Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study.

Microscale thermophoresis (MST), and the closely related Temperature Related Intensity Change (TRIC), are synonyms for a recently developed measurement technique in the field of biophysics to quantify biomolecular interactions, using the (capillary-based) NanoTemper Monolith and (multiwell plate-based) Dianthus instruments. Although this technique has been extensively used within the scientific community due to its low sample consumption, ease of use, and ubiquitous applicability, MST/TRIC has not enjoyed the unambiguous acceptance from biophysicists afforded to other biophysical techniques like isothermal titration calorimetry (ITC) or surface plasmon resonance (SPR). This might be attributed to several facts, e.g., that various (not fully understood) effects are contributing to the signal, that the technique is licensed to only a single instrument developer, NanoTemper Technology, and that its reliability and reproducibility have never been tested independently and systematically. Thus, a working group of ARBRE-MOBIEU has set up a benchmark study on MST/TRIC to assess this technique as a method to characterize biomolecular interactions. Here we present the results of this study involving 32 scientific groups within Europe and two groups from the US, carrying out experiments on 40 Monolith instruments, employing a standard operation procedure and centrally prepared samples. A protein-small molecule interaction, a newly developed protein-protein interaction system and a pure dye were used as test systems. We characterized the instrument properties and evaluated instrument performance, reproducibility, the effect of different analysis tools, the influence of the experimenter during data analysis, and thus the overall reliability of this method.

International society of sports nutrition position stand: ketogenic diets.

POSITION STATEMENT: The International Society of Sports Nutrition (ISSN) provides an objective and critical review of the use of a ketogenic diet in healthy exercising adults, with a focus on exercise performance and body composition. However, this review does not address the use of exogenous ketone supplements. The following points summarize the position of the ISSN.1. A ketogenic diet induces a state of nutritional ketosis, which is generally defined as serum ketone levels above 0.5 mM. While many factors can impact what amount of daily carbohydrate intake will result in these levels, a broad guideline is a daily dietary carbohydrate intake of less than 50 grams per day.2. Nutritional ketosis achieved through carbohydrate restriction and a high dietary fat intake is not intrinsically harmful and should not be confused with ketoacidosis, a life-threatening condition most commonly seen in clinical populations and metabolic dysregulation.3. A ketogenic diet has largely neutral or detrimental effects on athletic performance compared to a diet higher in carbohydrates and lower in fat, despite achieving significantly elevated levels of fat oxidation during exercise (~1.5 g/min).4. The endurance effects of a ketogenic diet may be influenced by both training status and duration of the dietary intervention, but further research is necessary to elucidate these possibilities. All studies involving elite athletes showed a performance decrement from a ketogenic diet, all lasting six weeks or less. Of the two studies lasting more than six weeks, only one reported a statistically significant benefit of a ketogenic diet.5. A ketogenic diet tends to have similar effects on maximal strength or strength gains from a resistance training program compared to a diet higher in carbohydrates. However, a minority of studies show superior effects of non-ketogenic comparators.6. When compared to a diet higher in carbohydrates and lower in fat, a ketogenic diet may cause greater losses in body weight, fat mass, and fat-free mass, but may also heighten losses of lean tissue. However, this is likely due to differences in calorie and protein intake, as well as shifts in fluid balance.7. There is insufficient evidence to determine if a ketogenic diet affects males and females differently. However, there is a strong mechanistic basis for sex differences to exist in response to a ketogenic diet.

Targeting protein-protein interactions and fragment-based drug discovery.

Protein-protein interactions (PPI) are integral to the majority of biological functions. Targeting these interactions with small molecule inhibitors is of increased interest both in academia as well as in the pharmaceutical industry, both for therapeutic purposes and in the search for chemical tools for basic science. Although the number of well-characterised examples is still relatively modest, it is becoming apparent that many different kinds of interactions can be inhibited using drug-like small molecules. Compared to active site targeting, PPI inhibition suffers from the particular problem of more exposed and less defined binding sites, and this imposes significant experimental challenges to the development of PPI inhibitors. PPI interfaces are large, up to thousands of square angstroms, and there is still debate as to what part of the interface one should target. We will review recent developments in the field of PPI inhibition, with emphasis on fragment-based methods, and discuss various factors one should take into account when developing small molecule inhibitors targeted at PPI interfaces.

