Leptospira. The acute disease is well-described, and, although it resembles other tropical diseases, it can be diagnosed through the use of serological and molecular methods. While the chronic renal disease, carrier state, and kidney fibrosis due to Leptospira infection in humans have been the subject of discussion by researchers, the mechanisms involved in these processes are still overlooked, and relatively little is known about the establishment and maintenance of the chronic status underlying this infectious disease. In this review, we highlight recent findings regarding the cellular communication pathways involved in the renal fibrotic process, as well as the relationship between renal fibrosis due to leptospirosis and CKD/CKDu.Leptospirosis is a neglected infectious disease caused by pathogenic species of the genus  Leptospira. These highly motile spirochetes, characterized by a helicoidal and thin shape, have two endoflagella that never cover the entire bacterial length, which is one of the most important and peculiar characteristics of these bacteria [Leptospirosis is an infectious and zoonotic disease caused by pathogenic bacteria of the genus bacteria ,2,3,4,5.bacteria ,7,8,9. Tbacteria ,11,12,13Natural disasters, rapid urbanization, and a lack of basic sanitization  are considered risk factors for the occurrence of the disease, both in high- and low-income countries—but mostly the latter ,14,15. PLeptospirosis-related fibrosis, as a sequela from acute disease or due to persistent chronic infection and maintenance of bacteria in proximal convoluted renal tubules, remains overlooked and may be more associated to chronic kidney disease (CKD) than previously considered ,16. KidnA few pathways have been described as inducers or enhancers of kidney fibrosis, such as the transforming growth factor-β1 (TGF-β1) ,23,24,25With that in mind, in this review, we describe the most important pathways involved in tissue fibrosis and summarize recent findings regarding leptospirosis-related renal fibrosis. Furthermore, we highlight the incidental concurrence of this infectious disease and the outbreak of CKDu globally. Leptospira researchers. As an overly complex process, kidney fibrosis in leptospirosis is approached in a stepwise manner for didactic purposes. First, we describe the pathways that are involved in fibrosis, the downstream effectors, and the outcomes of their activation, such as the epithelial–mesenchymal transition (EMT) and morphological and molecular patterns during this process and leptospiral infection. Then, we relate the pathways with the histological and gross alterations seen in kidney fibrosis during chronic leptospirosis. Finally, new approaches towards studying the relationship between leptospirosis and chronic kidney disease are discussed, as this subject has been a growing concern for nephrologists and As reviewed elsewhere ,42, TGF-Depending on the milieu and context of the cell, signaling by this pathway may have developmental effects during embryogenesis and promote tissue remodeling after injury but can also lead to fibrogenic effects when deregulated and overly activated ,43,44,45In addition to post-translational modification inhibitory mechanisms, this pathway has inhibitory Smads that are activated downstream and interact with other Smads, abolishing or diminishing the response to certain stimuli ,59,60. SIt also targets the TGFβ type-I receptor for ubiquitylation, leading to the receptor’s destruction by the proteasome, thus, decreasing the amount of receptors available in the plasma membrane and controlling the pathway’s activation . OverexpOther important TGF-β signaling pathway repressors are Sloan-Kettering Institute (Ski) and Ski novel (SnoN). Both of these have structural similarities, including a SAND domain , which interacts with Smad4 and represses the pathway after its activation . In a stIn line with a repressive role of these proteins on the TGF-β1 pathway, the ectopic expression of both in the cellular model rendered cells resistant to EMT and abolished the production of fibrosis markers, such as α-SMA . Still, Although the pathway has its own regulators and repressors, there are other cytokines and growth factors that may enhance, be activated by  ,29, or aHGF is a pleiotropic growth factor mainly secreted by mesenchymal cells after an injury or an inflammatory process ,72,73,74The exogenous administration of HGF in different animal models of fibrosis leads to less prominent or abolishes organ fibrosis in chronic kidney disease ,35,36, uAnother apparently simple but complex pathway involved in renal fibrosis is the Wnt/β-catenin pathway. It is simple, in that it has a few actors from its activation until the outcome, which are a Wnt-protein ligand, a frizzled receptor (and its co-receptors), and dephosphorylated β-catenin see . On the Drosophila melanogaster and a lack of anterior cerebellum in mice [Until the past decade, studies on the Wnt signaling pathway focused mainly on cancer and its pharmacological treatment, and embryonic development. Colorectal cancer has been directly associated with mutations on important Wnt positive- and negative-regulators, and mutations on the main actors of the pathway cause abnormalities in embryo development, such as a lack of wings in  in mice . Deregul in mice ,85,86,87In total, 19 Wnt ligands, which are glycoproteins that bind to the frizzled receptors to trigger activation, have been described ,88. The Then, stable dephosphorylated β-catenin is released from the complex, enters the nucleus, and interacts with LEF (lymphoid enhancer factor) and T-cell factor (TCF), leading to the expression of target genes of the pathway . As withIn a mouse model of kidney fibrosis caused by UUO, only three Wnt genes  are not upregulated, with a similar expression in sham mice. The other 16 Wnt genes are upregulated, following different patterns throughout the course of the experiment. Some of them are overexpressed within the first week, reach a peak, and then expression levels decline over the next seven days; others are upregulated within the first week and expression levels are sustained throughout the 14-day experiment; and, in others, the expression levels increase progressively during the entire experiment. There is also an increase in cytoplasmic and nuclear β-catenin during the course of the assay, further reinforcing that the canonical pathway is truly activated, with a final response of ECM production and kidney fibrosis evident at day 14 after UUO . Klotho is a membrane bound protein that has been implicated as a negative regulator of the Wnt/β-catenin pathway. In HK-2 cells, Klotho inhibition by si-RNA leads to a more evident EMT after TGF-β1 stimulation, showing that there is a crosstalk between both pathways. In an in vivo model of chronic allograft dysfunction, Klotho is downregulated by 24 weeks after transplantation . ReducedComplement system proteins play important roles in renal fibrosis. As part of the bloodstream’s innate immune system, the effectors and regulators of the classical, alternative, and lectin pathways of the complement system are mostly produced and secreted by the liver directly into the circulation, acting against pathogens and other threats in the bloodstream ,101,102.Classical pathway activation is mediated by the binding of the C1 complex (C1q/C1r/C1s) to immunoglobulins that are bound to target cell membranes. The lectin pathway is activated through the recognition of specific carbohydrates located on the pathogen surface and, finally, the alternative pathway, which is constantly activated at low levels and allows for the generation of C3b. All three pathways converge to a common, terminal pathway, leading to the assembly of the membrane attack complex (MAC), a