Toxicity Mechanisms of Nanoplastics on Crop Growth, Interference of Phyllosphere Microbes, and Evidence for Foliar Penetration and Translocation.

Despite the increasing prevalence of atmospheric nanoplastics (NPs), there remains limited research on their phytotoxicity, foliar absorption, and translocation in plants. In this study, we aimed to fill this knowledge gap by investigating the physiological effects of tomato leaves exposed to differently charged NPs and foliar absorption and translocation of NPs. We found that positively charged NPs caused more pronounced physiological effects, including growth inhibition, increased antioxidant enzyme activity, and altered gene expression and metabolite composition and even significantly changed the structure and composition of the phyllosphere microbial community. Also, differently charged NPs exhibited differential foliar absorption and translocation, with the positively charged NPs penetrating more into the leaves and dispersing uniformly within the mesophyll cells. Additionally, NPs absorbed by the leaves were able to translocate to the roots. These findings provide important insights into the interactions between atmospheric NPs and crop plants and demonstrate that NPs' accumulation in crops could negatively impact agricultural production and food safety.

Performance and micro-pacing in sprint cross-country skiing: A comparison of individual time-trial and head-to-head race formats.

This study compared performance strategies and sub-technique selection in cross-country skate skiing sprint races, specifically individual time-trial (ITT) and head-to-head (H2H) formats. Fourteen male cross-country skiers from the Chinese national team participated in the FIS-sanctioned sprint race day. GNSS and heart rate sensors recorded positioning, skiing speeds, heart rate, sub-technique usage, and skiing kinematics. Statistical parametric mapping (SPM) was used to determine the course positions (clusters) where instantaneous skiing speed was significantly associated with section time. One-way analyses of variance were used to examine differences between the ITT and H2H. H2H race speeds were 2.4 ± 0.2% faster than the ITT race (p < 0.05).Variations in sub-technique and skiing kinematics were observed between race formats, indicating different strategies and tactics employed by athletes. SPM identified specific clusters (primarily uphill) where the fastest athlete gained significant time over the slowest. The greatest time gains were associated with higher G3 sub-technique usage and longer G3 cycle length on steep uphill terrain (9-13% gradients). Integrating SPM analyses and sub-technique assessments can help optimise performance and tactics in sprint races. This study enhances our understanding of cross-country skiing dynamics and performance variations among elite competitors.

Phytotoxicity of polystyrene, polyethylene and polypropylene microplastics on tomato (Lycopersicon esculentum L.).

Despite the fact that microplastic pollution in terrestrial ecosystems has received increasing attention, there are few studies on the potential effects of different microplastics on terrestrial plants. In this study, the toxicity of polystyrene (PS), polyethylene (PE) and polypropylene (PP) microplastics with different concentrations (0, 10, 100, 500 and 1000 mg/L) to tomato (Lycopersicon esculentum L.) were studied by a hydroponic experiment. The results showed that the three microplastics had inhibitory effects on seed germination when the concentration was less than or equal to 500 mg/L, and the inhibition rate ranged from 10.1% to 23.6%. Interestingly, the inhibition effect was alleviated under 1000 mg/L microplastic treatment. Generally, PE was more toxic to seedling growth than PS and PP. Additionally, it was confirmed that microplastics could cause oxidative stress in plants, and PP was relatively less toxic to antioxidant enzymes than PS and PE. These results can provide a theoretical basis and data support for further investigation on the toxicity of microplastics to tomatoes, and contribute to understanding the type specificity of microplastics' toxic effects on plants.

Ultrathin Bimetal Color Filter Based on Plasmonic Nanostructure.

Plasmonic subtractive color filters through a nanostructured ultrathin Ag film have attracted intensive attention due to their good durability, high color tunability and high transmission. However, Ag film suffers from discontinuity when the thickness is below 15 nm, which limits the further increasement of transmission efficiency. Herein a bimetal ultrathin (~10 nm) subtractive color filter with one dimensional nanogratings was demonstrated and fabricated. By adding an embedded Al layer to suppress the formation of Ag islands, a smooth, continuous and reliable bimetal film was obtained. At the same time, the blue shift of transmission minimum was beneficial to overcome difficulty in nanostructure fabrication. This method also provided a new approach to tune the color by simply varying the thickness of Al layer. A broad palette of colors, including cyan, magenta and yellow, was attained in bimetal color filter with high transmission beyond 80%.

