Fluorescence detection of plant extracts that affect neuronal voltage-gated Ca2+ channels

Structurally novel compounds able to block voltage-gated Ca2+ channels (VGCCs) are currently being sought for the development of new drugs directed at neurological disorders. Fluorescence techniques have recently been developed to facilitate the analysis of VGCC blockers in a multi-well format. By utilising the small cell lung carcinoma cell line, NCI-H146, we were able to detect changes in intracellular Ca2+ concentration ([Ca2+]i) using a fluorescence microplate reader. NCI-H146 cells have characteristics resembling those of neuronal cells and express multiple VGCC subtypes, including those of the L-, N- and P-type. We found that K+-depolarisation of fluo-3 loaded NCI-H146 cells causes a rapid and transient increase in fluorescence, which was readily detected in a 96-well plate. Extracts of Australian plants, including those used traditionally as headache or pain treatments, were tested in this study to identify those affecting Ca2+ influx following membrane depolarisation of NCI-H146 cells. We found that E. bignoniiflora, A. symphyocarpa and E. vespertilio caused dose-dependent inhibition of K+-depolarised Ca2+ influx, with IC50 values calculated to be 234, 548 and 209 μg/ml, respectively. This data suggests an effect of these extracts on the function of VGCCs in these cells. Furthermore, we found similar effects using a fluorescence laser imaging plate reader (FLIPR) that allows simultaneous measurement of real-time fluorescence in a multi-well plate. Our results indicate that the dichloromethane extract of E. bignoniiflora and the methanolic extract of E. vespertilio show considerable promise as antagonists of neuronal VGCCs. Further analysis is required to characterise the function of the bioactive constituents in these extracts and determine their selectivity on VGCC subtypes.

Identification and simultaneous analysis of harmane, harmol, isovitexin and vitexin in Passiflora incarnata extracts with a novel HLPC method

A high performance liquid chromatographic method for the simultaneous analysis of two flavonoids (iso-vitexin and vitexin), and three indole alkaloids (harmane, harmine, and harmol) was developed. This method was then utilised to quantitate levels of these five constituents in methanolic extracts of Australian Passiflora incarnata. HPLC analysis was performed using a Waters™ Novapak C18 (150 × 4 mm, 4 μm) column, with a gradient solvent system of methanol-water-acetic acid. Detection was achieved by PDA UV (254 nm) and fluorescence (excitation 254 nm, emission 414 nm), utilising the external standard method to obtain quantification.

Inhibition of platelet aggregation and 5-HT release by extracts of Australian plants used traditionally as headache treatments

To identify potential migraine therapeutics, extracts of eighteen plants were screened to detect plant constituents affecting ADP induced platelet aggregation and [14C]5-hydroxytryptamine (5-HT) release. Extracts of the seven plants exhibiting significant inhibition of platelet function were reanalysed in the presence of polyvinyl pyrrolidone (PVP) to remove polyphenolic tannins that precipitate proteins. Two of these extracts no longer exhibited inhibition of platelet activity after removal of tannins. However, extracts of Crataegus monogyna, Ipomoea pes-caprae, Eremophila freelingii, Eremophila longifolia, and Asteromyrtus symphyocarpa still potently inhibited ADP induced human platelet [14C]5-HT release in vitro, with levels ranging from 62 to 95% inhibition. I. pes-caprae, and C. monogyna also caused significant inhibition of ADP induced platelet aggregation. All of these plants have been previously used as traditional headache treatments, except for C. monogyna which is used primarily for protective effects on the cardiovascular system. Further studies elucidating the compounds that are responsible for these anti-platelet effects are needed to determine their exact mechanism of action.

Simultaneous determination of aldrin, dieldrin, endrin, heptachlor, and p,p'-DDT in medicinal plant extracts using a novel high performance liquid chromatography method

A high performance liquid chromatographic method for the simultaneous determination of five organochlorine pesticides (aldrin, p,p’-DDT, dieldrin, endrin, and heptachlor) was developed. The method was used to determine the levels of these pesticides in medicinal plant samples. Analysis was carried out using a Merck LiChrospher 100 RP C18 (5 μm) column with a gradient solvent system of acetonitrile-water and PDA UV detection (224 nm). Quantification was carried out by the external standard method. The limit of detection for the utilized method was below the local legal limits (ANZFA) for similar plant materials for all 5 pesticides excepting endrin. Medicinal plant extracts were further analyzed by conventional GC-ECD and GC-NPD means using SPE and GPC cleanup as required.

