David J. Williams

The Global Snake Bite Initiative: an antidote for snake bite

Clinicians have for a long time witnessed the tragedy of injury, disability, and death from snake bite that is a daily occurrence in many parts of Africa, Asia, and Latin America. To many people living in these regions, including some of the world’s poorest communities, snake bite is an ever present occupational risk and environmental hazard, an additional penalty of poverty. Like malaria, dengue, tuberculosis, and parasitic diseases, the risk of snake bite is always present. Unlike many of these other public health risks, however, the burden of human suff ering caused by snake bite remains un-recognised, invisible, and unheard by the global public health community, forgotten by development agencies and governments alike. The problem is so underrated that it was only added to WHO’s list of neglected tropical diseases in April, 2009.Yet an estimated 5·4–5·5 million people are bitten by snakes each year,

Snakebite envenoming from a global perspective: Towards an integrated approach

Snakebite envenoming is a neglected public health challenge of compelling importance in many regions of the world, particularly sub-Saharan Africa, Asia, Latin America and Papua-New Guinea. Addressing the problem of snakebite effectively demands an integrated multifocal approach, targeting complex problems and involving many participants. It must comprise: (a) Acquisition of reliable information on the incidence and mortality attributable to snakebite envenoming, and the number of people left with permanent sequelae. (b) Improvements in production of effective and safe antivenoms, through strategies aimed at strengthening the technological capacity of antivenom manufacturing laboratories. (c) Increasing the capacity of low-income countries to produce specific immunogens(snake venoms) locally, and to perform their own quality control of antivenoms. (d) Commitments from regional producers to manufacture antivenoms for countries where antivenom production is not currently feasible. (e) Implementation of financial initiatives guaranteeing the acquisition of adequate volumes of antivenom at affordable prices in low-income countries. (f) Performance of collaborative studies on the safety and effectiveness of antivenoms assessed preclinically and by properly designed clinical trials. (g) Development of antivenom distribution programmes tailored to the real needs and epidemiological situations of rural areas in each country. (h) Permanent training programmes for health staff, particularly in rural areas where snakebites are frequent.(i) Implementation of programmes to support those people whose snakebites resulted in chronic disabilities. (j) Preventive and educational programmes at the community level, with the active involvement of local organizations and employing modern methods of health promotion. Such an integrated approach, currently being fostered by the Global Snake Bite Initiative of the International Society on Toxinology and by the World Health Organization, will help to alleviate the enormous burden of human suffering inflicted by snakebite envenoming.

Ending the drought: New strategies for improving the flow of affordable, effective antivenoms in Asia and Africa

The development of snake antivenoms more than a century ago should have heralded effective treatment of the scourge of snakebite envenoming in impoverished, mostly rural populations around the world. That snakebite still exists today, as a widely untreated illness that maims, kills and terrifies men, women and children in vulnerable communities, is a cruel anachronism. Antivenom can be an effective, safe and affordable treatment for snakebites, but apathy, inaction and the politicisation of public health have marginalised both the problem (making snakebite arguably the most neglected of all neglected tropical diseases) and its solution. For lack of any coordinated approach, provision of antivenoms has been pushed off the public health agenda, leading to an incongruous decline in demand for these crucial antidotes, excused and fed by new priorities, an absence of epidemiological data, and a poor regulatory framework. These factors facilitated the infiltration of poor quality products that degrade user confidence and undermine legitimate producers. The result is that tens of thousands are denied an essential life-saving medicine, allowing a toll of human suffering that is a summation of many individual catastrophes. No strategy has been developed to address this problem and to overcome the intransigence and inaction responsible for the global tragedy of snakebite. Attempts to engage with the broader public health community through the World Health Organisation (WHO), GAVI, and other agencies have failed. Consequently, the toxinology community has taken on a leadership role in a new approach, the Global Snakebite Initiative, which seeks to mobilise the resources, skills and experience of scientists and clinicians for whom venoms, toxins, antivenoms, snakes and snakebites are already fields of interest. Proteomics is one such discipline, which has embraced the potential of using venoms in bio-discovery and systems biology. The fields of venomics and antivenomics have recently evolved from this discipline, offering fresh hope for the victims of snakebites by providing an exciting insight into the complexities, nature, fundamental properties and significance of venom constituents. Such a rational approach brings with it the potential to design new immunising mixtures from which to raise potent antivenoms with wider therapeutic ranges. This addresses a major practical limitation in antivenom use recognised since the beginning of the 20th century: the restriction of therapeutic effectiveness to the specific venom immunogen used in production. Antivenomic techniques enable the interactions between venoms and antivenoms to be examined in detail, and if combined with functional assays of specific activity and followed up by clinical trials of effectiveness and safety, can be powerful tools with which to evaluate the suitability of current and new antivenoms for meeting urgent regional needs. We propose two mechanisms through which the Global Snakebite Initiative might seek to end the antivenom drought in Africa and Asia: first by establishing a multidisciplinary, multicentre, international collaboration to evaluate currently available antivenoms against the venoms of medically important snakes from specific nations in Africa and Asia using a combination of proteomic, antivenomic and WHO-endorsed preclinical assessment protocols, to provide a validated evidence base for either recommending or rejecting individual products; and secondly by bringing the power of proteomics to bear on the design of new immunising mixtures to raise Pan-African and Pan-Asian polyvalent antivenoms of improved potency and quality. These products will be subject to rigorous clinical assessment. We propose radically to change the basis upon which antivenoms are produced and supplied for the developing world. Donor funding and strategic public health alliances will be sought to make it possible not only to sustain the financial viability of antivenom production partnerships, but also to ensure that patients are relieved of the costs of antivenom so that poverty is no longer a barrier to the treatment of this important, but grossly neglected public health emergency.

