Induction of epithelial to mesenchymal transition in PMC42-LA human breast carcinoma cells by carcinoma-associated fibroblast secreted factors

Background
Breast carcinoma is accompanied by changes in the acellular and cellular components of the microenvironment, the latter typified by a switch from fibroblasts to myofibroblasts.


Methods
We utilised conditioned media cultures, Western blot analysis and immunocytochemistry to investigate the differential effects of normal mammary fibroblasts (NMFs) and mammary cancer-associated fibroblasts (CAFs) on the phenotype and behaviour of PMC42-LA breast cancer cells. NMFs were obtained from a mammary gland at reduction mammoplasty, and CAFs from a mammary carcinoma after resection.


Results
We found greater expression of myofibroblastic markers in CAFs than in NMFs. Medium from both CAFs and NMFs induced novel expression of α-smooth muscle actin and cytokeratin-14 in PMC42-LA organoids. However, although conditioned media from NMFs resulted in distribution of vimentin-positive cells to the periphery of PMC42-LA organoids, this was not seen with CAF-conditioned medium. Upregulation of vimentin was accompanied by a mis-localization of E-cadherin, suggesting a loss of adhesive function. This was confirmed by visualizing the change in active β-catenin, localized to the cell junctions in control cells/cells in NMF-conditioned medium, to inactive β-catenin, localized to nuclei and cytoplasm in cells in CAF-conditioned medium.


Conclusion
We found no significant difference between the influences of NMFs and CAFs on PMC42-LA cell proliferation, viability, or apoptosis; significantly, we demonstrated a role for CAFs, but not for NMFs, in increasing the migratory ability of PMC42-LA cells. By concentrating NMF-conditioned media, we demonstrated the presence of factor(s) that induce epithelial-mesenchymal transition in NMF-conditioned media that are present at higher levels in CAF-conditioned media. Our in vitro results are consistent with observations in vivo showing that alterations in stroma influence the phenotype and behaviour of surrounding cells and provide evidence for a role for CAFs in stimulating cancer progression via an epithelial-mesenchymal transition. These findings have implications for our understanding of the roles of signalling between epithelial and stromal cells in the development and progression of mammary carcinoma.

Determinants of native avian richness in suburban remnant vegetation : implications for conservation planning

While urban areas are increasingly recognized as having potential value for biodiversity conservation, the relationship between biodiversity and the structure and configuration of the urban landscape is poorly understood. In this study we surveyed birds in 39 remnant patches of native vegetation of various sizes (range 1–107 ha) embedded in the suburban matrix in Melbourne, Australia. The total richness of species within remnants was strongly associated with the size of remnants. Remnant-reliant species displayed a much stronger response to remnant area than matrix-tolerant species indicating the importance of large remnants in maintaining representative bird assemblages. Large remnants are important for other ecological groups of species including migratory species, ground foraging birds and canopy foraging birds. Other landscape (e.g. amount of riparian vegetation) and structural components (e.g. shrub cover) of remnants have a lesser role in determining the richness of individual remnants. This research provides conservation managers and planners with a hierarchical process to reserve design and management in order to conserve the highest richness of native species within urban areas. First of all, conservation efforts should preferentially focus on the retention of larger remnants of native vegetation. Second, where possible, riparian vegetation should be included within reserves or, where it is already present, should be carefully managed to ensure its integrity. Third, efforts should be focused at maintaining appropriate habitat and vegetation structure and complexity.

Diet of an eastern barn owl Tyto javanica on the Patho Plains, northern Victoria

A small sample of eastern Barn Owl Tyto javanica pellets, from native grasslands on the Patho Plains in northern Victoria in February 2007, contained the remains of 48 prey individuals: 38 Australian Plague Locusts Chortoicetes terminifera, nine house Mice Mus domesticus and one Fat-tailed dunnart Sminthopsis crassicaudata. Such a high proportion of locusts in the eastern Barn Owl's diet is noteworthy, and is discussed in the context of recent locust-spraying operations in the region.

Pollutants increase song complexity and the volume of the brain area HVC in a songbird

Environmental pollutants which alter endocrine function are now known to decrease vertebrate reproductive success. There is considerable evidence for endocrine disruption from aquatic ecosystems, but knowledge is lacking with regard to the interface between terrestrial and aquatic ecosystems. Here, we show for the first time that birds foraging on invertebrates contaminated with environmental pollutants, show marked changes in both brain and behaviour. We found that male European starlings (Sturnus vulgaris) exposed to environmentally relevant levels of synthetic and natural estrogen mimics developed longer and more complex songs compared to control males, a sexually selected trait important in attracting females for reproduction. Moreover, females preferred the song of males which had higher pollutant exposure, despite the fact that experimentally dosed males showed reduced immune function. We also show that the key brain area controlling male song complexity (HVC) is significantly enlarged in the contaminated birds. This is the first evidence that environmental pollutants not only affect, but paradoxically enhance a signal of male quality such as song. Our data suggest that female starlings would bias their choice towards exposed males, with possible consequences at the population level. As the starling is a migratory species, our results suggest that transglobal effects of pollutants on terrestrial vertebrate physiology and reproduction could occur in birds.

