Exploiting knowledge of mesenchymal stromal cells in vivo for bone disease therapy development

This thesis aims to demonstrate how understanding of the nature of MSCs in vivo can be used to guide bone disease therapy development. It first address the unpredictable MSC content of bone marrow (BM) aspirates used for therapy; then examines MSC enrichment using clinical grade immunomagnetic cell selection. Induction of osteogenesis was subsequently explored; by examining an induced membrane (IM) used for bone regeneration. Finally the potential of a janus kinase (JAK) inhibitor to block osteogenesis was assessed, by studying its effect on MSCs in vitro. Flow cytometry was used to enumerate cells expressing a CD45-/low CD271+ phenotype in BM aspirate and this was compared to colony forming unit fibroblast (CFU-F) content. MSCs were enriched from BM, enzymatically treated femoral heads (FH) and intramedullary canal aspirates and enumerated. The composition morphology, MSC content and differentiation potential of IM was compared to periosteum. The potential effects of JAK inhibition on MSC colony formation, expansion and differentiation potential were examined. The concentration of cells expressing a CD45-/low CD271+ phenotype strongly correlated with CFU-F concentration (R=0.812, p<0.001). Immunomagnetic cell selection resulted in an increase in the proportion of MSCs in BM, FH and intramedullary canal aspirates by 204, 14.1 and 291-fold respectively. The regenerative potential of periosteum and IM were comparable. JAK inhibition did not affect MSC growth, osteogenesis or chondrogenesis but caused an increase in adipogenesis at concentrations ≥100nM compared to controls (1.38 fold, p=0.041). Flow cytometry may be used to rapidly and accurately predict the MSC content of BM. Clinical grade immunomagnetic selection can substantially increase the purity of MSCs from bone cavities. The similarity, in terms of regenerative potential, of periosteum and IM gives insight into its use for bone regeneration. JAK inhibition did not affect in vitro osteogenesis but has potential to affect in vivo osteogenesis through stimulation of adipogenesis

A rapid biodiversity survey of Papua New Guinea's Manus and Mussau Islands

[Extract] Aim: The relatively remote islands of Manus and Mussau, located in the northern portion of the Bismark sea have been long identified as key biodiversity areas in Papua New Guinea and within greater Melanesia. Manus Island has long been known for its endemism and relatively intact forest, while Mussau Island, although relatively unstudied, has been recognised as an Endemic Bird Area. This report documents the findings of a series of rapid biodiversity surveys focusing on terrestrial flora and fauna, funded by the CEPF, encompassing four sites across the islands of Manus and Mussau; undertaken by a WCS led team of national and international taxonomic specialists in October 2014. The objective of these surveys was to investigate the biodiversity values of these areas. In conjunction with participatory community work conducted prior to, and following the surveys the wider WCS project aims to identify options for natural resource management in the region which addresses both community and biodiversity needs

The impacts of introduced house mice on the breeding success of nesting seabirds on Gough Island

Invasive species are the main threat to island biodiversity; seabirds are particularly vulnerable and are one of the most threatened groups of birds. Gough Island, a UNESCO World Heritage Site in the South Atlantic Ocean, is an Important Bird and Biodiversity Area, and one of the most important seabird colonies globally. Invasive House Mice Mus musculus depredate eggs and chicks of most seabird species on the island, but the extent of their impact has not been quantified. We used field data and bootstrapped normal distributions to estimate breeding success and the number of surviving chicks for 10 seabird species on Gough Island, and compared estimates with those of analogous species from predator-free islands. We examined the effects of season and nest-site location on the breeding success of populations on Gough Island, predicting that the breeding success of Gough birds would be lower than that of analogues, particularly among small burrow-nesting species. We also predicted that winter-breeding species would exhibit lower breeding success than summer-breeding species, because mice have fewer alternative food sources in winter; and below-ground nesters would have lower breeding success than surface nesters, as below-ground species are smaller so their chicks are easier prey for mice. We did indeed find that seabirds on Gough Island had low breeding success compared with analogues, losing an estimated 1 739 000 (1 467 000–2 116 000) eggs/chicks annually. Seven of the 10 focal species on Gough Island had particularly high chick mortality and may have been subject to intense mouse predation. Below-ground and winter breeders had lower breeding success than surface- and summer-breeders. MacGillivray's Prion Pachyptila macgillivrayi, Atlantic Petrel Pterodroma incerta and Tristan Albatross Diomedea dabbenena are endemic or near-endemic to Gough Island and are likely to be driven to extinction if invasive mice are not removed