Synthesis of complementary RNA by RNA-dependent RNA polymerases in plant extracts is independent of an RNA primer

RNA-dependent RNA polymerase (RDR) activities were readily detected in extracts from cauliflower and broccoli florets, Arabidopsis thaliana (L.) Heynh callus tissue and broccoli nuclei. The synthesis of complementary RNA (cRNA) was independent of a RNA primer, whether or not the primer contained a 3′ terminal 2′-O-methyl group or was phosphorylated at the 5′ terminus. cRNA synthesis in plant extracts was not affected by loss-of-function mutations in the DICER-LIKE (DCL) proteins DCL2, DCL3, and DCL4, indicating that RDRs function independently of these DCL proteins. A loss-of-function mutation in RDR1, RDR2 or RDR6 did not significantly reduce the amount of cRNA synthesis. This indicates that these RDRs did not account for the bulk RDR activities in plant extracts, and suggest that either the individual RDRs each contribute a fraction of polymerase activity or another RDR(s) is predominant in the plant extract. © CSIRO 2008.

Australian eco-pastoral poetry

Les Murray and Judith Wright are two Australian poets who are widely read as landscape poets. While this framing offers valuable insights into their work it often fails to bring the importance into a contemporary context or to recognise the long tradition Australia has had with , to use Leo Marx’ term, “the complex pastoral”. As Ruth Blair reminds us in her chapter “Hugging the Shore: The Green Mountains of South-East Queensland” in The Littoral Zone: Australian Contexts and their Writers it is accepted that North America has a tradition of the complex pastoral mode but it should be remembered that Australia also has a long history of this form. Both Judith Wright’s and Les Murray’s poetry encourages active campaigning for the environment .These Australian poets are eco-pastoral poets whose poetry encourages active reading rather than passive reflections. Their poetry speaks to the strong connection between the lived everyday landscape and the imagination of past, present and future. Their work is imbued with a strong sense of ecocritical awareness while at the same time drawing on pastoral conventions. These two Australian poets do not offer idealistic pastoral notions but rather reveal the complexities of lived human/nonhuman relationships. This paper will discuss these complexities and how poetry can be experienced as literature in action—ways for readers to connect with and negotiate with the land they inhabit. The research for this paper was, in part, drawn from the responses that local community library groups offered after reading the works of these poets. What became evident from this research was the way the poetry made the readers think not only of landscape as a place of refuge from the urban technological world but also as a contemporary place with connection to agency that motivates readers into active change.

Joy fear and poetry

Joy Fear and Poetry is an original performance work written, designed and directed by Natasha Budd in collaboration with 15 performers aged 7-12 years. It was performed in Brisbane as part of La Boite Theatre’s 2013 Indie Season. The production employs contemporary performance, postdramatic and constructivist methodologies to make an intervention into habituated patterns of positioning children in society. It embodies a model of practice that moves beyond participant empowerment toward a more nuanced process of co-artists creating intersubjective ‘composite texts’ (McCall 2011) for mainstream audiences. Joy Fear and Poetry experiments with techniques for performance making that create conditions conducive to authentic theatre making with children. These focus on dramaturgical, directorial and design strategies harnessed to maintain the performers’ focus, motivation and cognitive engagement within a reflexive, collaborative process.

Fresh Cuttings: A Celebration of Fiction and Poetry from UQP's Black Writing Series

As the first anthology of UQP's indigenous-authored books, Fresh Cuttings represents the very best of fiction and poetry publishing from UQP's Black Australian Writing series. An introduction by the editors and a biography of each author is included.

‘…souvlaki and a bit of lemon’: An Aboriginal Australian Greek Story.

In Victoria, Aboriginal peoples are collectively known as Koories (Koori History Website 2014). It’s a name that most people are comfortable with, even though each Koori will also hold their own specific tribal affiliations (Horton 1999). For example, the people of the Kulin nation are the Traditional Owners of the land that is now known by the English name of Melbourne. I am an Aboriginal Australian woman who originates from south-east Queensland (Brisbane/Ipswich). In south-east Queensland, some groups are collectively referred to as Murries...

Drops That Open Worlds : Image of Water in the Poetry of Euphrase Kezilahabi

Euphrase Kezilahabi on tansanialainen kirjailija, joka ensimmäisenä julkaisi swahilinkielisen vapaalla mitalla kirjoitetun runokokoelman. Perinteisessä swahilirunoudessa tiukat muotosäännöt ovat tärkeitä, ja teos synnytti kiivasta keskustelua. Runoteokset Kichomi ( Viilto , Kipu , 1974) ja Karibu Ndani ( Tervetuloa sisään , 1988) sekä Kezilahabin muu tuotanto voidaan nähdä uuden sukupolven taiteena. Kezilahabi on arvostettu runoilija, mutta hänen runojaan ei aiemmin ole käännetty englanniksi (yksittäisiä säkeitä lukuunottamatta), eikä juurikaan tutkittu yksityiskohtaisesti. Yleiskuvaan pyrkivissä lausunnoissa Kezilahabin runouden on hyvin usein määritelty olevan poliittista. Monet Kezilahabin runoista ottavatkin kantaa yhteiskunnallisiin kysymyksiin, mutta niiden pohdinta on kuitenkin runoissa vain yksi taso. Sen lisäksi Kezilahabin lyriikassa on paljon muuta ennen kartoittamatonta tämä tutkimus keskittyy veden kuvaan (the image of water). Kezilahabi vietti lapsuutensa saarella Victoria-järven keskellä, ja hänen vesikuvastonsa on rikasta. Tutkimuskysymyksenä on, mitä veden kuva runoteoksissa Kichomi ja Karibu Ndani esittää. Runojen analysoinnissa ja tulkinnassa on tarkasteltu myös sitä, miten äänteellinen taso osallistuu kuvien luomiseen. Tutkimuksen määritelmä kuvasta pohjautuu osittain Hugh Kennerin näkemykseen, jonka mukaan oleellista kuvassa on kirjaimellinen taso. Kennerin lähtökohtaan on yhdistetty John Shoptawin teoriaa, joka korostaa runon äänteellisen puolen tärkeyttä merkityksen muodostumisessa. Foneemien analyysissä vaikutteena on ollut Reuven Tsurin teoria. Analyysiosio osoittaa, että veden kuva edustaa ja käsittelee teoksissa lukuisia teemoja: elämää, kuolemaa, fyysistä vetovoimaa, runoutta, mielikuvitusta ja (ali)tajuntaa sekä moraalia. Veden kuvan tutkimuksen pohjalta on nähtävissä, että Kezilahabin filosofia asettuu elävä/kuollut- ja elämä/kuolema dikotomioiden ulkopuolelle.

