Combustion-derived particles are a significant concern due to their nanometric sizes and complex chemical compositions, which can negatively impact health. These particles contain polycyclic aromatic hydrocarbons (PAH) and their oxygenated derivatives (OPAH), which can dissolve in biological lung fluids. As a result, they form complex mixtures whose toxicity is often poorly studied. Therefore, evaluating inhalation bioaccessibility is crucial for accurately assessing the health risks associated with these particles. We determined the inhalation bioaccessibility fractions (IBAF) of 16 PAH and 5 OPAH from particles generated under controlled conditions. Several critical parameters were evaluated, including simulated lung fluids (SLF), extraction times, and solid-to-liquid (S/L) ratios. Our results indicated that eight compounds - NAP, ACY, FLU, 9-FLUO, PHE, ANT, FLT, PYR - were systematically extracted by the different SLF, namely Gamble's solution (GS), artificial lysosomal fluid (ALF), modified GS (MGS), and simulated epithelial lung fluid (SELF) after 24 h of extraction time with an S/L ratio of 1/2000. The evaluation of the toxicological response, based on cytotoxicity and the AhR-dependent pathway, demonstrated variability in the toxicological profiles, depending on the extraction conditions used. Notably, the extraction condition yielding a biological response equivalent to that assessed with the total particulate fraction was the use of MGS as the extraction fluid under our experimental conditions. From a toxicological perspective, these approaches appear relevant for assessing the toxicity mechanisms of PAH mixtures, closely mimicking the in vivo conditions found in the human respiratory tract. This study outlined that understanding whether the bioaccessible fraction of particulate-bound PAHs is a reliable predictor of toxicity is central to advancing inhalation risk assessment.

An accidental industrial fire happened in Rouen (France) in september 2019 from a plant specialized in the production of mineral oils and engine additives. A few days after, passive sampling of the air was performed on thermodesorption tubes, from 09/30/2019 to 10/07/2019 using Tenax (R) TA and Carbopack B&X sorbents. Two identical samplings were carried out 18 (2021) and 26 months (2022) after the fire in the same place to evaluate the urban and in-house background noises. Analyses were performed using a thermodesorption gas chromatograph coupled to a hybrid quadrupole Orbitrap mass spectrometer (TD-GC-Q-Orbitrap) in full scan mode. A nontargeted screening allowed to detect thousands of molecules (about 8000 on Tenax (R) TA and 3000 on Carbopack B&X sorbents) including a large number of hydrocarbons, alkylbenzenes and polycyclic aromatic hydrocarbons in the tubes collected in 2019. But such hydrocarbons were not specific of the fire and then were not identified as industrial fire markers, as their presence were detected in the samples collected in 2021 and/or 2022. However, 22 sulfur-containing compounds were considered as potential markers because they were only detected in the samples collected in 2019 or at significantly higher level than in 2021 and/or 2022 and were detected in samples collected during Laboratory fire of engine oils. Moreover, these compounds were annotated with good confidence level. Among them, the identification of 6 sulfur-containing compounds, 2-acetylthiophene, 2-propionylthiophene, 2,2 '-bithiophene, 3,3 '-bithiophene, thieno [3,2-b]thiophene and di-tert-butyl-disulfide, and one oxygencontaining compound, 1,3,5-trioxane, was confirmed by injection of corresponding standards.

