Agrobacterium rhizogenes-mediated transformation of Artemisia annua: production of transgenic plants

Transgenic hairy roots were induced in the leaves of Artemisia annua by treatment with the LBA 9402 strain of Agrobacterium rhizogenes. The axenic hairy root cultures were found to produce the sesquiterpenes artemisinic acid and arteannuin B. The hairy root cultures were observed to spontaneously regenerate into plantlets on solid hormone-free MS medium. The regenerated plants had phenotypic characteristics typical to the transformed plants. Among the plants of the age of one month in culture, the transgenic plant was bigger (2.643 g/plant) than the normal (0.856 g/plant). Both these kinds of in vitro plants carried sesquiterpenes-artemisinic acid and arteannuin B.     

Withanolide production by root cultures of Withania somnifera transformed with Agrobacterium rhizogenes

Transformed root cultures of Withania somnifera Dunal (Solanaceae) were obtained by infecting shoots cultured in vitro with Agrobacterium rhizogenes LBA 9402. They grew axenically in the absence of exogenous plant growth regulators in Murashige and Skoog's medium containing 3% (w/v) sucrose. The root cultures synthesized several withanolides of which withanolide D was isolated and identified. Transformed root (clone HR (1)) showed a growth index of 20.07 and a withanolide D yield of 0.30 mg g(-1) DW. The productivity of withanolide D in transformed roots (0.181 mg 1(-1) d(-1)) was higher than in untransformed root cultures (0.026 mg 1(-1) d(-1)).     

Plant cell cultures as a source of bioactive small molecules

Plants have provided some of our most important pharmaceuticals and are a continuing source of novel bioactive molecules for drug development. Cell suspension cultures of higher plants are a complementary source of novel chemistry with significant advantages over whole plants, including plasticity of expression and re-accessibility. With an emphasis on recent progress, the use of plant cell cultures for both new lead discovery and as a route to the production of known bioactives will be described. Developments in strategies that can be applied to facilitate future success in the area are discussed.     

Drugs as technologies of the self: Enhancement and transformation in LGBTQ cultures

The consumption of drugs has long been a mainstay of urban queer cultures and it is well-recognised that complex connections exist between sexual minoritisation and desires to chemically alter bodily experience. Yet despite evidence that rates of consumption are higher among LGBTQ populations, research exploring the gendered and sexual dynamics of these forms of consumption is limited and tends to frame such consumption as a response to stigma, marginalisation and discrimination. Against this dominant explanatory frame, this article explores the diverse experiences of LGBTQ consumers, and in so doing highlights both the pleasures and benefits of consumption, as well as potential risks and harms. Contributing to the growing body of ontopolitically oriented research that treats the materiality of drugs as emergent and contingent, we trace the ontologies of drugs, sexuality and gender that LGBTQ subjects generate through specific practices of consumption. Our analysis draws on qualitative interviews with 42 self-identified LGBTQ people from an Australian study designed to explore how sexual and gender-diverse minorities pursue particular drug effects to enhance or transform their experience of gender and/or sexuality. Our participants’ accounts illuminate how drug consumption materialises in relation to sex, desire and play where it enhances pleasure, facilitates transgression and increases endurance. In the context of gender variance, our findings suggest that drug use can transform gendered experience and enable the expression of non-normative gender identities, in the process challenging gender binarism. By considering the productive role of drugs in enacting queer identities, this article treats drugs as ‘technologies of the self’ (Foucault 1988) and explores how drug consumption, sex and gender shape each other across a range of settings. We conclude by reflecting on the implications of our findings for research and service provision, and suggest ways of engaging LGBTQ consumers in terms that address their diverse priorities and experiences.     

Utilization of methionine as a sulfhydryl source for metallothionein synthesis in rat primary hepatocyte cultures