membrane pore responsible for killing pathogens and/or target cells ,99,100. −/−), an increase in C1s/C1r proteases and diminished levels of C3 during kidney fibrosis were observed. The depletion of C1r in another mouse model causes an evident reduction in C3 production and activation. These findings indicate the relationship between C1r/C1s and C3 production and activation in the fibrotic kidney [In a model of kidney fibrosis induced by UUO C57Bl/6, mice knockout for C1qA and C3 genes have an obvious increase in the production and deposition of ECM proteins, augmented production of α-SMA, reduction of E-cadherin, and increased production and secretion of TGF-β1 from ex vivo pericytes, compared to sham mice. In mice depleted of C1q  to C3aR-, C5aR-, or C3aR/C5aR-deficient mice, the C3aR−/− mice show better renal function, less inflammatory cell infiltration, tubular dilatation, and atrophy, compared to C5aR−/−, C3aR/C5aR−/−, and WT transplanted mice. Interestingly, C5aR- and C3aR/C5aR-deficient mice show signs of renal failure, indicating that C3aR but not C5aR is involved in the poor outcome during renal complement activation. These findings highlight the involvement of the C3a/C3aR pathway in kidney inflammation, leading to subsequent kidney fibrosis [In a study of kidney transplantation from mice lacking the CR1-related gene/protein y  and C3  has been implicated in organ fibrosis, mainly by its capacity to activate the TGF-β1 signaling pathway [Despite its well-known role in maintaining blood volume, blood pressure, and Na pathway ,122,123.Angiotensin II (Ang II) directly binds to cellular receptors and activates important pathways . One of the described targets for Ang II is TLR4, known for its affinity for bacterial LPS. Renal dysfunction is characterized by augmented serum creatinine, albuminuria, and blood urea nitrogen, and kidney fibrosis hallmarks include increased collagens I/IV deposition and elevated levels of TGF-β1 and MMP9. Knocking down MD2—a TLR4 accessory protein that mediates binding of the ligand to its receptor—or blocking it with a small inhibitor molecule leads to a decrease in renal dysfunction and kidney fibrosis seen in wild type mice after the subcutaneous injection of Ang II. It also affects cytokine and chemokine production mediated by NF-κB and ERK pathways. Of note, all the effects observed upon Ang II administration are MD2 dependent, thus, indicating that Ang II signals using the TLR4/MD2 complex . The binding of Ang II to its type-1 receptor (AT1R) is apparently involved in the occurrence of EMT in HK-2 cells induced by high glucose medium, and mediated by the effector proteins mTOR and p70S6K. Silencing the receptor rescues the cells from EMT, characterized by overexpression of E-cadherin, diminished levels of α-SMA, and reduced expression of EMT core transcription factors . ConversAng II is capable of inducing secretion, and activating the latent form of TGF-β1, thus, having a pro-fibrotic role . Ang II Leptospira, as deep as it was possible for us to search, has not been addressed, but may be a possible subject of investigation regarding the sequelae from leptospiral infection. Although this pathway is well-described and associated to renal fibrosis in different pathological contexts ,126,127,Guidelines and definitions for research on epithelial–mesenchymal transition [As research involving EMT has grown in the past 20 years, it has become almost mandatory to elaborate guidelines to help researchers in formulating projects and to develop concepts that could aid in eliminating controversies regarding terms associated to EMT. In June 2020, an extensive committee, on behalf of The EMT International Association, published the ansition . The epithelial to mesenchymal transition is a commonly described phenomenon that takes place during three major events: embryogenesis, cancer, and fibrosis. It is described as a process that occurs after specific stimuli in epithelial cells, leading to the acquisition of mesenchymal phenotype markers and a loss of epithelial markers. Although it may seem to be a static and end-point event, there is a spectrum regarding epithelial to mesenchymal phenotype markers, and, in fibrosis, cells mostly undergo an incomplete EMT, retaining characteristics that resemble both epithelial and mesenchymal phenotypes . Although still controversial, there is evidence that the partial EMT of tubular cells participates in renal fibrosis indirectly, as EMT leads to an increased production of cytokines and chemokines, such as TGF-β1, that stimulate interstitial fibroblasts to produce more ECM proteins and, as such, attract more immune cells and enhance local inflammation ,131,132.Zeb1 and Zeb2, Snai1, and Snai2) are indicative of this process, pointing to the active participation of epithelial cells in the occurrence and progression of kidney fibrosis [In vitro and in vivo studies have also described the occurrence of EMT or partial EMT in tubular epithelial cells. The expression of mesenchymal markers , loss of epithelial markers , increased production of ECM proteins , and over- and down-regulation of specific genes  in kidney fibrosis is a controversial issue among EMT researchers. Using a mouse model expressing the Tomato fluorescent protein and UUO to induce fibrosis, it became clear that tubular cells undergo partial EMT, but do not delaminate and detach from the basement membrane to invade the interstitium, as less than 1% of interstitial cells were tdTomatotruction .Twist1 or Snail1 genes have 48.2% and 64.3% lesser tubular epithelial cells, respectively, that underwent EMT after UUO compared to wild type mice. In addition, those mice displayed less collagen deposition and better renal function. No signs of tubular epithelial cell detachment are seen both in wild type and knockout mice, indicating that these cells underwent partial EMT and that they are needed to induce α-SMA+ myofibroblasts. EMT in kidney fibrosis is associated with a lower expression of important solute transporters, such as aquaporin 1 and Na+/K+-ATPase, and suppressing the expression of core EMT-transcription factors may explain the better renal function in Twist1 or Snail1 KO mice [Mice lacking  KO mice . Zeb1 and Zeb2. Both transcription factors are overexpressed during fibrosis and EMT, and downregulate the expression of E-cadherin [EMT is under strict control in adult humans and animals ,131,136.cadherin ,138,139.cadherin . Leptospira disseminate through the bloodstream and reach target organs, especially the kidneys, liver, and lungs [During infection, pathogenic nd lungs . The kidLeptospira from the blood within the first few days [Leptospira has the ability to subvert and scape phagocytosis, invade renal convoluted tubules, and persist in this niche [Neutrophil-depleted mice have a higher leptospiral burden in the kidneys 15 days post-infection, which demonstrates that neutrophils help to eliminate few days ,143. Altis niche ,147,148.The transition of acute kidney injury (AKI) caused by leptospiral infection to chronic infection and, consequently, kidney fibrosis is still not completely understood, and many research gaps remain open. This is also applied to the transition from AKI to CKD of non-infectious cause . ContinuLeptospira in the convoluted tubules is responsible for the overactivation of inflammatory pathways, triggering pro-fibrotic signals. One month after infection, cellular infiltration in the kidneys is characterized by CD3-positive T-cells and CD11b-positive macrophages/monocytes, but no more neutrophils are observed [The sustained stimulus caused by the presence of observed . In a raobserved . The total healing of the kidney, with no signs of tubular necrosis or glomerular sclerosis, can also be observed. Activation of the NF-κB and NLRP3/IL-1β pathways are implicated in the recrudescence of signs in rat kidneys 180 days after injury, with a low-grade inflammation occurring together with activation of Angiotensin II, as well as collagen and fibronectin deposition in the interstitium. These findings indicate that sustained inflammatory stimulus may lead to kidney fibrosis after acute injury .