The widespread use of disposable masks has sparked concerns about potential exposure to harmful chemicals, including the already ubiquitous phthalates. It has been demonstrated that there is a causal relationship between the exposure to phthalates and the onset of respiratory diseases. Moreover, there is an increasing body of evidence, which suggests that mitochondria contribute to significant impact to the epithelial function in respiratory diseases. However, the specific effects induced by these compounds at the mitochondrial level in respiratory cells are unknown. The objective of this study was to identify phthalates in disposable face masks and to evaluate their effect on mitochondrial function in human bronchial epithelial cells (BEAS-2B). In order to identify and quantify the phthalates in face masks, an extraction method was developed and a phthalate quantification method using Gas Chromatography-Mass Spectrometry was employed. The analysis of the face masks sample revealed the presence of three major phthalates: di(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP) and diisobutyl phthalate (DiBP). To evaluate the effects of these phthalates on mitochondrial function, BEAS-2B were exposed to the phthalates individually and in a mixture of the three at concentrations ranging from 0.5 to 10 mg/L for 24 hours. Then extracellular flux analysis (Seahorse analysis), adenosine levels (LC-DAD) and lactate levels (LC-MS) were evaluated. The observed effects at the mitochondrial level varied according to the specific phthalates that were tested. The major dysfunctions were identified following exposure to DEHP, which exerted inhibitory effects on the basal and maximal oxygen consumption rate of BEAS-2B cells, as well as an alteration in oxidative phosphorylation and a reduction in the ATP/ADP ratio. This was accompanied by an increase in both extracellular lactate production and extracellular acidification rate. A shift towards a glycolytic phenotype was noted. Similar, yet more pronounced effects, were observed following exposure to the mixture of the three phthalates, which suggest that DEHP may contribute to the overall effect of the mixture. This study is the first to provide knowledge on the adverse effects of a mixture of phthalates on mitochondrial function in bronchial epithelial cells, demonstrating the capacity of some phthalates to induce a shift toward a glycolytic phenotype. These results highlight the potential risks of inhaled phthalates on mitochondrial function, predisposing to the development of respiratory diseases.

Coloured pyrotechnic smokes are frequently used in the military field and occasionally by civilians, but their health hazards have been little studied. The main concern could rise from inhalation of smoke particles. Our previous study showed that acute exposure to particles from a red signalling smoke (RSS) induced an antioxidant and inflammatory responses in small airway epithelial cells. The aim of this study was to further explore the toxicity of RSS particles at a more proximal level of the respiratory tract, using normal human bronchial epithelial cells grown at the Air-Liquid Interface. Acute exposure (24 h) induced an oxidative stress that persisted 24 h post-exposure, associated with particle internalization and epithelium morphological changes (cuboidal appearance and loss of cilia). Repeated exposures (4×16h) to RSS particles did not trigger oxidative stress but cell morphological changes occurred. Overall, this study provides a better overview of the toxic effects of coloured smoke particles.

Gasoline emissions contain high levels of pollutants, including particulate matter (PM), which are associated with several health outcomes. Moreover, due to the depletion of fossil fuels, biofuels represent an attractive alternative, particularly second-generation biofuels (B2G) derived from lignocellulosic biomass. Unfortunately, compared to the abundant literature on diesel and gasoline emissions, relatively few studies are devoted to alternative fuels and their health effects. This study aimed to compare the adverse effects of gasoline and B2G emissions on human bronchial epithelial cells. We characterized the emissions generated by propane combustion (CAST1), gasoline Surrogate, and B2G consisting of Surrogate blended with anisole (10%) (S+10A) or ethanol (10%) (S+10E). To study the cellular effects, BEAS-2B cells were cultured at air-liquid interface for seven days and exposed to different emissions. Cell viability, oxidative stress, inflammation, and xenobiotic metabolism were measured. mRNA expression analysis was significantly modified by the Surrogate S+10A and S+10E emissions, especially CYP1A1 and CYP1B1. Inflammation markers, IL-6 and IL-8, were mainly downregulated doubtless due to the PAHs content on PM. Overall, these results demonstrated that ultrafine particles generated from biofuels Surrogates had a toxic effect at least similar to that observed with a gasoline substitute (Surrogate), involving probably different toxicity pathways.

Pyrotechnic smokes are widely used in civilian and military applications. The major issue arise from the release of particles after smoke combustion but the health risks related to their exposure are poorly documented whereas toxicity of airborne particles on the respiratory target are very well known. Therefore, this study aimed to explore the in vitro toxicity of the particle fraction of different pyrotechnic smokes.Particles from a red signalling smoke (RSS), an hexachloroethane-based obscuring smoke (HC-OS) and an anti-intrusion smoke (AIS) were collected from the cloud. RSS particles displayed the highest organic fraction (quinones and polycyclic aromatic hydrocarbons) of the three samples characterized. AIS particles contained K and cholesterol derivatives. HC-OS particles were mainly metallic with very high concentrations of Al, Fe and Ca. Intrinsic oxidative potential of smoke particles was measured with two assays. Depletions of DTT by RSS particles was greater than depletion obtained with AIS and HC-OS particles but depletion of acid ascorbic (AA) was only observed with HC-OS particles. In vitro toxicity was assessed by exposing human small airway epithelial cells (SAEC) to various concentrations of particles. After 24 h of exposure, cell viability was not affected but significant modifications of mRNA expression of antioxidant (SOD-1 and -2, catalase, HO-1, NQO-1) and inflammatory markers (IL-6, IL-8, TNF-α) were observed and were dependent on smoke type. Particles rich in metal, such as HC-OS, induced a greatest depletion of AA and a greatest inflammatory response, whereas particles rich in organic compounds, such as RSS, induced a greatest DTT depletion and a greatest antioxidant response.In conclusion, the three smoke particles have an intrinsic oxidative potential and triggered a cell adaptive response. Our study improved the knowledge of particle toxicity of pyrotechnic smokes and scientific approach developed here could be used to study other type of particles.

