Microplastics affect membrane biofouling and microbial communities during gravity-driven membrane filtration of primary wastewater.

Recently, gravity-driven membrane (GDM) filtration has been adopted as an alternative solution for decentralized wastewater treatment due to easy installation and maintenance, reduced energy and operation cost, and low global warming impact. This study investigated the influence of microplastic size (0.5-0.8 µm and 40-48 µm) and amount (0.1 and 0.2 g/L) on the membrane performance and microbial community in GDM systems for primary municipal wastewater treatment. The results showed that dosing microplastics in the GDM systems led to 9-54% lower permeate flux than that in the control. This was attributed to more cake formation (up to 6.4-fold) with more deposition of extracellular polymeric substances (EPS, up to 1.5-fold) and divalent cations (up to 2.1-fold) in the presence of microplastics, especially with increasing microplastic amount or size. However, the dosed microplastics promoted formation of heterogeneous cake layers with more porous nature, possibly because microplastics created void space in the cake and also tended to bind with divalent cations to reduce EPS-divalent cations interactions. In the biofilm of the GDM systems, the presence of microplastics could lower the number of total species, but it greatly enhanced the abundance of certain dominant prokaryotes (Phenylobacterium haematophilum, Planctomycetota bacterium, and Flavobacteriales bacterium), eukaryotes (Stylonychia lemnae, Halteria grandinella, and Paramicrosporidium saccamoebae), and virus (phylum Nucleocytoviricota), as well as amino acid and lipid metabolic functions. Especially, the small-size microplastics at a higher dosed amount led to more variations of microbial community structure and microbial metabolic functions.

Perceptions and biomechanical effects of varying prosthetic ankle stiffness during uphill walking: A case series.

BACKGROUND: Prosthetic foot stiffness, which is typically invariable for commercially available prosthetic feet, needs to be considered when prescribing a prosthetic foot. While a biological foot adapts its function according to the movement task, an individual with lower limb amputation may be limited during more functionally demanding gait tasks by their conventional energy storing and return prosthetic foot. RESEARCH QUESTION: How do changes in prosthetic foot stiffness during incline walking affect biomechanical measures as well as perception of participants. METHODS: Kinetic and kinematic data were collected during incline walking, for five participants with trans-tibial amputation. A mixed model analysis of variance was used to analyse the effects of changing the stiffness during incline walking, using a novel variable-stiffness unit built on a commercially available prosthetic foot. Biomechanical results were also analysed on an individual level alongside the participant feedback, for a better understanding of the various strategies and perceptions exhibited during incline walking. RESULTS: Statistically significant effects were only observed on the biomechanical parameters directly related to prosthetic ankle kinematics and kinetics (i.e., peak prosthetic ankle dorsiflexion, peak prosthetic ankle power, dynamic joint stiffness during controlled dorsiflexion). Participant perception during walking was affected by changes in stiffness. Individual analyses revealed varied perceptions and varied biomechanical responses among participants. SIGNIFICANCE: While changes in prosthesis mechanical properties influenced the amputee's experience, minimal immediate effects were found with the overall gait pattern. The reported inter-participant variability may be due to the person's physical characteristics or habitual gait pattern, which may influence prosthesis function. The ability to vary prosthetic foot stiffness during the assessment phase of setting up a prosthesis could provide useful information to guide selection of the appropriate prosthetic device for acceptable performance across a range of activities.

Developing an Artificial Intelligence-Based Representation of a Virtual Patient Model for Real-Time Diagnosis of Acute Respiratory Distress Syndrome.

Acute Respiratory Distress Syndrome (ARDS) is a condition that endangers the lives of many Intensive Care Unit patients through gradual reduction of lung function. Due to its heterogeneity, this condition has been difficult to diagnose and treat, although it has been the subject of continuous research, leading to the development of several tools for modeling disease progression on the one hand, and guidelines for diagnosis on the other, mainly the "Berlin Definition". This paper describes the development of a deep learning-based surrogate model of one such tool for modeling ARDS onset in a virtual patient: the Nottingham Physiology Simulator. The model-development process takes advantage of current machine learning and data-analysis techniques, as well as efficient hyperparameter-tuning methods, within a high-performance computing-enabled data science platform. The lightweight models developed through this process present comparable accuracy to the original simulator (per-parameter R2 > 0.90). The experimental process described herein serves as a proof of concept for the rapid development and dissemination of specialised diagnosis support systems based on pre-existing generalised mechanistic models, making use of supercomputing infrastructure for the development and testing processes and supported by open-source software for streamlined implementation in clinical routines.