Electrospun N-Doped Hierarchical Porous Carbon Nanofiber with Improved Degree of Graphitization for High-Performance Lithium Ion Capacitor.

The lithium-ion capacitor (LIC) has been regarded as a promising device that combines the merits of lithium-ion batteries and supercapacitors, and that meets the requirements for both high energy and high power density. The development of advanced electrode materials is the key requirement. Herein, we report the bottom-up synthesis of activated carbon nanofiber (a-PANF) with a hierarchical porous structure and a high degree of graphitization. Electrospinning has been employed to prepare an interconnected fiber network with macropores, and ferric acetylacetonate has been introduced as both a mesopore-creating agent and a graphitic catalyst to increase the degree of graphitization. Furthermore, chemical activation enlarges the specific surface area by producing abundant micropores. Half-cell evaluation of the as-prepared a-PANF gave a discharge capacity of 80 mA h g-1 at 0.1 A g-1 within 2-4.5 V and no capacity fading after 1000 cycles at 2 A g-1 , which represents a significant improvement compared to conventional activated carbon (AC). Furthermore, an as-assembled LIC with a-PANF cathode and Fe3 O4 anode showed a superior energy density of 124.6 W h kg-1 at a specific power of 93.8 W kg-1 , which remained at 103.7 W h kg-1 at 4687.5 W kg-1 . This indicates promising application potential of a-PANF as an electrode material for efficient energy storage systems.

Effects of polystyrene, polyethylene, and polypropylene microplastics on the soil-rhizosphere-plant system: Phytotoxicity, enzyme activity, and microbial community.

The widespread presence of soil microplastics (MPs) has become a global environmental problem. MPs of different properties (i.e., types, sizes, and concentrations) are present in the environment, while studies about the impact of MPs having different properties are limited. Thus, this study investigated the effects of three common polymers (polystyrene, polyethylene, and polypropylene) with two concentrations (0.01% and 0.1% w/w) on growth and stress response of lettuce (Lactuca sativa L.), soil enzymes, and rhizosphere microbial community. Lettuce growth was inhibited under MPs treatments. Moreover, the antioxidant system, metabolism composition, and phyllosphere microbiome of lettuce leaves was also perturbed. MPs reduced phytase activity and significantly increased dehydrogenase activity. The diversity and structure of rhizosphere microbial community were disturbed by MPs and more sensitive to polystyrene microplastics (PSMPs) and polypropylene microplastics (PPMPs). In general, the results by partial least squares pathway models (PLS-PMs) showed that the presence of MPs influenced the soil-rhizosphere-plant system, which may have essential implications for assessing the environmental risk of MPs.

Mussel inspired 3D elastomer enabled rapid calvarial bone regeneration through recruiting more osteoprogenitors from the dura mater.

Currently, the successful healing of critical-sized calvarial bone defects remains a considerable challenge. The immune response plays a key role in regulating bone regeneration after material grafting. Previous studies mainly focused on the relationship between macrophages and bone marrow mesenchymal stem cells (BMSCs), while dural cells were recently found to play a vital role in the calvarial bone healing. In this study, a series of 3D elastomers with different proportions of polycaprolactone (PCL) and poly(glycerol sebacate) (PGS) were fabricated, which were further supplemented with polydopamine (PDA) coating. The physicochemical properties of the PCL/PGS and PCL/PGS/PDA grafts were measured, and then they were implanted as filling materials for 8 mm calvarial bone defects. The results showed that a matched and effective PDA interface formed on a well-proportioned elastomer, which effectively modulated the polarization of M2 macrophages and promoted the recruitment of dural cells to achieve full-thickness bone repair through both intramembranous and endochondral ossification. Single-cell RNA sequencing analysis revealed the predominance of dural cells during bone healing and their close relationship with macrophages. The findings illustrated that the crosstalk between dural cells and macrophages determined the vertical full-thickness bone repair for the first time, which may be the new target for designing bone grafts for calvarial bone healing.

Earthworms improve the rhizosphere micro-environment to mitigate the toxicity of microplastics to tomato (Solanum lycopersicum).