−/− mice have higher bacterial loads, greater susceptibility to infection, acute renal lesions, and more evident kidney fibrosis 90 days post-infection, with more tubulointerstitial collagen deposition than wild-type littermates. These findings point to a role of Daf1 in controlling the bacterial burden and inflammation during the acute phase, thus, helping to reduce chronic lesions and fibrosis [As described above, the complement is another important system activated during leptospirosis. Fluid-phase or membrane-associated negative complement regulatory proteins avoid overactivation of this system and, consequently, tissue damage ,101,105.fibrosis .L. interrogans serovars Manilae str. L495, Copenhageni str. Fiocruz L1-130, and Icterohaemorraghiae str. Verdun [Leptospira in cell culture and, as such, is associated with bacterial killing. Inducible nitric oxide synthase (iNOS) expression—and, consequently, NO production—is also associated with kidney fibrosis as the disease transitions from an acute to chronic state in C57BL/6J mice [Cytotoxicity mediated by nitric oxide (NO) is one of the mechanisms used by macrophages to control leptospiral infection. Use of the TLR2/NOD2 agonist CL429 increases NO production by mice peritoneal and bone marrow-derived macrophages when exposed to . Verdun . The NO /6J mice .Leptospira-related kidney disease [L. interrogans sorovar Copenhageni str. L1-130 have been investigated [Of note, the initial lesion and the type of cellular infiltrate play roles in the disease progression and, thus, contribute to the evolution to chronic and fibrotic  disease ,150,152.stigated . Althougstigated ,142,152.In brief, acute leptospirosis is characterized mainly by inflammatory and immune responses mediated by cells.Leptospira infection. Chronic leptospiral infection leads to kidney fibrosis, but the underlying mechanisms and pathways involved are still poorly elucidated. In the first part of this review, the potential pathways related to renal fibrosis were described. In this section, we will address the current knowledge on the findings related to renal fibrosis caused by Leptospira inoculation [Leptospira and fibrosis induction have begun to be elucidated [Hamsters and guinea pigs are considered good animal models for acute and severe leptospirosis, as both animals die within the first 5–10 days after culation ,153,154.ucidated ,152,155.Using wild-type and different knockout mouse lineages to understand which pathways may be activated during chronic leptospiral infection, Fanton d´Andon et al.  demonstrChronic leptospiral infection has been associated to fibrosis in many different models of mouse infection ,152,156.Leptospira outer membrane proteins induce the accumulation of ECM proteins in renal epithelial cells through activation of the TGF-β1/Smad3 pathway, thus, contributing to the evolution of fibrosis associated to chronic infection [L. santarosai serovar Shermani enhance the secretion of collagen types I and IV by HK-2 cells, and the process is mediated by the TGF-β1 pathway. nfection . Therefonfection , outer mBone marrow derived-macrophages can transdifferentiate into myofibroblasts—cells that are involved in the secretion of α-SMA. This transition is coordinated by the TGF-β1/Smad3 pathway, in a process known as the macrophage–myofibroblast transition (MMT). In chimeric mice that had their bone marrow depleted by radiation and reconstituted by exogenous GFP-expressing C57BL/6 bone marrow, GFP-positive myofibroblasts were observed in the kidneys after UUO. +Smad3+/+ and GFP+Smad3−/−. Mice lacking Smad3 did not present bone marrow-derived myofibroblasts after UUO surgery, thus, corroborating the involvement of TGF-β1/Smad3 in the MMT within the kidney [To ascertain that the MMT occurs, orchestrated by TGF-β1/Smad3, C57BL/6 mice were irradiated and reconstituted with bone marrow from GFPe kidney . L. interrogans serovar Copenhageni str. L1-130 and the role of leptospiral infection in progression of CKD, Chou et al. [To understand which pathways are involved in chronic infection and kidney fibrosis caused by u et al.  performeLeptospira infection is associated with CKD progression and may be an underlying cause of CKDu. Furthermore, as both Wnt and TGF-β1 pathways are enhanced in this model, it is suggested that they contribute to the progression of kidney fibrosis [Comparing orthologous genes from mice and humans with leptospiral infection plus nephrotoxic stimulus and CKDu, respectively, there is an overlap of enriched genes. These findings provide support for the hypothesis that fibrosis .L. interrogans serovar Manilae str. L495 were shown to produce greater amounts of fibronectin and collagen type IV, compared to non-infected cells. Morphological alterations, such as spindle shape, loss of cell–cell contact, cell grouping, and abundant ECM production, were also evident in infected HK-2 cells. Cellular and molecular mechanisms underlying this EMT process are under investigation. Further evidence of a fibrotic process triggered by infection with leptospires comes from recent in vitro and in vivo findings from our group. HK-2 cells infected with L. interrogans infection are shown in L. interrogans serovar Manilae str. L495  of Leptospira showed a few more areas of interstitial, and peritubular fibrosis (indicated by blue arrow heads) than those infected with less (1 × 106) Leptospira (indicated by black arrow heads). These findings show the importance of this serovar in causing acute disease as well as chronic and profibrotic infection.Collagen deposition was also seen in Periodic acid–Schiff  at 15 dai) impaired kidney function and fibrosis are present in almost every case of CKD, and (ii) the outcome of CKD is renal failure, requiring kidney transplantation. Highlighting those aspects is of utmost importance, as CKD patients demand constant care, such as dialysis, hospital care, and transplantation, thus, elevating healthcare expenditures [As CKD has become one of the most prevalent kidney diseases, due to systemic hypertension or diabetes, it is worth highlighting two major points:  [Another recent systematic review of observational studies has disclosed a correlation between leptospirosis and CKD. Poor renal function, characterized by a low estimated glomerular filtration rate (eGFR), is more prevalent in anti-er eGFR) .Leptospira antibodies and 13 were leptospiral 16S rRNA-positive by PCR from urine samples. Despite being asymptomatic, Leptospira molecular detection in those individuals evidence that the chronic carrier state is a reality. Although the likelihood of renal damage and kidney fibrosis was not assessed in this work, the presence of Leptospira in the kidneys may be a risk factor for the development of CKD [Leptospira shedding [In a study considering a Peruvian community, 189 out of 314 asymptomatic participants had high titers of anti-t of CKD . In a stshedding .Leptospira, showing that chronic leptospirosis may be a risk factor for the development of CKD [Mesoamerican nephropathy, a type of CKDu, has brought to light evidence that chronic leptospirosis could be a cause of CKD. This condition is highly prevalent in male sugarcane workers that are in close contact with flooded areas at least during a period of the year. It is proposed that leptospirosis may be one of the causes (but not the only) of Mesoamerican nephropathy, and that exposure to other potentially nephrotoxic conditions or substances may influence the occurrence of the disease . Recentlt of CKD .CKD/CKDu are important health problems that directly impact the quality of life and health systems all over the world. Despite recent advances in nephrology and conscientization campaigns, the searches for medical assistance and treatment are still insufficient. Assuming that leptospirosis may play a role in the development of CKD/CKDu, and considering the overlap of both conditions in Asia, Central and South America, and Sub-Saharan Africa, studies into the associated pathophysiology, cellular communication pathways, and treatment strategies to avoid or control the fibrotic process are needed. Leptospira infection, is a well-documented manifestation of the disease; however, studies on the mechanisms associated with the pathophysiology of leptospiral renal fibrosis are still scarce and deserve to be deepened. Deciphering the cellular communication pathways in Leptospira-induced kidney fibrosis may shed light on the specific actors involved in this process.As discussed throughout the text, kidney fibrosis and, consequently, CKD, as a sequela of


