Population Structure Based on Age-Class Distribution of \u3cem\u3eTapirus polkensis\u3c/em\u3e from the Gray Fossil Site Tennessee.

Individuals of Tapirus polkensis from the Gray Fossil Site exhibit an excellent level of preservation. Intact skulls collected from the site were arranged in a rough age class system separated into 7 categories based on the teeth present and amount of dental wear. Such an eruption series is useful for general comparisons amongst the individual tapirs; however, the classes do not represent an age in years due to a lack of data on living tapirs. Consequently, it is possible that some age classes may contain several years of a tapirs life, or comparatively only a few months. In this study I placed ages on individuals of T. polkensis from the Gray Fossil Site based on age data taken from The Baird\u27s Tapir Project of Costa Rica (Baird\u27s Tapirs, T. bairdii) ranging from several months to 7 years in age. As eruption data is only useful for aging tapirs up to 7 years, this study also took dental wear into account for adult tapirs. After aging all possible individuals in the sample, the sample was compared to other studies on perissodactyl population structure. Interestingly, the T. polkensis sample is remarkably similar to modern populations of Diceros bicornis

Polyampholytes as emerging macromolecular cryoprotectants

Cellular cryopreservation is a platform technology which underpins cell biology, biochemistry, biomaterials, diagnostics, and the cold chain for emerging cell-based therapies. This technique relies on effective methods for banking and shipping to avoid the need for continuous cell culture. The most common method to achieve cryopreservation is to use large volumes of organic solvent cryoprotective agents which can promote either a vitreous (ice free) phase or dehydrate and protect the cells. These methods are very successful but are not perfect: not all cell types can be cryopreserved and recovered, and the cells do not always retain their phenotype and function post-thaw. This Perspective will introduce polyampholytes as emerging macromolecular cryoprotective agents and demonstrate they have the potential to impact a range of fields from cell-based therapies to basic cell biology and may be able to improve, or replace, current solvent-based cryoprotective agents. Polyampholytes have been shown to be remarkable (mammalian cell) cryopreservation enhancers, but their mechanism of action is unclear, which may include membrane protection, solvent replacement, or a yet unknown protective mechanism, but it seems the modulation of ice growth (recrystallization) may only play a minor role in their function, unlike other macromolecular cryoprotectants. This Perspective will discuss their synthesis and summarize the state-of-the-art, including hypotheses of how they function, to introduce this exciting area of biomacromolecular science

Extracellular antifreeze protein significantly enhances the cryopreservation of cell monolayers

The cryopreservation of cells underpins many areas of biotechnology, healthcare, and fundamental science by enabling the banking and distribution of cells. Cryoprotectants are essential to prevent cold-induced damage. Here, we demonstrate that extracellular localization of antifreeze proteins can significantly enhance post-thaw recovery of mammalian cell monolayers cryopreserved using dimethyl sulfoxide, whereas they show less benefit in suspension cryopreservation. A type III antifreeze protein (AFPIII) was used as the macromolecular ice recrystallization inhibitor and its intra/extracellular locations were controlled by using Pep-1, a cell-penetrating peptide. Flow cytometry and confocal microscopy confirmed successful delivery of AFPIII. The presence of extracellular AFPIII dramatically increased post-thaw recovery in a challenging 2-D cell monolayer system using just 0.8 mg·mL–1, from 25% to over 60%, whereas intracellularly delivered AFPIII showed less benefit. Interestingly, the antifreeze protein was less effective when used in suspension cryopreservation of the same cells, suggesting that the cryopreservation format is also crucial. These observations show that, in the discovery of macromolecular cryoprotectants, intracellular delivery of ice recrystallization inhibitors may not be a significant requirement under “slow freezing” conditions, which will help guide the design of new biomaterials, in particular, for cell storage

Engineering cell surfaces by covalent grafting of synthetic polymers to metabolically-labeled glycans

Re-engineering mammalian cell surfaces enables modulation of their phenotype, function, and interactions with external markers and may find application in cell-based therapies. Here we use metabolic glycan labeling to install azido groups onto the cell surface, which can act as anchor points to enable rapid, simple, and robust “click” functionalization by the addition of a polymer bearing orthogonally reactive functionality. Using this strategy, new cell surface functionality was introduced by using telechelic polymers with fluorescence or biotin termini, demonstrating that recruitment of biomacromolecules is possible. This approach may enable the attachment of payloads and modulation of cell function and fate, as well as providing a tool to interface synthetic polymers with biological systems

Insights into the nature of plume-ridge interaction and outflux of H2O from the Galápagos Spreading Centre

The flow of high-temperature and compositionally-enriched material between mantle plumes and nearby spreading centres influences up to 30% of the global mid-ocean ridge system and represents a significant, but currently unconstrained, flux of volatiles out of the mantle. Here we present new analyses of H2O, F, Cl and S in basaltic glass chips from an archetypal region of plume-ridge interaction, the Galápagos Spreading Centre (GSC). Our dataset includes samples from the eastern GSC, on ridge segments that are strongly influenced by the adjacent Galápagos mantle plume, and complements published analyses of volatiles largely from the western GSC. We use forward models of mantle melting to investigate the role of solid and melt-phase transport from a lithologically heterogeneous (peridotite-pyroxenite) mantle in plume-ridge interaction along approximately 1000 km of the GSC. Our results indicate that the observed geochemical and geophysical variations cannot be recreated by models which only involve solid-state transfer of material between the Galápagos mantle plume and the GSC. Instead, we show that the geochemical and geophysical data from the GSC are well-matched by models that incorporate channelised flow of volatile-rich melts formed at high-pressures (>3 GPa) in the Galápagos plume stem to the GSC. In addition, our new models demonstrate that channelised flow of enriched, plume-derived melt can account for up to ~60% of the H2O outgassed from regions of the GSC which are most strongly influenced by the Galápagos mantle plume