Depressive disorder is characterized by disturbances in the hypothalamic-pituitary-thyroid (HPT) axis and in the metabolism of thyroid hormones (TH). The evidence for changes in TH levels is observed in human sera and cerebrospinal fluid as well as in animal model studies. Iodothyronine deiodinases (DIOs) type 1, 2 and 3 (DIO1, DIO2, DIO3) are important enzymes for the synthesis and determination of TH concentration. This study aims to examine the link between recurrent depressive disorders (rDD) and two functionally known polymorphisms DIO1a-C/T (rs11206244) and DIO1b-A/G (rs12095080) within the DIO1 gene encoding DIO1 and two polymorphisms DIO3-C/T (rs17716499), DIO3-A/C (rs7150269) within the DIO3 gene encoding DIO3.Both variants were genotyped in 254 rDD patients and 197 healthy subjects using polymerase chain reaction. Basic methods and statistical analyses were used to estimate genetic variants in the risk of the disease.No significant associations were found between the polymorphisms examined here and rDD. There were no significant associations between genotypes distribution and demographic/medical variables. Odds ratios (ORdis) and corresponding 95% confidence interval (95% CI) were calculated, for example: for CC genotype of DIO1a C/T (ORdis = 0.86, 95% CI: 0.59, 1.25).Functional variants within the DIO1 gene, which affect TH levels and polymorphisms in DIO3, are not confirmed to be associated with rDD. Nevertheless, considering previous data which indicate that the DIO1 gene is related to the depression, further studies on a larger sample size are recommended.

INTRODUCTION:Although the physiological role of the C-terminal hydrolase domain of the soluble epoxide hydrolase (sEH-H) is well investigated, the function of its N-terminal phosphatase activity (sEH-P) remains unknown. OBJECTIVES:This study aimed to assess in vivo the physiological role of sEH-P. METHODS:CRISPR/Cas9 was used to generate a novel knock-in (KI) rat line lacking the sEH-P activity. RESULTS:The sEH-P KI rats has a decreased metabolism of lysophosphatidic acids to monoacyglycerols. KI rats grew almost normally but with less weight and fat mass gain while insulin sensitivity was increased compared to wild-type rats. This lean phenotype was more marked in males than in female KI rats and mainly due to decreased food consumption and enhanced energy expenditure. In fact, sEH-P KI rats had an increased lipolysis allowing to supply fatty acids as fuel to potentiate brown adipose thermogenesis under resting condition and upon cold exposure. The potentiation of thermogenesis was abolished when blocking PPARγ, a nuclear receptor activated by intracellular lysophosphatidic acids, but also when inhibiting simultaneously sEH-H, showing a functional interaction between the two domains. Furthermore, sEH-P KI rats fed a high-fat diet did not gain as much weight as the wild-type rats, did not have increased fat mass and did not develop insulin resistance or hepatic steatosis. In addition, sEH-P KI rats exhibited enhanced basal cardiac mitochondrial activity associated with an enhanced left ventricular contractility and were protected against cardiac ischemia-reperfusion injury. CONCLUSION:Our study reveals that sEH-P is a key player in energy and fat metabolism and contributes together with sEH-H to the regulation of cardiometabolic homeostasis. The development of pharmacological inhibitors of sEH-P appears of crucial importance to evaluate the interest of this promising therapeutic strategy in the management of obesity and cardiac ischemic complications.

