Neem components as potential agents for cancer prevention and treatment

Azadirachta indica, also known as neem, is commonly found in many semi-tropical and tropical countries including India, Pakistan, and Bangladesh. The components extracted from neem plant have been used in traditional medicine for the cure of multiple diseases including cancer for centuries. The extracts of seeds, leaves, flowers, and fruits of neem have consistently shown chemopreventive and antitumor effects in different types of cancer. Azadirachtin and nimbolide are among the few bioactive components in neem that have been studied extensively, but research on a great number of additional bioactive components is warranted. The key anticancer effects of neem components on malignant cells include inhibition of cell proliferation, induction of cell death, suppression of cancer angiogenesis, restoration of cellular reduction/oxidation (redox) balance, and enhancement of the host immune responses against tumor cells. While the underlying mechanisms of these effects are mostly unclear, the suppression of NF-κB signaling pathway is, at least partially, involved in the anticancer functions of neem components. Importantly, the anti-proliferative and apoptosis-inducing effects of neem components are tumor selective as the effects on normal cells are significantly weaker. In addition, neem extracts sensitize cancer cells to immunotherapy and radiotherapy, and enhance the efficacy of certain cancer chemotherapeutic agents. This review summarizes the current updates on the anticancer effects of neem components and their possible impact on managing cancer incidence and treatment.     

The Integrated Effect of the Nematode-Bacteria Complex and Neem Plant Extracts Against Plutella Xylostella (L.) Larvae (Lepidoptera: Yponomeutidae) on Chinese Cabbage

The compatibility of the nematode-bacteria complex of Steinernema carpocapsae, Steinernema abbasi, and Heterorhabditis bacteriophora with neem plant extracts (seeds and leaves) to control diamondback moth, Plutella xylostella (L.) on Chinese cabbage plants was investigated under laboratory conditions. The initial treatment of Chinese plant leaves with neem plant extracts resulted in more susceptibility of P. xylostella to neem seeds extract than neem leaves extract. The combination studies resulted in a higher synergistic effect of nematode-bacteria complex of H. bacteriophora, S. abbasi and S. carpocapsae with neem seeds extract compared with that of S. abbasi with neem leaves extract. In contrast, the combination of nematode-bacteria complex of S. carpocapsae and H. bacteriophora with neem leaves extract gave an antagonistic effect. On the other hand, the time-mortality response was more reduced with neem seeds extract than with neem leaves extract, besides it was a concentration dependant factor. We conclude from these investigations that the combination of entomopathogenic nematodes with neem plant extracts could be of success in controlling P. xylostella on Chinese cabbage plants in the field.     

Ethanolic leaf extract of neem (Azadirachta indica) inhibits buccal pouch carcinogenesis in hamsters

We evaluated the chemopreventive effects of ethanolic neem leaf extract in the initiation and post-initiation phases of 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. The frequency of bone marrow micronuclei as well as the concentrations of lipid peroxides, ratio of reduced to oxidized glutathione (GSH/GSSG), and the activities of the GSH-dependent enzymes glutathione peroxidase (GPx) and glutathione-S-transferase (GST) in the buccal pouch, liver and erythrocytes were used as biomarkers of chemoprevention. All the hamsters painted with DMBA alone for 14 weeks developed buccal pouch carcinomas that showed diminished lipid peroxidation and enhanced antioxidant status associated with increased frequencies of bone marrow micronuclei. In the liver and erythrocytes of tumour-bearing animals, enhanced lipid peroxidation was accompanied by compromised antioxidant defences. Administration of ethanolic neem leaf extract effectively suppressed DMBA-induced HBP carcinogenesis as revealed by the absence of tumours in the initiation phase and reduced tumour incidence in the post-initiation phase. In addition, ethanolic neem leaf extract modulated lipid peroxidation and enhanced antioxidant status in the pouch, liver and erythrocytes and reduced the incidence of bone marrow micronuclei. The results of the present study, demonstrate that ethanolic neem leaf extract inhibits the development of DMBA-induced HBP tumours by protecting against oxidative stress.     

