Microplastics and nanoplastics: Source, behavior, remediation, and multi-level environmental impact.

Plastics introduced into the natural environment persist, degrade, and fragment into smaller particles due to various environmental factors. Microplastics (MPs) (ranging from 1 µm to 5 mm) and nanoplastics (NPs) (less than 1 µm) have emerged as pollutants posing a significant threat to all life forms on Earth. Easily ingested by living organisms, they lead to ongoing bioaccumulation and biomagnification. This review summarizes existing studies on the sources of MPs and NPs in various environments, highlighting their widespread presence in air, water, and soil. It primarily focuses on the sources, fate, degradation, fragmentation, transport, and ecotoxicity of MPs and NPs. The aim is to elucidate their harmful effects on marine organisms, soil biota, plants, mammals, and humans, thereby enhancing the understanding of the complex impacts of plastic particles on the environment. Additionally, this review highlights remediation technologies and global legislative and institutional measures for managing waste associated with MPs and NPs. It also shows that effectively combating plastic pollution requires the synergization of diverse management, monitoring strategies, and regulatory measures into a comprehensive policy framework.

Dural metastasis of prostate carcinoma mimicking intracranial hematoma: a case report and literature review.

Dural metastases of prostate adenocarcinoma are an extremely rare complication and may mimic intracranial hematoma. Preoperatively diagnosis may be difficult due to similarities in symptoms and radiological appearance. We present a 65-year-old man admitted to the ED with a history of headache, nausea, vomiting, vertigo, diplopia, as well as numbness of his left lower extremity. Past medical history confirmed metastatic prostate cancer disease. After computed tomography and contrast computed tomography, the consulting radiologist diagnosed a chronic subdural hematoma. After burr hole trephination and dural opening, tumorous mass was detected. Histopathologic samples were taken. Histopathological examination was consistent with metastatic adenocarcinoma of the prostate. Although rare, dural metastases need to be included in oncological patients presenting in the ED with symptoms and radiological imaging suggesting hematoma. Both neurooncological and neurosurgical consultations are essential in order to apply the best treatment strategy.

Genomic analysis reveals complex population structure within the smooth newt, Lissotriton vulgaris, in Central Europe.

Species with wide-range distributions usually display high genetic variation. This variation can be partly explained by historical lineages that were temporally isolated from each other and are back into secondary reproductive contact, and partly by local adaptations. The smooth newt (Lissotriton vulgaris) is one of the most widely distributed amphibians species across Eurasia and forms a species complex with a partially overlapping distribution and morphology. In the present study, we explored the population genomic structure of smooth newt lineages in the Carpathian Basin (CB) relying on single-nucleotide polymorphisms. Our dataset included new and previously published data to study the secondary contact zone between lineages in the CB and also tested for the barrier effect of rivers to gene flow between these lineages. We confirmed the presence of the South L. v. vulgaris Lineage distributed in Transdanubia and we provided new distribution records of L. v. ampelensis inhabiting the eastern territories of the CB. High genetic diversity of smooth newts was observed, especially in the North Hungarian Mountains and at the interfluves of the main rivers in the South with four distinct lineages of L. v. vulgaris and one lineage of L. v. ampelensis showing a low level of admixture with the spatially closest L. v. vulgaris lineage. Moreover, admixture detected at the interfluve of the main rivers (i.e. Danube and Tisza) suggested a secondary contact zone in the area. Finally, we found that the river Danube has a very weak effect on population divergence, while the river Tisza is a geographical barrier limiting gene flow between smooth newt lineages. As the range boundaries of L. v. vulgaris and L. v. ampelensis in the CB coincide with the river Tisza, our study underpins the influence of rivers in lineage diversification.

Introduction of a multiplex amplicon sequencing assay to quantify DNA methylation in target cytosine markers underlying four selected epigenetic clocks.