Outdoor public places and natural environments gained importance due to restrictions affecting access to sport facilities. Further research is needed to evaluate benefits and therapeutic values of outdoor nature for physical and mental health in times of a global pandemic to maintain resilient societies, as it might impact future active living and life satisfaction.The COVID-19 pandemic has dramatically impacted human lifestyles across the world. Lockdowns and home confinement decreased prior opportunities for everyday physical activity. To retrospectively assess how the Austrian population coped with these aspects of the crisis, we conducted a cross-sectional online survey from March to September 2021 using a structured questionnaire in German. In total, 1214 participants  living across Austria shared self-reported information on sociodemographic characteristics, indoor and outdoor physical activity, reasons for being outdoors, and life satisfaction before and after the emergence of the virus. As a result, overall indoor physical activity significantly decreased in a before–during COVID-19 crisis comparison, although exercising at home with online instructions increased by about 63%. Exercising outdoors increased overall, specifically in periurban forests and rural areas, both by about 9%. Life satisfaction decreased significantly by 19.7% ( Since March 2020, the outbreak of the newly detected severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causing an infectious disease soon classified as Coronavirus Disease 2019 (COVID-19) has changed the world [In this article, we focus on changes in active living during the COVID-19 pandemic, anticipating the multifaceted impacts on (semi)professional sports, fitness, and recreational active living. Sport mega-events such as the 2020 Olympic Games in Tokyo, the 2020 UEFA European Football Championship, and countless tournaments in many other sports such as golf, tennis, Formula 1 motor racing etc. were postponed or, more often, canceled. In most countries, indoor and also outdoor sport facilities such as gyms, fitness centers, and sport clubs were closed for months, or permanently . From a public health perspective, regular exercise is one of the most influential measures for health prevention. Physical activity improves overall quality of life, physical and mental health by positively affecting muscular and cardiorespiratory fitness, as well as bone and functional health, while reducing the risk of non-communicable disease, i.e., hypertension, coronary heart disease, stroke, metabolic syndrome, adiposity, various types of cancer, and depression ,5. Most Systematic gender differences in frequency and motivation for physical activity were observed in different studies. Exemplarily, Molanorouzi et al. found that females seem to be motivated by physical condition and appearance, whereas men have more interest in challenges and competition, which could influence the reasons of exercising . As a coIn times of urbanization and digitalization enforcing a sedentary lifestyle, people commonly spend too much time engaging in behaviors that consume low amounts of energy. Already in 2018, Guthold and colleagues described worldwide trends in insufficient physical activity, ultimately resulting in a range of chronic health conditions . RecentlAddressing the inherent conflict of goals between home confinement and active living, the impact of the pandemic on physical activity gained high interest during the last months . In the The impacts of the COVID-19 pandemic were on a global scale, with many nations following similar general paths in fighting the spread of the pandemic. However, the constantly adapted lockdown measures mirrored the different pandemic waves and were not identical in intensity and duration, not even in neighboring countries or even federal states. This warrants a region-specific evaluation on the respective population-wise perceptions of the national regulations in force during that period. In Austria, measures against the spread of the virus became compulsory on 16 March 2020, i.e., the first lockdown in a series of lockdowns in response to infection waves . Only thA rapidly increasing number of publications on societal and public health implications of the pandemic gradually add preliminary empirical data on the value of freely admissible urban and periurban public space, particularly green space, in taking up the role of commercial sport centers that are closed due to the pandemic . As far Physical activity has been one of the most important public health issues in modern civilizations already before the pandemic, which is legitimately often labelled as a public health crisis. We can also speak of a fitness crisis for a large part of the population, with many questions emerging concerning the risk–benefit ratio of physical activity in crowded public areas and indoor facilities alike. By retrospectively assessing behavioral and perceptive changes from an Austrian perspective, this report adds to the current literature on health and well-being effects of the pandemic on the Austrian population novel evidence from 2021, the second year of the crisis ,17,18. IThis non-representative, cross-sectional study aimed at identifying the self-reported prevailing perceptions regarding active living and life satisfaction in the context of the pandemic using an online questionnaire in German, targeting the German-speaking Austrian adult population. The online survey was pretested by 14 participants in February 2021 to review completeness and comprehensibility of the items. The adapted online survey was accessible barrier-free via the web-based survey tool SoSci Survey from March 8 until 8 September 2021 . During The cover page informed participants about the study aim, and their consent was implicitly obtained when completing the survey. Participants were informed that the period before the COVID-19 pandemic was defined as the time before 16 March 2020, when the first lockdown started in Austria . A furthUsing snowball sampling, the link was spread via commonly used social networks such as Facebook or WhatsApp of several public sports centers, organizations, clubs, and by private individuals such as national handball team members and trainers, and specifically as an online article by Sportunion Austria , and on The first part of the online survey collected data on socio-demographic characteristics age (in years), gender , residence , and education level . We further assessed ratings on frequency of physical activity in six different indoor  instructions) and outdoor  places. Each of