Using globally threatened pelagic birds to identify priority sites for marine conservation in the South Atlantic Ocean

The Convention on Biological Diversity aspires to designate 10% of the global oceans as Marine Protected Areas (MPAs), but so far, few MPAs protect pelagic species in the high seas. Transparent scientific approaches are needed to ensure that these encompass areas with high biodiversity value. Here we used the distribution of all globally threatened seabirds breeding in a centrally located archipelago (Tristan da Cunha) to provide guidance on where MPAs could be established in the South Atlantic Ocean. We combined year-round tracking data from six species, and used the systematic conservation-planning tool, 'Zonation', to delineate areas that would protect the largest proportion of each population. The areas used most intensively varied among species and seasons. Combining the sites used by all six species suggested that the most important areas of the South Atlantic are located south of South Africa, around the central South Atlantic between 30 degrees S and 55 degrees S, and near South America. We estimated that the longline fishing effort in these intensively used areas is around 11 million hooks on average each year, highlighting the need for improved monitoring of seabird bycatch rates and the enforcement of compliance with bird bycatch mitigation requirements by fisheries. There was no overlap between the identified areas and any of the existing MPAs in the South Atlantic. The conservation of these highly mobile, pelagic species cannot be achieved by single countries, but requires a multi-national approach at an ocean-basin scale, such as an agreement for the conservation of biodiversity beyond national jurisdiction under the United Nation Convention on the Law of the Sea

Rate of Decline of the Oriental White-Backed Vulture Population in India Estimated from a Survey of Diclofenac Residues in Carcasses of Ungulates

The non-steroidal anti-inflammatory drug diclofenac is a major cause of the rapid declines in the Indian subcontinent of three species of vultures endemic to South Asia. The drug causes kidney failure and death in vultures. Exposure probably arises through vultures feeding on carcasses of domesticated ungulates treated with the drug. However, before the study reported here, it had not been established from field surveys of ungulate carcasses that a sufficient proportion was contaminated to cause the observed declines. We surveyed diclofenac concentrations in samples of liver from carcasses of domesticated ungulates in India in 2004–2005. We estimated the concentration of diclofenac in tissues available to vultures, relative to that in liver, and the proportion of vultures killed after feeding on a carcass with a known level of contamination. We assessed the impact of this mortality on vulture population trend with a population model. We expected levels of diclofenac found in ungulate carcasses in 2004–2005 to cause oriental white-backed vulture population declines of 80–99% per year, depending upon the assumptions used in the model. This compares with an observed rate of decline, from road transect counts, of 48% per year in 2000–2003. The precision of the estimate based upon carcass surveys is low and the two types of estimate were not significantly different. Our analyses indicate that the level of diclofenac contamination found in carcasses of domesticated ungulates in 2004–2005 was sufficient to account for the observed rapid decline of the oriental white-backed vulture in India. The methods we describe could be used again to assess changes in the effect on vulture population trend of diclofenac and similar drugs. In this way, the effectiveness of the recent ban in India on the manufacture and importation of diclofenac for veterinary use could be monitored

Aging is associated with positive responding to neutral information but reduced recovery from negative information

Studies on aging and emotion suggest an increase in reported positive affect, a processing bias of positive over negative information, as well as increasingly adaptive regulation in response to negative events with advancing age. These findings imply that older individuals evaluate information differently, resulting in lowered reactivity to, and/or faster recovery from, negative information, while maintaining more positive responding to positive information. We examined this hypothesis in an ongoing study on Midlife in the US (MIDUS II) where emotional reactivity and recovery were assessed in a large number of respondents (N = 159) from a wide age range (36–84 years). We recorded eye-blink startle magnitudes and corrugator activity during and after the presentation of positive, neutral and negative pictures. The most robust age effect was found in response to neutral stimuli, where increasing age is associated with a decreased corrugator and eyeblink startle response to neutral stimuli. These data suggest that an age-related positivity effect does not essentially alter the response to emotion-laden information, but is reflected in a more positive interpretation of affectively ambiguous information. Furthermore, older women showed reduced corrugator recovery from negative pictures relative to the younger women and men, suggesting that an age-related prioritization of well-being is not necessarily reflected in adaptive regulation of negative affect