Analysis of Chest X-ray for COVID-19 Diagnosis as a Use Case for an HPC-Enabled Data Analysis and Machine Learning Platform for Medical Diagnosis Support.

The COVID-19 pandemic shed light on the need for quick diagnosis tools in healthcare, leading to the development of several algorithmic models for disease detection. Though these models are relatively easy to build, their training requires a lot of data, storage, and resources, which may not be available for use by medical institutions or could be beyond the skillset of the people who most need these tools. This paper describes a data analysis and machine learning platform that takes advantage of high-performance computing infrastructure for medical diagnosis support applications. This platform is validated by re-training a previously published deep learning model (COVID-Net) on new data, where it is shown that the performance of the model is improved through large-scale hyperparameter optimisation that uncovered optimal training parameter combinations. The per-class accuracy of the model, especially for COVID-19 and pneumonia, is higher when using the tuned hyperparameters (healthy: 96.5%; pneumonia: 61.5%; COVID-19: 78.9%) as opposed to parameters chosen through traditional methods (healthy: 93.6%; pneumonia: 46.1%; COVID-19: 76.3%). Furthermore, training speed-up analysis shows a major decrease in training time as resources increase, from 207 min using 1 node to 54 min when distributed over 32 nodes, but highlights the presence of a cut-off point where the communication overhead begins to affect performance. The developed platform is intended to provide the medical field with a technical environment for developing novel portable artificial-intelligence-based tools for diagnosis support.

Biocarriers facilitated gravity-driven membrane filtration of domestic wastewater in cold climate: Combined effect of temperature and periodic cleaning.

In this study, two lava stone biocarrier facilitated gravity-driven membrane (GDM) reactors were operated at ~8 °C and ~22 °C in parallel for treating primary wastewater effluent. Although the biocarrier reactor at 8 °C displayed less efficient removals of biodegradable organics than that at 22 °C, both GDM systems (without cleaning) showed comparable fouling resistance distribution patterns, accompanying with similar cake filtration constants and pore constriction constants by modelling simulation. Compared to the GDM at 8 °C, more foulants were accumulated on the GDM at 22 °C, but they presented similar soluble organics/inorganics contents and specific cake resistances. This indicated the cake layers at 22 °C may contain greater-sized foulants due to proliferation of both prokaryotes and eukaryotes, leading to a relatively less-porous nature. In the presence of periodic cleaning (at 50 °C), the cleaning effectiveness followed a sequence as ultrasonication-enhanced physical cleaning > two-phase flow cleaning > chemical-enhanced physical cleaning > physical cleaning, regardless of GDM operation temperature. However, significantly higher cake resistances were observed in the GDM system at 22 °C than those at 8 °C, because shear force tended to remove loosely-attached foulant layers and may compress the residual dense cake layer. The presence of periodic cleaning led to dissimilar dominant prokaryotic and eukaryotic communities in the cake layers as those without cleaning and in the lava stone biocarriers. Nevertheless, operation temperature did not influence GDM permeate quality, which met EU discharge standards.

Task dependent changes in mechanical and biomechanical measures result from manipulating stiffness settings in a prosthetic foot.

BACKGROUND: Adaptation of lower limb function to different gait tasks is inherently not as effective among individuals with lower limb amputation as compared to able-bodied individuals. Varying stiffness of a prosthetic foot may be a way of facilitating gait tasks that require larger ankle joint range of motion. METHODS: Three stiffness settings of a novel prosthetic foot design were tested for level walking at three speeds as well as for 7,5° incline and decline walking. Outcome measures, describing ankle range of motion and ankle dynamic joint stiffness were contrasted across the three stiffness settings. Standardized mechanical tests were done for the hindfoot and forefoot. FINDINGS: Dorsiflexion angle was incrementally increased with a softer foot and a faster walking speed / higher degree of slope. The concurrent dynamic joint stiffness exhibited a less systematic change, especially during INCLINE and DECLINE walking. The small difference seen between the stiffness settings for hindfoot loading limits analysis for the effects of stiffness during weight acceptance, however, a stiffer foot significantly restricted plantarflexion during DECLINE. INTERPRETATIONS: Varying stiffness settings within a prosthetic foot does have an effect on prosthetic foot dynamics, and differences are task dependent, specifically in parameters involving kinetic attributes. When considering the need for increased ankle range of motion while performing more demanding gait tasks, a foot that allows the users themselves to adjust stiffness according to the task at hand may be of benefit for active individuals, possibly enhancing the user's satisfaction and comfort during various daily activities.

