstrated that C. coggygria possesses diverse bioactive properties and immense utilization in medicine, health care, cosmetics and as health supplements.     

Native growth inhibitors from citrus shoots. Partition, bioassay, and characterization

A partition method has been devised to separate auxins, gibberellins, and their respective inhibitors in plant extracts. Inhibitors counteracting gibberellin activity have been detected by a modified barley endosperm bioassay. An inhibitor found in young citrus shoots counteracts both auxin and gibberellin activities and behaves during partition and ohromatography like abscisin II.     

Characterization of topoisomerase I and II activities in nuclear extracts during callogenesis in immature embryos of Zea mays

We have characterized the topoisomerase I and II activities in nuclear extracts from immature embryos of Zea mays and the effect of the treatment with 2,4-dichlorophenoxyacetic acid (2,4-D) and abscisic acid (ABA). These extracts were shown to be essentially devoid of protease and nuclease activities and they were tested for their ability to relax supercoiled DNA, unknotting P4 DNA and catenate circular duplex DNA under catalytic conditions. Unknotting and catenation reactions are strictly magnesium- and ATP-dependent, but not the relaxation of circular supercoiled DNA allowing the detection of both topoisomerase I and II activities. Two cytotoxic drugs, camptothecin, a plant alkaloid that inhibits cukaryotic topoisomerase I, and epipodophyllotoxin VM-26 (teniposide) that inhibits topoisomerase II, have been assayed in our extracts showing similar inhibitory effects on topoisomerase enzymes. Alkaline phosphatase treatment of nuclear extracts abolishes both topoisomerase activities. Nuclear extracts from embryos treated with 2,4-D showed 200% increase on topoisomerase II activity as compared with untreated ones, but only residual activity was detected in ABA-treated embryos. Nuclear extracts from hormone-treated and untreated embryos showed similar topoisomerase I activity with deviations of less than 25%. These differences are discussed in terms of possible post-translational modifications of the enzymes associated with the increase in proliferation activity of calli.     

Effects of anthocyanins and anthocyanin-rich extracts on the risk for cancers of the gastrointestinal tract

Anthocyanins are the largest group of water-soluble pigments in the plant kingdom. Anthocyanins are responsible for most of the red, blue, and purple colors of fruits, vegetables, flowers, and other plant tissues or products. In recent years, numerous studies have shown that anthocyanins display a wide range of biological activities. This review summarises recent literature evidence on the association of anthocyanins and anthocyanin-rich extracts consumption with the risk for gastrointestinal tract cancer, concentrating on the results from in vivo animal model tumor systems, as well as data from human epidemiological studies. Potential cancer chemopreventive activities of anthocyanins were revealed from in vitro studies. In vivo animal model tumor systems showed that dietary anthocyanins inhibit cancers of the gastrointestinal tract. Some epidemiological studies have revealed protective effects of anthocyanins consumption on gastrointestinal cancer risk in humans. Pharmacokinetic data indicate that absorption of anthocyanins into the bloodstream of rodents and humans is minimal, suggesting that they may have little efficacy in tissues other than the gastrointestinal tract and skin. Future studies should be undertaken to determine if the anticancer effects of anthocyanins are due to the parent compounds and/or to their metabolites.     

Metabolic Activities in Extracts of Mesophyll and Bundle Sheath Cells of Panicum miliaceum (L.) in Relation to the C(4) Dicarboxylic Acid Pathway of Photosynthesis

The activities of certain enzymes related to the carbon assimilation pathway in whole leaves, mesophyll cell extracts, and bundle sheath extracts of the C₄ plant Panicum miliaceum have been measured and compared on a chlorophyll basis. Enzymes of the C₄ dicarboxylic acid pathway—phosphoenolpyruvate carboxylase and NADP-malic dehydrogenase—were localized in mesophyll cells. Carbonic anhydrase was also localized in mesophyll cell extracts. Ribose 5-phosphate isomerase, ribulose 5-phosphate kinase, and ribulose diphosphate carboxylase—enzymes of the reductive pentose phosphate pathway—were predominantly localized in bundle sheath extracts. High activities of aspartate and alanine transaminases and glyceraldehyde-3-P dehydrogenase were found about equally distributed between the photosynthetic cell types. P. miliaceum had low malic enzyme activity in both mesophyll and bundle sheath extracts.     

