Intrinsic Ball Retrieving in Wolf Puppies Suggests Standing Ancestral Variation for Human-Directed Play Behavior.
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ABSTRACT:
SUBMITTER: Hansen Wheat C
PROVIDER: S-EPMC7033638 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Ancestral sequence reconstruction and resurrection provides useful information for protein engineering, yet its alliance with directed evolution has been little explored. In this study, we have resurrected several ancestral nodes of fungal laccases dating back ?500 to 250 million years. Unlike modern laccases, the resurrected Mesozoic laccases were readily secreted by yeast, with similar kinetic parameters, a broader stability, and distinct pH activity profiles. The resurrected Agaricomycetes laccase carried 136 ancestral mutations, a molecular testimony to its origin, and it was subjected to directed evolution in order to improve the rate of 1,3-cyclopentanedione oxidation, a ?-diketone initiator commonly used in vinyl polymerization reactions.IMPORTANCE The broad variety of biotechnological uses of fungal laccases is beyond doubt (food, textiles, pulp and paper, pharma, biofuels, cosmetics, and bioremediation), and protein engineering (in particular, directed evolution) has become the key driver for adaptation of these enzymes to harsh industrial conditions. Usually, the first requirement for directed laccase evolution is heterologous expression, which presents an important hurdle and often a time-consuming process. In this work, we resurrected a fungal Mesozoic laccase node which showed strikingly high heterologous expression and pH stability. As a proof of concept that the ancestral laccase is a suitable blueprint for engineering, we performed a quick directed evolution campaign geared to the oxidation of the ?-diketone 1,3-cyclopentanedione, a poor laccase substrate that is used in the polymerization of vinyl monomers.
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Project description:BackgroundGroin pain is a very common injury in multidirectional sports such as soccer, ice hockey, rugby and Australian football. Long-standing adductor-related groin pain is a persistent clinical condition and a frequent complaint in athletes involved in sports that require multiplanar movement patterns (change of direction, high-speed sprinting and kicking). To date, the lack of rehabilitation guidelines and return-to-play criteria makes this clinical entity difficult to manage. The aim of the present Delphi was to suggest, based on opinion and practical experience of a panel of experts, potential criteria that could be taken into consideration by clinicians in the RTP decision-making process in athletes suffering from long-standing adductor-related groin pain.MethodsThirty two out of 40 experts participated to a 3-Round Delphi questionnaire. In round 1, open-ended and closed questions about 9 different sections (palpation, flexibility, strength, patient-reported outcome measures, imaging, intersegmental control, performance tests, sport-specific skills, training load) were proposed to investigate return to play evaluation criteria used by each expert. Responses were analysed and coded to produce round 2 questionnaire that investigated only the sections and the items that reached the cut-off value (≥ 70%). Round 3 questionnaire was based on sections and items that reached cut-off value in previous rounds and experts rated their agreement for return to play criteria with a 5-point Likert Scale. Descriptive statistics enabled interpretation of consensus.ResultsHigh participation rate (80%) and response rate across the 3 rounds (100%) were recorded. 6 sections reached positive consensus in round 1, 1 section reached negative consensus. In round 2 positive consensus was confirmed only for 3 sections and negative consensus for 1 section. In round 3, positive agreement was established for strength (3 items), performance tests (3 items) and sport-specific skills (2 items) sections. Negative consensus was confirmed for imaging section.ConclusionExperts agreed that strength, performance tests and sport-specific skills can be used to support RTP decision, while imaging cannot be used. These findings could be useful in assisting clinicians in the RTP decision making.
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