A modern method of multiple working hypotheses to improve inference in ecology.

Science provides a method to learn about the relationships between observed patterns and the processes that generate them. However, inference can be confounded when an observed pattern cannot be clearly and wholly attributed to a hypothesized process. Over-reliance on traditional single-hypothesis methods (i.e. null hypothesis significance testing) has resulted in replication crises in several disciplines, and ecology exhibits features common to these fields (e.g. low-power study designs, questionable research practices, etc.). Considering multiple working hypotheses in combination with pre-data collection modelling can be an effective means to mitigate many of these problems. We present a framework for explicitly modelling systems in which relevant processes are commonly omitted, overlooked or not considered and provide a formal workflow for a pre-data collection analysis of multiple candidate hypotheses. We advocate for and suggest ways that pre-data collection modelling can be combined with consideration of multiple working hypotheses to improve the efficiency and accuracy of research in ecology.

Lessons and recommendations from three decades as an NSF REU site: A call for systems-based assessment.

For more than 30 years, the US National Science Foundation's Research Experiences for Undergraduates (REU) program has supported thousands of undergraduate researchers annually and provides many students with their first research experiences in field ecology or evolution. REUs embed students in scientific communities where they apprentice with experienced researchers, build networks with their peers, and help students understand research cultures and how to work within them. REUs are thought to provide formative experiences for developing researchers that differ from experiences in a college classrooms, laboratories, or field trips. REU assessments have improved through time but they are largely ungrounded in educational theory. Thus, evaluation of long-term impacts of REUs remains limited and best practices for using REUs to enhance student learning are repeatedly re-invented. We describe how one sociocultural learning framework, cultural-historical activity theory (CHAT), could be used to guide data collection to characterize the effects of REU programs on participant's learning in an educationally meaningful context. CHAT embodies a systems approach to assessment that accounts for social and cultural factors that influence learning. We illustrate how CHAT has guided assessment of the Harvard Forest Summer Research Program in Ecology (HF-SRPE), one of the longest-running REU sites in the United States. Characterizing HF-SRPE using CHAT helped formalize thoughts and language for the program evaluation, reflect on potential barriers to success, identify assessment priorities, and revealed important oversights in data collection.

Identification of proteins associated with adhesive prints from Holothuria dofleinii Cuvierian tubules.

Cuvierian tubules are expelled as a defence mechanism against predators by various species within the family Holothuridae. When the tubules are expelled, they become sticky almost immediately and ensnare the predator. The mechanism of this rapid adhesion is not clear, but proteins on the surface of the expelled tubules are widely believed to be involved. This study has examined such proteins from Holothuria dofleinii, sourced from adhesive prints left on glass after the removal of adhered tubules. Gel electrophoresis showed that seven strongly staining protein bands were consistently present in all samples, with molecular masses ranging from 89 to 17 kDa. N-terminal sequence data was obtained from two bands, while others seemed blocked. Tandem mass spectrometry-based sequencing of tryptic peptides derived from individual protein bands indicated that the proteins were unlikely to be homopolymers. PCR primers designed using the peptide sequences enabled us to amplify, clone and sequence cDNA segments relating to four gel bands; for each, the predicted translation product contained other peptide sequences observed for that band that had not been used in primer design. Database searches using the peptide and cDNA-encoded sequences suggest that two of the seven proteins are novel and one is a C-type lectin, while-surprisingly-at least three of the other four are closely related to enzymes associated with the pentose phosphate cycle and glycolysis. We discuss precedents in which lectins and metabolic enzymes are involved in attachment and adhesion phenomena.

Characterization of a new Bacillus thuringiensis endotoxin, Cry47Aa, from strains that are toxic to the Australian sheep blowfly, Lucilia cuprina.

Sixteen isolates of Bacillus thuringiensis, derived from various soil samples collected in Australia, are highly toxic to larvae of the sheep blowfly (Lucilia cuprina). The toxin gene from one of the strains (CAA890) was cloned by genome walking, and sequencing of the cloned fragments revealed a new cry gene, encoding a protein of 1134 amino acid residues, with a theoretical molecular mass of 139,209Da. Based on the amino acid sequence comparison with known Cry delta-endotoxins, the gene was designated cry47Aa. Homology modelling based on known crystal structures of the Cry toxins reveals the differences to be located in the loops of domain II in the putative toxin-receptor binding surfaces between Cry47Aa and the dipteran active Cry2Aa. We also showed that the cry47Aa gene is present in the other isolates that are highly toxic to the sheep blowfly.

Increased site 1 affinity improves biopotency of porcine growth hormone. Evidence against diffusion dependent receptor dimerization.

Based on phage display optimization studies with human growth hormone (GH), it is thought that the biopotency of GH cannot be increased. This is proposed to be a result of the affinity of the first receptor for hormone far exceeding that which is required to trap the hormone long enough to allow diffusion of the second receptor to form the ternary complex, which initiates signaling. We report here that despite similar site 1 kinetics to the hGH/hGH receptor interaction, the potency of porcine GH for its receptor can be increased up to 5-fold by substituting hGH residues involved in site 1 binding into pGH. Based on extensive mutations and BIAcore studies, we show that the higher potency and site 1 affinity of hGH for the pGHR is primarily a result of a decreased off-rate associated with residues in the extended loop between helices 1 and 2 that interact with the two key tryptophans Trp104 and Trp169 in the receptor binding hot spot. Our mutagenic analysis has also identified a second determinant (Lys165), which in addition to His169, restricts the ability of non-primate hormones to activate hGH receptor. The increased biopotency of GH that we observe can be explained by a model for GH receptor activation where subunit alignment is critical for effective signaling.