Delivery of short hairpin RNA in the neotropical brown stink bug, Euschistus heros, using a composite nanomaterial.

The response of insects to orally delivered double-stranded RNA ranges widely among taxa studied to date. Long dsRNA does elicit a response in stink bugs but the dose required to achieve an effect is relatively high compared to other insects such Colorado potato beetle or western corn rootworm. Improving the delivery of dsRNA to stink bugs will improve the likelihood of using RNA-based biocontrols for the management of these economically important pests. Short hairpin RNA (shRNA) is a useful molecule with which to test improvements in the delivery of double stranded RNA in the neotropical brown stink bug, Euschistus heros, since shRNA alone does not elicit a clear effect like that for long dsRNA. Here, we show for the first time the oral delivery of shRNA triggering RNA interference (RNAi) in E. heros using 4 nm cerium oxide nanoparticles (CeO2 NPs) coated with diethylamioethyl dextran (Dextran-DEAE) as a carrier. We identified particle properties (coating composition and degree of substitution, hydrodynamic diameter, and zeta potential) and shRNA loading rates (Ce:shRNA mass ratio) that resulted in successful transcript reduction or RNAi. When the Z-average diameter of CeO2 Dextran-DEAE-shRNA NP complex was less than 250 nm and the zeta potential was in the 15-25 mV range (Ce:shRNA mass ratio of 0.7:1), significant mortality attributed to RNAi was observed with a shRNA concentration in feeding solution of 250 ng/µl. The degradation of the targeted troponin transcript by NP-delivered shRNA was equivalent to that observed with long dsRNA, while naked shRNA transcript reduction was not statistically significant. Elemental mapping by synchrotron X-ray fluorescence microprobe confirmed uptake and distribution of Ce throughout the body with the highest concentrations found in gut tissue. Taken together, our results suggest that a nanoparticle delivery system can improve the delivery of RNA-based biocontrols to E. heros, and therefore its attractiveness as an application in the management of this important pest in soybean production.

Volatile profile of Echinacea purpurea plants after in vitro endophyte infection.

The differences in volatile profile of Echinacea purpurea plants not-inoculated (EpC) and inoculated with their endophytes from roots (EpR) and stem/leaves (EpS/L) were analysed and compared by GC-FID/GC-MS in an in vitro model. Non-terpenes and sesquiterpene hydrocarbons were the most abundant classes with an opposite behaviour of EpS/L showing a decreased emission of sesquiterpenes and an increase of non-terpene derivatives. The main compounds obtained from EpS/L were (Z)-8-dodecen-1-ol and 1-pentadecene, while germacrene D and ß-caryophyllene were the key compounds in EpC and EpR. For the first time, this work indicates that bacterial endophytes modify the aroma profiles of infected and non-infected in vitro plants of the important medicinal plant E. purpurea. Therefore, our model of infection could permit to select endophytic strains to use as biotechnological tool in the production of medicinal plants enriched in volatile bioactive compounds.

Chemical composition of essential oil from plants of abandoned mining site of Elba island.

The essential oil composition of three spontaneous species growing in an abandoned mining of Elba island was analyzed by GC-MS. A total of 194 compounds were identified representing 73.7-100% of the whole oil composition. The essential oils of Cistus salvifolius and Dittrichia viscosa from this site showed different profiles in comparison with those from not polluted area, where oxygenated sesquiterpenes were the main class. Volatiles from D. viscosa growing in ex-mining area presented 10-epi-γ-eudesmol and α-eudesmol as main compounds while ß-caryophyllene and limonene were the main ones in not polluted area. Ambroxide and ambrial were the most important compounds in the essential oil from C. salvifolius harvested in polluted area while nonanal and tridecanal were the main compounds in control samples. Oxygenated monoterpenes were the most abundant class from both Lavandula stoechas samples, with fenchone and camphor as main compounds.

Chemical composition and antifungal activity of essential oils from four Asteraceae plants grown in Egypt.

The objective of the present paper was the assessment of the chemical composition of the essential oils from four Asteraceae species with a considerable food, medicinal, and agricultural value, collected in Egypt, together with their in vitro inhibitory activity against molds and yeasts. The essential oil of Launaea cornuta flowers was also evaluated for the first time, but because of its very low yield (<0.01%), no antifungal test was performed.

Efficient Binding, Protection, and Self-Release of dsRNA in Soil by Linear and Star Cationic Polymers.