Predicting spatio-temporal distribution for eastern Australian birds using birds Australia's atlas data : survey method, habitat and seasonal effects

Faunal atlases are landscape-level survey collections that can be used for describing spatial and temporal patterns of distribution and densities. They can also serve as a basis for quantitative analysis of factors that may influence the distributions of species. We used a subset of Birds Australia’s Atlas of Australian Birds data (January 1998 to December 2002) to examine the spatio-temporal distribution patterns of 280 selected species in eastern Australia (17–37°S and 136–152°E). Using geographical information systems, this dataset was converted into point coverage and overlaid with a vegetation polygon layer and a half-degree grid. The exploratory data analysis involved calculating species-specific reporting rates spatially, per grid and per vegetation unit, and also temporally, by month and year. We found high spatio-temporal variability in the sampling effort. Using generalised linear models on unaggregated point data, the influences of four factors – survey method and month, geographical location and habitat type – were analysed for each species. When counts of point data were attributed to grid-cells, the total number of species correlated with the total number of surveys, while the number of records per species was highly variable. Surveys had high interannual location fidelity. The predictive values of each of the four factors were species-dependent. Location and habitat were correlated and highly predictive for species with restricted distribution and strong habitat preference. Month was only of importance for migratory species. The proportion of incidental sightings was important for extremely common or extremely rare species. In conclusion, behaviour of species differed sufficiently to require building a customized model for each species to predict distribution. Simple models were effective for habitat specialists with restricted ranges, but for generalists with wide distributions even complex models gave poor predictions.

Effects of sublethal fenitrothion ingestion on cholinesterase inhibition, standard metabolism, thermal preference, and prey-capture ability in the Australian central bearded dragon (Pogona vitticeps, Agamidae)

The central bearded dragon (Pogona vitticeps) is a medium-sized lizard that is common in semiarid habitats in Australia and that potentially is at risk of fenitrothion exposure from use of the chemical in plague locust control. We examined the effects of single sublethal doses of this organophosphate (OP; low dose = 2.0 mg/kg; high dose = 20 mg/kg; control = vehicle alone) on lizard thermal preference, standard metabolic rate, and prey-capture ability. We also measured activities of plasma total cholinesterase (ChE) and acetylcholinesterase before and at 0, 2, 8, 24, 120, and 504 h after OP dosing. Predose plasma total ChE activity differed significantly between sexes and averaged 0.66 ± 0.06 and 0.45 ± 0.06 μmol/min/ml for males and females, respectively. Approximately 75% of total ChE activity was attributable to butyrylcholinesterase. Peak ChE inhibition reached 19% 2 h after OP ingestion in the low-dose group, and 68% 8 h after ingestion in high-dose animals. Neither OP doses significantly affected diurnal body temperature, standard metabolic rate, or feeding rate. Plasma total ChE levels remained substantially depressed up to 21 d after dosing in the high-dose group, making this species a useful long-term biomonitor of OP exposure in its habitat.

Diacetone alcohol, a dispersant solvent, contributes to acute toxicity of a fipronil-based insecticide in a passerine bird

Fipronil, a phenyl pyrazole pesticide, is aerially applied in eastern Australia to control locust outbreaks, usually as “Adonis 3UL Insecticide^®” (BASF), an ultra low (UL) volume formulation containing 0.3% active pesticide. We tested the toxicities of technical-grade fipronil, the Adonis 3UL formulation and its components in zebra finch, a native bird at risk of exposure in locust control regions. We estimated oral-dose LD50 by the Up-and-Down method. Under laboratory conditions, we identified unexpectedly high toxicities due exclusively to diacetone alcohol (DAA), a solvent making up 12.5% of the Adonis 3UL formulation. In contrast, finches were asymptomatic when exposed to 0.3% technical grade fipronil dissolved in a minimum amount of acetone. Depending upon the behaviour and persistence of DAA under field conditions, this formulation of Adonis 3UL may pose a far greater threat to the health of small birds and possibly other vertebrates than expected for fipronil alone.