Microplastics (MPs) are widespread in agricultural soil, potentially threatening soil environmental quality and plant growth. However, toxicological research on MPs has mainly been limited to individual components (such as plants, microbes, and animals), without considering their interactions. Here, we examined earthworm-mediated effects on tomato growth and the rhizosphere micro-environment under MPs contamination. Earthworms (Eisenia fetida) mitigated the growth-inhibiting effect of MPs on tomato plant. Particularly, when exposed to environmentally relevant concentrations (ERC, 0.02% w/w) of MPs, the addition of earthworms significantly (p < 0.05) increased shoot and root dry weight by 12-13% and 13-14%, respectively. MPs significantly reduced (p < 0.05) soil ammonium (NH4+-N) (0.55-0.69 mg/kg), nitrate nitrogen (NO3--N) (7.02-8.65 mg/kg) contents, and N cycle related enzyme activities (33.47-42.39 µg/h/g) by 37.7-50.9%, 22.6-37.2%, and 34.2-48.0%, respectively, while earthworms significantly enhanced (p < 0.05) inorganic N mineralization and bioavailability. Furthermore, earthworms increased bacterial network complexity, thereby enhancing the robustness of the bacterial system to resist soil MPs stress. Meanwhile, partial least squares modelling showed that earthworms significantly influenced (p < 0.01) soil nutrients, which in turn significantly affected (p < 0.01) plant growth. Therefore, the comprehensive consideration of soil ecological composition is important for assessing MPs ecological risk.

Type-dependent effects of microplastics on tomato (Lycopersicon esculentum L.): Focus on root exudates and metabolic reprogramming.

Much attention has been paid to the prevalence of microplastics (MPs) in terrestrial systems. MPs have been shown to affect the physio-biochemical properties of plants. Different MPs may have distinctive behaviors and diverse effects on plant growth. In the present study, the effects of polystyrene (PS), polyethylene (PE), and polypropylene (PP) MPs on physio-biochemical properties, root exudates, and metabolomics of tomato (Lycopersicon esculentum L.) under hydroponic conditions were investigated. Our results show that MPs exposure has adverse effects on tomato growth. MPs exposure had a significant type-dependent effect (p < 0.001) on photosynthetic gas parameters, chlorophyll content, and antioxidant enzyme activities. After exposure to MPs, the content of low molecular weight organic acids in tomato root exudates was significantly increased, which was considered as a strategy to alleviate the toxicity of MPs. In addition, MPs treatment significantly changed the metabolites of tomato root and leaf. Metabolic pathway analysis showed that MPs treatment had a great effect on amino acid metabolism. We also found that plants exposed to PS and PP MPs produced more significant metabolic reprogramming than those exposed to PE MPs. This study provides important implications for the mechanism studies on the toxic effect of various MPs on crops and their future risk assessment.

Single and combined toxicity of polystyrene nanoplastics and arsenic on submerged plant Myriophyllum verticillatum L.

The contamination of nanoplastics (NPs) and heavy metals (HM) in water bodies has caused widespread concern, while their effects on submerged plants are poorly reported. Polystyrene nanoplastics (PSNPs) and arsenic (As) were used to assess their toxicity on Myriophyllum verticillatum L. via the orthogonal experiments. PSNPs significantly reduced the accumulation of As (17.24%-66.67%) in plant. Single As and high As-PSNPs treatments significantly inhibited plant growth, with a maximum reduction of 70.09% in the growth rate. The mineral nutrient content was significantly affected by PSNPs and As treatments. The antioxidant system was significantly inhibited, which was more pronounced in the roots. Similar findings were observed for soluble protein and soluble sugar. Some organic acids and amino acids showed down-regulation at high concentrations of As, leading to a decrease in the content of the mineral element and down-regulation of antioxidant enzyme synthesis. Furthermore, PSNPs could alleviate As toxicity under 0.1 mg/L As treatment but exacerbate As toxicity at 1 mg/L As dose. This study has important implications for the study of submerged plants exposed to co-contamination of microplastics and heavy metals, as well as the possible ecological risk assessment in freshwater.

Toxicity effects of polystyrene nanoplastics and arsenite on Microcystis aeruginosa.