Age validation, growth and mortality rates of the tropical snappers (Pisces: Lutjanidae) Lutjanus adetii (Castelnau, 1873) and L. quinquelineatus (Bloch, 1790) from the central Great Barrier Reef, Australia

The age and growth of Lutjanus adetii and L. quinquelineatus from the central Great Barrier Reef were determined from studies of annuli in sectioned otoliths (sagittae). The period of annulus formation was validated by oxytetracycline labelling of externally tagged fishes. For L. adetii, validation was obtained from tagged fishes that were recaptured after a minimum of 12 months at liberty, the first time this has been achieved for a Lutjanus species. A single opaque and translucent zone was formed once a year, with the opaque band (annulus) being formed during the winter months. Otolith weight was strongly correlated with age for both species. There was significant differential growth between the sexes in length-at-age and weight-at-age for both species, with males growing larger than females. The oldest individuals found were a male L. adetii of 24 years of age and a female L. quinquelineatus of 31 years of age. The shape of the growth curves were steep for the first few years and then became asymptotic. The annual instantaneous rate of natural mortality (M) was 0.235 for L. adetii and 0.154 for L. quinquelineatus, representing an annual survivorship of 79% and 86%, respectively. The protracted longevity and low natural mortality rates imply that both L. adetii and L. quinquelineatus are vulnerable to overfishing despite their small size.

Breaking processes in three-dimensional bonded granular materials with general shapes

The paper presents an extension to the spheropolyhedra method for the simulation of granular materials comprising particles of general shapes with bonding. A bonding, cement, or cohesion model for particles sharing common faces is introduced. The bonding force is elastic and has a strain-based breaking threshold for modelling fracture. An initial study is conducted based on the Brazilian tensile test to check how the parameters of the proposed model affect the principal variables measured in this test. Afterwards, solid cubic blocks are then subjected to a triaxial test to explore the mathematical macroscopic failure model. It is found that the peak strength envelope is the product of the superposition of frictional and fracture failure mechanisms. The fracture failure is mainly produced by an avalanche of broken cohesive bonds. The intensity of the avalanche exhibits a power law distribution, as reported in previous studies. The method allows for random divisions of solid bodies without any pre-existing internal voids. It offers a natural, effective tool to model, simulate and study fragmentation processes in 3D

Snake venomics of two poorly known Hydrophiinae: Comparative proteomics of the venoms of terrestrial Toxicocalamus longissimus and marine Hydrophis cyanocinctus

The venom proteomes of Toxicocalamus longissimus and Hydrophis cyanocinctus, a fossorial and a marine species, respectively, of the Hydrophiinae genus of Elapidae, were investigated by Edman degradation of RP-HPLC isolated proteins, and de novo MS/MS sequencing of in-gel derived tryptic peptide ions. The toxin arsenal of T. longissimus is made up of 1-2 type-I PLA(2) molecules, which account for 6.5% of the venom proteins, a minor PIII-SVMP (1.4% of the venom toxins), and ~20 members of the 3FTx family comprising 92% of the venom proteome. Seventeen proteins (5 type-I PLA(2)s and 12 3FTxs) were found in the venom of H. cyanocinctus. Three-finger toxins and type-I PLA(2) proteins comprise, respectively, 81% and 19% of its venom proteome. The simplicity of the H. cyanocinctus venom proteome is highlighted by the fact that only 6 venom components (3 short-chain neurotoxins, two long-chain neurotoxins, and one PLA(2) molecule) exhibit relative abundances >5%. As expected from its high neurotoxin abundance, the LD(50) for mice of H. cyanocinctus venom was fairly low, 0.132μg/g (intravenous) and 0.172μg/g (intraperitoneal). Our data indicate that specialization towards a lethal cocktail of 3FTx and type-I PLA(2) molecules may represent a widely adopted trophic solution throughout the evolution of Elapidae. Our results also points to a minimization of the molecular diversity of the toxin arsenal of the marine snake Hydrophis cyanocinctus in comparison to the venom proteome of its terrestrial relatives, and highlight that the same evolutionary solution, economy of the toxin arsenal, has been convergently adopted by different taxa in response to opposite selective pressures, loss and gain of neurotoxicity.