Effects of population size on reproductive success of the endangered and endemic species Primula merrilliana

The reproductive ability related to the population size of the endangered and endemic species Primula merrilliana Schltr.was investigated.In 26 populations observed,only four contain more than 500 flowering individuals,whereas most of them(53.8%) consist of less than 100 flowering individuals.Though the ratio of pin and thrum plants keeps its balance at 1:1 for all populations,the frequency of pin and thrum flowers was significantly biased in most small populations.Population size strongly affected reproductive success; plants in small populations produced significantly fewer flowers and fruits per plant and fewer seeds per fruit,and therefore fewer seeds per plant.The floral density was another important factor that influenced the reproductive success of P.merrilliana,because four main reproductive success parameters (fruits per plant,seeds per fruit,seeds per plant,and the proportion of flowers setting fruit) were all positively correlated with floral density.The size of plants and the number of leaves per plant (measure of habitat quality) were not influenced by the variation of population size,suggesting that the reduced fecundity in small populations may not be a consequence of lower habitat quality,inbreeding depression and pollen limitation as a result of less attractiveness in small populations are therefore likely explanations for these patterns.     

Seasonal variation of diet and food availability in a group of Sichuan snub-nosed monkeys in Shennongjia Nature Reserve, China

We studied the diet and food availability of a group of Sichuan snub-nosed monkeys for 14 months (July 2003 to September 2004, except for February) in the Shennongjia Nature Reserve, China. This species is primarily a lichen eater, with lichens (Usneaceae) accounting for 43.28% of feeding records (n=3,452). Other food types in the diet were young leaves (28.71%), fruits or seeds (14.57%), buds (5.36%), mature leaves (3.51%), herbs (2.09%), bark (1.36%), and flowers (1.13%). The monkeys used 23 plant species. Their diet showed a complicated seasonal variation: the monthly diet varied from primarily lichens in November-April, to a mixture of leaves and lichens in May-July, to a mixture of fruits or seeds and lichens in August-October (the latter depended on annual fruit and seed availability). The proportion of fruits or seeds in the diet was negatively correlated with that of lichens, which suggests that the monkeys prefer fruits or seeds to lichens when all of these items are available. The fruit or seed availability varied greatly between the two study years. The proportion of lichens, young leaves, flowers, and fruits or seeds in the diet was positively associated with their availability. The monkeys appeared to be selective feeders. They preferred 10 tree species for plant parts, and nine tree species for lichens. The selection index of tree species for lichens was positively related to lichen coverage per branch on tree species, demonstrating that the monkeys preferred tree species with abundant lichens, as well as dead trees for lichens. The results suggest that dead-tree harvesting in the reserve could significantly reduce the quality of habitat for these monkeys, and should therefore be prohibited. Connus controversa, Cerasus discadenia, Salix willichiana, and Malus halliana should be conserved as top priority species because the monkeys preferred them for both their vegetative parts and the lichens that grow on them.     

Modulatory effects of garlic and neem leaf extracts on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced oxidative stress in Wistar rats

The effects of garlic and neem leaf extracts on lipid peroxidation and antioxidant status during administration of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), a carcinogenic nitrosamine were evaluated in male Wistar rats. Extracts of garlic and neem leaf were administered orally for five consecutive days before intraperitoneal injection of MNNG. Enhanced lipid peroxidation in the stomach, liver and circulation of MNNG-treated rats was accompanied by a significant decrease in glutathione (GSH) and the activities of glutathione peroxidase (GPx), glutathione-S-transferase (GST) and gamma glutamyl transpeptidase (GGT). Administration of garlic and neem leaf extracts significantly decreased the formation of lipid peroxides and enhanced the levels of antioxidants and detoxifying enzymes in stomach, the primary target organ for MNNG, as well as in the liver and circulation. The results of the present study suggest that garlic and neem may exert their protective effects by modulating lipid peroxidation and enhancing the levels of GSH and GSH-dependent enzymes.     