these activities were separately assessed before and during the COVID-19 pandemic using a Likert-scale format: 1: never, 2: ≤1 h per week, 3: 1 h per week, 4: 1–2 h per week, 5: 3–5 h per week, and 6: ≥5 h per week. Moreover, participants rated six reasons for being outdoors  before and during the COVID-19 pandemic, using a 5-point Likert-scale format. The response options were: 1: strongly disagree, 2: disagree, 3: neutral, 4: agree, and 5: strongly agree. The last part of the questionnaire included the three items of the standardized Short Life Satisfaction Questionnaire for Lockdowns (SLSQL) to assess participants’ life satisfaction before and during the COVID-19 pandemic . The resp < 0.05. We used descriptive statistics to report categorical data as absolute and relative frequencies, and continuous data as mean and standard deviation (SD). All statistical analyses were performed using the commercial statistical software SPSS Statistics for Windows, Version 27.0 . We employed paired samples T tests to calculate total as well as separately for females and males in scored responses between the before and during COVID-19 period. We summarized the three items of the SLSQL before and during the COVID-19 pandemic, respectively, to generate the total score Life Satisfaction for each of these two periods. We determined internal consistency for the three items of the SLSQL before and during the COVID-19 crisis, respectively, using Cronbach’s alpha. We further calculated the effect sizes to determine the magnitude of the change score using Cohen’s d and interpreted them as small , moderate , or large , as well as lower and upper 95% confidence intervals (CI). Statistical significance was set at n = 691, 56.9%) compared to males , 32.5% in Lower Austria (n = 394), and the rest in the remaining parts of the country . Further, 15.9% (n = 193) of participants reported having a primary, 41.2% (n = 500) a secondary, and 42.9% (n = 521) a tertiary education level. Average age of study participants was mean 36.92 years . Average completion time of the survey was mean 6.0 min SD 2.2.The web link to the online survey was accessed 3161 times; 1486 participants started and 1243 of those fully completed the survey (83.7% completion rate). We excluded three data sets from people indicating to be of diverse gender, as we were interested in potential differences between males and females, and 26 data sets from participants living abroad, as we were interested in circumstances caused by the Austrian lockdowns, 12 from Germany, seven from Italy, three from Switzerland, and one person each from Luxembourg, the United Kingdom, Unites States of America, and Mongolia. Thus, the final sample included 1214 participants, with a higher proportion of females (p < 0.001), with a moderate effect size . In contrast, we found an overall statistically significant average increase of 5.29% from mean 2.28 (SD 0.90) before to mean 2.40 (SD 0.95) during COVID-19 for the frequency of outdoor sports facility use .p < 0.001) in the frequency of indoor sports activity at a fitness center (49.6%), a sport club (53.3%), and a school/university sports center (29.3%). In contrast, we found increased indoor sports activities at home independently as well as with online instructions (25.7 vs. 62.5%) and respective differences among the female and male groups . Compared to males, females showed a strong increase for exercising at home with online instructions (42.75% vs. 73.81%). We found a small decrease before vs. during the COVID-19 pandemic in the frequency of outdoor sports activity in public green spaces (4.3%), which was only statistically significant in males (p = 0.018). We found a small increase in the frequency of outdoor sports activity in urban areas (4.8%), which was also only statistically significant in males (p = 0.007). Participants reported small increases of slightly over 9% in the frequency of outdoor sports activity in periurban forests and also rural areas, with statistically significant differences among the gender groups .Specifically, we revealed large decreases before vs. during the COVID-19 period and gender differences , with a small effect size . The top three changes before vs. during the COVID-19 pandemic were identified for mental health (8.6%), distraction (7.8%), and urge to move (6.7%). Whereas all reasons statistically significantly increased in females, males only showed a statistically significant increase in the ratings for “recreation” and “social contacts”.As shown in p < 0.001, d = 0.79). We found highest changes in the total score of I1 with −25.8%, and observed statistically significant decreases in the total, and also in the female and the male samples for all items, with all p values from paired samples T tests smaller than 0.001. The total score Life Satisfaction was lower in males compared to females but with a higher change in males compared to females (−20.0 versus −19.4%).Statistical analysis showed that the total score of SLSQL, i.e., Life Satisfaction, decreased statistically significantly by 19.7% during compared to before the COVID-19 period  during an unprecedented time during a global pandemic. As the epidemiological situation changed, and still changes, dynamically and unpredictably, the answers given by the respondents in April might differ from those in July. However, this retrospective study focused on self-reported data comparing a time period before the first lockdown measures in March 2020 to a time period after this event. There is the possibility of unmeasured confounding associated with opinions on the COVID-19 pandemic such as lack of interest or even resistance to acknowledge the crisis. Most participants were highly educated and rather young, with an average age of about 37 years, limiting generalizability of the results and representativeness of our study population to the entire population. To further elucidate reasons for differences in the perceptions of gender in active living and life satisfaction of the Austrian population during the COVID-19 pandemic, further research using mixed-methods studies and longitudinal study designs should be conducted.Regular physical activity is an important public health issue, with its positive effects on physical and mental health being recognized since ancient times. Despite the importance of sports, our non-representative cross-sectional online study revealed decreased physical activity and life satisfaction during the COVID-19 pandemic in a quite large sample of Austrian inhabitants. Further, outdoor experience gained popularity, although to a lesser extent than anticipated, whereas home-based online classes boomed due to the closure of indoor sports facilities and other venues of social life.As overall active living decreased during the crisis, public health messages should highlight the value of outdoor sports and nature contact for physical and mental health. Further, sport programs in outdoor environments should be supported to enlarge the current options for regular physical activity and prevent eventual post-pandemic health disparities. These multisector measurements require the cooperation among different stakeholders  and substantial financial support for hygiene concepts to support sports clubs and reduce fear and actual as well as perceived infection risk.While recognizing the importance of restricting mobility to contain the COVID-19 pandemic, a bundle of nation-wide behavioral strategies for preventing global acute and chronic physical and mental health effects during lockdowns is key. We strongly recommend designing, implementing, and evaluating national programs for safe outdoor or home-based approaches for regularly and repetitively disrupting physical inactivity in all strata of the population.