Physiological assessment of avian exposure to fipronil, a new-generation pesticide

Fipronil, a phenyl pyrazole pesticide, is aerially applied in semi-arid and agricultural areas of Australia to control locust outbreaks. Locust populations build to plague proportions when rainfall occurs in late winter and spring, promoting early vegetation growth. These conditions also attract breeding birds. Over 100 species have been observed coincident with locust control operations. Avian exposure to fipronil occurs via direct contact and by ingesting contaminated insects or seeds. Avian toxicity information demonstrates there is high species-specific variability in fipronil sensitivity in the few avian species studied. There is no research, however, explaining this variability, nor is there research regarding physiological or behavioural sub-lethal effects on avian species. This makes it extremely difficult to predict the toxicity of fipronil on unstudied species at high risk of exposure. Our research aims to resolve this lack of essential information in two ways: firstly we examine whether fipronil has identifiable sublethal effects in exposed birds and their offspring that compromise population health, and secondly evaluate avian metabolism of fipronil in selected species to gain insight into the mechanisms underlying variation in species sensitivity. Our results provide critically needed information for evaluating field effects of locust-control spraying in Australia.

Patterns of use and exchange of genetic resources of the striped catfish Pangasianodon hypophthalmus (Sauvage 1878)

The present paper reviews the use and exchange of genetic resources of the migratory freshwater fish Pangasianodon hypophthalmus (Sauvage 1878) (the striped or sutchi catfish). This species is naturally distributed in the Mekong River and Chao Phraya River basins, and is cultured in several countries, but current production occurs predominantly in the Mekong Delta, Vietnam. Catfish aquaculture in Vietnam has evolved from extensive systems using wild-caught seed to an intensified farming system that is entirely dependent on hatchery-produced seed. Genetic improvement programmes on catfish have started in Vietnam, but are still in their infancy. Genetic studies have revealed several subpopulations of the species. Apart from selective breeding and the production of hybrids with closely related species, no other technologies have been applied to improve the performance of catfish. The use and exchange of P. hypophthalmus genetic resources have brought benefits to rural communities. Aquaculture development of catfish has evolved from being seen as an exploitation of natural resources to an activity that can reduce pressure on wild fish populations. Management of aquaculture stocks need to be rationalised to minimise the potential impacts it might cause to wild populations.

Will Wallace’s Line save Australia from avian influenza?

Australia is separated from the Asian faunal realm by Wallace's Line, across which there is relatively little avian migration. Although this does diminish the risk of high pathogenicity avian influenza of Asian origin arriving with migratory birds, the barrier is not complete. Migratory shorebirds, as well as a few landbirds, move through the region on annual migrations to and from Southeast Asia and destinations further north, although the frequency of infection of avian influenza in these groups is low. Nonetheless,high pathogenicity H5N1 has recently been recorded on the island of New Guinea in West Papua in domestic poultry. This event increases interest in the movements of birds between Wallacea in eastern Indonesia, New Guinea, and Australia, particularly by waterbirds. There are frequent but irregular movements of ducks, geese, and other waterbirds across Torres Strait between New Guinea and Australia, including movements to regions in which H5N1 has occurred in the recent past. Although the likelihood of avian influenza entering Australia via an avian vector is presumed to be low, the nature and extent of bird movements in this region is poorly known. There have been five recorded outbreaks of high pathogenicityavian influenza in Australian poultry flocks, all of the H7 subtype. To date, Australia is the only inhabited continent not to have recorded high pathogenicity avian influenza since 1997, and H5N1 has never been recorded. The ability to map risk from high pathogenicity avian influenza to Australia is hampered by the lack of quantitative data on the extent of bird movements between Australia and its northern neighbors.Recently developed techniques offer the promise to fill this knowledge gap.

The role of wild birds in the transmission of avian influenza for Australia : an ecological perspective

Waterbirds, particularly Anatidae, are natural reservoirs for low-pathogenic avian influenza and have been implicated as the primary source of infection in outbreaks of highly pathogenic avian influenza. An understanding of the movements of birds and the ecology of avian influenza viruses within the wild bird population is essential in assessing the risks to human health and production industries. Marked differences in the movements of Australian birds from those of the Northern Hemisphere emphasises the danger of generalising trends of disease prevalence to Australian conditions. Populations of Anatidae in Australia are not migratory, as they are in the Northern Hemisphere, but rather display typical nomadic traits, sometimes moving large distances across continental Australia in response to flooding or drought. There is little known regular interchange of anatids between Australia and Asia. In contrast, species such as shorebirds and some seabirds are annual migrants to Australia along recognised flyways from breeding grounds in the Northern Hemisphere. Movement into Australia by these species mainly occurs into the north-west and along the east coast over the Pacific Ocean. These species primarily arrive during the Australian spring and form large aggregations along the coastline and on inland wetlands. Other Australian migratory species (passerines, bee-eaters, dollar-birds, cuckoos, doves) regularly move to and from Asia through the Torres Strait Islands. The disease status of these birds is unknown. The movements of some species, particularly anatids and ardeids, which have ranges including Australia and regions where the virus is known to occur, have been poorly studied and there is potential for introduction of avian influenza subtypes via this route. Avian influenza viruses are highly unpredictable and can undergo reassortment to more pathogenic forms. There is insufficient knowledge of the epidemiology and transmission of these viruses in Australia and broad-scale surveillance of wild birds is logistically difficult. Long-term studies of anatids that co-habit with Charadriiformes are recommended. This would provide an indication of the spatial and temporal patterns of subtypes entering Australia and improve our understanding of the ecology of endemic viruses. Until such time as these data become available, Australia's preparedness for avian influenza must focus on biosecurity at the wild bird–poultry interface.

