Breaching the kinetic barrier to in vitro somatic stem cell propagation

ABSTRACT: Here we have reviewed the conventional definitions and fundamental characteristics of the two basic types of stem cells, embryonic stem cells (ESCs) and somatic stem cells (SSCs). By taking into account the often-overlooked asymmetric cell kinetics of SSCs, we consider the evidence that should SSCs retain these growth kinetics in vitro, a natural kinetic barrier to SSC propagation exists. Recent discoveries showing that the tumor suppressor gene p53 can act as a regulator of asymmetric cell kinetics provide a target pathway for in vitro SSC propagation strategies

CFP and YFP, but Not GFP, Provide Stable Fluorescent Marking of Rat Hepatic Adult Stem Cells

The stable expression of reporter genes in adult stem cells (ASCs) has important applications in stem cell biology. The ability to integrate a noncytotoxic, fluorescent reporter gene into the genome of ASCs with the capability to track ASCs and their progeny is particularly desirable for transplantation studies. The use of fluorescent proteins has greatly aided the investigations of protein and cell function on short-time scales. In contrast, the obtainment of stably expressing cell strains with low variability in expression for studies on longer-time scales is often problematic. We show that this difficulty is partly due to the cytotoxicity of a commonly used reporter, green fluorescent protein (GFP). To avoid GFP-specific toxicity effects during attempts to stably mark a rat hepatic ASC strain and, therefore, obtain stable, long-term fluorescent ASCs, we evaluated cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP), in addition to GFP. Although we were unable to derive stable GFP-expressing strains, stable fluorescent clones (up to 140 doublings) expressing either CFP or YFP were established. When fluorescently marked ASCs were induced to produce differentiated progeny cells, stable fluorescence expression was maintained. This property is essential for studies that track fluorescently marked ASCs and their differentiated progeny in transplantation studies

A non-invasive approach to estimate the energetic requirements of an increasing seabird population in a perturbed marine ecosystem

This is the author accepted manuscript. The final version is available from Springer Nature via the DOI in this record.There is a growing desire to integrate the food requirements of predators living in marine ecosystems impacted by humans into sustainable fisheries management. We used non- invasive video-recording, photography and focal observations to build time-energy budget models and to directly estimate the fish mass delivered to chicks by adult greater crested terns Thalasseus bergii breeding in the Benguela ecosystem. Mean modelled adult daily food intake increased from 140.9 g·d−1 of anchovy Engraulis capensis during incubation to 171.7g·d−1 and 189.2 g·d−1 when provisioning small and large chicks, respectively. Modelled prey intake expected to be returned to chicks was 58.3 g·d−1 (95% credible intervals: 44.9–75.8 g·d−1) over the entire growth period. Based on our observations, chicks were fed 19.9 g·d−1 (17.2–23.0 g·d−1) to 45.1 g·d−1 (34.6–58.7 g·d−1 25 ) of anchovy during early and late provisioning, respectively. Greater crested terns have lower energetic requirements at the individual (range: 15–34%) and population level (range: 1–7%) than the other Benguela endemic seabirds that feed on forage fish. These modest requirements – based on a small body size and low flight costs – coupled with foraging plasticity have allowed greater crested terns to cope with changing prey availability, unlike the other seabirds species using the same exploited prey base.Our research was supported by a Department of Science and Technology-National Research Foundation Centre of Excellence grant to the FitzPatrick Institute of African Ornithology, the Leiden Conservation Foundation (RBS) and our institutes. Robben Island Museum provided logistical support and access to the tern colonies

The costs of kleptoparasitism: a study of mixed-species seabird breeding colonies

