A homozygous genome‐edited Sept2‐EGFP fibroblast cell line

Septins are a conserved, essential family of GTPases that interact with actin, microtubules, and membranes and form scaffolds and diffusion barriers in cells. Several of the 13 known mammalian septins assemble into nonpolar, multimeric complexes that can further polymerize into filamentous structures. While some GFP‐coupled septins have been described, overexpression of GFP‐tagged septins often leads to artifacts in localization and function. To overcome this ubiquitous problem, we have here generated a genome‐edited rat fibroblast cell line expressing Septin 2 (Sept2) coupled to enhanced green fluorescent protein (EGFP) from both chromosomal loci. We characterize these cells by genomic polymerase chain reaction (PCR) for genomic integration, by western blot and reverse transcriptase‐PCR for expression, by immunofluorescence and immunoprecipitation for the colocalization of septins with one another and cellular structures and for complex formation of different septins. By live cell imaging, proliferation and migration assays we investigate proper function of septins in these cells. We find that EGFP is incorporated into both chromosomal loci and only EGFP‐coupled Sept2 is expressed in homozygous cells. We find that endogenous Sept2‐EGFP exhibits expression levels, localization and incorporation into cellular septin complexes similar to the wt in these cells. The expression level of other septins is not perturbed and cell division and cell migration proceed normally. We expect our cell line to be a useful tool for the cell biology of septins, especially for quantitative biology

An evaluation of large carnivore translocations into free-range environments in Namibia

Around the world, large carnivores are involved in human wildlife conflict by killing livestock or compromising peoples’ safety. This results in widespread lethal persecution that contributes to carnivore population declines. Alternatively, translocation of so-called ‘problem animals’ is an often-used approach to resolve conflict non-lethally. However, translocations are rarely assessed in terms of their capacity to reduce conflict or their biological and financial implications. This study evaluates the efficacy of this strategy by investigating 22 intensively monitored translocations that were carried out into free-range environments in Namibia between 2008 and 2012. Translocations involved 23 cheetahs (Acinonyx jubatus) (plus 10 dependent offspring), six leopards (Panthera pardus) and one brown hyaena (Parahyaena brunnea). Translocation objectives included conflict mitigation and the rehabilitation of confiscated, indiscriminately trapped or orphaned individuals. Study animals were released at an average distance of 404.3 km (47 – 816 km) following captive periods ranging from 1 – 1,184 days. Using survival, livestock predation and homing as key measures of translocation success one year post-release, out of 27 individuals with known outcomes, 44.4% were successful. Success was higher for leopards (67%) than for cheetahs (40%), which were particularly unsuccessful if they habituated to human presence during prolonged captivity. Human-induced mortality accounted for most deaths in year one (58%; 10 cheetahs, one leopard, and the hyaena). Translocation success did not differ significantly by sex (M: 39%; F: 50%) or between hard (47%) and soft releases (40%). Regardless of species, release mode and recipient area size, all carnivores displayed extensive post-release movements ranging beyond protected area boundaries, but only two cheetahs returned to their capture site. Most animals that survived the first year successfully reproduced (five leopard cubs, 14 cheetah cubs) and settled into permanent ranges. Following an initial period of orientation and exploration, the ecology of translocated large carnivores reflects that of resident conspecifics. Only three case studies resulted in post-release conflict, but translocations did not resolve conflict on source properties permanently, leading to repeat requests for carnivore removals by those land managers. A farmland survey (26,090 km2, n = 221 respondents) demonstrated that conflict with, and persecution of, large carnivores remain widespread, suggesting a high potential demand for carnivore translocations. However, release area suitability modelling across Namibia’s protected area network showed that only a few public or private reserves can potentially accommodate individual cheetahs and leopards. Translocations are also costly, with a total expense of $80,681 in this study ($269 – $7,559 per individual). The main cost factor was tracking technology (56$5,983 (adjusted median) and 65% of all costs were recuperated from public support. Translocations can successfully conserve individual carnivores and help supplement low-density populations locally. However, due to its limited success, associated costs, and a high degree of variability in terms of outcomes, the strategy is not a feasible standard response to human-carnivore conflict. It is best reserved as a last-resort tool for the selective management of few individuals from highly endangered species. Where it is necessary, rigorous candidate and recipient area selection can improve outcomes significantly. Wildlife managers should predominantly aim at improving tolerance of large carnivores in unprotected, multi-use landscapes, thereby reducing the number of indiscriminately captured animals