BACKGROUND: DNA methylation analysis has proven to be a powerful tool for age assessment. However, the implementation of epigenetic age prediction in diagnostics or routine forensic casework requires appropriate laboratory methods. In this study, we aimed to compare the performance of large-scale DNA methylation analysis protocols that show promise in terms of accuracy, throughput, multiplexing capacity, and high sensitivity. RESULTS: The protocols were designed to target a predefined panel of 161 genomic CG/CA sites from four known estimators of epigenetic age-related parameters, optimized and validated using artificially methylated controls or blood samples. We successfully targeted 96% of these loci using two enrichment protocols: Ion AmpliSeq™, an amplicon-based method integrated with Ion Torrent S5, and SureSelectXT Methyl-Seq, a hybridization-based method followed by MiSeq FGx sequencing. Both protocols demonstrated high accuracy and robustness. Although hybridization assays have greater multiplexing capabilities, the best overall performance was observed for the amplicon-based protocol with the lowest variability in DNA methylation at 25 ng of starting DNA, mean observed marker coverage of ~ 6.7 k reads, and accuracy of methylation quantification with a mean absolute difference between observed and expected methylation beta value of 0.054. The Ion AmpliSeq method correlated strongly with genome-scale EPIC microarray data (R = 0.91) and showed superiority in terms of methylation measurement accuracy. Method-to-method bias was accounted for by the use of linear transformation, which provided a highly accurate prediction of calendar age with a mean absolute error of less than 5 years for the VISAGE and Hannum age clocks used. The pace of aging (PoAm) and the mortality risk score (MRS) estimators included in our panel represent next-generation clocks, were found to have low to moderate correlations with the VISAGE and Hannum models (R < 0.75), and thus may capture different aspects of epigenetic aging. CONCLUSIONS: We propose a laboratory tool that allows the quantification of DNA methylation in cytosines underlying four different clocks, thus providing broad information on epigenetic aging while maintaining a reasonable number of CpG markers, opening the way to a wide range of applications in forensics, medicine, and healthcare.

Synthesis, thermal evolution and magnetic investigations of SBA-15 silica functionalized with anchored iron phosphonate molecules.

In the current work, we report on the synthesizing of a series of novel nanocomposite materials obtained by functionalizing the SBA-15 silica matrix with anchored iron phosphonate molecules and the following thermal treatment. The obtained results reveal the formation of a unique amorphic layer of Fe-based compounds on the surface of silica walls of SBA-15 channels as a result of the organic groups' decomposition after moderate thermal treatment. Due to their unique structure, represented in an active Fe-containing amorphous coating spread over a large surface area, these materials are of great interest for their potential applications in fields such as catalysis, adsorption, and non-linear optics. The obtained materials remain amorphous, preserving the SBA-15 mesoporous structure up to temperatures of approximately 800 °C, after which the partial melting of the silica backbone is observed with the simultaneous formation of nanocrystals inside the newly-formed glassy mass. All obtained materials were characterized using such techniques as thermogravimetry, transmission and scanning electron microscopy combined with energy dispersive x-ray spectroscopy mapping, Raman spectroscopy, N2sorption analysis, x-ray diffraction, x-ray photoelectron spectroscopy, Mössbauer spectroscopy, and SQUID measurements.

Long COVID syndrome after SARS-CoV-2 survival in patients with pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension.

Pulmonary arterial hypertension (PAH) and chronic thromboembolic pulmonary hypertension (CTEPH) patients have a more severe COVID-19 course than the general population. Many patients report different persistent symptoms after SARS-CoV-2 infection. The aim of our study is to analyze the prevalence of long COVID-19 symptoms and assess if COVID-19 affects pulmonary hypertension (PH) prognosis. PAH/CTEPH patients who survived COVID-19 for at least 3 months before visiting the PH centers were included in the study. The patients were assessed for symptoms in acute phase of SARS-CoV-2 infection and persisting in follow-up visit, WHO functional class, 6-min walk distance, NT-proBNP concentration. The COMPERA 2.0 model was used to calculate 1-year risk of death due to PH at baseline and at follow-up. Sixty-nine patients-54 (77.3%) with PAH and 15 (21.7%) with CTEPH, 68% women, with a median age of 47.5 years (IQR 37-68)-were enrolled in the study. About 17.1% of patients were hospitalized due to COVID-19 but none in an ICU. At follow-up (median: 155 days after onset of SARS-CoV-2 symptoms), 62% of patients reported at least 1 COVID-19-related symptom and 20% at least 5 symptoms. The most frequently reported symptoms were: fatigue (30%), joint pain (23%), muscle pain (17%), nasal congestion (17%), anosmia (13%), insomnia (13%), and dyspnea (12%). Seventy-two percent of PH patients had a low or intermediate-low risk of 1-year death due to PH at baseline, and 68% after COVID-19 at follow-up. Over 60% of PAH/CTEPH patients who survived COVID-19 suffered from long COVID-19 syndrome, but the calculated 1-year risk of death due to PH did not change significantly after surviving mild or moderate COVID-19.