Despite the increasing research on the fate of nanoplastics (NPs, <100 nm) in freshwater systems, little is known about the joint toxic effects of metal(loid)s and NPs modified with different functional groups on microalgae. Here, we explored the joint toxic effects of two types of polystyrene NPs [one modified with a sulfonic acid group (PSNPs-SO3H), and one without this functional group (PSNPs)] and arsenic (As) on the microalgae Microcystis aeruginosa. The results highlighted that PSNPs-SO3H showed a smaller hydrodynamic diameter and greater potential to adsorb positively charged ions than PSNPs, contributing to the more severe growth inhibition, while both of them produced oxidative stress. Metabolomics further revealed that the fatty acid metabolism of the microalgae was significantly up-regulated under both NPs exposure, while PSNPs-SO3H down-regulated the tricarboxylic acid cycle (TCA cycle) of the microalgae. As uptake by algae was significantly reduced by 82.58 % and 59.65 % in the presence of 100 mg/L PSNPs and PSNPs-SO3H, respectively. The independent action model showed that the joint toxicity of both NPs with As was assessed as antagonistic. In addition, PSNPs and PSNPs-SO3H had dissimilar effects on the composition of the microalgae extracellular polymeric substances (EPS), resulting in different uptake and adsorption of As, thereby affecting the physiology and biochemistry of algae. Overall, our findings propose that the specific properties of NPs should be considered in future environmental risk assessments.

Single and combined effects of polystyrene nanoplastics and Cd on submerged plants Ceratophyllum demersum L.

Nanoplastics (NPs) and heavy metals are widely distributed in aquatic ecosystem, posing a potential threat to ecosystem function. Submerged macrophytes play an important role in water purification and maintaining ecological functions. However, the coupled effects of NPs and cadmium (Cd) on submerged macrophytes physiology and the mechanisms involved are still unclear. Here, the potential effects of single and co-Cd/PSNPs exposure on Ceratophyllum demersum L. (C. demersum) were explored. Our results showed that NPs aggravated the inhibition of Cd on plant growth ate (a decrease of 35.54 %), reduced chlorophyll synthesis (a decrease of 15.84 %), and disrupted the antioxidant enzyme system (a decrease of 25.07 % on SOD activity) of C. demersum. Massive PSNPs adhered to the surface of C. demersum when exposed to co-Cd/PSNPs while they did not adhere when exposed to single-NPs. The metabolic analysis further demonstrated that co-exposure down-regulated plant cuticle synthesis and that Cd exacerbated the physical damage and shadowing effects of NPs. In addition, co-exposure upregulated pentose phosphate metabolism, leading to the accumulation of starch grains. Furthermore, PSNPs reduced Cd enrichment capacity of C. demersum. Our results unraveled distinct regulatory networks for submerged macrophytes exposed to single and composite of Cd and PSNPs, providing a new theoretical basis for assessing the risks of heavy metals and NPs in the freshwater environment.

The interfacial interaction between Dechlorane Plus (DP) and polystyrene nanoplastics (PSNPs): An overlooked influence factor for the algal toxicity of PSNPs.

As pollution has attracted attention due to its wide distribution. An environmental concern that may be overlooked is that NPs additives are easily released into the environment due to their physical combination with NPs. However, the knowledge gaps still exist about the interfacial reactions of NPs and the additives (e.g. flame retardants) and the joint ecological effect. In the present study, fourier transform infrared (FTIR) spectrometer coupled with 2D correlation spectroscopy (2D-COS) analysis revealed the interfacial reactions between polystyrene nanoplastics (PSNPs) and Dechlorane Plus (DP). Results showed that carbon­oxygen bonds and carbon­chlorine bonds were the important binding sites during adhesion and DP could reduce the colloidal stability. Single and joint ecological effects of PSNPs and DP on the microalgae Chlorella vulgaris were further deliberated. Reduced photosynthetic efficiency (reduced Fv/Fm by 0.03 %), higher growth inhibition (16.15 %) and oxidative damage (increased ROS by 152 %) were observed in algae under co-exposure. Notably, DP could significantly increase the attachment of PSNPs to the surface of the algae. Metabolomics further revealed that co-exposure significantly down-regulated amino acid metabolism and tricarboxylic acid cycle (TCA) cycle, and up-regulated fatty acid metabolism. The present study provides new insights into the risk assessment of NPs in aquatic environment by investigating the interfacial reaction mechanism and combined ecotoxicity of NPs and additives.

Effects of co-modified biochar immobilized laccase on remediation and bacterial community of PAHs-contaminated soil.