Variable stiffness foot design and validation.

Energy storing and returning prosthetic feet are commonly prescribed. Research has demonstrated advantages to use these types of prosthetic feet. However, their stiffness in the sagittal plane is fixed and cannot adapt to different walking tasks and user preference. In this paper, we propose a novel prosthetic foot design capable of modulating its stiffness in the sagittal plane. The Variable Stiffness Ankle unit (VSA) is mounted on a commercially available prosthetic foot. The stiffness of the foot is adjusted with a lightweight servo motor controlled wirelessly. The stiffness change is accomplished by moving the supports points on the glass fiber leaf spring of the VSA ankle unit. We described the design and characterized changes in ankle stiffness using a mechanical test bench. A novel method was used to capture mechanical test data using a six degree of freedom load cell, allowing us to contrast mechanical and biomechanical data. A transtibial unilateral amputee performed level ground walking on an instrumented treadmill. The VSA prosthetic foot exhibited ankle stiffness change in the mechanical test bench. Ankle stiffness changes were also confirmed during the biomechanical analysis. Future work will involve additional subjects. The VSA prosthetic foot could improve user satisfaction and help prosthetist to fine tune prosthetic feet during fittings.

GPCR Genes as Activators of Surface Colonization Pathways in a Model Marine Diatom.

Surface colonization allows diatoms, a dominant group of phytoplankton in oceans, to adapt to harsh marine environments while mediating biofoulings to human-made underwater facilities. The regulatory pathways underlying diatom surface colonization, which involves morphotype switching in some species, remain mostly unknown. Here, we describe the identification of 61 signaling genes, including G-protein-coupled receptors (GPCRs) and protein kinases, which are differentially regulated during surface colonization in the model diatom species, Phaeodactylum tricornutum. We show that the transformation of P. tricornutum with constructs expressing individual GPCR genes induces cells to adopt the surface colonization morphology. P. tricornutum cells transformed to express GPCR1A display 30% more resistance to UV light exposure than their non-biofouling wild-type counterparts, consistent with increased silicification of cell walls associated with the oval biofouling morphotype. Our results provide a mechanistic definition of morphological shifts during surface colonization and identify candidate target proteins for the screening of eco-friendly, anti-biofouling molecules.

Functional joint center of prosthetic feet during level ground and incline walking.

Energy storage and returning prosthetic feet do not provide a well-defined articulation point compared to the human ankle. Calculation of user relevant parameters, such as ankle power, requires such a joint center point when using traditional mechanical models. However, shortcomings of current calculation methods result in some errors. The aim of this case study was to compare conventional ankle joint calculations to a functional joint center (FJC) using data collected on a roll-over test machine and in a motion lab during dissimilar walking tasks. Three prosthetic feet were evaluated on a roll-over test machine. Then, two trans-tibial amputees were each fitted with the same three prosthetic feet matching their weight and activity category. Kinematic data were collected during walking on level ground, as well as up and down a slope. The FJC during the stance phase of gait was calculated for each test method and compared with outcomes using conventional methods. The location of the FJC was generally anterior and inferior to the estimated anatomical joint position. Importantly, the FJC location varied for the different prosthetic feet and was task dependent as per the three gait conditions. This was reflected in different ankle angles and moments of FJC calculations compared to conventional methods for level ground walking. Differences in the calculated FJC between conditions represented the variations in prosthetic foot deformation, and explained how this parameter is influenced by the prosthetic's stiffness. For level ground walking, calculated FJC location between human subject testing and machine evaluation were strongly correlated. Both stiffness and task dependent demands of the prosthetic foot should be considered during testing. The FJC of elastic ankles can serve as a parameter for characterization and differentiation between various prosthetic foot designs and be an important parameter for prosthetic foot designers to consider. As the position of the FJC is dependent on the design and task, it is a more informative measure of the prosthetic foot's response to the user's needs. Furthermore, prosthetists could use this metric in clinical practice to better appreciate amputee feedback and perception. FJC provides an alternative center during calculation of ankle power using standard methods.

Combined artificial high-silicate medium and LED illumination promote carotenoid accumulation in the marine diatom Phaeodactylum tricornutum.