Improper ferroelastic phase transition in a hydrogen-bonded metallocyanide-based (azetidinium)2(H3O)[Co(CN)6] framework.

A hydrogen-bonded metallocyanide-based (AZE)2(H3O)[Co(CN)6] framework was obtained by introducing a hydronium cation to the A2B(1)B(2)CN6-type structure. Simple K+/H3O+ substitution exchange the three-dimensional structure of the double-perovskite into two-dimensional layers containing open inorganic cages with a strongly discontinuous improper ferroelastic phase transition.

Litter per liter - Lakes' morphology and shoreline urbanization index as factors of microplastic pollution: Study of 30 lakes in NE Poland.

Human activity and anthropopression play key roles in contaminating aquatic environments with microplastics (MPs). The lakes of northeastern Poland provide a wide range of freshwater ecosystems differing in morphology, hydrology and ecology. In this study, we investigate 30 lakes during summer stagnation, considering their varying levels of anthropogenization of the catchment area and taking into account increased tourist activity. MPs were found in each of the studied lakes in the range from 0.27 MPs/L to 1.57 MPs/L, and the average value was 0.78 ± 0.42 MPs/L. The features of the MPs were evaluated, including size, form and color (most frequent: 4-5 mm (35.0 %); fragments (36.7 %); blue (30.6 %), respectively). Gradual accumulation of MPs has been observed in the lakes that comprise the hydrological sequence. The amount of sewage produced by wastewater treatment plants was considered in the study area. Statistically significant differences were found between lakes with different surface areas and shoreline lengths, where lakes with the highest and smallest values were notably more polluted with MPs than lakes in the medium range (F = 34.64, p < .0001 and F = 5.96, p < .01, respectively). The study presents an easy-to-obtain shoreline urbanization index (SUI) that is particularly useful for lakes with heavily transformed catchments in terms of hydrology. A significant correlation between the MP concentration and SUI, reflecting the degree of direct catchment anthropopression (r = +0.4282; p < .05), was observed. The analysis of the human impact on shoreline conversions and development should also pique the interest of other researchers as a potential indicator of MP pollution.

DIC-Enhanced Identification of Bodner-Partom Model Parameters for Bitumen Binder.

Bitumen binder is a component of asphalt mixtures that are commonly used as the materials constituting the upper layers of a pavement's structure. Its main role is to cover all the remaining constituents (aggregate, filler and other possible additives) and create a stable matrix, in which they are embedded due to the adhesion forces. The long-term performance of bitumen binder is crucial to the holistic behavior of the layer made of the asphalt mixture. In this study, we use the respective methodology to identify the parameters of the well-established Bodner-Partom material model. For the purposes of its parameters identification, we carry out a number of the uniaxial tensile tests with different strain rates. The whole process is enhanced with a digital image correlation (DIC) to capture the material response in a reliable way and to provide deeper insight into the experiment results. The obtained model parameters were used to compute numerically the material response using the Bodner-Partom model. Good agreement between the experimental and numerical results was observed. The maximum error for the elongation rates equal to 6 mm/min and 50 mm/min is of order of 10%. The novel aspects of this paper are as follows: the application of the Bodner-Partom model to the bitumen binder analysis and the DIC-enhancement of the laboratory experiment.

The Incidence and Risk Factors for Allogeneic Blood Transfusions in Pediatric Spine Surgery: National Data.

(1) Background: Pediatric spinal surgery is a blood-intensive procedure. In order to introduce a rational blood management program, identifying the risk factors for transfusions is mandatory. (2) Methods: Data from the national database covering the period from January 2015 to July 2017 were analyzed. The available data included the demographics, characteristics of the surgeries performed, length of stay, and in-house mortality. (3) Results: The total number of patients used for the analysis was 2302. The primary diagnosis was a spinal deformity (88.75%). Most fusions were long, with four levels or more (89.57%). A total of 938 patients received a transfusion; thus, the transfusion rate was 40.75%. The present study identified several risk factors; the most significant was a number of levels fused greater than 4 (RR 5.51; CI95% 3.72-8.15; p < 0.0001), followed by the deformity as the main diagnosis (RR 2.69; CI95% 1.98-3.65; p < 0.0001). These were the two most significant factors increasing the odds of a transfusion. Other factors associated with an increased risk of transfusion were elective surgery, the female sex, and an anterior approach. The mean length of stay in days was 11.42 (SD 9.93); this was greater in the transfused group (14.20 vs. 9.50; p < 0.0001). (4) Conclusions: The rate of transfusions in pediatric spinal surgery remains high. A new patient blood management program is necessary to improve this situation.