Evolutionary factors affecting the cross-species utility of newly developed microsatellite markers in seabirds

Microsatellite loci are ideal for testing hypotheses relating to genetic segregation at fine spatio-temporal scales. They are also conserved among closely related species, making them potentially useful for clarifying interspecific relationships between recently diverged taxa. However, mutations at primer binding sites may lead to increased non-amplification, or disruptions that may lead to decreased polymorphism in non-target species. Furthermore, high mutation rates and constraints on allele size may also lead, with evolutionary time, to an increase in convergently evolved allele size classes, biasing measures of interspecific genetic differentiation. Here, we used next-generation sequencing to develop microsatellite markers from a shotgun genome sequence of the sub-Antarctic seabird, the thin-billed prion (Pachyptila belcheri), that we tested for cross-species amplification in other Pachyptila and related sub-Antarctic species. We found that heterozygosity decreased and the proportion of non-amplifying loci increased with phylogenetic distance from the target species. Surprisingly, we found that species trees estimated from interspecific FST provided better approximations of mtDNA relationships among the studied species than those estimated using DC, even though FST was more affected by null alleles. We observed a significantly non-linear second order polynomial relationship between microsatellite and mtDNA distances. We propose that the loss of linearity with increasing mtDNA distance stems from an increasing proportion of homoplastic allele size classes that are identical in state, but not identical by descent. Therefore, despite high cross-species amplification success and high polymorphism among the closely related Pachyptila species, we caution against the use of microsatellites in phylogenetic inference among distantly related taxa

Pollution, habitat loss, fishing and climate change as critical threats to penguins