BACKGROUND: Diatoms, which can accumulate large amounts of carotenoids, are a major group of microalgae and the dominant primary producer in marine environments. Phaeodactylum tricornutum, a model diatom species, acquires little silicon for its growth although silicon is known to contribute to gene regulation and play an important role in diatom intracellular metabolism. In this study, we explored the effects of artificial high-silicate medium (i.e. 3.0 mM sodium metasilicate) and LED illumination conditions on the growth rate and pigment accumulation in P. tricornutum, which is the only known species so far that can grow without silicate. It's well known that light-emitting diodes (LEDs) as novel illuminants are emerging to be superior monochromatic light sources for algal cultivation with defined and efficient red and blue lights. RESULTS: Firstly, we cultivated P. tricornutum in a synthetic medium supplemented with either 0.3 mM or 3.0 mM silicate. The morphology and size of diatom cells were examined: the proportion of the oval and triradiate cells decreased while the fusiform cells increased with more silicate addition in high-silicate medium; the average length of fusiform cells also slightly changed from 14.33 µm in 0.3 mM silicate medium to 12.20 µm in 3.0 mM silicate medium. Then we cultivated P. tricornutum under various intensities of red light in combination with the two different levels of silicate in the medium. Higher biomass productivity also achieved in 3.0 mM silicate medium than in 0.3 mM silicate medium under red LED light irradiation at 128 µmol/m2/s or higher light intensity. Increasing silicate reversed the down-regulation of fucoxanthin and chlorophyll a under high red-light illumination (i.e. 255 µmol/m2/s). When doubling the light intensity, fucoxanthin content decreased under red light but increased under combined red and blue (50:50) lights while chlorophyll a content reduced under both conditions. Fucoxanthin accumulation and biomass productivity increased with enhanced red and blue (50:50) lights. CONCLUSION: High-silicate medium and blue light increased biomass and fucoxanthin production in P. tricornutum under high light conditions and this strategy may be beneficial for large-scale production of fucoxanthin in diatoms.

Sugar-stimulated CO2 sequestration by the green microalga Chlorella vulgaris.

To convert waste CO2 from flue gases of power plants into value-added products, bio-mitigation technologies show promise. In this study, we cultivated a fast-growing species of green microalgae, Chlorella vulgaris, in different sizes of photobioreactors (PBRs) and developed a strategy using small doses of sugars for enhancing CO2 sequestration under light-emitting diode illumination. Glucose supplementation at low levels resulted in an increase of photoautotrophic growth-driven biomass generation as well as CO2 capture by 10% and its enhancement corresponded to an increase of supplied photon flux. The utilization of urea instead of nitrate as the sole nitrogen source increased photoautotrophic growth by 14%, but change of nitrogen source didn't compromise glucose-induced enhancement of photoautotrophic growth. The optimized biomass productivity achieved was 30.4% higher than the initial productivity of purely photoautotrophic culture. The major pigments in the obtained algal biomass were found comparable to its photoautotrophic counterpart and a high neutral lipids productivity of 516.6 mg/(L·day) was achieved after optimization. A techno-economic model was also developed, indicating that LED-based PBRs represent a feasible strategy for converting CO2 into value-added algal biomass.

Dataset on economic analysis of mass production of algae in LED-based photobioreactors.

The data presented in this article are related to the research article entitled "Sugar-stimulated CO2 sequestration by the green microalga Chlorella vulgaris" (Fu et al., 2019) [1]. The data describe a rational design and scale-up of LED-based photobioreactors for producing value-added algal biomass while removing waste CO2 from flu gases from power plants. The dataset were created from growth rate experiments for biomass production including direct biomass productivity data, PBR size and setup parameters, medium composition as well as indirect energy cost and overhead in Iceland. A complete economic analysis is formed through a cost breakdown as well as PBR scalability predictions.

Chemical Mutagenesis and Fluorescence-Based High-Throughput Screening for Enhanced Accumulation of Carotenoids in a Model Marine Diatom Phaeodactylum tricornutum.