Thorough postmortem investigations of fatalities following vaccination with coronavirus disease 2019 (COVID-19) vaccines are of great social significance. From 11.03.2021 to 09.06.2021, postmortem investigations of 18 deceased persons who recently received a vaccination against COVID-19 were performed. Vaxzevria was vaccinated in nine, Comirnaty in five, Spikevax in three, and Janssen in one person. In all cases, full autopsies, histopathological examinations, and virological analyses for the severe acute respiratory syndrome coronavirus 2 were carried out. Depending on the case, additional laboratory tests  and neuropathological examinations were conducted. In 13 deceased, the cause of death was attributed to preexisting diseases while postmortem investigations did not indicate a causal relationship to the vaccination. In one case after vaccination with Comirnaty, myocarditis was found to be the cause of death. A causal relationship to vaccination was considered possible, but could not be proven beyond doubt. VITT was found in three deceased persons following vaccination with Vaxzevria and one deceased following vaccination with Janssen. Of those four cases with VITT, only one was diagnosed before death. The synopsis of the anamnestic data, the autopsy results, laboratory diagnostic examinations, and histopathological and neuropathological examinations revealed that VITT was the very likely cause of death in only two of the four cases. In the other two cases, no neuropathological correlate of VITT explaining death was found, while possible causes of death emerged that were not necessarily attributable to VITT. The results of our study demonstrate the necessity of postmortem investigations on all fatalities following vaccination with COVID-19 vaccines. In order to identify a possible causal relationship between vaccination and death, in most cases an autopsy and histopathological examinations have to be combined with additional investigations, such as laboratory tests and neuropathological examinations. Vaccination with potent and well-tolerated vaccines against coronavirus disease 2019 (COVID-19) is considered an effective measure to contain the coronavirus pandemic. At the same time, the vaccines are intended to prevent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections, or at least severe or fatal COVID-19 infections. So far, four COVID-19 vaccines have been approved by the European Medical Agency (EMA), all encoding the spike protein antigen of SARS-CoV-2. Those are two mRNA-based vaccines: BNT162b2  and mRNA1273 ; and two recombinant vector-based vaccines: adenovirus type 26 vector COVID-19 Vaccine Janssen , and the recombinant chimpanzee adenoviral vector vaccine ChAdOx1 nCoV-19 . As of June 30, 2021, a total of 74,871,502 vaccinations have been administered in Germany, including 54,898,640 with Comirnaty, 11,570,155 with Vaxzevria, 6,471,052 with Spikevax, and 1,931,655 with Janssen. A total of 30,986,128 people were fully vaccinated . Due to Since end of February 2021, venous thromboses at unusual sites including cerebral venous thrombosis (CVT) and/or splanchnic vein thrombosis in combination with moderate to severe thrombocytopenia occurred in several hundred individuals typically 4 to 30 days after vaccination with Vaxzevria and Janssen , 55–57. Besides VITT, other immune syndromes were reported following COVID-19 vaccination including thrombotic thrombocytopenic purpura/hemolytic uremic syndrome, (auto-)immune thrombocytopenia or Guillain-Barré Syndrome. Furthermore, peri-/myocarditis was reported as a rare event following vaccination with mRNA-based vaccines .In Germany, the Paul Ehrlich Institute is responsible for the registration of adverse events after vaccination. The Paul Ehrlich Institute summarizes all reports it receives, irrespective of a causal relationship with vaccination, and regularly issues safety reports . The PauA total of 25 fatal hemorrhages, 24 of which were cerebral, were reported after vaccination with Vaxzevria. A total of 27 fatal hemorrhages occurred after Comirnaty vaccination, 18 of which were cerebral hemorrhages. A total of 2 fatal hemorrhages were reported after Spikevax vaccination.There were 5 deaths reported with myocarditis ranging from 1 to 50 days after vaccination with Corminaty. All 5 individuals had preexisting cardiovascular diseases that were potential causes of death. There was one patient who died 2 days after Vaxzevria vaccination from septic shock, acute renal failure, and myocarditis. Anaphylactic reactions were reported in 293 cases. Among these, 94 cases had the highest level of diagnostic safety. No fatal anaphylaxis was reported .In cases of deaths with a temporal relation to vaccinations, autopsies together with additional investigations can provide an important contribution for clarifying the causal relationship between vaccination and death. While numerous studies have been published on postmortem investigation of COVID-19 , 60, 62,From 11.03.2021 to 09.06.2021, postmortem investigations of 18 persons who recently received a vaccination against COVID-19 were performed on behalf of the public prosecutor's offices in Bielefeld, Detmold, and Münster. The catchment area of these public prosecutor’s offices comprises around 3.24 million people, which accounts for about 4% of Germany’s population , 59.The following data were available for evaluation: case histories provided by the investigating authorities, medical records for in-hospital deaths, autopsy protocols, and reports on additional investigations .The autopsies were performed in accordance with the guidelines of the German Society of Legal Medicine for forensic autopsies . In addiThe following data were gathered in a registry: age and gender of the deceased, vaccine, 1st or 2nd vaccination, time interval between vaccination and death, place of death, findings of the autopsy and the additional investigations, cause of death, and the assessment of the possible causal relationship between vaccination and death.Within the study period, postmortem investigations were performed in 18 fatalities  in temporal relationship to COVID-19 vaccination. Table The mean age of the deceased was 62.6 years (age range: 32–91 years). Vaxzevria was vaccinated in nine, Comirnaty in five, Spikevax in three, and Janssen in one person. The time interval between the last vaccination and death ranged from a few minutes (case 17) up to 14 days (case 9). Case 17 involved a person who collapsed in a vaccination center immediately after vaccination; prompt resuscitation efforts were unsuccessful. A total of 12 deaths occurred at home, 4 deaths at a hospital, 1 death at a vaccination center, and 1 death at work.In 13 deceased, the cause of death was attributed to preexisting diseases while postmortem investigations did not indicate a causal relationship to