Indoor Air Quality in Passivhaus Dwellings: A Literature Review.

Indoor air quality (IAQ) is a critical consideration in airtight buildings that depend on mechanical ventilation, such as those constructed to the Passivhaus standard. While previous reviews of IAQ on Passivhaus-certified buildings foccused on offices, this study examines residential buildings. A summary of data collection methods and pollutant concentrations is presented, followed by a critical discussion of the impact of Passivhaus design strategies on IAQ. This review indicates that IAQ in Passivhaus-certified dwellings is generally better than in conventional homes, but both occupant behaviour and pollution from outdoor sources play a significant role in indoor concentrations. Moreover, there are differences in data collection and reporting methods. Many of the available studies depend on short-term IAQ monitoring of less than two weeks, making it difficult to determine the longer impact of housing design on IAQ and occupants' well-being. There is also a lack of studies from non-European countries. Future research should focus on investigating associations between IAQ and Passivhaus design strategies in hot and humid climates, where evidence is particularly lacking. Further effort is also required to investigate potential links between occupant's perception of IAQ and physical exposure to indoor pollution. Finally, the lack of homogeneous monitoring and reporting methods for IAQ studies needs to be addressed.

The role of the turn in beta-hairpin formation during WW domain folding.

The folding of WW domains is rate limited by formation of a beta-hairpin comprising residues from strands 1 and 2. Residues in the turn of this hairpin have reported Phi-values for folding close to 1 and have been proposed to nucleate folding. High Phi-values do not necessarily imply that the energetics of formation are a driving force for initiating folding. We demonstrate by NMR studies and molecular dynamics simulations that the first turn of the hYAP, FBP28, and PIN1 WW domains is structurally dynamic and solvent exposed in the native and folding transition states. It is, therefore, unlikely that the formation of the beta-turn per se provides the energetic driving force for hairpin folding. It is more likely that the turn acts as an easily formed hinge that facilitates the formation of the hairpin; it is a nucleus as defined by the nucleation-condensation mechanism whereby a diffuse nucleus is stabilized by associated interactions.

Occupant Interactions and Effectiveness of Natural Ventilation Strategies in Contemporary New Housing in Scotland, UK.

The need to reduce carbon emissions and fuel poverty has led to increased building envelope air tightness, intended to reduce uncontrolled ventilation heat losses. Ventilation strategies in dwellings still allow the use of trickle ventilators in window frames for background ventilation. The extent to which this results in "healthy" Indoor Air Quality (IAQ) in recently constructed dwellings was a concern of regulators in Scotland. This paper describes research to explore this. First a review of literature was conducted, then data on occupant interactions with ventilation provisions (windows, doors, trickle vents) gathered through an interview-based survey of 200 recently constructed dwellings, and measurements made on a sample of 40 of these. The main measured parameter discussed here is CO2 concentration. It was concluded after the literature review that 1000 ppm absolute was a reasonable threshold to use for "adequate" ventilation. The occupant survey found that there was very little occupant interaction with the trickle ventilators e.g., in bedrooms 63% were always closed, 28% always open, and in only 9% of cases occupants intervened to make occasional adjustments. In the measured dwellings average bedroom CO2 levels of 1520 ppm during occupied (night time) hours were observed. Where windows were open the average bedroom CO2 levels were 972 ppm. With windows closed, the combination of "trickle ventilators open plus doors open" gave an average of 1021 ppm. "Trickle ventilators open" gave an average of 1571 ppm. All other combinations gave averages of 1550 to 2000 ppm. Ventilation rates and air change rates were estimated from measured CO2 levels, for all dwellings calculated ventilation rate was less than 8 L/s/p, in 42% of cases calculated air change rate was less than 0.5 ach. It was concluded that trickle ventilation as installed and used is ineffective in meeting desired ventilation rates, evidenced by high CO2 levels reported across the sampled dwellings. Potential implications of the results are discussed.