Landscape genetics reveals contrasting patterns of connectivity in two newt species (Lissotriton montandoni and L. vulgaris).

Ecologically distinct species may respond to landscape changes in different ways. In addition to basic ecological data, the extent of the geographic range has been successfully used as an indicator of species sensitivity to anthropogenic landscapes, with widespread species usually found to be less sensitive compared to range-restricted species. In this study, we investigate connectivity patterns of two closely related but ecologically distinct newt species - the range-restricted, Lissotriton montandoni and the widespread, L. vulgaris - using genomic data, a highly replicated setting (six geographic regions per species), and tools from landscape genetics. Our results show the importance of forest for connectivity in both species, but at the same time suggest differential use of forested habitat, with L. montandoni and L. vulgaris showing the highest connectivity at forest-core and forest-edges, respectively. Anthropogenic landscapes (i.e., higher crop- or urban-cover) increased resistance in both species, but the effect was one to three orders of magnitude stronger in L. montandoni than in L. vulgaris. This result is consistent with a view of L. vulgaris as an ecological generalist. Even so, currently, the negative impact of anthropogenic landscapes is mainly seen in connectivity among L. vulgaris populations, which show significantly stronger isolation and lower effective sizes relative to L. montandoni. Overall, this study emphasizes how habitat destruction is compromising genetic connectivity not only in endemic, range-restricted species of conservation concern but also in widespread generalist species, despite their comparatively lower sensitivity to anthropogenic landscape changes.

3D printing of bio-instructive materials: Toward directing the cell.

Fabrication of functional scaffolds for tissue engineering and regenerative medicine applications requires material systems with precise control over cellular performance. 3D printing is a powerful technique to create highly complex and multicomponent structures with well-defined architecture and composition. In this review paper, we explore extrusion-based 3D printing methods (EBP, i.e., Near Field Electrospinning (NFES), Melt Electrowriting (MEW), Fused Deposition Modeling (FDM), and extrusion bioprinting) in terms of their ability to produce scaffolds with bio-instructive properties. These material systems provide spatio-temporal guidance for cells, allowing controlled tissue regeneration and maturation. Multiple physical and biochemical cues introduced to the EBP scaffolds are evaluated in their ability to direct cell alignment, proliferation, differentiation, specific ECM production, and tissue maturation. We indicate that the cues have different impacts depending on the material system, cell type used, or coexistence of multiple cues. Therefore, they must be carefully chosen based on the targeted application. We propose future directions in bio-instructive materials development, including such concepts as metamaterials, hybrid living materials, and 4D printing. The review gathers the knowledge essential for designing new materials with a controlled cellular response, fabrication of advanced engineered tissue, and developing a better understanding of cell biology, especially in response to the biomaterial.

Neuropeptide Y and its receptors in prostate cancer: associations with cancer invasiveness and perineural spread.

PURPOSE: Neuropeptide Y (NPY) is a pleiotropic peptide, which is involved in many biological mechanisms important in regulation of cell growth and survival. The aim of this study was a comprehensive analysis of the NPY system in prostate pathology. METHODS: The study was based on immunohistochemical analysis of NPY and its receptors, Y1R, Y2R and Y5R, in tissue samples from benign prostate (BP), primary prostate cancer (PCa) and PCa bone metastases. Tissue microarray (TMA) technique was employed, with analysis of multiple cores from each specimen. Intensity of the immunoreactivity and expression index (EI), as well as distribution of the immunostaining in neoplastic cells and stromal elements were evaluated. Perineural invasion (PNI) and extraprostatic extension (EPE) were areas of special interests. Moreover, a transwell migration assay on the LNCaP PCa cell line was used to assess the chemotactic properties of NPY. RESULTS: Morphological analysis revealed homogeneous membrane and cytoplasmic pattern of NPY staining in cancer cells and its membrane localization with apical accentuation in BP glands. All elements of the NPY system were upregulated in pre-invasive prostate intraepithelial neoplasia, PCa and metastases. EI and staining intensity of NPY receptors were significantly higher in PCa then in BP with correlation between Y2R and Y5R. The strength of expression of the NPY system was further increased in the PNI and EPE areas. In bone metastases, Y1R and Y5R presented high expression scores. CONCLUSION: The results of our study suggest that the NPY system is involved in PCa, starting from early stages of its development to disseminated states of the disease, and participates in the invasion of PCa into the auto and paracrine matter.

