Screening of Indian plants for biological activity: Part XIV

Alcoholic extracts of 300 botanically identified plant materials from 275 plant species have been tested for various biological activities including chemotherapeutic and pharmacological screenings. Biological activities have been observed in 111 extracts. Follow-up studies have been carried out in some plants with confirmed activity. The active principles and results of these studies are reported.     

Antioxidant and antitopoisomerase activities in plant extracts of some Colombian flora from La Marcada Natural Regional Park

Many plants have been used to treat some diseases and infections since time immemorial, and this potential has been exploited by the pharmaceutical industry in the search of new analgesic, anticarcinogenic and antimicrobial agents, among other active agents. In order to contribute with bioprospection studies on the Colombian flora, 35 extracts from 13 plant species belonging to seven families (Apocynaceae, Cactaceae, Costaceae, Eremolepidaceae, Passifloraceae, Solanaceae and Urticaceae) were collected from La Marcada Natural Regional Park (LMNRP), Colombia. Dichloromethane, n-hexane and aqueous-methanol crude extracts were prepared and evaluated for their activity against Saccharomyces cerevisiae RS322N, R52Y and RS321 strains in the yeast mutant assay and their antioxidant capacity through the DPPH test. The dichloromethane extract from Myriocarpa stipitata (Urticaceae) showed moderate inhibitory activity against the three S. cerevisiae strains tested. The capacity of the dichloromethane extract from M. stipitata to inhibit the enzyme topoisomerase I and to cause DNA damage was inferred from these results. In the DPPH assay, the n-hexane crude extract from Costus sp. (Costaceae) showed good antioxidant activity (48%); in addition, the crude dichloromethane and aqueous-methanol extracts from Rhipsalis micrantha (Cactaceae) showed moderate antioxidant activity with percentage of 29 and 21%, respectively.     

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.     

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.     

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.     

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.     

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。     

Screening of some plants used in the Brazilian folk medicine for the treatment of infectious diseases

Extracts of 13 Brazilian medicinal plants were screened for their antimicrobial activity against bacteria and yeasts. Of these, 10 plant extracts showed varied levels of antibacterial activity. Piper regnellii presented a good activity against Staphylococus aureus and Bacillus subtilis, a moderate activity on Pseudomonas aeruginosa, and a weak activity against Escherichia coli. Punica granatum showed good activity on S. aureus and was inactive against the other standard strains. Eugenia uniflora presented moderate activity on both S. aureus and E. coli. Psidium guajava,Tanacetum vulgare, Arctium lappa, Mikania glomerata, Sambucus canadensis, Plantago major and Erythrina speciosa presented some degree of antibacterial activity. Spilanthes acmella, Lippia alba, and Achillea millefolium were considered inactive. Five of the plant extracts presented compounds with Rf values similar to the antibacterial compounds visible on bioautogram. Of these, three plants belong to the Asteraceae family. This may mean that the same compounds are responsible for the antibacterial activity in these plants. Anticandidal activity was detected in nine plant extracts (P. guajava, E. uniflora, P. granatum, A. lappa, T. vulgare, M. glomerata, L. alba, P. regnellii, and P. major). The results might explain the ethnobotanical use of the studied species for the treatment of various infectious diseases.     

Teleost fish spermatozoa contain a cytosolic protein factor that induces calcium release in sea urchin egg homogenates and triggers calcium oscillations when injected into mouse oocytes

Established studies in a variety of organisms including amphibians, fish, ascidians, nemerteans, echinoderms, mammals, and even a species of flowering plant, clearly demonstrate that an increase in intracellular egg calcium is crucial to the process of egg activation at fertilization. In echinoderms, egg activation appears to involve an egg phospholipase C gamma (PLCgamma). However, numerous studies in mammalian species suggest that calcium is released from internal egg stores at fertilization by a sperm-derived cytosolic protein factor. Recent studies in the mouse have identified this sperm-derived factor as being a novel sperm-specific PLC isoform with distinctive properties (PLCzeta). Homologues of PLCzeta have since been isolated from human and cynomolgus monkey sperm. In addition, sperm factor activity has been detected in non-mammalian species such as chicken, Xenopus, and a flowering plant. Here we report evidence for the existence of a similar sperm-derived factor in a commercially important species of teleost fish, the Nile tilapia Oreochromis niloticus (L). Using an established bioassay for calcium release, the sea urchin egg homogenate, we demonstrate that protein extracts obtained from tilapia spermatozoa exhibit PLC activity similar to that seen in mammalian sperm extracts, and also induce calcium release when added directly to the homogenate. Further, tilapia sperm extracts induced calcium oscillations when injected into mouse oocytes.     

