Transcriptional profiling of growth perturbations of the human malaria parasite Plasmodium falciparum

Functions have yet to be defined for the majority of genes of Plasmodium falciparum, the agent responsible for the most serious form of human malaria. Here we report changes in P. falciparum gene expression induced by 20 compounds that inhibit growth of the schizont stage of the intraerythrocytic development cycle. In contrast with previous studies, which reported only minimal changes in response to chemically induced perturbations of P. falciparum growth, we find that ~59% of its coding genes display over three-fold changes in expression in response to at least one of the chemicals we tested. We use this compendium for guilt-by-association prediction of protein function using an interaction network constructed from gene co-expression, sequence homology, domain-domain and yeast two-hybrid data. The subcellular localizations of 31 of 42 proteins linked with merozoite invasion is consistent with their role in this process, a key target for malaria control. Our network may facilitate identification of novel antimalarial drugs and vaccines.

Transient responses of a wetting film to mechanical and electrical perturbations

This article reports real-time observations and detailed modeling of the transient response of thin aqueous films bounded by a deformable surface to external mechanical and electrical perturbations. Such films, tens to hundreds of nanometers thick, are confined between a molecularly smooth mica plate and a deformable mercury/electrolyte interface on a protuberant drop at a sealed capillary tube. When the mercury is negatively charged, the water forms a wetting film on mica, stabilized by electrical double layer forces. Mechanical perturbations are produced by driving the mica plate toward or by retracting the mica plate from the mercury surface. Electrical perturbations are applied to change the electrical double layer interaction between the mica and the mercury by imposing a step change of the bias voltage between the mercury and the bulk electrolyte. A theoretical model has been developed that can account for these observations quantitatively. Comparison between experiments and theory indicates that a no-slip hydrodynamic boundary condition holds at the molecularly smooth mica/electrolyte surface and at the deformable mercury/electrolyte interface. An analysis of the transient response based on the model elucidates the complex interplay between disjoining pressure, hydrodynamic forces, and surface deformations. This study also provides insight into the mechanism and process of droplet coalescence and reveals a novel, counterintuitive mechanism that can lead to film instability and collapse when an attempt is made to thicken the film by pulling the bounding mercury and mica phases apart.

Robust non-fragile control for offshore steel jacket platform with nonlinear perturbations

This study is concerned with the design of a non-fragile controller for an offshore steel jacket platform with nonlinear perturbations. The delay-dependent sufficient conditions are derived in terms of linear matrix inequalities based on suitable Lyapunov–Krasovskii functional, the second-order reciprocally convex approach and the lower bound lemma. The results indicate asymptotic stability of the offshore steel jacket platform utilizing the proposed non-fragile controller. Besides that, robust stability conditions are derived for an uncertain offshore platform subject to the non-fragile controller. A numerical example is given to illustrate the effectiveness of the proposed theoretical results.

Explicit invariant solutions associated with nonlinear atmospheric flows in a thin rotating spherical shell with and without west-to-east jets perturbations

A class of non-stationary exact solutions of two-dimensional nonlinear Navier–Stokes (NS) equations within a thin rotating spherical shell were found as invariant and approximately invariant solutions. The model is used to describe a simple zonally averaged atmospheric circulation caused by the difference in temperature between the equator and the poles. Coriolis effects are generated by pseudoforces, which support the stable west-to-east flows providing the achievable meteorological flows. The model is superimposed by a stationary latitude dependent flow. Under the assumption of no friction, the perturbed model describes zonal west-to-east flows in the upper atmosphere between the Ferrel and Polar cells. In terms of nonlinear modeling for the NS equations, two small parameters are chosen for the viscosity and the rate of the earth’s rotation and exact solutions in terms of elementary functions are found using approximate symmetry analysis. It is shown that approximately invariant solutions are also valid in the absence of the flow perturbation to a zonally averaged mean flow.

Long-term flowering patterns of melliferous Eucalyptus (Myrtaceae) species

The flowering patterns of 28 Victorian melliferous (honey-producing) eucalypts were investigated by using long-term observations of highly experienced, commercial apiarists. Frequency, timing, duration and intensity of flowering were determined, as were spatial differences within and among species. Data were obtained by face-to-face interviews with 25 Victorian apiarists, each of whom had operated a minimum of 350 hives for a minimum of 30 years. Flowering frequency ranged from 1 to 7 years, and most species flowered once every 2–4 years. Long-term flowering frequency, timing and duration were reported as constant, although short-term perturbations could occur. Most melliferous species flowered during spring and summer for a period of 3 months or more. Only few species had shorter flowering periods. Information provided by apiarists compared well with available published information (e.g. flowering period reported in field guides) and revealed a reliable, largely untapped source of long-term data, the use of which could benefit many ecological research endeavours.