Double stranded RNA (dsRNA) exhibits severe degradation within 3 days in live soil, limiting its potential application in crop protection. Herein we report the efficient binding, protection, and self-release of dsRNA in live soil through the usage of a cationic polymer. Soil stability assays show that linear poly(2-(dimethylamino)ethyl acrylate) can delay the degradation of dsRNA by up to 1 week while the star shaped analogue showed an increased stabilization of dsRNA by up to 3 weeks. Thus, the architecture of the polymer can significantly affect the lifetime of dsRNA in soil. In addition, the hydrolysis and dsRNA binding and release profiles of these polymers were carefully evaluated and discussed. Importantly, hydrolysis could occur independently of environmental conditions (e.g., different pH, different temperature) showing the potential for many opportunities in agrochemicals where protection and subsequent self-release of dsRNA in live soil is required.

Plant-endophytes interaction influences the secondary metabolism in Echinacea purpurea (L.) Moench: an in vitro model.

The influence of the interaction(s) between the medicinal plant Echinacea purpurea (L.) Moench and its endophytic communities on the production of alkamides is investigated. To mimic the in vivo conditions, we have set up an infection model of axenic in vitro E. purpurea plants inoculated with a pool of bacterial strains isolated from the E. purpurea stems and leaves. Here we show different alkamide levels between control (not-inoculated) and inoculated plants, suggesting that the alkamide biosynthesis may be modulated by the bacterial infection. Then, we have analysed the branched-chain amino acids (BCCA) decarboxylase gene (GenBank Accession #LT593930; the enzymatic source for the amine moiety formation of the alkamides) expression patterns. The expression profile shows a higher expression level in the inoculated E. purpurea tissues than in the control ones. These results suggest that the plant-endophyte interaction can influence plant secondary metabolism affecting the therapeutic properties of E. purpurea.

Evolution of Volatile Emission in Rhus coriaria Organs During Different Stages of Growth and Evaluation of the Essential Oil Composition.

Rhus coriaria, also known as Sumac, has been traditionally used in many countries as spice, condiment, dying agent, and medicinal herb. The chemical composition of essential oils (EOs) and the volatile emissions from different organs of this species collected in Sicily (Italy) were analyzed by gas chromatography-flame ionization detection and gas chromatography/mass spectrometry. Monoterpene and sesquiterpene hydrocarbons were the most abundant class in the volatile emissions with ß-caryophyllene and α-pinene were the main constituents in the majority of the examined samples. The EO composition was characterized by high amount of monoterpene and sesquiterpene hydrocarbons together with diterpenes. The main compounds in the EO obtained from the leaves and both stages of fruit maturation were cembrene and ß-caryophyllene, while α-pinene and tridecanoic acid were the key compounds in the flower EO. All the data were submitted to multivariate statistical analysis showing many differences among the different plant parts and their ontogenetic stages.

Traditional Mediterranean plants: characterization and use of an essential oils mixture to treat Malassezia otitis externa in atopic dogs.

Several plants extracts from Mediterranean countries are traditionally employed in skin troubles both in humans and in animals. Malassezia pachydermatis is a lipophylic yeast responsible for otitis externa and dermatitis in dogs and for cutaneous and systemic disease in humans. Five mixtures of essential oils obtained from Mediterranean plants (Citrus paradisi, Salvia sclarea, Ocimum basilicum, Rosmarinus officinalis, Citrus limon, Anthemis nobilis, Lavandula hybrida and Thymus vulgaris) provided with antifungal and/or anti-inflammatory action assayed in vitro, were tested in vivo versus M. pachydermatis to treat once daily for 2 weeks 25 atopic dogs with Malassezia otitis externa. Mixture composed by C. limon 1%, S. sclarea 0,5%, R. officinalis 1%, A. nobilis 0,5% yielded excellent results in all treated dogs. Despite of clinical resolution after all treatments the number of blastospores did not decrease. This study confirms recent findings suggesting a multifactorial alternative approach for the management of canine Malassezia otitis.

Aroma Profile of Rubus ulmifolius Flowers and Fruits During Different Ontogenetic Phases.

The chemical composition of spontaneous volatile emission from Rubus ulmifolius flowers and fruits during different stages of development was evaluated by HS-SPME-GC/MS. In total, 155 chemical compounds were identified accounting 84.6 - 99.4% of whole aroma profile of flowers samples and 92.4 - 96.6% for fruit samples. The main constituents were α-copaene, ß-caryophyllene, germacrene D, (E,E)-α-farnesene, 1,7-octadien-3-one,2-methyl-6-methylene, tridecane, (E)-2-hexenol acetate, (E)-3-hexenol acetate and cyperene. The results give a chemotaxonomic contribution to the characterization of the VOCs emitted from flowers and fruits during their ontogenic development.

Phosphonium Polymethacrylates for Short Interfering RNA Delivery: Effect of Polymer and RNA Structural Parameters on Polyplex Assembly and Gene Knockdown.