Considering the stability and economy of immobilized enzymes, this study prepared co-modified biochar immobilized laccase product named Fe3O4@NaBC@GA@LC via orthogonal experimental design and explored its possibility of remediating polycyclic aromatic hydrocarbons (PAHs) contaminated soil in steel plants. Compared with the free laccase treatment, the relative activity of Fe3O4@NaBC@GA@LC remained 60 % after 50 days of incubation at room temperature. The relative activity of Fe3O4@NaBC@GA@LC could still retain nearly 80 % after five reuses. In the process of simulating the PAHs-contaminated site treatment experiment in Hangzhou Iron and steel plant, immobilized laccase exhibited efficient adsorption and degradation performances and even the removal rate of 5-ring PAHs reached more than 90 % in 40 days, resulting in improving urease activity and dehydrogenase in the soil and promoted the growth of a PAH degrading bacteria (Massilia). Our results further explained the efficient degradation effects of Fe3O4@NaBC@GA@LC on PAHs, which make it a promising candidate for PAHs-contaminated soil remediation.

Three typical microplastics affect the germination and growth of amaranth (Amaranthus mangostanus L.) seedlings.

Microplastics (MPs) have been a global emerging contaminant and have aroused wide public concern. Currently, it is still unknown the phytotoxicity effect of MPs on amaranth (Amaranthus mangostanus L.). This study investigated the early responses of amaranth by exposing its seeds to suspensions of polystyrene (PS), polyethylene (PE), and polypropylene (PP) MPs. We observed the effects of MPs on seed germination and growth of amaranth, especially on the oxidative damage in amaranth roots. Impacts of MPs on the germination and growth of amaranth varied with the type, concentration, and particle size of MPs. PE MPs and PP MPs inhibited the shoot extension of amaranth, while the root length under PP MPs treatment was generally shorter than that under PS MPs and PE MPs. The accumulation of H2O2 in amaranth roots increased with the rising of MPs concentration. Compared with the control, a little number of dead cells were found in the roots of amaranth under high MPs treatment. It is noteworthy that only under 100 mg/L PP treatment, the amaranthus seedlings root cells were disorganized, due to the reactive oxygen species (ROS) damage in the roots. These findings provide essential information to assess the phytotoxicity of MPs in agricultural products, and provide insights into the underlying mechanisms of the observed phytotoxicity.

A retrospective study of the accuracy of Invisalign Progress Assessment with clear aligners.

The objective of this study was to detective the accuracy of model superimposition and automatic analysis for upper and lower dentition width in Invisalign Progress Assessment during the process of clear aligners. 19 cases were included in this study. Pre-treatment dental cast (T0) and post-treatment dental cast after staged treatment (T1) were available for three-dimensional model superimposition. Subsequently, movements of maxillary teeth in the horizontal plane (cross-section) after staged treatment and width of upper and lower dentition were measured by three-dimensional model superimposition in the real world and Invisalign Progress Assessment separately. Consequently, the data collected from these two methods were compared. In Invisalign Progress Assessment, movements of maxillary teeth in the horizontal plane after staged treatment was 2.31 (1.59,3.22) [median (upper quartile, lower quartile)] millimeter (mm), while in three-dimensional model superimposition, the result was 1.79 (1.21,3.03) mm. The difference between the two groups is significant (P < 0.05). Intercanine width upper, intermolar width upper, intercanine width lower, and intermolar width lower were 36.55 ± 2.76 mm, 56.98 ± 2.62 mm, 28.16 ± 1.85 mm, 53.21 ± 2.72 mm separately in Invisalign Progress Assessment and were 36.48 ± 2.78 mm, 56.89 ± 2.58 mm, 28.05 ± 1.85 mm, 53.16 ± 2.64 mm separately in three-dimensional model analysis, which was no significant difference among these groups (P > 0.05). The data from Invisalign Progress Assessment was not in parallel with what was achieved from model superimposition with palate as a reference completely. The accuracy of model superimposition in Invisalign Progress Assessment needs further investigation, whereas the accuracy of model analysis in Invisalign Progress Assessment was accurate. Thereby, results from Invisalign Progress Assessment should be interpreted with caution by the orthodontist in the clinic.

Broadband absorption enhancement in a-Si:H thin-film solar cells sandwiched by pyramidal nanostructured arrays.