Ultrafine particles represent a growing concern in the public health community but their precise role in many illnesses is still unknown. This lack of knowledge is related to the experimental difficulty in linking their biological effects to their multiple properties, which are important determinants of toxicity. Our aim is to propose an interdisciplinary approach to study fine (FP) and ultrafine (UFP) particles, generated in a controlled manner using a miniCAST (Combustion Aerosol Standard) soot generator used with two different operating conditions (CAST1 and CAST3). The chemical characterization was performed by an untargeted analysis using ultra-high resolution mass spectrometry. In conjunction with this approach, subsequent analysis by gas chromatography–mass spectrometry (GC–MS) was performed to identify polycyclic aromatic hydrocarbons (PAH). CAST1 enabled the generation of FP with a predominance of small PAH molecules, and CAST3 enabled the generation of UFP, which presented higher numbers of carbon atoms corresponding to larger PAH molecules. Healthy normal human bronchial epithelial (NHBE) cells differentiated at the air-liquid interface (ALI) were directly exposed to these freshly emitted FP and UFP. Expression of MUC5AC, FOXJ1, OCLN and ZOI as well as microscopic observation confirmed the ciliated pseudostratified epithelial phenotype. Study of the mass deposition efficiency revealed a difference between the two operating conditions, probably due to the morphological differences between the two categories of particles. We demonstrated that only NHBE cells exposed to CAST3 particles induced upregulation in the gene expression of IL-8 and NQO1. This approach offers new perspectives to study FP and UFP with stable and controlled properties.

Plasticizers are chemicals in high demand, used in a wide range of commercial products. Human are exposed through multiple pathways, from numerous sources, to multiple plasticizers. This is a matter of concern, as it may contribute to adverse health effects. The vascular system carries plasticizers throughout the body and therefore can interact with the endothelium. The aim of the study was to evaluate the in vitro toxicity on endothelial cells by considering the individual and the mixture effects of bis-(2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DINP) or bis-(2-ethylhexyl) terephthalate (DEHT). In this study, their cytotoxicity on HMEC-1 cells was evaluated on cell function (viability, cell counting, total glutathione and intracellular adenosines) and mitochondrial function (mitochondrial respiration). Results showed cellular physiological perturbations induced with all the condition tested, excepted for DEHT. Plasticizers induced a cytotoxicity by targeting mitochondrial respiration, depleting mitochondrial ATP production and increasing glycolytic metabolism. Additionally, delayed effects were observed between the cellular and the mitochondrial parameters. These results suggest that endothelial cells could go through a metabolic adaptation to face plasticizer-induced cellular stress, to effectively maintain their cellular processes. This study provides additional information on the adverse effects of plasticizers on endothelial cells.

Air pollution has significant health effects worldwide, and airborne particles play a significant role in these effects. Ultrafine particles (UFPs) have an aerodynamic diameter of 0.1 μm or less, can penetrate deep into the respiratory tree, and are more toxic due to their large specific surface area, which should adsorb organic compounds. The aim of this study is to show the toxicological effects of UFPs with high organic content at low dose on BEAS-2B cells through at air-liquid interface (ALI) exposure using a Vitrocell® technology and a miniCAST (Combustion Aerosol Standard) generator. In conjunction with this approach, chemical analysis of particles and gas phase was performed to evaluate the presence of polycyclic aromatic hydrocarbons (PAHs). Chemical analyses confirmed the presence of PAHs in UFPs. With this experimental setup, exposure of the BEAS-2B cells induced neither cytotoxicity nor mitochondrial dysfunction. However, an increase of oxidative stress was observed, as assessed through Nrf2, NQO1, HO-1, CuZnSOD, MnSOD, and Catalase gene expression, together with significant induction of genes related to xenobiotic metabolism CYP1A1 and CYP1B1. Negative regulation of inflammatory genes expression (IL-6 and IL-8) was present three hours after the exposition to the UFPs. Taken together, this experimental approach, using repeatable conditions, should help to clarify the mechanisms by which organic UFPs induce toxicological effects.

Air pollution has significant health effects worldwide and airborne particles play a significant role in these effects. Ultrafine particles (UFPs) have an aerodynamic diameter of 0.1 μm or less and can penetrate deep into the respiratory tree and reach other organs such as the liver, the heart and the brain. UFPswould be more toxic due to their specific surface area which would absorb more organic compounds. The objective of this study is to show the toxicological effects of ultrafine particles with an important organic compounds content on Beas-2B model and by using an exposure at the air-liquid interface in order to keep their microphysic characteristics.