Constitutive activation of zebrafish Stat5 expands hematopoietic cell populations in vivo

Objective
Constitutive activation of Stat5 has been observed in a variety of malignancies, particularly myeloid leukemias. To directly investigate the in vivo consequences of Stat5 perturbation, we expressed constitutively active forms in zebrafish.
Methods
We generated mutants of the zebrafish stat5.1 protein (N646H, H298R/N714F, and N714F) based on previously identified constitutively active mutants of murine Stat5a. The in vitro properties of these mutants were determined using phosphorylation-specific antibodies and luciferase reporter assays, and their in vivo effects were analyzed through microinjection of zebrafish embryos.
Results
Two of these stat5.1 mutants (N646H and H298R/N714F) showed increased tyrosine phosphorylation and transactivation activity compared to the wild-type protein. Expression of either mutant led to a range of hematological perturbations, which were more pronounced for the H298R/N714F mutant. Interestingly, expression of wild-type also produced generally similar phenotypes. Further analysis showed that expression of the H298R/N714F mutant led to increased numbers of early and late myeloid cells, erythrocytes, and B cells. Some nonhematopoietic developmental perturbations were also observed, but these were equally prominent with wild-type or mutant forms.
Conclusion
These data implicate Stat5 activity as a direct critical regulator of hematological cell proliferation, suggesting a causal role for constitutively-active Stat5 in the etiology of hematological malignancies.

Ontogeny of diving behaviour in the Australian sea lion: trials of adolescence in a late bloomer

1.Foraging behaviours of the Australian sea lion (Neophoca cinerea) reflect an animal working hard to exploit benthic habitats. Lactating females demonstrate almost continuous diving, maximize bottom time, exhibit elevated field metabolism and frequently exceed their calculated aerobic dive limit. Given that larger animals have disproportionately greater diving capabilities, we wanted to examine how pups and juveniles forage successfully.
2.Time/depth recorders were deployed on pups, juveniles and adult females at Seal Bay Conservation Park, Kangaroo Island, South Australia. Ten different mother/pup pairs were equipped at three stages of development (6, 15 and 23 months) to record the diving behaviours of 51 (nine instruments failed) animals.
3. Dive depth and duration increased with age. However, development was slow. At 6 months, pups demonstrated minimal diving activity and the mean depth for 23-month-old juveniles was only 44 ± 4 m, or 62% of adult mean depth.
4. Although pups and juveniles did not reach adult depths or durations, dive records for young sea lions indicate benthic diving with mean bottom times (2·0 ± 0·2 min) similar to those of females (2·1 ± 0·2 min). This was accomplished by spending higher proportions of each dive and total time at sea on or near the bottom than adults. Immature sea lions also spent a higher percentage of time at sea diving.
5. Juveniles may have to work harder because they are weaned before reaching full diving capability. For benthic foragers, reduced diving ability limits available foraging habitat. Furthermore, as juveniles appear to operate close to their physiological maximum, they would have a difficult time increasing foraging effort in response to reductions in prey. Although benthic prey are less influenced by seasonal fluctuations and oceanographic perturbations than epipelagic prey, demersal fishery trawls may impact juvenile survival by disrupting habitat and removing larger size classes of prey. These issues may be an important factor as to why the Australian sea lion population is currently at risk.

The three R`s of resilience: redundancy, requisite variety and resources

This paper offers a way of operationalizing resilience according to the sub-systems that comprise a business (technical, social and economic), the kinds of perturbations that might impact a business (local, national and global) and the criteria that determine resilient capacity (redundancy, requisite variety and resources). When a business system has incorporated redundancy, developed requisite variety and adequately monitors its resources, we might then conclude that such a business is a resilient business. The model offered here is theoretical, and is yet to undergo empirical scrutiny. Empirical modeling will enable us to ascertain the strength of a business's internal characteristics against different levels/kinds of external perturbation. Sensitivity analysis of this kind will lead to more in-depth understanding of the dynamics that generate resilient businesses in a complex world.