Diatoms are a major group of unicellular algae that are rich in lipids and carotenoids. However, sustained research efforts are needed to improve the strain performance for high product yields towards commercialization. In this study, we generated a number of mutants of the model diatom Phaeodactylum tricornutum, a cosmopolitan species that has also been found in Nordic region, using the chemical mutagens ethyl methanesulfonate (EMS) and N-methyl-N'-nitro-N-nitrosoguanidine (NTG). We found that both chlorophyll a and neutral lipids had a significant correlation with carotenoid content and these correlations were better during exponential growth than in the stationary growth phase. Then, we studied P. tricornutum common metabolic pathways and analyzed correlated enzymatic reactions between fucoxanthin synthesis and pigmentation or lipid metabolism through a genome-scale metabolic model. The integration of the computational results with liquid chromatography-mass spectrometry data revealed key compounds underlying the correlative metabolic pathways. Approximately 1000 strains were screened using fluorescence-based high-throughput method and five mutants selected had 33% or higher total carotenoids than the wild type, in which four strains remained stable in the long term and the top mutant exhibited an increase of 69.3% in fucoxanthin content compared to the wild type. The platform described in this study may be applied to the screening of other high performing diatom strains for industrial applications.

Adaptive Laboratory Evolution for Enhanced Carotenoid Production in Microalgae.

In order to produce natural pigments with competitive prices, algal strains employed in industrial production need to be improved for increasing the productivity of valuable metabolites, thereby reducing the overall production cost. Adaptive laboratory evolution (ALE) is a traditional method for strain improvement, which has been effectively utilized in bacteria and fungi. With the growing interest in algal biotechnology, attempts have recently been put forward to improve microalgal strains with ALE approach. This chapter describes a stepwise adaptive evolution strategy that enhances carotenoid yield from microalgae.

Quantitative time-course metabolomics in human red blood cells reveal the temperature dependence of human metabolic networks.

The temperature dependence of biological processes has been studied at the levels of individual biochemical reactions and organism physiology (e.g. basal metabolic rates) but has not been examined at the metabolic network level. Here, we used a systems biology approach to characterize the temperature dependence of the human red blood cell (RBC) metabolic network between 4 and 37 °C through absolutely quantified exo- and endometabolomics data. We used an Arrhenius-type model (Q10) to describe how the rate of a biochemical process changes with every 10 °C change in temperature. Multivariate statistical analysis of the metabolomics data revealed that the same metabolic network-level trends previously reported for RBCs at 4 °C were conserved but accelerated with increasing temperature. We calculated a median Q10 coefficient of 2.89 ± 1.03, within the expected range of 2-3 for biological processes, for 48 individual metabolite concentrations. We then integrated these metabolomics measurements into a cell-scale metabolic model to study pathway usage, calculating a median Q10 coefficient of 2.73 ± 0.75 for 35 reaction fluxes. The relative fluxes through glycolysis and nucleotide metabolism pathways were consistent across the studied temperature range despite the non-uniform distributions of Q10 coefficients of individual metabolites and reaction fluxes. Together, these results indicate that the rate of change of network-level responses to temperature differences in RBC metabolism is consistent between 4 and 37 °C. More broadly, we provide a baseline characterization of a biochemical network given no transcriptional or translational regulation that can be used to explore the temperature dependence of metabolism.

Mannose and fructose metabolism in red blood cells during cold storage in SAGM.

BACKGROUND: Alternate sugar metabolism during red blood cell (RBC) storage is not well understood. Here we report fructose and mannose metabolism in RBCs during cold storage in SAGM and the impact that these monosaccharides have on metabolic biomarkers of RBC storage lesion. STUDY DESIGN AND METHODS: RBCs were stored in SAGM containing uniformly labeled 13 C-fructose or 13 C-mannose at 9 or 18 mmol/L concentration for 25 days. RBCs and media were sampled at 14 time points during storage and analyzed using ultraperformance liquid chromatography-mass spectrometry. Blood banking quality assurance measurements were performed. RESULTS: Red blood cells incorporated fructose and mannose during cold storage in the presence of glucose. Mannose was metabolized in preference to glucose via glycolysis. Fructose lowered adenosine triphosphate (ATP) levels and contributed little to ATP maintenance when added to SAGM. Both monosaccharides form the advanced glycation end product glycerate. Mannose activates enzymes in the RBC that take part in glycan synthesis. CONCLUSIONS: Fructose or mannose addition to RBC SAGM concentrates may not offset the shift in metabolism of RBCs that occurs after 10 days of storage. Fructose and mannose metabolism at 4°C in SAGM reflects their metabolism at physiologic temperature. Glycerate excretion is a measure of protein deglycosylation activity in stored RBCs. No cytoprotective effect was observed upon the addition of either fructose or mannose to SAGM.

ReconMap: an interactive visualization of human metabolism.