Gloves represent an essential feature for hand protection because it is a requirement in the professional framework to comply with both hand hygiene standards and the principles of good laboratory practice. Despite their wide use, there is a knowledge gap regarding their composition, including phthalates. The purpose of the present study was to develop two orthogonal methods, GC–MS and HPLC–DAD, for the screening of plasticizers in gloves. Performances of these two methods were compared in terms of ease of use, number of analyzed plasticizers, and sample preparation. The two methods were validated and applied for the identification and quantification of plasticizers in ten gloves made with different materials (vinyl, nitrile, latex, and neoprene). Results revealed the presence of three main ones: DEHP, DEHT, and DINP. Additionally, the contents of plasticizers were extremely variable, depending on the glove material. As expected, the results point out a predominant use of plasticizers in vinyl gloves with an amount that should be of concern. While DEHP is classified as a toxic substance for reproduction 1B, it was, however, quantified in the ten different glove samples studied. This study provides new data regarding the plasticizers’ content in protective gloves, which could be useful for risk assessment.

Abstract Aims In post‐menopausal women, incidence of heart failure with preserved ejection fraction is higher than in men. Hormonal replacement therapies did not demonstrate benefits. We tested whether the non‐steroidal mineralocorticoid receptor antagonist finerenone limits the progression of heart failure in ovariectomized (OVX) mice with metabolic disorders. Methods and results Ovariectomy was performed in 4‐month‐old mice, treated or not at 7 months old for 1 month with finerenone (Fine) 1 mg/kg/day. Left ventricular (LV) cardiac and coronary endothelial functions were assessed by echocardiography, catheterization, and myography. Blood pressure was measured by plethysmography. Insulin and glucose tolerance tests were performed. Exercise capacity and spontaneous activity were measured on treadmill and in combined indirect calorimetric cages equipped with voluntary running wheel. OVX mice presented LV diastolic dysfunction without modification of ejection fraction compared with controls (CTL), whereas finerenone improved LV filling pressure (LV end‐diastolic pressure, mmHg: CTL 3.48 ± 0.41, OVX 6.17 ± 0.30**, OVX + Fine 3.65 ± 0.55†, **P < 0.01 vs. CTL, †P < 0.05 vs. OVX) and compliance (LV end‐diastolic pressure–volume relation, mmHg/RVU: CTL 1.65 ± 0.42, OVX 4.77 ± 0.37***, OVX + Fine 2.87 ± 0.26††, ***P < 0.001 vs. CTL, ††P < 0.01 vs. OVX). Acetylcholine‐induced endothelial‐dependent relaxation of coronary arteries was impaired in ovariectomized mice and improved by finerenone (relaxation, %: CTL 86 ± 8, OVX 38 ± 3**, OVX + Fine 83 ± 7††, **P < 0.01 vs. CTL, ††P < 0.01 vs. OVX). Finerenone improved decreased ATP production by subsarcolemmal mitochondria after ovariectomy. Weight gain, increased blood pressure, and decreased insulin and glucose tolerance in OVX mice were improved by finerenone. The exercise capacity at race was diminished in untreated OVX mice only. Spontaneous activity measurements in ovariectomized mice showed decreased horizontal movements, reduced time spent in a running wheel, and reduced VO2 and VCO2, all parameters improved by finerenone. Conclusions Finerenone improved cardiovascular dysfunction and exercise capacity after ovariectomy‐induced LV diastolic dysfunction with preserved ejection fraction.

A new HPLC method for the simultaneous quantitative analysis of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) was developed and validated. ATP, ADP, and AMP were extracted from human bronchial epithelial cells with a rapid extraction procedure and separated with a C18 column (3 × 150 mm, 2.7 µm) using isocratic elution with a mobile phase consisting of 50 mM of potassium hydrogen phosphate (pH 6.80). The absorbance was monitored at 254 nm. The calibration curves were linear in 0.2 to 10 µM, selective, precise, and accurate. This method allowed us to quantify the nucleotides from two cell models: differentiated NHBE primary cells grown at the air–liquid interface (ALI) and BEAS-2B cell line. Our study highlighted the development of a sensitive, simple, and green analytical method that is faster and less expensive than other existing methods to measure ATP, ADP, and AMP and can be carried out on 2D and 3D cell models.