内蒙古产柽柳和多枝柽柳α-葡萄糖苷酶抑制活性

首次利用体外α-葡萄糖苷酶抑制模型以96微孔板法,对内蒙古产2种柽柳属植物不同溶剂提取物进行活性评价,并与阳性对照Acarbose比较,发现6种提取物均有较好的α-葡萄糖苷酶抑制作用,远远强于阳性对照Acarbose(IC50=1103.01μg·mL-1)的抑制活性。结果显示,同一植物不同溶剂提取物相比较,两者石油醚提取物α-葡萄糖苷酶抑制活性不及乙酸乙酯和正丁醇提取物;不同植物同一溶剂的提取物抑制活性也不同,6种提取物中,多枝柽柳的正丁醇和柽柳的乙酸乙酯提取物抑制活性最高(IC50=13.36和17.35μg·mL-1)。所有提取物对α-葡萄糖苷酶活性的抑制效果均很好,且多枝柽柳抑制活性整体上较柽柳好,具有良好的潜在开发价值。     

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.     

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.     

The use of herbarium specimens in DNA phylogenetics: Evaluation and improvement

During the last few years we have been confronted with the need to use herbarium specimens in the molecular phylogeny studies, since it is generally difficult to obtain living material of some rare species. Ancient DNA has been sequenced, and there are also reports on successful DNA amplification from herbarium specimens. However, it is not easy to obtain amplified DNA from the first herbarium sample tested. In this paper, experiments are described about trials of DNA amplification from two to 151-year-old herbarium specimens of plant species we needed for our projects. Of the 17 herbarium samples tested only two allowed DNA amplification under standard DNA isolation conditions. Different types of PCR inhibiting activities were demonstrated in DNA extracts. In some of the extracts there was extremely low concentration of template with satisfactory quality. In some instances, PCR inhibiting activities were successfully removed by treating them either with insoluble polyvinylpyrrolidone or by adding bovine serum albumin (BSA) to the amplification mixture. However, some PCR-inhibiting activities were resistant to the treatments described above. When the concentration of template was very low, a second PCR amplification with internal primers was necessary to increase the amount of DNA for sequencing. Nevertheless, contamination of either DNA extract or amplification mixture were sometimes observed, and consequently precautions were taken to minimize them. Finally, successful amplification was obtained in eight samples out of the 17 examined.     

Seasonal variation in antimicrobial and phytochemical properties of frequently used medicinal bulbous plants from South Africa

The growing popularity of traditional medicine and the unrestricted collection of medicinal plants from the wild have put many of the slow growing bulbous plant species at the risk of over-exploitation and extinction in South Africa. This study was aimed at comparing the phytochemical composition and biological (antibacterial and anticandidal) activities of bulb and leaf extracts of Tulbaghia violacea, Hypoxis hemerocallidea, Drimia robusta and Merwilla plumbea between spring, summer, autumn and winter seasons, with the view of promoting the use of leaves, as a conservation strategy. Antibacterial and anticandidal activities of petroleum ether (PE), dichloromethane (DCM), 80% ethanol and water extracts of bulbs and leaves were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae bacteria and the fungus Candida albicans using the microdilution bioassay. Spectrophotometric methods were used to evaluate saponin and phenolic compositions for the four seasons. Leaf and bulb extracts exhibited comparable anticandidal activity (MIC < 1 mg/ml) in all the plant species in all seasons. Only ethanol and water extracts of H. hemerocallidea corms (autumn and winter) showed correspondingly good fungicidal activity amongst the bulbs tested. Antibacterial activity was fairly comparable between bulbs and leaves with at least one extract of each plant species showing some good MIC values in most of the seasons. The best antimicrobial activities were recorded in winter and autumn seasons, with MIC values as low as 0.2 mg/ml in the DCM bulb extracts of T. violacea (winter) against K. pneumoniae and S. aureus. The amounts of total phenolic compounds in all plant samples were generally higher in spring compared to the other seasons. Condensed tannin, gallotannin and flavonoid levels, depending on the sample, were either higher in spring or winter except for H. hemerocallidea (corm) which had higher gallotannin levels in autumn. Total saponin levels were higher in winter in all plant samples. Although variation was observed in the phytochemical concentrations between the bulbs and leaves of each plant species, their antimicrobial activities were fairly comparable. Leaves may be used as substitutes for bulbs in the treatment of bacterial and fungal ailments.     