Evaluation of Movement Restriction of Spinal Orthoses Using Inertial Measurement Units.

Despite the frequent use of orthopedic braces or spine stabilizers in diseases such as kyphosis, lordosis, and scoliosis, as well as in the case of injuries and rehabilitation after surgeries, there is no clear evidence of their proper stabilization of the spine while carrying out daily activities. This study sought to assess the spine's mobility while wearing three different orthopedic braces while performing basic tasks. Ten healthy subjects were enrolled. Three Inertial Measurement Units (IMUs) were attached superficially along the spine at approximate levels: cervical (C7), between thoracic (T8) and lumbar (L3), and sacrum. The angle between sensors was monitored to provide data on the sagittal profile. In addition, the displacement of the spine's longitudinal axis was measured (rotation). There are three types of orthopedic braces: the semi-rigid Hohmann corset, the Jewett brace, and the Thoracolumbar Fixed Spinal Orthosis (TLSO). Four tasks were monitored: standing, sitting, walking, and picking up an item from the floor with one hand. All braces provided a similar level of stability in both the sagittal plane and rotational axis while lifting an object. On the other hand, while walking and sitting, the TLSO was the only orthosis providing a statistically significant rigidity in the sagittal plane. When performing a more voluntary task, the measured rigidity of softer braces was significantly increased when compared with more involuntary tasks. A certain degree of motion restriction with spinal orthoses may come from the feedback pressure, which stimulates paraspinal muscles to contract and thus increases the overall rigidity of the trunk.

Effects of ambient temperature during the nestling stage on a stress indicator in nestling pied flycatchers Ficedula hypoleuca.

Long-term and short-term changes in ambient temperature can cause stress in birds, leading to changes in the level of hematological parameters. The H:L ratio (heterophil-to-lymphocyte ratio) is a hematological index that allows for the assessment of the stress induced by environmental changes, including weather conditions. In this paper, we examined the influence of temperatures and the sum of precipitation on the health of nestling pied flycatchers (Ficedula hypoleuca) by using the H:L ratio reflecting the body's response to stress. All examined temperature indicators influenced the H:L ratio, yet the average value of daily minimum temperature during the first 12 days of nestling life had the strongest influence, maximum temperature had the weakest effect, while precipitation had no significant influence. Our research indicates that even a small increase in temperature caused a stress reaction in nestling pied flycatchers, which was reflected by an increase in the H:L ratio. The increase in the stress index (H:L ratio) was probably a result of poor weather conditions (precipitation, low temperature), which prevented the adult birds from actively foraging and properly feeding the nestlings.

Emerging infectious diseases of amphibians in Poland: distribution and environmental drivers.

Emerging infectious diseases are a threat to biodiversity and have taken a large toll on amphibian populations worldwide. The chytrid fungi Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal), and the iridovirus Ranavirus (Rv), are of concern as all have contributed to amphibian declines. In central and eastern Europe, their geographical and host distributions and main environmental drivers determining prevalence are poorly known. We screened over 1000 amphibians from natural and captive populations in Poland for the presence of Bd, Bsal and Rv. In wild amphibian populations, we found that Bd is widespread, present in 46 out of 115 sampled localities as well as 2 captive colonies, and relatively common with overall prevalence at 14.4% in 9 species. We found lower prevalence of Rv at 2.4%, present in 11 out of 92 sampling sites, with a taxonomic breadth of 8 different amphibian species. Bsal infection was not detected in any individuals. In natural populations, Pelophylax esculentus and Bombina variegata accounted for 75% of all Bd infections, suggesting a major role for these 2 species as pathogen reservoirs in Central European freshwater habitats. General linear models showed that climatic as well as landscape features are associated with Bd infection in Poland. We found that higher average annual temperature constrains Bd infection, while landscapes with numerous water bodies or artificial elements (a surrogate for urbanization) increase the chances of infection. Our results show that a combination of climatic and landscape variables may drive regional and local pathogen emergence.