Metallothioneins (MT) contain a high concentration of cysteine which bind heavy metals. Exposure of liver cells to metals induces the synthesis of MT and thus causes the cells to draw upon their sulfhydryl (SH) pools. The utilization of methionine as compared with that of cysteine as a source of SH for the synthesis of MT has not been shown. Therefore, studies were designed to determine whether methionine, in addition to cysteine, serves as an SH donor for Zn-induced synthesis of MT in rat hepatocyte cultures. Hepatocytes were able to synthesize only low levels of MT when the concentration of both amino acids was extremely low; however, when either of the amino acids was present at a high concentration, production of MT was independent of the other amino acid concentration. Subsequently, induction of MT was compared in four media: complete (0.5 mM methionine, 0.5 mM cysteine), Met (0.5 mM methionine), Cys (0.5 mM cysteine), and SH free (-SH). Somewhat higher concentrations of MT were produced by the hepatocytes in the Met than in the Cys media and no differences were observed between the Met and the complete media. By contrast, GSH synthesis was much more dependent on methionine than on cysteine for its synthesis. Incorporation studies with 35S-labeled cysteine and methionine indicated that lower concentrations of MT found in hepatocytes in the Cys media may be due to less accumulation of cysteine by the hepatocytes. Cellular accumulation of cysteine was initially rapid and then reached a plateau, whereas the rate for methionine accumulation was more constant and eventually obtained higher cellular levels. To provide additional evidence for the role of methionine in MT production, a known inhibitor of the cystathionine pathway, DL-propargylglycine (PPG), was added to each of the four media. Reductions in MT levels were not observed in the cells cultured in the complete and Cys media; however, a 95% reduction was observed in the cells cultured in the Met media. In summary, the present results suggest that both cysteine and methionine can serve as a SH source for MT synthesis, and that the availability of SH in most culture mediums would not limit the synthesis of MT. Whereas methionine is a much better SH source than cysteine for GSH synthesis in hepatocyte cultures, it is only slightly better for MT synthesis.     

Thebaine from root cultures of Papaver bracteatum

Root cultures derived from cell suspension cultures of PAPAVER BRACTEATUM were shown to produce thebaine (yield 0.03%).     

Phytochemical study of the bark of some plants of the Elaeagnaceae family as a natural source of β-carboline indole alkaloids

We have studied the qualitative and quantitative composition of β-carboline indole alkaloids that have been isolated for the first time from bark of Hippopha? rhamnoides L., Elaeagnus angustifolia L., E. orientalis L., E. umbellata Thunb., E. multiflora Thunb., and E. argentea Pursh. occurring in Russia. Results of a phytochemical study showed that the bark of all these plants contains β-carboline alkaloids and that the first four of them can be promising sources of β-carboline alkaloids for the creation of drugs. Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 42, No. 11, pp. 27–29, November, 2008.     

Genetic transformation of Salvia austriaca by Agrobacterium rhizogenes and diterpenoid isolation

Hairy roots of Salvia austriaca Jacq. transformed with Agrobacterium rhizogenes strain A4 were obtained and transgenic status of the roots was confirmed by polymerase chain reaction (PCR) using rolB and rolC specific primers. The root cultures growing in half-strength Gamborg (1/2 B5) liquid medium supplemented with sucrose (30 g L(-1)) under light conditions (photoperiod: 16 h light/8 h dark) were examined for their ability to produce diterpenoids. From n-hexane extract the abietane-type diterpenoids royleanone, 15-deoxyfuerstione and taxodione were isolated and identified. This is the first report on the genetic transformation of S. austriaca.     

Agrobacterium tumefaciens-mediated genetic transformation of the cardenolide-producing plant Digitalis minor L

A repeatable transformation system has been established for Digitalis minor using Agrobacterium tumefaciens. Leaf explants from 30-day-old seedlings were inoculated with either EHA105 (carrying the nptII and gusA genes) or AGL1 (with the bar and gusA genes) strains. Among the tested factors influencing T-DNA transfer to plants, the EHA105 strain and the addition of acetosyringone to the co-culture medium increased transformation. The highest transformation efficiency (8.4 %) was obtained when freshly isolated explants, soaked in a bacterial suspension with an OD550 of 0.9, were subcultured on selection medium after a 4-day co-culture with the bacteria. Evidence of stable transgene integration was obtained by PCR, growth on media selective for nptII or bar genes, and expression of the gusA gene. Southern hybridisation, performed in six plants, provided information about the number of inserts. More than 200 transgenic plants were recovered from 65 independent explants. Thirty of these plants were successfully established in soil. This is the first report on transgenic Digitalis spp plants using an A. tumefaciens-mediated leaf disc transformation procedure.     

Dorsal root ganglia sensory neuronal cultures: a tool for drug discovery for peripheral neuropathies

Background: Peripheral neuropathies affect many people worldwide and are caused by or associated with a wide range of conditions, both genetic and acquired. Current therapies are directed at symptomatic control because no effective regenerative treatment exists. The primary challenge is that mechanisms that lead to distal axonal degeneration, a common feature of all peripheral neuropathies, are largely unknown. Objective/methods: To address the role and specific characteristics of dorsal root ganglia (DRG) derived sensory neuron culture system as a useful model in evaluating the pathogenic mechanisms of peripheral neuropathies and examination and validation of potential therapeutic compounds. A thorough review of the recent literature was conducted and select examples of the use of DRG neurons in different peripheral neuropathy models were chosen to highlight the utility of these cultures. Conclusion: Many useful models of different peripheral neuropathies have been developed using DRG neuronal culture and potential therapeutic targets have been examined, but so far none of the potential therapeutic compounds have succeeded in clinical trials. In recent years, the focus has changed to evaluation of axon degeneration as the primary outcome measure advocating a drug development strategy starting with phenotypic drug screening, followed by validation in primary complex co-cultures and animal models.     