Singing as a handicap : the effects of food availability and weather on song output in the Australian reed warbler Acrocephalus australis

Bird song is generally regarded as a sexually selected trait, and may represent a reliable handicap signal under at least certain conditions. Females may use the degree of male song production as a reliable cue to male condition or territory quality. We investigated the effect of supplementary feeding on song output in the migratory Australian reed warbler Acrocephalus australis. We experimentally increased the food availability on alternate days, and recorded several weather variables. We measured song rate and song length independently. Supplementary fed birds sang more on feeding days than on non-feeding days, while control birds did not show this effect. Song output was not significantly associated with any of the weather variables examined. Our results indicate that singing has the potential to serve as a reliable handicap signal to territorial food availability irrespective of the prevailing weather conditions. We discuss the role of energetic constraints and behavioural flexibility on the signaling function of song.

Fishways in Australia

In Australian regulated rivers, dams and weirs have contributed to serious declines in the distribution and abundance of migratory fish. This thesis has evaluated four fishways in Victoria, New South Wales and Queensland and provided new information for 51 migratory fish species. Fishways will be crucial for management and conservation of biodiversity.

Migrant housing in the city and the village : from Melbourne to Zavoj

This paper will discuss the kinds of communities that evolve through historical practices of migration. The migrant house is associated with a new architecture that hod appeared in the cities of immigration of the new worlds (Melbourne, Toronto, Chicago). It is perceived as a stereotypical symbolisation of immigrants from Southern European origins that had arrived in the decades following the Second World War. The appearance of houses built by returning migrants in sites of origin suggests other traiectories, other modes of travel, and other forms of community. Central to the thesis of this paper is the testimony of two types of migrant houses. The study draws on theories of migration that address the site of departure, the site of arrival, and the question and conflict of return which is at the centre of the migrant's imaginary. This study will examine the migrant houses in the village of emigration (Zavoj in Macedonia), migrant houses built by returning emigrants. A study of the two houses of migration implicates a set of networks, forces, relations, circumscribing a large global geopolitical and cultural field that questions our understandings of diaspora, the binary structure of dwelling/travelling, and the fabric and fabrication of community. In addition, the paper will explore the notion of house as an imaginary landscape, a psychic geography narrated through migratory travels.

Identification of significant shorebird areas : thresholds and criteria

Aim Conservation managers designate significant areas for shorebirds based on imperfect data. Significant wetlands for migratory shorebirds have usually been identified on the basis of whether they exceed certain thresholds, defined either by total abundance (usually 20,000 waterbirds) or percentage of a population (usually 1.0%). We evaluate the performance of existing criteria and determine if lowering thresholds would improve shorebird conservation without adding unreasonable numbers of significant sites.

Location Australia.

Methods We evaluated the best available data, which is used by managers to designate significant areas, to describe the effect of lowering thresholds on the number of significant sites identified and the number of shorebirds these sites support using a range of thresholds in existing criteria. We also investigated factors which may explain interspecific differences evident when lowering thresholds.

Results When the threshold for total abundance was lowered from 20,000 to 2000 shorebirds, an additional 45 shorebird areas, holding 65% more shorebirds, were identified. When thresholds for the percentage of a population criterion were lowered from 1.0 to 0.1%, an additional 86 shorebird areas were identified which held 29% more shorebirds. The proportion of a species population counted within wetlands identified as significant by the application of criteria varied widely between species. The percentage of population criterion always identified a network of areas that included more individuals of each species than the total abundance criterion at all threshold levels tested. The percentage of species populations found in networks of significant areas showed greater increase as thresholds were lowered for species that were abundant, widespread and well represented at existing thresholds.

Main conclusions Our results suggest lowering thresholds will substantially increase the number of shorebirds in identified significant areas. However, some species will remain under-represented, partly because of interspecific differences in distribution and inadequate sampling of some shorebird habitats.