A global perspective of aquaculture in the new millennium

In modern times, not many primary industries have consistently recorded high yearly growth over a period of two decades. Aquaculture has sustained a global growth, continues to grow, and is expected to increasingly fill the shortfall in aquatic food products resulting from static or declining capture fisheries and population increase well into the year 2025. Its further growth and development will have to occur under a different socio-economic milieu in the new millennium. The basic paradigm changes will be from an increased production at almost any cost, to a sustainable increase in production with minimal environmental perturbations. Despite such paradigm changes, aquaculture will increasingly contribute to food security, poverty alleviation and social equity. The contribution of aquaculture to world food supply of aquatic products has been increasing over the past 10 years, in comparison to capture fisheries, growing from 15 to 28 percent of total production between 1988 and 1997. As the bulk of aquaculture is rural and subsistence, it plays a major role as a provider of direct and indirect employment to the rural poor and, thereby, to poverty alleviation. In many developing countries, aquaculture provides opportunities for diversification on agriculture farms and productive use to otherwise idle land during certain seasons. The main cause for the upsurge in the sector has been the transformation of aquaculture from an “art” form to a “science”. This brought many advantages, ranging from less dependence on wild stock to the development of techniques that optimized yields, such as polyculture, or enabled the achievement of high yields with low inputs. Two major developments also enabled the sector to maintain growth momentum, appropriate institutional frameworks and concerted research and development. Regions or continents have many commonalities. These include the predominance of finfish among the cultivated species, and the predominance of species that feed lower in the food chain, although shrimp, which does not naturally feed high in the trophic level but is mostly reared on artificial feed, has become a significant culture commodity. Notable differences, however, include the fact that all regions, except Africa and the countries of the former USSR, have recorded a significant increase in per capita production between 1984 and 1997. While Asia continues to dominate world aquaculture in overall tonnage, as well as in every major commodity, South America has registered a very high (72.8 percent) average annual growth between 1984 and 1997. The global and regional trends over the last 20 years in the sector from a number of perspectives, such as production trends, contribution of aquaculture to aquatic food consumption etc., are evaluated. Based on these different trends and in the light of changing socio-economic conditions globally, and in particular, in developing nations, the potential changes in the sector in the new millennium are highlighted. Finally, projections are made for the next 20 years, where opportunities, constraints and strategies for achieving the targets are presented and discussed.

The interaction of lipophilic drugs with intestinal fatty acid-binding protein

Intestinal fatty acid-binding protein (I-FABP) is a small protein that binds long-chain dietary fatty acids in the cytosol of the columnar absorptive epithelial cells (enterocytes) of the intestine. The binding cavity of I-FABP is much larger than is necessary to bind a fatty acid molecule, which suggests that the protein may be able to bind other hydrophobic and amphipathic ligands such as lipophilic drugs. Herein we describe the binding of three structurally diverse lipophilic drugs, bezafibrate, ibuprofen (both R- and S-isomers) and nitrazepam to I-FABP. The rank order of affinity for I-FABP determined for these compounds was found to be R-ibuprofen {approx} bezafibrate > S-ibuprofen >> nitrazepam. The binding affinities were not directly related to aqueous solubility or partition coefficient of the compounds; however, the freely water-soluble drug diltiazem showed no affinity for I-FABP. Drug-I-FABP interaction interfaces were defined by analysis of chemical shift perturbations in NMR spectra, which revealed that the drugs bound within the central fatty acid binding cavity. Each drug participated in a different set of interactions within the cavity; however, a number of common contacts were observed with residues also involved in fatty acid binding. These data suggest that the binding of non-fatty acid lipophilic drugs to I-FABP may increase the cytosolic solubility of these compounds and thereby facilitate drug transport from the intestinal lumen across the enterocyte to sites of distribution and metabolism.