Preparation of a porphyrinic bis(pyridyl aldehyde) and its supramolecular complexes

Shape-specific molecular assemblies require the preparation of the constituent building blocks with the necessary properties to bias exclusive formation of the proposed structures. In this work, a novel linear porphyrin dialdehyde was synthesised and used to assemble a supramolecular grid via Cu(I) heteroleptic phenanthroline/pyridyl imine complexation, and a tetrahedral cage via Fe(II) pyridyl imine coordination.We gratefully acknowledge funding from the Consejo Nacional de Ciencia y Tecnologia (CONACYT) Mexico (MAAG) and the Cambridge NanoDTC (CRG).This is the author accepted manuscript. The final version is available from the Royal Society of Chemistry via http://dx.doi.org/10.1039/C5CC06399

In situ impedance measurements on postmortem porcine brain

The paper presents an experimental study where the distinctness of grey and white matter of an in situ postmortem porcine brain by impedance measurements is investigated. Experimental conditions that would allow to conduct the same experiment on in vivo human brain tissue are replicated

Evaluation of a conflict-related brown hyaena translocation in central Namibia

The distribution of the brown hyaena (Hyaena brunnea) in southern Africa overlaps widely with commercial livestock ranching.As a direct result,both perceived and confirmed conflict with farmers occurs and hyaenas are trapped for lethal control or translocation.We studied the outcomes of a conflict-related brown hyaena translocation in Central Namibia involving a subadult female – the first reported GPS-monitored translocation of this species. The animal was moved 63 km from the conflict site and after exploratory movements settled into a new home range incorporating resident conspecifics. The hyaena caused no further conflict and did not return home to its original capture site where livestock depredation ceased. The hyaena was killed in a road accident five months after release. We assess and review our results (and brown hyaena translocations in general) with respect to species ecology, previous translocations as well as monitoring data from resident conspecifics.We provide supporting information that individual hyaenas can be translocated successfully but emphasize that decisions need to be made case-specifically considering the age, sex and social status of the animals. We highlight the importance of brown hyaena sociality when considering translocation as a management tool.Chester Zoo, U.K., and Colchester Zoo, U.K., for funding the GPS satellite transponders. The Brown Hyena Research Project acknowledges support from Namdeb Diamond Corporation and Nedbank’s Go Green Fund.http://www.sawma.co.zaam201

Lions at the Gates: Trans-disciplinary Design of an Early Warning System to Improve Human-Lion Coexistence

Across Africa, lions (Panthera leo) are heavily persecuted in anthropogenic landscapes. Trans-disciplinary research and virtual boundaries (geofences) programmed into GPS-tracking transmitters offer new opportunities to improve coexistence. During a 24-month pilot study (2016–2018), we alerted communities about approaching lions, issuing 1,017 alerts to four villages and 19 cattle posts. Alerts reflected geofence breaches of nine lions (2,941 monitoring days) moving between Botswana's Okavango Delta and adjacent agro-pastoral communities. Daily alert system costs per lion were US$18.54, or$5,460.24 per GPS deployment (n = 13). Alert-responsive livestock owners mainly responded by night-kraaling of cattle (68.9%), significantly reducing their losses (by $124.61 annually), whereas losses of control group and non-responsive livestock owners remained high ($317.93 annually). Community satisfaction with alerts (91.8%) was higher than for compensation of losses (24.3%). Study lions spent 26.3% of time monitored in geofenced community areas, but accounted for 31.0% of conflict. Manual alert distribution proved challenging, static geofences did not appropriately reflect human safety or the environment's strong seasonality that influenced cattle predation risk, and tracking units with on-board alert functions often failed or under-recorded geofence breaches by 27.9%. These insufficiencies prompted the design of a versatile and autonomous lion alert platform with automated, dynamic geofencing. We co-designed this prototype platform with community input, thereby incorporating user feedback. We outline a flexible approach that recognizes conflict complexity and user community heterogeneity. Here, we describe the evolution of an innovative Information and Communication Technologies-based (ICT) alert system that enables instant data processing and community participation through interactive interfaces on different devices. We highlight the importance of a trans-disciplinary co-design and development process focussing on community engagement while synthesizing expertise from ethnography, ecology, and socio-informatics. We discuss the bio-geographic, social, and technological variables that influence alert system efficacy and outline opportunities for wider application in promoting coexistence and conservation