the vaccination. In one case (case 6), myocarditis was found to be the cause of death . In case 14, the cause of death was established in hospital, being CVT and cerebral hemorrhage with hypoxic brain damage. The diagnosis was confirmed by postmortem investigations. A second patient who died in a hospital (case 12) suffered a severe anaphylactic shock during narcosis induction, resulting in hypoxic brain damage. This patient was vaccinated 12 days before she developed the shock symptomatic. She received unfractionated heparin after the adverse event. The serum-activated platelets were stronger in presence of PF4 than in presence of PF4/heparin complexes. This suggests that the patient had true VITT antibodies related to vaccination, rather than HIT antibodies related to later heparin therapy. The autopsy revealed multiple thrombi, including in the cerebral venous sinuses. Neuropathological examination found these to be fresh thrombi that must have formed after the onset of hypoxic brain damage  antibodies against a specific antigen, which occurs within minutes or up to 4 h after exposure to the vaccine . The medThe scientific literature on vaccine-related deaths is vast. However, a majority of this literature is based on data from passive voluntary reporting systems. In contrast, the forensic literature on fatal anaphylaxis following vaccination is surprisingly scarce. In a recent review, Palmiere et al.  found siFor forensic pathologists, diagnosis of anaphylaxis is challenging. Postmortem investigations should include full autopsy, histology, immunohistochemistry, and postmortem biochemistry. Macroscopical findings may include cutaneous swelling, upper airway edema, and hyperinflation of the lungs with mucus plugging , but areTo our knowledge, no anaphylaxis-related deaths have been reported following COVID-19 vaccination. In all deceased persons investigated in the present study who died within 1 day of vaccination, there was no evidence of an anaphylactic event. In particular in case 17, in which an anaphylactic shock was initially suspected due to the short time interval between vaccination and death, an anaphylactic reaction could be largely excluded by the autopsy as well as postmortem biochemistry and histopathological examinations.VITT is characterized by thrombocytopenia, combined with thrombosis in most cases. Thrombosis can occur in both arterial and, more common, venous system. However, a distinctive feature of VITT is thrombosis in unusual locations. These include CVT, as well as splanchnic venous thrombosis , 55, 56.Cases have been reported to be typically identified between 4 and 30 days after vaccination with either of two currently EMA-approved vector vaccines Vaxzevria and Janssen . One sinIn initial reporting, female sex and younger age were proposed possible risk factors, but this may simply have reflected the demographics of the first populations to be vaccinated, i.e., often young female health care workers , 48, 55.Death with VITT appears to occur very rarely. Up to 31.05.2021, 21 patients had been reported to the Paul Ehrlich Institute who had died of VITT in temporal relationship with a Vaxzevria vaccination . In our In the clinical setting, the diagnosis of VITT is made by detecting high titers of anti-PF4/heparin antibodies with an antigen-binding assay (ELISA) for PF4/heparin antibodies in combination with positive functional platelet activation assays such as the PF4 induced platelet activation test (PIPA) and the heparin induced platelet activation test (HIPA). The detection of thrombocytopenia is done by determining the platelet count in the blood , 44.Postmortem, a valid determination of the platelet count is not possible if a VITT is suspected, since numerous processes take place in the blood compared to the living, which have a falsifying effect on the number of countable platelets . HoweverOur study shows that an autopsy and platelet laboratory testings are not sufficient to clarify a possible causal relationship between VITT and death. Indeed, it could only be decided in the synopsis of the anamnestic data, the autopsy results, laboratory diagnostic examinations, and histopathological and neuropathological examinations that VITT was very probably the cause of death in only two of our four cases. In the other two cases, no neuropathological correlate of VITT explaining death was found, while possible causes of death emerged that were not necessarily attributable to VITT.The first reports concerning cases of myocarditis temporally linked to the vaccination with the mRNA vaccines from Comirnaty and Spikevax came from the Israeli Ministry of Health and the U.S. Department of Defense . To dateViruses are commonly the cause of myocarditis. In addition to other infectious agents, however, drugs, toxins, or autoimmune diseases can trigger myocarditis. In some cases, the etiology cannot be determined at all . MyocardSymptoms of myocarditis following other vaccinations have been reported after 1 week at the earliest. Due to the considerably earlier occurrence of myocarditis after COVID-19 vaccinations, the 1st vaccination, rather than the 2nd vaccination, is suspected to possibly trigger the autoimmune processes leading to myocarditis .Regarding our investigated case of myocarditis, the period between the 2nd vaccination with a COVID-19 vaccine and the time of death was 11 h, which is also quite short. A causal relationship to vaccination was considered possible, but could not be proven beyond doubt.The postmortem confirmation of myocarditis requires histologic examination of the heart and fulfillment of the Dallas criteria , 6. KytöThe results of our study demonstrate the necessity of postmortem investigations on all fatalities following vaccination with COVID-19 vaccines. In 13 out of 18 cases, deaths were attributed to preexisting pathological changes. In these cases, extensive postmortem investigations did not reveal any evidence for known complications of vaccination with COVID-19 vaccines. In one case, myocarditis was determined to be the cause of death. A causal relationship to vaccination was considered possible, but could not be proven beyond doubt. In four cases, there was macromorphological evidence of VITT, which was confirmed by laboratory tests. Of those four, only one was diagnosed before death. Additional neuropathologic and histopathologic examinations revealed that VITT was the very likely cause of death in only two of the four cases. In order to identify a possible causal relationship between vaccination and death, in most cases an autopsy and histopathological examinations have to be combined with additional investigations, such as neuropathological examinations and laboratory diagnostics including platelet laboratory testings. This procedure requires close collaboration between several medical disciplines.