A qualitative study of clinicians’ experience of a clinical trial for displaced distal radius fractures

Aims: The aim of this study was to explore clinicians’ experience of a paediatric randomized controlled trial (RCT) comparing surgical reduction with non-surgical casting for displaced distal radius fractures. Methods: Overall, 22 staff from 15 hospitals who participated in the RCT took part in an interview. Interviews were informed by phenomenology and analyzed using thematic analysis. Results: Analysis of the findings identified the overarching theme of “overcoming obstacles”, which described the challenge of alleviating staff concerns about the use of non-surgical casting and recruiting families where there was treatment uncertainty. In order to embed and recruit to the Children’s Radius Acute Fracture Fixation Trial (CRAFFT), staff needed to fit the study within clinical practice, work together, negotiate treatment decisions, and support families. Conclusion: Recruiting families to this RCT was challenging because staff were uncertain about longer-term patient outcomes, and the difficulties were exacerbated by interdisciplinary tensions. Strong family and clinician beliefs, coupled with the complex nature of emergency departments and patient pathways that differed site-by-site, served as barriers to recruitment. Cementing a strong research culture, and exploring families’ treatment preferences, helped to overcome recruitment obstacles

Polyproline is a minimal antifreeze protein mimetic and enhances the cryopreservation of cell monolayers

Tissue engineering, gene therapy, drug screening and emerging regenerative medicine therapies are fundamentally reliant on high-quality adherent cell culture, but current methods to cryopreserve cells in this format can give low cell yields and requires large volumes of solvent 'antifreezes'. Herein we report polyproline is a minimum (bio)synthetic mimic of antifreeze proteins, which is accessible by solution, solid phase and recombinant methods. We demonstrate that polyproline has ice recrystallization inhibition activity linked to its amphipathic helix and that it enhances the DMSO- cryopreservation of adherent cell lines. Polyproline may be a versatile additive in the emerging field of macromolecular cryoprotectants

Pollen derived macromolecules serve as a new class of ice-nucleating cryoprotectants

Cryopreservation of biological material is vital for existing and emerging biomedical and biotechnological research and related applications, but there remain significant challenges. Cryopreservation of cells in sub-milliliter volumes is difficult because they tend to deeply supercool, favoring lethal intracellular ice formation. Some tree pollens are known to produce polysaccharides capable of nucleating ice at warm sub-zero temperatures. Here we demonstrated that aqueous extractions from European hornbeam pollen (pollen washing water, PWW) increased ice nucleation temperatures in 96-well plates from approximate to - 13 degrees C to approximate to - 7 degrees C. Application of PWW to the cryopreservation of immortalized T-cells in 96-well plates resulted in an increase of post-thaw metabolic activity from 63.9% (95% CI [58.5 to 69.2%]) to 97.4% (95% CI [86.5 to 108.2%]) of unfrozen control. When applied to cryopreservation of immortalized lung carcinoma monolayers, PWW dramatically increased post-thaw metabolic activity, from 1.6% (95% CI [- 6.6 to 9.79%]) to 55.0% (95% CI [41.6 to 68.4%]). In contrast to other ice nucleating agents, PWW is soluble, sterile and has low cytotoxicity meaning it can be readily incorporated into existing cryopreservation procedures. As such, it can be regarded as a unique class of cryoprotectant which acts by inducing ice nucleation at warm temperatures

Being recovered: a qualitative study of parents’ experience of their child’s recovery up to a year after a displaced distal radius fracture

Aims The aim of this study was to explore parents’ experience of their child’s recovery, and their thoughts about their decision to enrol their child in a randomized controlled trial (RCT) of surgery versus non-surgical casting for a displaced distal radius fracture. Methods A total of 20 parents of children from 13 hospitals participating in the RCT took part in an interview five to 11 months after injury. Interviews were informed by phenomenology and analyzed using thematic analysis. Results Analysis of the findings identified the theme “being recovered”, which conveyed: 1) parents’ acceptance and belief that their child received the best treatment for them; 2) their memory of the psychological impact of the injury for their child; and 3) their pride in how their child coped with their cast and returned to activities. The process of recovery was underpinned by three elements of experience: accepting the treatment, supporting their child through challenges during recovery, and appreciating their child’s resilience. These findings extend our framework that highlights parents’ desire to protect their child during early recovery from injury, by making the right decision, worrying about recovery, and comforting their child. Conclusion By one year after injury, parents in both treatment groups considered their child “recovered”. They had overcome early concerns about healing, the appearance of the wrist, and coping after cast removal. Greater educational support for families during recovery would enable parents and their child to cope with the uncertainty of recovery, particularly addressing the loss of confidence, worry about reinjury, and the appearance of their wrist