Identification and molecular cloning of novel trypsin inhibitors from the dermal venom of the Oriental fire-bellied toad (Bombina orientalis) and the European yellow-bellied toad (Bombina variegata).

The structural diversity of polypeptides in amphibian skin secretion probably reflects different roles in dermal regulation or in defense against predators. Here we report the structures of two novel trypsin inhibitor analogs, BOTI and BVTI, from the dermal venom of the toads, Bombina orientalis and Bombina variegata. Cloning of their respective precursors was achieved from lyophilized venom cDNA libraries for the first time. Amino acid alignment revealed that both deduced peptides, consisting of 60 amino acid residues, including 10 cysteines and the reactive center motif, -CDKKC-, can be affirmed as structural homologs of the trypsin inhibitor from Bombina bombina skin.

Hylaranins: prototypes of a new class of amphibian antimicrobial peptide from the skin secretion of the oriental broad-folded frog, Hylarana latouchii

Amphibian skin secretions contain a broad spectrum of biologically active compounds, particularly antimicrobial peptides, which are considered to constitute a first line of defence against bacterial infection. Here we describe the identification of two prototype peptides representing a novel structural class of antimicrobial peptide from the skin secretion of the oriental broad-folded frog, Hylarana latouchii. Named hylaranin-L1 (GVLSAFKNALPGIMKIIVamide) and hylaranin-L2 (GVLSVIKNALPGIMRFIAamide), both peptides consist of 18 amino acid residues, are C-terminally amidated and are of unique primary structures. Their primary structures were initially deduced by MS/MS fragmentation sequencing from reverse-phase HPLC fractions of skin secretion that demonstrated antimicrobial activity. Subsequently, their precursor-encoding cDNAs were cloned from a skin secretion-derived cDNA library and their primary structures were confirmed unequivocally. Synthetic replicates of both peptides exhibited broad-spectrum antimicrobial activity with mean inhibitory concentrations (MICs) of 34 µM against Gram-negative Escherichia coli, 4.3 µM against Gram-positive Staphylococcus aureus and 4–9 µM against the yeast, Candida albicans. Both peptides exhibited little haemolytic activity (<6 %) at the MICs for S. aureus and C. albicans. Amphibian skin secretions thus continue to provide novel antimicrobial peptide structures that may prove to be lead compounds in the design of new classes of anti-infection therapeutics.

Amphibian skin peptides and their corresponding cDNAs from single lyophilized secretion samples: identification of novel brevinins from three species of Chinese frogs

Brevinins are peptides of 24 amino acid residues, originally isolated from the skin of the Oriental frog, Rana brevipoda porsa, by nature of their microbicidal activity against a wide range of Gram-positive and Gram-negative bacteria and against strains of pathogenic fungi. cDNA libraries were constructed from lyophilized skin secretion of three, unstudied species of Chinese frog, Odorrana schmackeri, Odorrana versabilis and Pelophylax plancyi fukienensis, using our recently developed technique. In this report, we describe the “shotgun” cloning of novel brevinins by means of 3'-RACE, using a “universal” degenerate primer directed towards a highly conserved nucleic acid sequence domain within the 5'-untranslated region of previously characterized frog skin peptide cDNAs. Novel brevinins, deduced from cloned cDNA open-reading frames, were subsequently identified as mature peptides in the same samples of respective species skin secretions. Bioinformatic analysis of both prepro-brevinin nucleic acid sequences and translated open-reading frame amino acid sequences revealed a highly conserved signal peptide domain and a hypervariable anti-microbial peptide-encoding domain. The experimental approach described here can thus rapidly provide robust structural data on skin anti-microbial peptides without harming the donor amphibians.

Molecular cloning of mRNA from toad granular gland secretion and lyophilized skin: identification of Bo8 - a novel prokineticin from Bombina orientalis

Prokineticins are small (8 kDa), biologically active secretory proteins whose primary structures have been highly conserved throughout the Animal Kingdom. Representatives have been identified in the defensive skin secretions of several amphibians reflecting the immense structural/functional diversity of polypeptides in such. Here we describe the identification of a prokineticin homolog (designated Bo8) from the skin secretion of the Oriental fire-bellied toad (Bombina orientalis). Full primary structural characterization was achieved using a combination of direct Edman microsequencing, mass spectrometry and cloning of encoding skin cDNA. The latter approach employed a recently described technique that we developed for the cloning of secretory peptide cDNAs from lyophilized skin secretion, and this was further extended to employ lyophilized skin as the starting material for cDNA library construction. The Bo8 precursor was found to consist of an open-reading frame of 96 amino acid residues consisting of a putative 19-residue signal peptide followed by a single 77-residue prokineticin (Mr = 7990 Da). Amino acid substitutions in skin prokineticins from the skin secretions of bombinid toads are confined to discrete sites affording the necessary information for structure/activity studies and analog design.

Lividins: Novel antimicrobial peptide homologs from the skin secretion of the Chinese Large Odorous frog, Rana (Odorrana) livida: Identification by “shotgun” cDNA cloning and sequence analysis.