Cognitive transformation as a marker of resilience

Individuals often report positive, transformative changes in response to adversity. Cognitive transformation involves a turning point in a person's life characterized by: (1) the recognition that coping with adversity resulted in new opportunities; and, (2) the reevaluation of the experience from one that was primarily traumatic or threatening to one that is growth-promoting. Cognitive transformation often signifies enhanced adaptation to adverse circumstances, and thus, is a marker of resilience. The present study examined the relationship of cognitive transformation to indicators of resilience among 35 acutely bereaved young adults and a nonbereaved comparison group. Findings strongly supported the hypothesis that transformation predicts resilience, and may reduce one's risk trajectory to enhance adaptation. Results are discussed in terms of their implications for research on resilience, and on recovery from acute or chronic adverse circumstances, including addiction.     

Identification of the alkaloids and anthraquinones in Cinchona ledgeriana callus cultures

The isolation of quinoline alkaloids, indole alkaloids and anthraquinones present in callus cultures initiated from different parts of Cinchona ledgeriana plants is described. From leaves of C. ledgeriana a new minor alkaloid was isolated and identified as 10-methoxycinchonamine.     

粉防己生物碱化学成分的分离与鉴定

目的对防己科千金藤属植物粉防己(Stephania tetrandraS.Moore)的生物碱化学成分进行研究。方法采用反复硅胶柱色谱法、Sephadex LH-20柱色谱法等进行分离纯化,并通过光谱分析鉴定其化学结构。结果分离得到7个生物碱类化合物,分别鉴定为粉防己碱(tetrandrine,1)、防己诺林碱(fangchinoline,2)、2′-N-chloromethyltetrandrine(3)、氧化防己碱(oxofangchirine,4)、粉防己碱D盐酸盐(fenfangjine D hydrochloride,5)、荷苞牡丹碱((+)-dicentrine,6)、tazopsine(7)。结论化合物7为首次从该属植物中分离得到,化合物6为首次从该植物中分离得到。     

Biodiversity as a source of anticancer drugs

Natural Products have been the most significant source of drugs and drug leads in history. Their dominant role in cancer chemotherapeutics is clear with about 74% of anticancer compounds being either natural products, or natural product-derived. The biodiversity of the world provides a resource of unlimited structural diversity for bioprospecting by international drug discovery programs such as the ICBGs and NCDDGs, the latter focusing exclusively on anticancer compounds. However, many sources of natural products remain largely untapped. Technology is gradually overcoming the traditional difficulties encountered in natural products research by improving access to biodiverse resources, and ensuring the compatibility of samples with high throughput procedures. However, the acquisition of predictive biodiversity remains challenging. Plant and organism species may be selected on the basis of potentially useful phytochemical composition by consulting ethnopharmacological, chemosystematic, and ecological information. On the conservation/political front, the Convention on Biological Diversity (CBD) is allaying the anxiety surrounding the notion of biopiracy, which has defeated many attempts to discover and develop new natural products for human benefit. As it becomes increasingly evident and important, the CBD fosters cooperation and adaptation to new regulations and collaborative research agreements with source countries. Even as the past inadequacies of combinatorial chemistry are being analyzed, the intrinsic value of natural products as a source of drug leads is being increasingly appreciated. Their rich structural and stereochemical characteristics make them valuable as templates for exploring novel molecular diversity with the aim of synthesizing lead generation libraries with greater biological relevance. This will ensure an ample supply of starting materials for screening against the multitude of potentially "druggable" targets uncovered by genomics technologies. Far from being mutually exclusive, biodiversity and genomics should be the driving force of drug discovery in the 21st century.     

Alcoholic beverages as a source of estrogens

Alcoholic beverages contain not only alcohol but also numerous other substances (i.e., congeners) that may contribute to the beverages' physiological effects. Plants used to produce alcoholic beverages contain estrogenlike substances (i.e., phytoestrogens). Observations that men with alcoholic cirrhosis often show testicular failure and symptoms of feminization have suggested that alcoholic beverages may contain biologically active phytoestrogens as congeners. Biochemical analyses have identified several phytoestrogens in the congeners of bourbon, beer, and wine. Studies using subjects who produced no estrogen themselves (i.e., rats whose ovaries had been removed and postmenopausal women) demonstrated that phytoestrogens in alcoholic beverage congeners exerted estrogenlike effects in both animals and humans. Those effects were observed even at moderate drinking levels.