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Rigorous scientific review of research protocols is critical to making funding decisions, and to the protection of both human and non-human research participants. Given the increasing complexity of research designs and data analysis methods, quantitative experts, such as biostatisticians, play an essential role in evaluating the rigor and reproducibility of proposed methods. However, there is a common misconception that a statistician’s input is relevant only to sample size/power and statistical analysis sections of a protocol. The comprehensive nature of a biostatistical review coupled with limited guidance on key components of protocol review motived this work. Members of the Biostatistics, Epidemiology, and Research Design Special Interest Group of the Association for Clinical and Translational Science used a consensus approach to identify the elements of research protocols that a biostatistician should consider in a review, and provide specific guidance on how each element should be reviewed. We present the resulting review framework as an educational tool and guideline for biostatisticians navigating review boards and panels. We briefly describe the approach to developing the framework, and we provide a comprehensive checklist and guidance on review of each protocol element. We posit that the biostatistical reviewer, through their breadth of engagement across multiple disciplines and experience with a range of research designs, can and should contribute significantly beyond review of the statistical analysis plan and sample size justification. Through careful scientific review, we hope to prevent excess resource expenditure and risk to humans and animals on poorly planned studies.
Introduction
Supercooling ability is a critical component among the suite of adaptations contributing to subzero temperature-tolerance of insects, whether they follow freeze-tolerance or freeze-avoidance strategies. Supercooling points (SCP, nucleation temperature, or crystallization temperature) of insects and other terrestrial arthropods range tremendously, from −2 °C to −100 °C or lower. Supercooling is affected by a number of factors, including the volume and water content of the organism, and the ability of the body surface to prevent inoculative freezing by external ice. However, the topics of this review, ice nucleators and antifreeze proteins, are often of critical importance. Antifreezes can be both small-molecular-mass solutes, such as polyhydroxyl alcohols that depress the freezing point of water on a strictly colligative basis, and high-molecular-mass molecules such as antifreeze proteins that suppress freezing by a non-colligative mechanism. Freeze-tolerant species often exhibit high SCPs (above −10 °C) and have selected for extracellular ice nucleators, while freeze-avoiding insects generally have selected against ice nucleators and for antifreezes, allowing them to supercool below ambient temperatures to which they are exposed over the winter. This review will attempt to provide a broad update on ice nucleators, antifreeze proteins and related adaptations in insects and other arthropods, primarily from the standpoint of how they function in organisms to promote winter survival.
Protein ice nucleators
Ice nucleators (INs) limit supercooling by organizing water into an ice-like structure, the embryo crystal, that promotes freezing at a temperature higher than that where ice would otherwise form (Knight, 1967).
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