Synthetic polymers containing quaternary phosphonium salts are an emerging class of materials for the delivery of oligo/polynucleotides. In this work, cationic phosphonium salt-containing polymethacrylates and their corresponding ammonium analogues were synthesized by reversible addition-fragmentation chain transfer polymerization. Both the nature of the charged heteroatom (N vs P) and the length of the spacer separating the cationic units along the polymer backbone (oxyethylene vs trioxyethylene) were systematically varied. Polymers efficiently bound short interfering RNA (siRNA) at N(+)/P(-) or P(+)/P(-) ratios of 2 and above. At a 20:1 ratio, small polyplexes (Rh: 4-15 nm) suitable for cellular uptake were formed that displayed low cytotoxicity. While siRNA polyplexes from both ammonium and phosphonium polymers were efficiently internalized by green fluorescent protein (GFP)-expressing 3T3 cells, no knockdown of GFP expression was observed. However, 65% Survivin gene knockdown was observed when siRNA was replaced with novel, multimerized long interfering RNA in HeLa cells, demonstrating the importance of RNA macromolecular architecture on RNA-mediated gene silencing.

Experiences of self-management support from GPs among Australian ethnically diverse diabetes patients: a qualitative study.

Ethnically diverse diabetes patients face significant challenges in diabetes self-management ranging from cultural expectations to inequalities in health care provision. This study explored the experiences of ethnically diverse patients with diabetes attending group diabetes education in receiving self-management support from GPs. An approach based on phenomenology was used to analyse participants' experiences in self-management support across three group interviews comprising 28 Australian ethnically diverse diabetes patients: Arabic-speaking group (n = 11), English-speaking group (n = 9) and Vietnamese-speaking group (n = 8). Two themes emerged related to the poor quality of information to support self-management and challenges in negotiating traditional consultation styles. In particular, participants believed they knew more about diabetes self-management than their GPs but felt unable to influence consultation style and communicate their changing needs in self-management support. The health care and information needs of ethnically diverse patients continue to be marginalised within health systems. This small exploratory study highlights the need for further research to illuminate interactions between ethnically diverse diabetes patients and GPs in supporting long-term diabetes self-management.

Chemical composition and antioxidant activity of the essential oil of Tabernaemontana catharinensis A. DC. leaves.

The essential oils obtained by hydrodistillation from the leaves of Tabernaemontana catharinensis had their composition analysed by GC-MS. A total of 18 substances were identified, consisting of a complex mixture of sesquiterpenes (83.52%), monoterpenes (5.46%) and triterpenes (4.56%). The main components in the oil were ß-caryophyllene (56.87%), α-cadinol (12.52%), 8S,13-cedran-diol (5.41%), α-terpineol (3.99%), ß-eudesmol (2.54%), caryophyllene oxide (2.51%) and ethyl iso-allocholate (2.03%) along with ß-cubebene, γ-cadinene, cubenol, 1,8-cineol, o-cymene, curcumenol, spathulenol, friedeline and ß-sitosterol as minor constituents. An antioxidant property was tested with the oil obtained by means of 1,1-diphenyl-2-picrylhydrazyl assay; the oil presented interesting radical scavenging activity. To the best of our knowledge, this is the first study of the composition and antioxidant activity of essential oil from the T. catharinensis collected from Brazil.

Mammary gland selective excision of c-jun identifies its role in mRNA splicing.

The c-jun gene regulates cellular proliferation and apoptosis via direct regulation of cellular gene expression. Alternative splicing of pre-mRNA increases the diversity of protein functions, and alternate splicing events occur in tumors. Here, by targeting the excision of the endogenous c-jun gene within the mouse mammary epithelium, we have identified its selective role as an inhibitor of RNA splicing. Microarray-based assessment of gene expression, on laser capture microdissected c-jun(-/-) mammary epithelium, showed that endogenous c-jun regulates the expression of approximately 50 genes governing RNA splicing. In addition, genome-wide splicing arrays showed that endogenous c-jun regulated the alternate exon of approximately 147 genes, and 18% of these were either alternatively spliced in human tumors or involved in apoptosis. Endogenous c-jun also was shown to reduce splicing activity, which required the c-jun dimerization domain. Together, our findings suggest that c-jun directly attenuates RNA splicing efficiency, which may be of broad biologic importance as alternative splicing plays an important role in both cancer development and therapy resistance.

Specific issues, exact locations: case study of a community mapping project to improve safety in a disadvantaged community.

ISSUE ADDRESSED: Safety is a health issue and a significant concern in disadvantaged communities. This paper describes an example of community-initiated action to address perceptions of fear and safety in a suburb in south-west Sydney which led to the development of a local, community-driven research project. METHODS: As a first step in developing community capacity to take action on issues of safety, a joint resident-agency group implemented a community safety mapping project to identify the extent of safety issues in the community and their exact geographical location. Two aerial maps of the suburb, measuring one metre by two metres, were placed on display at different locations for four months. Residents used coloured stickers to identify specific issues and exact locations where crime and safety were a concern. RESULTS: Residents identified 294 specific safety issues in the suburb, 41.9% (n=123) associated with public infrastructure, such as poor lighting and pathways, and 31.9% (n=94) associated with drug-related issues such as drug activity and discarded syringes. CONCLUSIONS: Good health promotion practice reflects community need. In a very practical sense, this project responded to community calls for action by mapping resident knowledge on specific safety issues and exact locations and presenting these maps to local decision makers for further action.