This is the author accepted manuscript. The final version is available from OUP via the DOI in this record.Mixed-species assemblages are common in nature, providing mutual benefits to associating species including anti-predator advantages or resource facilitation. However, associating with other species may also impose costs through kleptoparasitism (food theft). Identification of these costs, and how they vary when different species breed alongside one another, is essential to understand the payoffs of mixed-species assemblages. We explore the costs of kleptoparasitism for greater crested terns Thalasseus bergii provisioning offspring at a single-species colony, where individuals suffer kleptoparasitism from conspecifics, and at a mixed colony where terns breed alongside Hartlaub’s gulls Chroicocephalus hartlaubii and are vulnerable to both intra and interspecific kleptoparasitism. Gull presence likely contributes to increases in both kleptoparasitic attacks and the proportion of prey lost or stolen during provisioning, relative to the single-species colony. Provisioning adults suffered additional energetic costs in response to gull kleptoparasitism, requiring more attempts to deliver prey, taking longer to do so, and swallowing more prey (to the detriment of their offspring). Gulls also appear to increase the duration of tern vulnerability to kleptoparasitism, because they continued to steal food from adults and chicks after precocial chicks left the nest, when intraspecific kleptoparasitism is negligible. Terns breeding in a mixed colony, therefore, suffer direct and indirect costs through decreased provisioning and increased provisioning effort, which may ultimately affect reproductive success, resulting in colony decline where kleptoparasitism is frequent. This study illustrates how forming a mixed-species seabird breeding assemblage has costs as well as benefits, potentially fluctuating between a parasitic and a mutualistic relationship.This work was supported by a Department of Science and TechnologyCentre of Excellence grant to the FitzPatrick Institute of African Ornithology. R.B.S.  was supported by a fellowship from the Leiden Conservation Foundation. This research was approved by SANParks (CONM1182), the Department of Environmental Affairs (RES2013/24, RES2014/83, and RES2015/65) and the animal ethics committee of the University of Cape Town (2013/V3/TC)

EVALUASI PELAKSANAAN PROGRAM NASIONAL AGRARIA DAN PENDAFTARAN TANAH SISTEMATIS LENGKAP DALAM PEMBANGUNAN WILAYAH DESA KINABUHUTAN, KECAMATAN LIKUPANG BARAT, KABUPATEN MINAHASA UTARA

This study aims to evaluate the implementation of : (1) the national agrarian program certification (PRONA), and (2) complete systematic land registration (PTSL). The research used descriptive qualitative approach. This research was conducted in Kinabuhutan Village, West Likupang Sub-district, North Minahasa District, North Sulawesi Province, that conducted from June 2017 to January 2018. The sample consisted of 23 respondents for PRONA and 21 respondents for PTSL. Sample determination was done by using purposive sampling technique. Data collection methods used are Observation, Interview and Documentation. Data analysis technique used was descriptive qualitative. The results showed that: (1) Implementation of land certification through PRONA program has been run in accordance with the stages of activities determined according to regulations. However, there are problems in the implementation of: (i) proof of ownership (ownership rights) whether in the form of ownership based on sale and purchase or grant or inheritance, (ii) land tenure status as cultivator causing not knowing land history from first holder right until last owner and sign the upper limit of their land which is the main requirement in the management of the certificate. (2) The implementation of a Complete Systematic Soil System (PTSL) program is in accordance with the stages regulated. There are, however, shortcomings in which members of the community who have selected to participate in the PTSL program cannot show evidence for to proof of ownership, at the time of measurement there are not available at the site,and there is also no way of indicating the boundaries of the land because it has no boundary markers. The problem encountered in the implementation of both programs is the limited number of officers who have the competence as land measurement officers

Bottom-up effects of a no-take zone on endangered penguin demographics.