Cumulative human impacts across the world’s oceans are considerable. We therefore examined a single model taxonomic group, the penguins (Spheniscidae), to explore how marine species and communities might be at risk of decline or extinction in the southern hemisphere. We sought to determine the most important threats to penguins and to suggest means to mitigate these threats. Our review has relevance to other taxonomic groups in the southern hemisphere and in northern latitudes, where human impacts are greater. Our review was based on an expert assessment and literature review of all 18 penguin species; 49 scientists contributed to the process. For each penguin species, we considered their range and distribution, population trends, and main anthropogenic threats over the past approximately 250 years. These threats were harvesting adults for oil, skin, and feathers and as bait for crab and rock lobster fisheries; harvesting of eggs; terrestrial habitat degradation; marine pollution; fisheries bycatch and resource competition; environmental variability and climate change; and toxic algal poisoning and disease. Habitat loss, pollution, and fishing, all factors humans can readily mitigate, remain the primary threats for penguin species. Their future resilience to further climate change impacts will almost certainly depend on addressing current threats to existing habitat degradation on land and at sea. We suggest protection of breeding habitat, linked to the designation of appropriately scaled marine reserves, including in the High Seas, will be critical for the future conservation of penguins. However, large-scale conservation zones are not always practical or politically feasible and other ecosystem-based management methods that include spatial zoning, bycatch mitigation, and robust harvest control must be developed to maintain marine biodiversity and ensure that ecosystem functioning is maintained across a variety of scales.Los impactos humanos acumulativos a lo largo de los océanos del planeta son considerables. Por eso examinamos un solo modelo de grupo taxonómico, los pingüinos (Sphenischidae), para explorar cómo las especies y las comunidades marinas pueden estar en riesgo de disminuir o de extinguirse en el hemisferio sur. Buscamos determinar la amenaza más importante para los pingüinos y sugerir métodos para mitigar estas amenazas. Nuestra revisión tiene relevancia para otros grupos taxonómicos en el hemisferio sur y en las latitudes norteñas, donde los impactos humanos son mayores. Nuestra revisión se basó en una evaluación experta y una revisión de literaratura de las 18 especies de pingüinos; 49 científicos contribuyeron al proceso. Para cada especie de pingüino, consideramos su rango y distribución, tendencias poblacionales y las principales amenazas antropogénicas en aproximadamente los últimos 250 años. Estas amenazas fueron la captura de adultos para obtener aceite, piel y plumas y el uso como carnada para la pesca de cangrejos y langostas: la recolección de huevos; la degradación del hábitat terrestre; la contaminación marina; la pesca accesoria y la competencia por recursos; la variabilidad ambiental y el cambio climático; y el envenenamiento por algas tóxicas y enfermedades. La pérdida de hábitat, la contaminación y la pesca, todos factores que los humanos pueden mitigar, siguen siendo las amenazas principales para las especies de pingüinos. Su resiliencia futura a más impactos por cambio climático dependerá certeramente de que nos enfoquemos en las amenazas actuales a la degradación de hábitats existentes en tierra y en el mar. Sugerimos que la protección de hábitats de reproducción, en conjunto con la designación de reservas marinas de escala apropiada, incluyendo alta mar, será crítica para la conservación futura de los pingüinos. Sin embargo, las zonas de conservación a gran escala no son siempre prácticas o políticamente viables, y otros métodos de manejo basados en ecosistemas que incluyen la zonificación espacial, la mitigación de captura accesoria, y el control fuerte de captura deben desarrollarse para mantener la biodiversidad marina y asegurar que el funcionamiento de los ecosistemas se mantenga a lo largo de una variedad de escalas.Fil: Trathan, Phil N.. British Antartic Survey; Reino UnidoFil: Garcia Borboroglu, Jorge Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Nacional Patagónico; ArgentinaFil: Boersma, P. Dee. University of Washington; Estados UnidosFil: Bost, Charles André. Centre d´Etudes Biologiques de Chizé; FranciaFil: Crawford, Robert J. M.. Department of Environmental Affairs; SudáfricaFil: Crossin, Glenn T.. Dalhousie University Halifax; CanadáFil: Cuthbert, Richard. Royal Society for the Protection of Birds; Reino UnidoFil: Dann, Peter. Phillip Island Nature Parks; AustraliaFil: Davis, Lloyd Spencer. University Of Otago; Nueva ZelandaFil: de la Puente, Santiago. Universidad Cayetano Heredia; PerúFil: Ellenberg, Ursula. University Of Otago; Nueva ZelandaFil: Lynch, Heather J.. Stony Brook University; Estados UnidosFil: Mattern, Thomas. University Of Otago; Nueva ZelandaFil: Pütz, Klemens. Antarctic Research Trust; AlemaniaFil: Seddon, Philip J.. University Of Otago; Nueva ZelandaFil: Trivelpiece, Wayne. Southwest Fisheries Science Center; Estados UnidosFil: Wienecke, Bárbara. Australian Antarctic Division; Australi

Observed controls on resilience of groundwater to climate variability in sub-Saharan Africa

Groundwater in sub-Saharan Africa supports livelihoods and poverty alleviation1,2, maintains vital ecosystems, and strongly influences terrestrial water and energy budgets. Yet the hydrological processes that govern groundwater recharge and sustainability—and their sensitivity to climatic variability—are poorly constrained4. Given the absence of firm observational constraints, it remains to be seen whether model-based projections of decreased water resources in dry parts of the region4 are justified. Here we show, through analysis of multidecadal groundwater hydrographs across sub-Saharan Africa, that levels of aridity dictate the predominant recharge processes, whereas local hydrogeology influences the type and sensitivity of precipitation–recharge relationships. Recharge in some humid locations varies by as little as five per cent (by coefficient of variation) across a wide range of annual precipitation values. Other regions, by contrast, show roughly linear precipitation–recharge relationships, with precipitation thresholds (of roughly ten millimetres or less per day) governing the initiation of recharge. These thresholds tend to rise as aridity increases, and recharge in drylands is more episodic and increasingly dominated by focused recharge through losses from ephemeral overland flows. Extreme annual recharge is commonly associated with intense rainfall and flooding events, themselves often driven by large-scale climate controls. Intense precipitation, even during years of lower overall precipitation, produces some of the largest years of recharge in some dry subtropical locations. Our results therefore challenge the ‘high certainty’ consensus regarding decreasing water resources in such regions of sub-Saharan Africa. The potential resilience of groundwater to climate variability in many areas that is revealed by these precipitation–recharge relationships is essential for informing reliable predictions of climate-change impacts and adaptation strategies