A new thin-film solar cell structure with a broadband absorption enhancement is proposed. The active a-Si:H film is sandwiched by two periodic pyramidal structured layers. The upper dielectric pyramidal layer acts as matching impedance by gradual change of the effective refractive index to enhance the absorption of the active layer in the short wavelength range. The lower metallic pyramidal layer traps light by the excitation of Fabry-Perot (FP) resonance, waveguide (WG) resonance and surface plasmon (SP) mode to enhance the absorption in the long wavelength range. With the cooperation of the two functional layers, a broadband absorption enhancement is realized. The structure parameters are designed by the cavity resonance theory, which shows that the results are accordant with the finite-difference time-domain (FDTD) simulation. By optimizing, the absorption of the sandwich structure is enhanced up to 48% under AM1.5G illumination in the 350-900 nm wavelength range compared to that of bare thin-film solar cells.

Surface-plasmon-polaritons-assisted nanolithography with dual-wavelength illumination for high exposure depth.

We propose a new direct writing nanolithography approach using a plasmonic focusing device and a nano silver mirror with dual-wavelength illumination for high exposure depth. Arrays of pyramid aperture are used to focus the incident light beams into 80 nm light spots. The pyramid combined with a thin silver film coated on the substrate constructs a surface plasmon polaritons (SPP) coupling cavity, which amplifies the intensity of the light field in it by SPP effect and resonance. The transmission depth of the standing wave formed by forward and reflected light could reach hundreds of nanometers. Two lasers with different wavelengths are used as illumination sources to homogenize the light field through complementation between the two standing waves. Simulation results show by using 355 nm and 441 nm wavelengths, a space of 44 nm at the bottom of the photoresist could be obtained after exposure and development. The feature size of resist patterns could be further scaled down, depending on the optimization of parameters of photoresist exposure and development, illumination wavelengths, etc.

Kochia scoparia L., a newfound candidate halophyte, for phytoremediation of cadmium-contaminated saline soils.

In recent years, heavy metal pollution in saline soil is increasingly severe due to the rapid development of industry and agriculture. Halophytes can survive at higher concentrations of salt and heavy metal, which make them suitable candidates for the phytoremediation of heavy metals in saline soils. In the present study, the halophyte plant Kochia scoparia (L.) Schrad. seedlings were exposed to different doses of Cd (0, 5, 10, 30 mg/kg) and NaCl (0, 200, 400, 800 mM) to explore its tolerance and phytoremediation ability for Cd. There was no significant toxic effect of Cd on the K. scoparia seedlings. NaCl reduced the biomass of K. scoparia compared with the control, but did not show any visible toxic symptom. Furthermore, Cd accumulation in K. scoparia is mainly distributed in the shoot; especially when exposed to low-Cd (5 mg/kg) treatment, the accumulation of Cd in the shoots was up to 5.42-22.25 mg/kg, which was 3.18-53.4 times of that in the roots. Moreover, the contents of glutathione and oxalate in plants increased gradually with the increase of NaCl concentration. Under the treatment of 800 mM NaCl without Cd, the content of glutathione reached the highest 51.21 µg/g, and the proportion of oxalate reached the highest 28.76% under the treatment of 30 mg/kg Cd with 400 Mm NaCl. Finally, we also found the significant alterations of cadmium chemical forms in rhizosphere soil with the addition of NaCl. Overall, K. scoparia could be an efficient and valuable candidate for the phytoextraction of low-Cd (5 mg/kg)-contaminated saline soil.

Analysis of a Skating Time-Trial Competition and Associated Performance-Determinants in Cross-Country Skiers.

Purpose: To examine the contributions of time in different terrains and sub-technique distribution to overall time-trial performance, as well as the relationships of laboratory and field-based performance determinants in cross-country skiers. Methods: Fourteen male XC skiers were monitored during a 10 km (3 × 3.3 km) skating time-trial competition. On separate days, the skiers performed body composition assessments, laboratory tests while roller-ski skating and a 3 km uphill skating field test. Results: Time in uphill terrain was most strongly correlated with overall performance (r = 0.99, p < 0.01). G2 and G3 were the predominant sub-techniques (61% of overall time) with more use of G2 on lap three compared to lap one (p < 0.05). Body mass and lean mass were inversely correlated with overall and uphill performance (r = -0.60-0.75, all p < 0.05). VO2 at 4 mmol·L-1, VO2peak and TTE while roller-ski skating in the laboratory and the 3 km uphill skating field test correlated with overall performance (r = -0.66-0.85, all p < 0.05). Conclusions: Time in uphill terrain was the main contributor to overall performance, and G3 and G2 the most used sub-techniques with increased utilization of G2 throughout the competition. VO2peak and TTE while roller-ski skating in the laboratory and performance in an uphill skating field test had the strongest associations with time-trial performance.