Evaluation of neem (Azadirachta indica L.) products for the control of root-knot nematode of chilli pepper (Capsicum annum L.)

Chilli pepper is an important vegetable in the diet of Ghanaians but there are some constraints militating against increased production notable among which is nematode infestation. A study was therefore carried out at the plant house of the Faculty of Agriculture, University for Development Studies, Nyankpala to investigate the efficacy of neem products against root-knot nematode of chilli pepper (Capsicum annum L.). The treatments which included 20 g neem leaf powder, 20 g neem cake, 20 g neem seed powder and control were arranged in a completely randomised design with four replications. Neem products significantly reduced (p < 0.05) nematode population, with 20 g of neem leaf powder recording the highest reduction. Similarly, plants treated with neem leaf powder produced a significantly lower (p < 0.05) number of root galls compared to the other treatments. Neem leaf powder could be used as a nematicide for nematodes control as a replacement for synthetic nematicides.     

Increased insect resistance in transgenic wheat stably expressing trypsin inhibitor CMe

Proteinase inhibitors have been proposed to function as plant defence agents against herbivorous pests. We have introduced the barley trypsin inhibitor CMe (BTI-CMe) into wheat (Triticum aestivum L.) by biolistic bombardment of cultured immature embryos. Of the 30 independent transgenic wheat lines selected, 16 expressed BTI-CMe. BTI-CMe was properly transcribed and translated as indicated by northern and western blot, with a level of expression in transgenic wheat seeds up to 1.1% of total extracted protein. No expression was detected in untransformed wheat seeds. Functional integrity of BTI-CMe was confirmed by trypsin inhibitor activity assay. The significant reduction of the survival rate of the Angoumois grain moth (Sitotroga cerealella, Lepidoptera: Gelechiidae), reared on transgenic wheat seeds expressing the trypsin inhibitor BTI-CMe, compared to the untransformed control confirmed the potential of BTI-CMe for the increase of insect resistance. However, only early-instar larvae were inhibited in transgenic seeds and expression of BTI-CMe protein in transgenic leaves did not have a significant protective effect against leaf-feeding insects.     

非木材森林产品利用的生态影响

非木材森林产品指来自森林的非木材植物产品,如果实、种子、枝叶、树皮、树胶等,在维持当地居民生活以及森林生物多样性保护中扮演着重要角色.非木材森林产品利用对植物个体、种群、群落和生态系统均会产生影响,这不仅取决于植物的利用部位、生物学或生态学特征,而且与资源的利用方式密切相关.本文从植物生物学和生态学角度综述了非木材森林产品利用影响的研究进展,讨论了研究中存在的问题和今后的研究方向.     

Aqueous fruit extracts of Azadirachta indica induce systemic acquired resistance in barley against Drechslera graminea

The study reported here primarily focuses on whether fruit extracts of Azadirachta indica Juss. (Neem) can induce systemic acquired resistance (SAR) in Hordeum vulgare against Drechslera graminea. A single leaf from each of the 1-month-old seedlings grown in 50 pots was treated with neem extract. Seven samples were collected at 12-h intervals for estimation of salicylic acid (SA) content and activities of phenylalanine ammonia lyase (PAL) and peroxidase (PO). Disease incidence was recorded on uninoculated leaves after 2 weeks and on newly emerged leaves after 3 weeks of inoculation of spores of the pathogen. Treatment of single leaf of barley seedlings with aqueous fruit extract of neem could protect the untreated and later emerging leaves of these seedlings from infection by leaf stripe pathogen. The concentration of SA and activities of PAL and PO were significantly higher in untreated leaves of seedlings given a single leaf treatment with neem fruit extract. The results show that neem fruit extract induced SAR in barley seedlings against D. graminea. The results of the study are significant for developing an environment-friendly biocide, which could induce SAR in crop plants leading to efficient management of pathogens     