Precipitation of the antigens of the causative agents of plague, glanders and melioidosis using aqueous saline extracts of various plant species

The interaction of the aqueous saline extracts of 55 plant species with the antigens of the causative agents of plague, glanders and melioidosis in the reaction of immunodiffusion (RID) in gel has been studied. The aqueous saline extracts obtained from the seeds of 12 plant species have been revealed to possess precipitating capacity. At the same time these extract have been found to ensure, as a rule, RID with several antigens under test.     

In vitro evaluation of antibacterial,anticollagenase, and antioxidant activities of hop components (Humulus lupulus) addressing acne vulgaris

Seven naturally derived components from hop plant (Humulus lupulus L.) extracts were tested for evaluation of biological activities affecting acne vulgaris. Five strains, Propionibacterium acnes, Staphylococcus epidermidis, Staphylococcus aureus, Kocuria rhizophila and, Staphylococcus pyogenes, were selected as the main acne-causing bacteria. Hop extracts xanthohumol and the lupulones showed strong inhibitory activities against all of the strains. Although hydrogenated derivatives did not show the same level of activity, naturally occurring xanthohumol, humulones, and lupulones all showed moderate to strong anticollagenase inhibitory activities. Antioxidant capacity was also evaluated with seven different methods based on different reactive oxygen species. Xanthohumol showed the highest activity in total oxygen radical absorbance capacity as well as singlet oxygen absorbance capacity.     

Ecdysteroid agonist and antagonist activities in species of the Solanaceae

Previously, it has been shown that certain withanolides from Iochroma gesnerioides (Solanaceae) possess ecdysteroid antagonistic activity. Phytoecdysteroids (agonists) are widely distributed in the plant world, but solanaceous species have not been extensively examined for their presence. We have now surveyed 128 species of solanaceous plants for the presence of ecdysteroid agonist and antagonist activities using the Drosophila melanogaster B(II) cell line bioassay. Only weak antagonistic activity was associated with a few of the methanolic extracts, including those from species known to contain high levels of withanolides. Therefore, the major withanolides are inactive per se, but they may be activated after ingestion by invertebrate predators. Several extracts possessed ecdysteroid agonist activity as a consequence of the presence of phytoecdysteroids. Phytoecdysteroid-accumulating species are at least as common in the Solanaceae as they are in plants in general. Preliminary characterization of the identities of the phytoecdysteroids present in the most active extracts has been performed by hplc separations on normal- and reversed-phase systems in conjunction with ecdysteroid-specific radioimmunoassay and bioassay. Each of the phytoecdysteroid-accumulating species examined (Browallia speciosa, Nierembergia hippomanica var violacea, N. solanacea and Solanum nigrum) contain a cocktail of ecdysteroids, of which 20-hydroxyecdysone and polypodine B (5beta,20-dihydroxyecdysone) are major components.     

Ecological phytochemistry of Cerrado (Brazilian savanna) plants

The Cerrado (the Brazilian savanna) is one of the vegetation formations of great biodiversity in Brazil and it has experienced strong deforestation and fragmentation. The Cerrado must contain at least 12,000 higher plant species. We discuss the ecological relevance of phytochemical studies carried out on plants from the Cerrado, including examples of phytotoxicity, antifungal, insecticidal and antibacterial activities. The results have been classified according to activity and plant family. The most active compounds have been highlighted and other activities are discussed. A large number of complex biochemical interactions occur in this system. However, only a small fraction of the species has been studied from the phytochemical viewpoint to identify the metabolites responsible for these interactions.     

6种陆生植物提取物对白脊藤壶无节幼体的毒杀活性比较

对夹竹桃科（Apocynaceae）的红花夹竹桃（Nerium indicum Mill．）和黄花夹竹桃（Thevetia peruviana K．Schum．）、百合科（Li｜iaceae）的洋葱（Allium cepa L．）和大蒜（A．sativum L．）、楝科（Meliaceae）的苦楝（Melia azedarach L．）和印楝（Azadirachta indica A．Juss．）等6种陆生植物的不同溶剂提取物对白脊藤壶（Balanus albicostatus Pilsbry）无节幼体的毒杀作用进行了比较研究。结果表明,供试的楝科和夹竹桃科的4种植物提取物对白脊藤壶无节幼体的毒杀活性高于洋葱和大蒜;不同提取物的毒杀活性与植物的种类及提取部位（叶、花）以及活性成分的类型有关,用非极性有机溶剂提取的脂溶性提取物的毒杀活性高于极性溶剂提取物。处理12h,0．3％印楝素乳油的乙酸乙酯及95％乙醇稀释液的毒杀活性最高,LC50仅为0．33和3．01g·mL^-1,可作为防治海洋污损生物的植物资源进行进一步的开发利用。