Polysaccharide microarrays for high-throughput screening of transglycosylase activities in plant extracts

Polysaccharide transglycosylases catalyze disproportionation of polysaccharide molecules by cleaving glycosidic linkages in polysaccharide chains and transferring their cleaved portions to hydroxyl groups at the non-reducing ends of other polysaccharide or oligosaccharide molecules. In plant cell walls, transglycosylases have a potential to catalyze both cross-linking of polysaccharide molecules and grafting of newly arriving polysaccharide molecules into the cell wall structure during cell growth. Here we describe a polysaccharide microarray in form of a glycochip permitting simultaneous high-throughput monitoring of multiple transglycosylase activities in plant extracts. The glycochip, containing donor polysaccharides printed onto nitrocellulose-coated glass slides, was incubated with crude plant extracts, along with a series of fluorophore-labelled acceptor oligosaccharides. After removing unused labelled oligosaccharides by washing, fluorescence retained on the glycochip as a result of transglycosylase reaction was detected with a standard microarray scanner. The glycochip assay was used to detect transglycosylase activities in crude extracts from nasturtium (Tropaeolum majus) and mouse-ear cress (Arabidopsis thaliana). A number of previously unknown saccharide donor-acceptor pairs active in transglycosylation reactions that lead to the formation of homo- and hetero-glycosidic conjugates, were detected. Our data provide experimental support for the existence of diverse transglycosylase activities in crude plant extracts.     

Fat metabolism in higher plants. Characterization of plant acyl-ACP and acyl-CoA hydrolases.

Avocado mesocarp extracts contain both acyl-CoA and acyl-acyl carrier protein (ACP) hydrolase activities. These activities have been separated by a sequence of ammonium sulfate fractionations, DEAE-cellulose, and hydroxyapatite column chromatography. Two distinct acyl-CoA hydrolase fractions and one acyl-ACP hydrolase fraction were obtained. The acyl-ACP hydrolase fraction was essentially free of acyl-CoA hydrolase activity, had a pH optimum of 9.5 and a molecular weight of 70–80,000 based on sucrose density gradient centrifugation and gel filtration chromatography. Substrate specificity studies showed that lauroyl-ACP, myristoyl-ACP, palmitoyl-ACP, and stearoyl-ACP were slowly hydrolyzed but oleoyl-ACP was rapidly hydrolyzed to free fatty acid. These results suggest a possible role for acyl-ACP hydrolase as one component of a switching system which allows, indirectly, acyl transfer from ACP to CoA derivatives in plant cells.     

Mushrooms, tumors, and immunity: an update

There is significant interest in the use of mushrooms and/or mushroom extracts as dietary supplements based on theories that they enhance immune function and promote health. To some extent, select mushrooms have been shown to have stimulatory action on immune responsiveness, particularly when studied in vitro. However, despite their widespread use for potential health benefits, there is a surprising paucity of epidemiologic and experimental studies that address the biologic activities of mushrooms after oral administration to animals or humans. There have been a number of studies that have addressed the ability of mushrooms to modulate mononuclear cell activation and the phenotypic expression of cytokines and their cognate receptors. There have also been a number of attempts to determine antitumor activities of mushrooms. Such studies are important because many of the components of mushrooms do potentially have significant biologic activity. All data, however, should be tempered by the possibility that there are toxic levels of metals, including arsenic, lead, cadmium, and mercury as well as the presence of radioactive contamination with 137Cs. In this review, we will present the comparative biology with respect to both immunological and antitumor activities of mushroom extracts and also highlight the need for further evidence-based research.     

A potential antioxidant resource: Endophytic fungi from medicinal plants

Medicinal plants and their endophytes are important resources for discovery of natural products. Several previous studies have found a positive correlation between total antioxidant capacity (TAC) and total phenolic content (TPC) of many medicinal plant extracts. However, no information is available on whether such a relationship also exists in their endophytic fungal metabolites. We investigated the relationship between TAC and TPC for 292 morphologically distinct endophytic fungi isolated from 29 traditional Chinese medicinal plants. The antioxidant capacities of the endophytic fungal cultures were significantly correlated with their total phenolic contents, suggesting that phenolics were also the major antioxidant constituents of the endophytes. Some of the endophytes were found to produce metabolites possessing strong antioxidant activities. Several bioactive constituents from the fungal cultures and host plant extracts were identified. This investigation reveals that the metabolites produced by a wide diversity of endophytic fungi in culture can be a potential source of novel natural antioxidants.     

Anti-herpes simplex virus activities of crude water extracts of Thai medicinal plants.

A number of Thai medicinal plants, recommended as remedies for herpesvirus infection and have been used in primary health care were investigated for their intracellular activities against herpes simplex viruses (HSV). Centella asiatica L., Maclura cochinchinensis Cornor, and Mangifera indica L. contained both anti-HSV-1 and -2 activities, as determined by plaque inhibition assay. An inhibition of the production of infectious HSV-2 virions from infected Vero cells could also be demonstrated. Combinations of each of these reconstituted extracts with 9-(2-hydroxyethoxymethyl) guanosine (acyclovir; ACV) resulted either in subadditive, additive, or synergistic interaction, against HSV-2, depending on the dose of ACV used; mixture of C. asiatica and M. indica exerted an additive effect in a similar assay. Furthermore, the inhibitory effects of these plant extracts were also substantiated by flow cytometric analysis of virus-specific antigens in the infected cells. The active constituent present in C. asiatica extract was determined to be asiaticoside while in M. indica was mangiferin. Thus, these data suggest therapeutic potential of these plant extracts.