Odorous frogs of the sub-genus Odorrana are of oriental distribution, and are so called due to the foul smell of their defensive skin secretions released from specialized skin glands following stress or predator attack. Here we report the application of a “shotgun” skin secretion cDNA library cloning technique which can rapidly expedite identification of secretion bioactive peptides. From a library constructed from the skin secretion of the Large Chinese Odorous frog, Rana (Odorrana) livida, we have identified four novel peptides whose primary structures were deduced initially from cloned precursors. Subsequently, mature peptides were located in and structurally characterized from reverse phase HPLC fractions of skin secretion. Named lividins 1–4, these were found to be structural homologs of known antimicrobial peptide families from Rana frogs. Rapid identification of novel peptides can thus be rapidly achieved using this non-invasive, non-destructive technology and the extensive similarities revealed between antimicrobial peptide precursor organization and nucleic acid sequences would lend support to the hypothesis that they have a common ancestral origin.

“ Humanitarian Competition: An Intellectual History in Perspective”.

Competition has become the mantra for survival in a globalised world where meaningful existence is fraught with demands, which go beyond the material to the immaterial ‘byte-size’. This has been exemplified by our obsession with illusions of immediate fame and fortune. This paper contextualises and extends the debate about the role of competition in general. Here the four major myths of competition are explored and deconstructed, from a Darwinian perspective to a more demonstrably engaged perspective on ‘capabilities’ (Sen, 1999). The second section deals particularly with the key debates, theories that influenced Tsunesaburo Makiguchi’s seminal ideas of ‘humanitarian competition’ in 1903. The final part of the paper seeks to decipher the relevance of the key ideas of ‘humanitarian competition’ as proposed by Dr Daisaku Ikeda in his 2009 peace proposal. Here the transition from competition to cooperation is explored by tying together the key principles of global coexistence enunciated by both Makiguchi and Ikeda in the context of expanding spiritual influence by the forces of culture, morality and virtue. To engage with humanitarian competition calls for a major shift from hard power to soft power, from subordination to one of engagement. In other words this concept advances the Buddhist principle of peaceful co-existence, or Panchsheel, as a norm for human behaviour of love, kindness, sacrifice and peace through cooperation, where equality and mutual benefit are critical. Humanitarian competition provides the essential framework to establish a new world order as highlighted by both Makiguchi and Ikeda.

Every Situation has its Rhythm: Acoustic Mimesis and Sonic Exoticism in Tambours Sur La Digue.

This article examines the soundscapes of Ariane Mnouchkine’s Tambours Sur La Digue and explores the concept of acoustic mimesis located in the performance as a dramaturgical strategy to create, aurally, an imagined Far East. In Tambours, mimesis is the performative principle exemplified by the presentation of the mise en scène, and most distinctly Mnouckine’s decision to adapt the Japanese performance tradition of Bunraku through a process of 'reversed' mimicry (in which human bodies simulate the wooden marionettes of the Japanese style). Mimesis pervades the acoustemologies of the performance as it is heard in the extracted sounds, styles, and rhythms of Asian musical modes and movements that consequently become dislocated from context; the sounds become imitated, iconicised and exoticised as sonic signatures as they reify the Orientalist spectacle. The 'oriental' soundscape, reverberating with exotic overtones, becomes the means by which the production creates an imaginary Orient – one in which the Orient Other is silenced, and is resounded only through the musical sensibilities of the Occidental Self.

"From Indomania to Indophobia":Thomas De Quincey's Providential Orientalism

Thomas De Quincey’s terrifying oriental nightmares, reported to sensational acclaim in his Confessions of an English Opium-Eater (1821), have become a touchstone of romantic imperialism in recent studies of the literature of the period (Leask 1991; Barrell 1992 et al). De Quincey’s collocation of “all creatures, birds, beasts, reptiles, all trees and plants, usages and appearances, that are found in all tropical regions” in the hypnagogic hallucinations that characterized what he called “the pains of opium” seems to anticipate neatly Said’s theory of orientalism, whereby the orient was supplied by the west with “a mentality, a genealogy, an atmosphere,” the attitudinal basis as he argues for the continuing march of imperialism from the late eighteenth century. Yet, as Thomas Trautmann (1997) has pointed out, orientalist scholarship based in India and led by the influential Asiatic Society of Bengal in the late eighteenth century was extremely enthusiastic about Indian classical antiquity. The early orientalist scholarship posited ethnic, linguistic, cultural and religious links between Europe and India, while recognizing the greater antiquity of Indian civilization. This favourable attitude (which Trautmann calls “Indomania”) was overtaken in the nineteenth century by disavowal of that scholarship and repugnance (which he calls “Indophobia”), influenced by utilitarian and evangelical attitudes to colonialism. De Quincey’s lifespan covers this crucial period of change. My paper examines his evangelical upbringing and interest in biblical and orientalist scholarship to suggest his anxious investment in these modes of thinking. I will suggest that the bizarre orientalist fusions of his dreams can be better understood in the context of changing attitudes to the imperialism during the period. An examination of his work provides a far more dynamic understanding of the processes of orientalism than the binary model suggested by Said. The transformation implied from imperial scholarship to governance, I will suggest, is not irrelevant to a world which continues to pull apart on various grounds of race and ethnicity, and reflects on our own role in the academy today.