Stability theory and numerical analysis of non-autonomous dynamical systems

The development and use of cocycles for analysis of non-autonomous behaviour is a technique that has been known for several years. Initially developed as an extension to semi-group theory for studying rion-autonornous behaviour, it was extensively used in analysing random dynamical systems [2, 9, 10, 12]. Many of the results regarding asymptotic behaviour developed for random dynamical systems, including the concept of cocycle attractors were successfully transferred and reinterpreted for deterministic non-autonomous systems primarily by P. Kloeden and B. Schmalfuss [20, 21, 28, 29]. The theory concerning cocycle attractors was later developed in various contexts specific to particular classes of dynamical systems [6, 7, 13], although a comprehensive understanding of cocycle attractors (redefined as pullback attractors within this thesis) and their role in the stability of non-autonomous dynamical systems was still at this stage incomplete. It was this purpose that motivated Chapters 1-3 to define and formalise the concept of stability within non-autonomous dynamical systems. The approach taken incorporates the elements of classical asymptotic theory, and refines the notion of pullback attraction with further development towards a study of pull-back stability arid pullback asymptotic stability. In a comprehensive manner, it clearly establishes both pullback and forward (classical) stability theory as fundamentally unique and essential components of non-autonomous stability. Many of the introductory theorems and examples highlight the key properties arid differences between pullback and forward stability. The theory also cohesively retains all the properties of classical asymptotic stability theory in an autonomous environment. These chapters are intended as a fundamental framework from which further research in the various fields of non-autonomous dynamical systems may be extended. A preliminary version of a Lyapunov-like theory that characterises pullback attraction is created as a tool for examining non-autonomous behaviour in Chapter 5. The nature of its usefulness however is at this stage restricted to the converse theorem of asymptotic stability. Chapter 7 introduces the theory of Loci Dynamics. A transformation is made to an alternative dynamical system where forward asymptotic (classical asymptotic) behaviour characterises pullback attraction to a particular point in the original dynamical system. This has the advantage in that certain conventional techniques for a forward analysis may be applied. The remainder of the thesis, Chapters 4, 6 and Section 7.3, investigates the effects of perturbations and discretisations on non-autonomous dynamical systems known to possess structures that exhibit some form of stability or attraction. Chapter 4 investigates autonomous systems with semi-group attractors, that have been non-autonomously perturbed, whilst Chapter 6 observes the effects of discretisation on non-autonomous dynamical systems that exhibit properties of forward asymptotic stability. Chapter 7 explores the same problem of discretisation, but for pullback asymptotically stable systems. The theory of Loci Dynamics is used to analyse the nature of the discretisation, but establishment of results directly analogous to those discovered in Chapter 6 is shown to be unachievable. Instead a case by case analysis is provided for specific classes of dynamical systems, for which the results generate a numerical approximation of the pullback attraction in the original continuous dynamical system. The nature of the results regarding discretisation provide a non-autonomous extension to the work initiated by A. Stuart and J. Humphries [34, 35] for the numerical approximation of semi-group attractors within autonomous systems. . Of particular importance is the effect on the system's asymptotic behaviour over non-finite intervals of discretisation.

Semi-hyperbolic mappings in Banach spaces

The definition of semi-hyperbolic dynamical systems generated by Lipschitz continuous and not necessarily invertible mappings in Banach spaces is presented in this thesis. Like hyperbolic mappings, they involve a splitting into stable and unstable spaces, but a slight leakage from the strict invariance of the spaces is possible and the unstable subspaces are assumed to be finite dimensional. Bi-shadowing is a combination of the concepts of shadowing and inverse shadowing and is usually used to compare pseudo-trajectories calculated by a computer with the true trajectories. In this thesis, the concept of bi-shadowing in a Banach space is defined and proved for semi-hyperbolic dynamical systems generated by Lipschitz mappings. As an application to the concept of bishadowing, linear delay differential equations are shown to be bi-shadowing with respect to pseudo-trajectories generated by nonlinear small perturbations of the linear delay equation. This shows robustness of solutions of the linear delay equation with respect to small nonlinear perturbations. Complicated dynamical behaviour is often a consequence of the expansivity of a dynamical system. Semi-hyperbolic dynamical systems generated by Lipschitz mappings on a Banach space are shown to be exponentially expansive, and explicit rates of expansion are determined. The result is applied to a nonsmooth noninvertible system generated by delay differential equation. It is shown that semi-hyperbolic mappings are locally φ-contracting, where -0 is the Hausdorff measure of noncompactness, and that a linear operator is semi-hyperbolic if and only if it is φ-contracting and has no spectral values on the unit circle. The definition of φ-bi-shadowing is given and it is shown that semi-hyperbolic mappings in Banach spaces are φ-bi-shadowing with respect to locally condensing continuous comparison mappings. The result is applied to linear delay differential equations of neutral type with nonsmooth perturbations. Finally, it is shown that a small delay perturbation of an ordinary differential equation with a homoclinic trajectory is ‘chaotic’.