Motivation: A genome-scale reconstruction of human metabolism, Recon 2, is available but no interface exists to interactively visualize its content integrated with omics data and simulation results. Results: We manually drew a comprehensive map, ReconMap 2.0, that is consistent with the content of Recon 2. We present it within a web interface that allows content query, visualization of custom datasets and submission of feedback to manual curators. Availability and Implementation: ReconMap can be accessed via http://vmh.uni.lu , with network export in a Systems Biology Graphical Notation compliant format released under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. A Constraint-Based Reconstruction and Analysis (COBRA) Toolbox extension to interact with ReconMap is available via https://github.com/opencobra/cobratoolbox . Contact: ronan.mt.fleming@gmail.com.

Photo-Oxidative Stress-Driven Mutagenesis and Adaptive Evolution on the Marine Diatom Phaeodactylum tricornutum for Enhanced Carotenoid Accumulation.

Marine diatoms have recently gained much attention as they are expected to be a promising resource for sustainable production of bioactive compounds such as carotenoids and biofuels as a future clean energy solution. To develop photosynthetic cell factories, it is important to improve diatoms for value-added products. In this study, we utilized UVC radiation to induce mutations in the marine diatom Phaeodactylum tricornutum and screened strains with enhanced accumulation of neutral lipids and carotenoids. Adaptive laboratory evolution (ALE) was also used in parallel to develop altered phenotypic and biological functions in P. tricornutum and it was reported for the first time that ALE was successfully applied on diatoms for the enhancement of growth performance and productivity of value-added carotenoids to date. Liquid chromatography-mass spectrometry (LC-MS) was utilized to study the composition of major pigments in the wild type P. tricornutum, UV mutants and ALE strains. UVC radiated strains exhibited higher accumulation of fucoxanthin as well as neutral lipids compared to their wild type counterpart. In addition to UV mutagenesis, P. tricornutum strains developed by ALE also yielded enhanced biomass production and fucoxanthin accumulation under combined red and blue light. In short, both UV mutagenesis and ALE appeared as an effective approach to developing desired phenotypes in the marine diatoms via electromagnetic radiation-induced oxidative stress.

Developing diatoms for value-added products: challenges and opportunities.

As a major primary producer in marine environments, diatoms have been considered as promising feedstocks for their applications in functional foods, bioactive pharmaceuticals, and cosmetics. This review focuses on the biotechnology potential of diatoms for value-added products like carotenoids. The impact of abiotic environmental stresses, such as intensity and quality of incident light, nutrient deficiency and silicon depletion, on diatoms has been examined to determine key factors that affect the growth performance and the accumulation of valuable compounds. Previous studies suggested that adaptive evolution could be an efficient method to improve the diatom productivity of valuable compounds. Light emitting diode (LED)-based photobioreactors were introduced and proposed as a promising new technology for producing quality products from diatoms. Currently available molecular biology tools were also summarized and discussed in relation to their application in the production of carotenoids and other valuable products. Taken together, systems biology and synthetic biology approaches have the potential to address the challenges faced while working toward the industrial application of diatoms.

Metabolomic analysis of platelets during storage: a comparison between apheresis- and buffy coat-derived platelet concentrates.

BACKGROUND: Platelet concentrates (PCs) can be prepared using three methods: platelet (PLT)-rich plasma, apheresis, and buffy coat. The aim of this study was to obtain a comprehensive data set that describes metabolism of buffy coat-derived PLTs during storage and to compare it with a previously published parallel data set obtained for apheresis-derived PLTs. STUDY DESIGN AND METHODS: During storage we measured more than 150 variables in 8 PLT units, prepared by the buffy coat method. Samples were collected at seven different time points resulting in a data set containing more than 8000 measurements. This data set was obtained by combining a series of standard quality control assays to monitor the quality of stored PLTs and a deep coverage metabolomics study using liquid chromatography coupled with mass spectrometry. RESULTS: Stored PLTs showed a distinct metabolic transition occurring 4 days after their collection. The transition was evident in PLT produced by both production methods. Apheresis-derived PLTs showed a clearer phenotype of PLT activation during early days of storage. The activated phenotype of apheresis PLTs was accompanied by a higher metabolic activity, especially related to glycolysis and the tricarboxylic acid cycle. Moreover, the extent of the activation differed between bags resulting in interbag variability in the storage lesion of apheresis-prepared PLTs. This may be related to donor-related polymorphism. CONCLUSION: This study demonstrated two discrete metabolic phenotypes in stored PLTs prepared with both apheresis and buffy coat methods. PLT activation occurs during the first metabolic phenotype and might lead to a low reproducibility of the apheresis PCs.