Background: The effects of a dietary supplementation with the vegetable omega-3 alpha-linolenic acid (ALA) on cardiovascular homeostasis are unclear. In this context. it would be interesting to assess the effects of camelina oil. Objective: This study aimed to assess the cardiovascular and metabolic effects of camelina oil in hypertensive patients with metabolic syndrome. Methods: In a double-blind, placebo-controlled randomized study, treated essential hypertensive patients with metabolic syndrome received, during 6 mo, either cyclodextrin-complexed camelina oil containing approximate to 1.5 g ALA/d (n = 40) or an isocaloric placebo (n = 41), consisting of the same quantity of cyclodextrins and wheat starch. Anthropometric data, plasma lipids. glycemia, insulinemia. creatininemia. TBARs, high-sensitivity C-reactive protein, and n-3, n-6, and n-9 fatty acids in erythrocyte membranes were measured. Peripheral and central blood pressures, arterial stiffness, carotid intima-media thickness, and brachial artery endothelium-dependent flow-mediated dilatation (FMD) and endothelium-independent dilatation were assessed. Results: Compared with placebo. camelina oil increased ALA (mean +/- SD: 0 +/- 0.04 compared with 0.08 +/- 0.06%. P <0.001), its elongation product EPA (0 +/- 0.5 compared with 0.16 +/- 0.65%, P <0.05), and the n-9 gondoic acid (GA; 0 f 0.04 compared with 0.08 +/- 0.04%, P <0.001). No between-group difference was observed for cardiovascular parameters. Ilowever, changes in FMD were associated with the magnitude of changes in EPA (r = 0.26, P = 0.03). Compared with placebo. camelina oil increased fasting glycemia (-0.2 +/- 0.6 compared with 0.3 +/- 0.5 mmol/L. P <0.001) and HOMA-IR index (-0.8 +/- 25 compared with 0.5 +/- 0.9. P <0.01). without affecting plasma lipids, or inflammatory and oxidative stress markers. Changes in HOMA-IR index were correlated with the magnitude of changes in GA (r = 0.32, P <0.01). Nutritional intake remained similar between groups. Conclusion: ALA supplementation with camelina oil did not improve vascular function but adversely affected glucose metabolism in hypertensive patients with metabolic syndrome. Whether this adverse effect on insulin sensitivity is related to GA enrichment, remains to be elucidated.

Biodiesel is considered as a valuable and less toxic alternative to diesel. However, cellular and molecular effects of repeated exposure to biodiesel emissions from a recent engine equipped with a diesel particle filter (DPF) remain to be characterized. To gain insights about this point, the lung transcriptional signatures were analyzed for rats (n = 6 per group) exposed to filtered air, 30% rapeseed biodiesel (B30) blend or reference diesel (RFO), upstream and downstream a DPF, for 3 weeks (3 h/day, 5 days/week). Genomic analysis revealed a modest regulation of gene expression level (lower than a 2-fold) by both fuels and a higher number of genes regulated downstream the DPF than upstream, in response to either RF0 or to B30 exhaust emissions. The presence of DPF was found to notably impact the lung gene signature of rats exposed to B30. The number of genes regulated in common by both fuels was low, which is likely due to differences in concentrations of regulated pollutants in exhausts, notably for compound organic volatiles, polycyclic aromatic hydrocarbons, NO or NOx. Nevertheless, we have identified some pathways that were activated for both exhaust emissions, such as integrin-, IGF-1- and Rac-signaling pathways, likely reflecting the effects of gas phase products. By contrast, some canonical pathways relative to "oxidative phosphorylation" and "mitochondrial dysfunction" appear as specific to B30 exhaust emission; the repression of transcripts of mitochondrial respiratory chain in lung of rats exposed to B30 downstream of DPF supports the perturbation of mitochondria function. This study done with a recent diesel engine (compliant with the European IV emission standard) and commercially-available fuels reveals that the diesel blend composition and the presence of an after treatment system may modify lung gene signature of rats repeatedly exposed to exhaust emissions, however in a rather modest manner. (C) 2020 Published by Elsevier Ltd.