Deterring rodent seed‐predation using seed‐coating technologies

With many degraded environments undergoing restoration efforts, there is a growing need for the optimization of direct seeding practices. Seeds planted on wildlands are often consumed by rodents, leading to reduced plant establishment. Coating seeds in rodent aversive products may prevent seed‐predation. We tested 10 seed‐coating formulations containing products expected to deter rodents, namely: ghost and cayenne pepper powders; essential oils from bergamot, neem, and pine; methyl‐nonyl‐ketone, anthraquinone, activated carbon, beta‐cyclodextrin, and a blank coating containing no rodent deterrents to serve as a control treatment. Each treatment was applied to Pseudoroegneria spicata (bluebunch wheatgrass) seeds. These seeds germinated similarly to uncoated control seeds unless the coating contained methyl‐nonyl‐ketone which reduced germination. When seeds were offered to Ord's kangaroo rats (Dipodomys ordii), they strongly avoided the treatments in favor of uncoated control seeds. Notably, the blank coating, lacking active ingredients, still elicited 99% avoidance. However, these results indicated behavior when alternative food sources are readily available, a scenario rare in nature. To address this, a second feeding experiment was conducted to observe D. ordii's behavior under calorie‐restricted conditions. D. ordii were subjected to a fast period, then offered only one treatment. Under these conditions, many subjects chose to consume coated seeds, but to a lesser degree than subjects offered control seeds. Seeds coated in ghost pepper, neem oil, and activated carbon reduced consumption by 47–50%. Given these lab results, we would expect these treatments to increase native plant establishment following the direct seeding of wildlands by protecting seeds from rodent predation.     

Systemic effects of neem on western flower thrips,Frankliniella occidentalis (Thysanoptera: Thripidae)

The systemic effects of neem on the western flower thrips, Frankliniella occidentalis (Pergande), were investigated in laboratory trials using green bean, Phaseolus vulgaris L., in arena and microcosm experiments. In arena experiments, systemic effects of neem against western flower thrips larvae on primary bean leaves were observed with maximum corrected mortality of 50.6%. In microcosm experiments using bean seedlings, higher efficacy in the control of western flower thrips were observed with soil applications of neem on a substrate mixture (i.e., Fruhstorfer Erde, Type P, and sand) in a 1:1 ratio (93% corrected mortality) compared with application on the commercial substrate only (76% corrected mortality). However, longer persistence of neem was observed with soil application on the commercial substrate, which showed effects against thrips for up to 6 d after application. In addition to systemic effects observed on all foliage-feeding stages of western flower thrips, mortality on contact and repellent effects were observed on soil-inhabiting stages after soil applications of neem. Finally, bean seedlings grown from seeds pregerminated for 3 d in neem emulsion were also toxic to western flower thrips.     

Two types of aspartic proteinases from buckwheat seed--gene structure and expression analysis

Two types of aspartic proteinase (AP) genes have been isolated from the cDNA library of developing buckwheat seeds. Analysis of their sequences showed that one of these, FeAP9, resembled the structure and shared high homology with the so-called typical plant APs characterized by the presence of a plant-specific insert (PSI), an element unique among APs. The other cDNA, FeAPL1, encoded an AP-like protein lacking that domain. Different expression profiles were observed for FeAP9 and FeAPL1. FeAPL1 mRNAs were restricted to the seeds only, whereas FeAP9 mRNAs were also present in the other plant tissues - leaves, roots, and flowers. Higher levels of FeAP9 were observed in senescent leaves compared with green leaves. The differential expression pattern of these two unique APs raises the interesting possibility that these proteinases have unique substrate specificity and may have different roles in plant development and other physiological processes.     