Effect of low dose irradiation on composition of tropical fruits and vegetables

Abstract Capsicums (green and red), cucumbers, custard apples, lemons, lychees, mandarins, mangoes, nectarines, papaws, peaches, persimmons, and zucchinis were irradiated at 0, 75, or 300 Gy in replicated factorial experiments. Commodities were analyzed shortly after irradiation and again after 3 to 4 weeks of storage at 1–7°C for soluble solids, pH, titratable, acidity, internal color, total vitamin C, dehydroascorbic acid, organic acids, and sugars. Significant (p

Investigating the Effect of Cyclic Loading on the Indirect Tensile Strength of Rocks

This paper presents the results of laboratory experiments during the investigation of the stress–strain characteristics of Brisbane tuff disc specimens under diametral compressive cyclic loading. Two different cyclic loading methods were used: namely, sinusoidal cyclic loading and cyclic loading with increasing mean level. The first method applied the S–N curve approach to the indirect tensile strength (ITS) of rock specimens for the first time in the literature, and the second method investigated the effect of increasing cyclic loading on the ITS of rock specimens. The ITS of Brisbane tuff disc specimens was measured using the Brazilian tensile strength test. The reduction in ITS was found to be 33% with sinusoidal loading tests, whereas increasing cyclic loading caused a maximum reduction of 37%. It is believed that the fracturing under cyclic loading starts at contact points between strong grains and weak matrices, and that contact points at grain boundaries are the regions of stress concentration (i.e., indenters). Transgranular cracks emanate from these regions and intergranular cracks sometimes pass through the contact points. Once cracking begins, there is a steady progression of damage and a general ‘loosening’ of the rock, which is a precursor to the formation of intergranular cracks.

Comparative proteomic analysis of the venom of the taipan snake, Oxyuranus scutellatus, from Papua New Guinea and Australia: Role of neurotoxic and procoagulant effects in venom toxicity

The venom proteomes of populations of the highly venomous taipan snake, Oxyuranus scutellatus, from Australia and Papua New Guinea (PNG), were characterized by reverse-phase HPLC fractionation, followed by analysis of chromatographic fractions by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. Proteins belonging to the following seven protein families were identified in the two venoms: phospholipase A(2) (PLA(2)), Kunitz-type inhibitor, metalloproteinase (SVMP), three-finger toxin (3FTx), serine proteinase, cysteine-rich secretory proteins (CRISP), and coagulation factor V-like protein. In addition, C-type lectin/lectin-like protein and venom natriuretic peptide were identified in the venom of specimens from PNG. PLA(2)s comprised more than 65% of the venoms of these two populations. Antivenoms generated against the venoms of these populations showed a pattern of cross-neutralization, corroborating the immunological kinship of these venoms. Toxicity experiments performed in mice suggest that, at low venom doses, neurotoxicity leading to respiratory paralysis represents the predominant mechanism of prey immobilization and death. However, at high doses, such as those injected in natural bites, intravascular thrombosis due to the action of the prothrombin activator may constitute a potent and very rapid mechanism for killing prey.

Epithiospecifier protein activity in broccoli: the link between terminal alkenyl glucosinolates and sulphoraphane nitrile.

The chemical nature of the hydrolysis products from the glucosinolate-myrosinase system depends on the presence or absence of supplementary proteins, such as epithiospecifier proteins (ESPs). ESPs (non-catalytic cofactors of myrosinase) promote the formation of epithionitriles from terminal alkenyl glucosinolates and as recent evidence suggests, simple nitriles at the expense of isothiocyanates. The ratio of ESP activity to myrosinase activity is crucial in determining the proportion of these nitriles produced on hydrolysis. Sulphoraphane, a major isothiocyanate produced in broccoli seedlings, has been found to be a potent inducer of phase 2 detoxification enzymes. However, ESP may also support the formation of the non-inductive sulphoraphane nitrile. Our objective was to monitor changes in ESP activity during the development of broccoli seedlings and link these activity changes with myrosinase activity, the level of terminal alkenyl glucosinolates and sulphoraphane nitrile formed. Here, for the first time, we show ESP activity increases up to day 2 after germination before decreasing again to seed activity levels at day 5. These activity changes paralleled changes in myrosinase activity and terminal alkenyl glucosinolate content. There is a significant relationship between ESP activity and the formation of sulforaphane nitrile in broccoli seedlings. The significance of these findings for the health benefits conferred by eating broccoli seedlings is briefly discussed.