Targeting bacteria causing otitis media using nanosystems containing nonspherical gold nanoparticles and ceragenins.

Aim: To evaluate the antibacterial and antibiofilm activity of ceragenin-conjugated nonspherical gold nanoparticles against the most common agents of otitis media. Methods: Minimal inhibitory and bactericidal concentrations and colony-counting assays, as well as colorimetric and fluorimetric methods, were used to estimate the antibacterial activity of compounds in phosphate-buffered saline and human cerumen. The nanosystems' biocompatibility and ability to decrease IL-8 release was tested using keratinocyte cells. Results: The tested compounds demonstrated strong antimicrobial activity against planktonic and biofilm cultures at nontoxic doses due to the induction of oxidative stress followed by the damage of bacterial membranes. Conclusion: This study indicates that ceragenin-conjugated nonspherical gold nanoparticles have potential as new treatment methods for eradicating biofilm-forming pathogens associated with otitis media.

Lay abstract Middle-ear infections can be painful and cause hearing difficulties. If untreated, they can lead to hearing loss. These infections are usually treated with antibiotic drugs. However, the microbes causing the infection can gain drug resistance. This article reports research into a new way of delivering antibiotics to kill the microbes and the communities they form (biofilms). The authors developed tiny gold particles loaded with the antimicrobial drug ceragenin and tested the drug-loaded particles on three common middle-ear infection-causing bacteria. Compared with ceragenin alone, the ceragenin-loaded particles were better at killing the bacteria and their biofilm communities.

Peanut-Shaped Gold Nanoparticles with Shells of Ceragenin CSA-131 Display the Ability to Inhibit Ovarian Cancer Growth In Vitro and in a Tumor Xenograft Model.

Gold nanoparticles-assisted delivery of antineoplastics into cancerous cells is presented as an effective approach for overcoming the limitations of systemic chemotherapy. Although ceragenins show great potential as anti-cancer agents, in some tumors, effective inhibition of cancer cells proliferation requires application of ceragenins at doses within their hemolytic range. For the purpose of toxicity/efficiency ratio control, peanut-shaped gold nanoparticles (AuP NPs) were functionalized with a shell of ceragenin CSA-131 and the cytotoxicity of AuP@CSA-131 against ovarian cancer SKOV-3 cells and were then analyzed. In vivo efficiency of intravenously and intratumorally administered CSA-131 and AuP@CSA-131 was examined using a xenograft ovarian cancer model. Serum parameters were estimated using ELISA methods. Comparative analysis revealed that AuP@CSA-131 exerted stronger anti-cancer effects than free ceragenin, which was determined by enhanced ability to induce caspase-dependent apoptosis and autophagy processes via reactive oxygen species (ROS)-mediated pathways. In an animal study, AuP@CSA-131 was characterized by delayed clearance and prolonged blood circulation when compared with free ceragenin, as well as enhanced anti-tumor efficiency, particularly when applied intratumorally. Administration of CSA-131 and AuP@CSA-131 prevented the inflammatory response associated with cancer development. These results present the possibility of employing non-spherical gold nanoparticles as an effective nanoplatform for the delivery of antineoplastics for the treatment of ovarian malignancy.

Evaluation of a Fatigue Detector Using Eye Closure-Associated Indicators Acquired from Truck Drivers in a Simulator Study.

This paper presents a camera-based prototype sensor for detecting fatigue and drowsiness in drivers, which are common causes of road accidents. The evaluation of the detector operation involved eight professional truck drivers, who drove the truck simulator twice-i.e., when they were rested and drowsy. The Fatigue Symptoms Scales (FSS) questionnaire was used to assess subjectively perceived levels of fatigue, whereas the percentage of eye closure time (PERCLOS), eye closure duration (ECD), and frequency of eye closure (FEC) were selected as eye closure-associated fatigue indicators, determined from the images of drivers' faces captured by the sensor. Three alternative models for subjective fatigue were used to analyse the relationship between the raw score of the FSS questionnaire, and the eye closure-associated indicators were estimated. The results revealed that, in relation to the subjective assessment of fatigue, PERCLOS is a significant predictor of the changes observed in individual subjects during the performance of tasks, while ECD reflects the individual differences in subjective fatigue occurred both between drivers and in individual drivers between the 'rested' and 'drowsy' experimental conditions well. No relationship between the FEC index and the FSS state scale was found.