Integrated approach for the management of Meloidogyne javanica on eggplant using oil cakes and biocontrol agents

Investigations were carried out to evaluate the efficacy of biocontrol agents (Paecilomyces lilacinus and Cladosporium oxysporum) and/or oil cakes of castor, linseed, groundnut, mahua and neem in the management of root knot nematode, Meloidogyne javanica infecting eggplant under glasshouse conditions. All the treatments effectively suppressed the nematode population and kept the infection at significantly low level. Individual treatment of P. lilacinus was more effective than C. oxysporum in controlling M. javanica, whereas among oil cakes individual treatment of neem was more effective in the management of M. javanica followed by linseed cake, castor cake, groundnut cake and mahua cake. However, the efficacy of biocontrol agents increased in the presence of oil cakes. The highest improvement in plant growth and best protection against M. javanica was obtained by the integration of P. lilacinus with groundnut cake followed by neem cake, linseed cake, castor cake and mahua cake. On the other hand the integration of C. oxysporum with neem cake followed by groundnut cake, linseed cake, castor cake and mahua cake gave the best results in managing M. javanica on eggplant.     

Cleistogamy in Scutellaria indica (Labiatae): effective mating system and population genetic structure

Scutellaria indica is a perennial herb with both chasmogamous (CH) and cleistogamous (CL) flowers on the same plant in some populations, and only CL flowers in other populations. Actual seed production by CH and CL flowers was investigated in populations of S. indica. The average seed set of CL flowers was 19 times higher than CH flowers, indicating much greater fertilization success. The CL seeds were also significantly heavier than the CH seeds. However, the resource cost of producing a CH flower was much higher than that of producing a CL flower. The CH flower was approximately seven times larger, and its pollen/ovule ratio was approximately five times higher than flowers. The level and pattern of genetic diversity at both allozyme and random amplified polymorphic DNA (RAPD) levels were consistent with a predominantly selfing system in the species. The average amount of within-population genetic variation was extremely low (A = 1.025, P = 2.36%, HO = 0.001 and HE = 0.008 based on allozyme data, and P = 8.94% and HE = 0.03 based on RAPD data). At the species level, the estimates of total gene diversity (HT) were 0. 101 based on allozyme data and 0.139 based on RAPD data. A very high level of genetic differentiation occurred between populations (allozyme GST = 0.92 and RAPD GST = 0.81). Genetic drift coupled with predominant cleistogamous selfing apparently played the major role in determining the population genetic structure in S. indica. Although the features associated with CH and CL flower and seed production seem to be sufficient for the evolution of complete cleistogamy in S. indica, random fixation of alternative alleles for dimorphic or complete cleistogamy in small populations could maintain the multiple strategy of chasmogamous and cleistogamous reproduction in the species.     

Why is there variation in mean seed size among plants within single populations? test of the fertilization efficiency hypothesis

We tested the fertilization efficiency hypothesis, which attempts to explain mean seed size variation among plants within single populations, by comparing the patterns of seed size variation between chasmogamous (CH) flowers and cleistogamous (CL) flowers in Impatiens noli-tangere and Viola grypoceras, respectively. The fertilization efficiency hypothesis predicts that larger plants produce larger seeds if the number of pollen grains captured by a plant increases with increased allocation of resources to its attractive structures (e.g., corolla and nectar), but with diminishing gains. Thus, seed size should depend on plant size in seeds from CH flowers because of the diminishing gains of capturing pollen in these flowers, whereas seed size should not depend on plant size in seeds from CL flowers because CL flowers need not capture outcross pollen. We found significant positive correlations between mean seed size per plant and plant size for seeds from CH flowers in both species. However, there was no significant positive correlation between these two factors for seeds from CL flowers of both species. The results of the present investigations were thus consistent with the fertilization efficiency hypothesis.     