Role of leptin receptors in the development of obesity

The focus of this dissertation was leptin and the leptin receptor, and the role of these genes (OB and OB-R) in the development of obesity and type 2 diabetes in humans and Psammomys obesus, a polygenic rodent model of obesity and type 2 diabetes. Studies in humans showed that circulating leptin concentrations were positively associated with adiposity, and independently associated with circulating insulin and triglyceride concentrations. Analysis of two leptin receptor sequence polymorphisms in a Caucasian Australian population and a population of Nauruan males, with very high prevalence rates of obesity, showed no associations between sequence variation within the OB-R gene and obesity- or diabetes-related phenotypic measures. In addition, these two OB-R polymorphisms were not associated with longitudinal changes in body mass or composition in either of the populations examined. A unique analysis of the effects of multiple gene defects in the Nauruan population, demonstrated that the presence of sequence alterations in both the OB and OB-R genes were associated with insulin resistance. Psammomys obesus is regarded as an excellent rodent model in which to study the development of obesity and type 2 diabetes in humans. Examination of circulating leptin concentrations in Psammomys revealed that, as in humans, leptin concentrations were associated with adiposity, and independently associated with circulating insulin concentrations. This animal model was utilised to examine expression of OB-R, and the regulation of expression of this gene after dietary manipulation. OB-R is known to have several isoforms, and in particular, OB-RA and OB-RB gene expression were examined. OB-RB is the main signalling isoform of the leptin receptors. It has a long intracellular domain and has previously been shown to play an important role in energy balance and body weight regulation in rodents and humans. OB-RA is a much shorter isoform of OB-R, and although it lacks the long intracellular domain necessary to activate the JAK/STAT pathway, OB-RA is also capable of signalling, although to a lesser degree than OB-RB. OB-RA is found to be expressed almost ubiquitously throughout the body, and this isoform may be involved in transport of leptin into the cell, although its role remains unclear. OB-RA and OB-RB were both found to be expressed in a large number of tissues in Psammomys obesus. Interestingly, obese Psammomys were found to have lower levels of expression of OB-RA and OB-RB in the hypothalamus, compared to lean animals. This finding raises the possibility that decreased leptin signalling in the brain of obese, hyperleptinemic Psammomys obesus may contribute to the leptin resistance previously described in this animal model. However, the primary defect is unclear, as alternatively, increased circulating leptin concentrations may lead to down-regulation of leptin receptors. The effect of fasting on leptin concentrations and gene expression of OB-RA and OB-RB was also examined. A 24-hour fast resulted in no change in body weight, but a reduction in circulating leptin concentrations, and an increase in hypothalamic OB-RB gene expression in lean Psammomys. In obese animals, fasting again did not alter body weight, but resulted in an increase in both circulating leptin concentrations and hypothalamic OB-RB gene expression. In the liver, fasting resulted in a large increase in OB-RA gene expression in both lean and obese animals. These results highlighted the fact that regulation of leptin receptor gene expression in polygenic models of obesity and type 2 diabetes is complex, and not solely under the control of circulating leptin concentrations. Sucrose-feeding is an established method of inducing obesity and type 2 diabetes in rodents, and this experimental paradigm was utilised to examine the effects of longer term perturbations of energy balance on the leptin signalling pathway in Psammomys obesus. Addition of a 5% sucrose solution to the diet of lean and obese Psammomys resulted in increased body weight in both groups of animals, however only obese Psammomys showed increased fat mass and the development of type 2 diabetes. The changes in body mass and composition with sucrose-feeding were accompanied by decreased circulating leptin concentrations in both groups of animals, as well as a range of changes in leptin receptor gene expression. Sucrose-feeding increased hypothalamic OB-RB gene expression in obese Psammomys only, while in the liver there was evidence of a reduction in OB-RA and OB-RB gene expression in both lean and obese animals. The direct effects of sucrose on the leptin signalling pathway are unclear, however it is possible to speculate that the effect of sucrose to decrease leptin concentrations may have been involved in the exacerbation of obesity and the development of type 2 diabetes in obese Psammomys, From these studies, it appears that sequence variation in the OB and OB-R genes is unlikely to be a major factor in the etiology of obesity in human populations. The ability to examine regulation of expression of these genes in Psammomys obesus, however, has demonstrated that the effects of nutritional modifications on leptin receptor gene expression need closer attention. The role of the OB and OB-R genes in metabolism and the development of type 2 diabetes also warrants further examination, with particular attention on the differential effects of dietary modifications on leptin receptor gene expression across a range of tissues.