GP and community nurse co-location in a disadvantaged community.

People living in socioeconomically disadvantaged communities have a high burden of disease but often receive 'inverse care'. We explored a model of general practitioner and community nurse co-location in a disadvantaged community in south-west Sydney, Australia. Co-location resulted in increased referrals from doctors to the community nurse, including an increase in referrals related to psychosocial issues. This small study suggests integrated primary health care might have an impact on specialised state-based psychosocial health services.

An ethnographic process evaluation of a community support program with Sudanese refugee women in western Sydney.

ISSUE ADDRESSED: Through the humanitarian entrant program, a rapidly growing community of Sudanese refugees are resettling in Australia. Although the impact of pre-migration trauma upon refugee mental health is well established, there is a paucity of research exploring the impact of post-migration sociocultural factors. Women are often the most marginalised, which affects their mental health and ability to adjust in resettlement. METHODS: An ethnographic study was conducted to evaluate a Sudanese women's group exercise program designed from a community development strengths-based model. Qualitative analysis of a group interview and ethnographic process evaluation techniques enabled a deeper understanding of the perspectives of Sudanese women in Australia and their resettlement difficulties and needs. RESULTS: Participants (n=12) viewed the program positively due to educational components and the opportunity for respite. Transport and childminding support were seen as vital. Interview and evaluation processes were perceived as ineffectual. Key stressors raised include: acculturation, housing difficulties, developing language skills, lack of employment opportunities and family separation. CONCLUSION: It is vital that any attempts to address these issues are sustainable, aiming to empower the women and promote their existing strengths and resilience techniques. Research specific to cultural and ethnic groups of refugee women in an Australian context enables tailoring of appropriate support services, but can be tiresome for participants.

Labour market initiatives: potential settings for improving the health of people who are unemployed.

ISSUE ADDRESSED: Unemployment is detrimental to health. The Unemployment and Health Project in South Western Sydney sought to work with labour market programs to improve the health, particularly, mental health, of unemployed people. This paper describes the experiences of the Project. METHODS: Phase one commenced in 1995 and involved consultation with the majority of Skillshares (labour market programs under the Labor government) in south-western Sydney to identify potential areas of action. Phase two commenced in 1998 and involved the development of a brief cognitive behaviour therapy intervention that was delivered in Job Network Settings (the next generation of labour market programs under the Liberal government). RESULTS: The cognitive behaviour therapy intervention has been successful in improving mental health in five small scale trials but the intervention has proved difficult to scale up and evaluate comprehensively. Generating more general interest in improving the health of unemployed people through the Job Network has also been difficult. This is related to different understanding and valuing of evidence, a highly volatile context, lack of shared core business by the health and employment sectors, and the changing nature of work in Australia. CONCLUSIONS: There are theoretical and practical reasons why it is difficult for labour market programs to be a setting for improving the health of unemployed people. However, the reach of labour market programs into the high risk groups warrants more attention by mental health promotion programs.

A better model of diabetes self-management? Interactions between GP communication and patient self-efficacy in self-monitoring of blood glucose.

OBJECTIVE: The aim of this exploratory study was to investigate the interaction between patient self-efficacy and GP communication in explaining diabetes self-management in a disadvantaged region of Sydney, Australia. METHODS: This study was undertaken in South West Sydney with the Fairfield Division of General Practice. We used a cross-sectional survey design to assess patients' self-reported beliefs and behaviours about diabetes self-management. We used hierarchical multiple linear regression to test for interaction effects in diabetes self-management, following tests for clustering using multilevel modeling. RESULTS: Of those eligible for survey, 105 patients completed the telephone survey (72%). There was a significant interaction between diabetes self-efficacy and GP communication in blood glucose testing; high-ratings of GP communication enhanced self-monitoring of blood glucose when patient self-efficacy was high but impeded self-monitoring of blood glucose when self-efficacy was low. There were no significant interaction effects for the general diet or exercise scales. CONCLUSION: This exploratory study suggests a complex relationship between patient self-efficacy and GP communication in self-monitoring of blood glucose. It is likely optimal diabetes self-management behaviours are produced by a fit between high patient self-efficacy and high quality GP communication. PRACTICE IMPLICATIONS: There is a risk that GPs who are sensitive to their patients' low self-efficacy in self-monitoring of blood glucose may step in and take over the monitoring role and inadvertently reduce self-management.