Plant phenolics as inhibitors of mutational and precarcinogenic events

Initiation of chemical carcinogenesis involves the intracellular formation of a highly reactive electrophile that can attack many chemical nucleophiles in the cell, including DNA, a process that seems to be a central mechanism of initiation. Competing chemical nucleophiles in the cell, such as endogenous glutathione, can act as protecting or blocking agents against the attack on DNA. There are chemical substances in our food supply that may act as anticarcinogens or antimutagens by blocking or trapping ultimate carcinogen electrophiles in a nucleophilic chemical reaction, to form innocuous products. A continuous input of these substances could serve as an additional buffer against DNA damage, supplementing the endogenous systems qualitatively and quantitatively. Certain plant phenolics can be effective inhibitors of chemical mutagens and (or) carcinogens. Tetrapyrroles and porphyrins, both plant and animal, can also act as blocking agents. Both plant phenolics and porphyrins are primarily active against aromatic carcinogens as inhibitors of mutagenesis in in vitro systems. Plant phenolics have also demonstrated inhibiting activity against aromatic chemically induced carcinogenesis.     

Identification and partial characterization of a highly active and stable phospholipase D from Brassica juncea seeds

Phospholipase D (PLD) activity has been identified in some new plant sources i.e. Brassica juncea (mustard) seeds, Zingibar officinale (ginger) rhizomes and Azadirachta indica (neem) leaves with the aim of identifying PLDs that possess high catalytic activity and stability. PLD from mustard seeds (PLD(ms)) exhibited the highest PLD specific activity, which was highly pH and temperature tolerant. PLD(ms) unlike many plant PLDs exhibited high thermal stability. The activity of PLD(ms) is optimum in the millimolar concentration of calcium ions and is independent of phosphatidylinositol-4,5-bisphosphate (PIP2). An active and stable enzyme like PLD(ms) may be utilized in the lipid industry.     

Diet and nutrition in wild mongoose lemurs (Eulemur mongoz) and their implications for the evolution of female dominance and small group size in lemurs

Data collected on the feeding behavior, food intake, and chemical analyses of plant foods were used to document seasonal variation in diet and nutrition in Eulemur mongoz in northwestern Madagascar. E. mongoz conforms to the general Eulemur dietary pattern, with a predominantly frugivorous diet supplemented mainly by leaves, flowers, and nectar. Phytochemical analysis revealed high water contents in all the main plant foods; mature fruit and flowers contained the most water-soluble carbohydrates; immature leaves were richest in protein and essential amino acids; the limiting amino acids in all plant foods were methionine and cystine; ash (mineral) content was highest in petioles and mature leaves; crude lipid content was highest in seeds; and crude fiber content was indistinguishable between immature and mature fruit and leaves. High-fiber foods were eaten during both seasons; the wet season diet was dominated by high-energy foods (mature fruit, nectar, and seeds), while the dry season diet contained foods high in energy (mature fruit and flowers) and high in protein (immature leaves) and minerals (mature leaves and petioles). However, nutrient intake did not vary between seasons, implying that nutrient requirements are met throughout the year. These results suggest we draw more conservative conclusions when interpreting dietary variability in the absence of chemical analysis, and also draw into question the idea that nutritional stress is a factor in the timing of reproduction in lemurs and, by extension, is linked to the prevalence of female dominance and small group size in lemurs.     

Quinolizidine alkaloid profiles of Lupinus varius orientalis, L. albus albus, L. hartwegii, and L. densiflorus

Alkaloid profiles of two Lupinus species growing naturally in Egypt (L. albus albus [synonym L. termis], L. varius orientalis) in addition to two New World species (L. hartwegii, L. densiflorus) which were cultivated in Egypt were studied by capillary GLC and GLC-mass spectrometry with respect to quinolizidine alkaloids. Altogether 44 quinolizidine, bipiperidyl and proto-indole alkaloids were identified; 29 in L. albus, 13 in L. varius orientalis, 15 in L. hartwegii, 6 in L. densiflorus. Some of these alkaloids were identified for the first time in these plants. The alkaloidal patterns of various plant organs (leaves, flowers, stems, roots, pods and seeds) are documented. Screening for antimicrobial activity of these plant extracts demonstrated substantial activity against Candida albicans, Aspergillus flavus and Bacillus subtilis.