"Four Ways of Writing the City" : St. Petersburg-Leningrad as a Metaphor in the Poetry of Joseph Brodsky

The present study discusses the theme of St. Petersburg-Leningrad in Joseph Brodsky's verse works. The chosen approach to the evolving im-age of the city in Brodsky's poetry is through four metaphors: St. Petersburg as "the common place" of the Petersburg Text, St. Petersburg as "Paradise and/or Hell", St. Petersburg as "a Utopian City" and St. Petersburg as "a Void". This examination of the city-image focusses on the aspects of space and time as basic categories underlying the poet's poetic world view. The method used is close reading, with an emphasis on semantical interpretation. The material consists of eighteen poems dating from 1958 to 1994. Apart from investigating the spatio-temporal features, the study focusses on exposing and analysing the allusions in the scrutinised works to other texts from Russian and Western belles lettres. Terminology (introduced by Bakhtin and Yury Lotman, among others) concerning the poetics of space in literature is employed in the present study. Conceptions originating from the paradigm of possible worlds are also used in elucidating the position of fictional and actual chronotopes and heroes in Brodsky's poetry. Brodsky's image of his native city is imbued with intertextual linkings. Through reminiscences of the "Divine Comedy" and Russian modernists, the city is paralleled with Dante's "lost and accursed" Florence, as well as with the lost St. Petersburg of Mandel'shtam and Akhmatova. His city-image is related to the Petersburg myth in Russian literature through their common themes of death and separation as well as through the merging of actual realia with the fictional worlds of the Petersburg Text. In his later poems, when his view of the city is that of an exiled poet, the city begins to lose its actual world referents, turning into a mental realm which is no longer connected to any particular geographical location or historical time. It is placed outside time. The native city as the homeland in its entirety is replaced by another existence created in language.

Patterns of Transcendence : Classical Myth in Marina Tsvetaeva's Poetry of the 1920s

This study examines the position and meaning of Classical mythological plots, themes and characters in the oeuvre of the Russian Modernist poet Marina Tsvetaeva (1892-1941). The material consists of lyric poems from the collection Posle Rossii (1928) and two longer lyrical tragedies, Ariadna (1924) and Fedra (1927). These works are examined in the context of Russian Modernism and Tsvetaeva s own poetic development, also taking into account the author s biography, namely, her correspondence with Boris Pasternak. Tsvetaeva s appropriations of the myths enter into a dialogue with the Classical tradition and with the earlier Russian and Western literary manifestations of the source material. Her Classical texts are inextricably linked with her own authorial myth, they are used to project both her ideas about poetry as well as the authored self of her poems. An important context for Tsvetaeva s application of the Classical myths is the concept of the Platonic ladder of Eros. This plot evokes the process of transcendence of the mortal subject into the immaterial realm and is applied by the author as an extended metaphor of the poet s birth. Emphasizing the dialectical movement between the earthly and the divine, Tsvetaeva s Classical personae foreground various positions of the individual between these two realms. By means of kaleidoscopic reformulations of similar metaphors and concepts, Tsvetaeva s mythological poems illustrate the poet s position between the material and the immaterial and the various consequences of this dichotomy on the creative mission. At the heart of Tsvetaeva s appropriation of the Sibyl, Phaedra, Eurydice and Ariadne is the tension between the body and disembodiment. The two lyrical tragedies develop the dichotomous worldview further, nevertheless emphasizing the dual perspective of the divine and the earthly realms: immaterial existence is often evaluated from a material perspective and vice versa. The Platonic subtext is central for Ariadna, focussing on Theseus development from an earthly hero to a spiritual one. Fedra concentrates on Phaedra s divinely induced physical passion, which is nevertheless evoked in a creative light.