This is the author accepted manuscript. The final version is available from The Royal Society via the DOI in this record.Marine no-take zones can have positive impacts for target species and are increasingly important management tools. However, whether they indirectly benefit higher order predators remains unclear. The endangered African penguin (Spheniscus demersus) depends on commercially exploited forage fish. We examined how chick survival responded to an experimental 3-year fishery closure around Robben Island, South Africa, controlling for variation in prey biomass and fishery catches. Chick survival increased by 18% when the closure was initiated, which alone led to a predicted 27% higher population compared with continued fishing. However, the modelled population continued to decline, probably because of high adult mortality linked to poor prey availability over larger spatial scales. Our results illustrate that small no-take zones can have bottom-up benefits for highly mobile marine predators, but are only one component of holistic, ecosystem-based management regimes.We thank the Earthwatch Institute, Bristol Zoological Society, Leiden Conservation Foundation and National Research Foundation

Individuality, kin similarity and experimental playback of contact calls in cooperatively breeding riflemen

Riflemen/tītipounamu (Acanthisitta chloris) are kin-based cooperatively breeding birds, which appear able to recognise their relatives. Here, we investigate the potential for vocalisations to act as recognition cues in riflemen. We identified an appropriate contact call and recorded it at the nest from 19 adult riflemen. Measurements of call characteristics were individually repeatable. In addition, call similarity was significantly correlated with relatedness among all birds and among males. Thus, in principle, these contact calls contain sufficient information for individual recognition of familiar kin, and some assessment of relatedness between unfamiliar birds. To test whether riflemen responded differently to calls of kin, we broadcast calls of relatives and non-relatives as separate treatments in a playback experiment. Focal birds rarely responded aggressively or affiliatively, and their tendency to do so was unrelated to treatment. We conclude that zip calls are suitable kin recognition cues, but whether they are used as such remains unknown

Vulnerability of northern gannets to offshore wind farms; seasonal and sex-specific collision risk and demographic consequences

There is a pressing need to quantify the risks of renewable energy developments such as offshore wind farms for protected populations. However, assessments are often based on incomplete data, or fail to consider variation in risk between sexes and at different times of year. We tracked northern gannets foraging from the world's largest colony (Bass Rock, Scotland) across five consecutive breeding seasons. We examine how seasonal and sex differences in behaviour affect the collision risk from planned and operational wind farms within their foraging range and assess the likely consequences for long-term population viability. Both sexes made shorter trips during chick-rearing than prior to chick-hatching, spent a greater proportion of time within wind farm sites and had an eight times greater potential collision risk during chick-rearing. Females made longer trips than males at both these times of year, flew higher and spent more time within wind farm sites, leading to three times greater collision risk for females. After accounting for the potential additional mortality from collisions, and assuming that the death of a parent also led to the loss of its offspring, the breeding population was projected to increase by 3.57% (95% CI: 2.16–5.15%) per year, compared with 6.56% (95% CI: 4.59–8.73%) in the absence of turbines, suggesting a negligible effect on population viability. However, additional mortality could result in greater immigration from neighbouring colonies, potentially affecting their viability and highlighting a need for research within a metapopulation framework to assess the impacts of offshore wind developments on vulnerable species across multiple connected sites

A Resource for Discovering Specific and Universal Biomarkers for Distributed Stem Cells

Specific and universal biomarkers for distributed stem cells (DSCs) have been elusive. A major barrier to discovery of such ideal DSC biomarkers is difficulty in obtaining DSCs in sufficient quantity and purity. To solve this problem, we used cell lines genetically engineered for conditional asymmetric self-renewal, the defining DSC property. In gene microarray analyses, we identified 85 genes whose expression is tightly asymmetric self-renewal associated (ASRA). The ASRA gene signature prescribed DSCs to undergo asymmetric self-renewal to a greater extent than committed progenitor cells, embryonic stem cells, or induced pluripotent stem cells. This delineation has several significant implications. These include: 1) providing experimental evidence that DSCs in vivo undergo asymmetric self-renewal as individual cells; 2) providing an explanation why earlier attempts to define a common gene expression signature for DSCs were unsuccessful; and 3) predicting that some ASRA proteins may be ideal biomarkers for DSCs. Indeed, two ASRA proteins, CXCR6 and BTG2, and two other related self-renewal pattern associated (SRPA) proteins identified in this gene resource, LGR5 and H2A.Z, display unique asymmetric patterns of expression that have a high potential for universal and specific DSC identification