Multi-walled carbon nanotubes (CNTs) were added to provide electrical conductivity to bio-based polymer blends with improved toughness . A preliminary study including three different Pebax grades was carried out to select the grade and the composition that would best improve the impact properties of PA410. Thus, tough multiphasic PA/Pebax/CNT nanocomposites (NCs) with enhanced electrical conductivity were obtained. The CNTs were added either: (1) in the form of pristine nanotubes or (2) in the form of a PA6-based masterbatch. Hence, PA410/Pebax/CNT ternary NCs and PA410/PA6/Pebax/CNT quaternary NCs were obtained, respectively, up to a CNT content of 1 wt%. The ternary and quaternary NCs both showed similar mechanical and electrical properties. The electrical percolation threshold decreased with respect to previously studied corresponding NCs without Pebax, i.e., PA410/CNT and PA410/PA6/CNT, due to the partial volume exclusion effect of Pebax over the CNTs that were dispersed mainly in the PA matrix; materials with percolation concentrations as low as 0.38 wt% were obtained. With respect to mechanical properties, contrary to the NCs without Pebax, all the PA/Pebax/CNT NCs showed a ductile behavior and impact strength values that were from three to five-fold higher than that of the pure PA410. Polyamides are one of the most widely used families of engineering polymers due to their good mechanical and thermal properties; their applications include the electrical, electronics and automobile industries ,2,3. PolSimilar to traditional polyamides, bio-polyamides also have outstanding mechanical properties , but theTo date, the elastomers used to improve the toughness of brittle polymers have generally been petrochemical, and very few bio-based or biodegradable elastomers have been studied. However, the study of bio-based elastomers has proven to be very interesting, considering that impact properties can be enhanced without sacrificing the renewable nature of the material. In this sense, the partially bio-based polyether-block-amide copolymers (Pebax) series is one of the most studied groups of bio-based thermoplastic elastomers ,21,22,23c) is achieved, i.e., when a three-dimensional network of interconnected nanotubes is formed in the polymeric matrix. Moreover, it is widely known that CNTs can improve the mechanical properties of polymeric materials. For this reason, the combination of thermoplastic elastomers and CNTs is of particular interest. The simultaneous addition of an elastomer and CNTs in a polymeric system can bring toughness and electrical conductivity, respectively; at the same time, the combination is very interesting from an engineering point of view. Moreover, the addition of an immiscible phase in a polymeric matrix can reduce the electrical percolation concentration due to a volume exclusion effect, thus, reducing the amount of CNTs needed to attain electrical conductivity. Gonzalez et al. [Furthermore, polymer nanocomposites (NCs) based on polyamide and carbon nanotubes (CNTs) have attracted much interest in recent decades ,26,27,28z et al. , for insz et al.  observedz et al. ,33,34.In this study, once the Pebax grade and composition that would best improve the properties of the pure PA410 was established, and in order to combine the benefits of adding Pebax and the advantages provided by CNTs, multiphasic PA/Pebax/CNT NCs were prepared. Two alternative processing routes were employed in preparing the multiphasic NCs. First, CNTs were added in pristine form, thus, obtaining PA410/Pebax/CNT NCs. Then, they were added in the form of a PA6/CNT masterbatch, which has previously been proven an efficient procedure to add CNTs to PA410 [® Q150-D, kindly provided by DSM .The PA410 used in this work was EcoPaXX® Rnew® 25R53 SP 01, 40R53 SP 01, and 63R53 SP 01). They were provided by Arkema  and consist of flexible polytetramethylene oxide (PTMO) and rigid polyamide 11 (PA11) constituents [In this study, we used three grades of commercially available and partially bio-based Pebax elastomers  The PA410/Pebax2 system shows full immiscibility with some level of interfacial interaction between both components, probably related to the presence of polyamide groups in the matrix and the elastomer. (2) The crystalline structure of PA410 slightly changed on the blends, showing slightly lower lastomer ,39,40 anlastomer ,43,44 blAs a result of both preliminary studies, PA410/Pebax2 70/30 composition was selected as the base composition for the preparation of the CNT-filled nanocomposites.TM . Additionally, a PA6-based MWCNT masterbatch, also manufactured by Nanocyl  and commercialized as PlasticylTM PA 1503, containing 15 wt% of the aforementioned NC7000TM CNTs was also used to obtain the PA410-based NCs.The pristine carbon nanotubes used were Nanocyl NC7000TM PA 1503 masterbatch were dried in a dry air dehumidifier  for 48–72 h at 80 °C. The Pebax elastomers were dried in an air-circulation oven at 60 °C for 24 h. Unfilled CNT-free PA410/Pebax blends were obtained by melt mixing at 260 °C using a Collin ZK 25T SCD 15 Teach-Line corotating twin-screw extruder-kneader with a screw rotation speed of 200 rpm . The diameter and length-to-diameter ratio of the screws were 25 mm and 18, respectively. The CNT-filled PA410/Pebax/CNT ternary NCs were prepared by direct melt mixing of the pristine CNTs and the quaternary PA410/PA6/Pebax/CNT NCs were prepared using the masterbatch, which were also obtained under the same conditions using a Collin ZK25 corotating twin screw extruder-kneader . The diameter and length-to-diameter ratio of the screws were, in this case, 25 mm and 30, respectively. 3/s, 3 s, and 15 s. Postprocessing humidity absorption was avoided by keeping the specimens in a desiccator. Extruded materials were pelletized after cooling in a water bath. After drying again, they were injection molded in a Battenfeld PLUS 350/75 reciprocating screw injection molding machine  with a press closing force of 350 kN. Tensile  and impact  specimens were obtained. The diameter of the screw in the plasticizing unit was 25 mm and the L/D ratio was 14. The melt and mold temperatures were, respectively, 260 °C and 30 °C. The injection speed, and pressure-holding and cooling times were, respectively, 42 cmElectrical conductivity measurements were performed using standard circular sheets . These sheets were obtained by hot pressing using a Collin P200E hydraulic press . Processing temperature and pressure were set at 260 °C and 130 bar, respectively. The molding process was carried out following this procedure: (1) preheating or plasticizing ; (2) compression ; (3) cooling under pressure (6 min). Therefore, the only property of the materials in the study that was not measured using injection molded tensile specimens was the conductivity. Although injection molding and hot-pressing techniques can be expected to produce different CNT dispersion levels, the methodology is still valid when the objective of the measurements is to compare the electrical conductivity of the materials obtained using different CNTs and preparation methods.mT, cT) were determined, respectively, from the maxima of the corresponding peaks during the heating and cooling scans, and the melting and crystallization enthalpies were determined from the areas under each of these peaks. The crystallinity of PA410 was calculated from the melting and cold crystallization enthalpies, considering an enthalpy of a 100% crystalline  that provided the loss tangent (tanδ) against temperature was employed. The single cantilever bending mode was selected and constant heating rate and frequency were set at, respectively, 4 °C/min and 1 Hz. The studied temperature interval ranged from −100 °C to 150 °C. Thermal properties related to melting and crystallization were studied by DSC using a Perkin-Elmer DSC-7 calorimeter  calibrated using an indium standard as a reference. First heating scan was performed from 30 °C to 300 °C at 20 °C/min, and then cooled at the same rate. The melting and crystallization temperatures  after gold coating. A Hitachi S-2700 electron microscope  was used at an accelerating voltage of 15 kV.The nanostructure was analyzed by transmission electron microscopy (TEM). The samples were obtained from injection-molded specimens which were ultrathin sectioned at ~100 nm using a Leica EMFC 6 ultramicrotome equipped with a diamond knife. The micrographs were taken in a Tecnai G2 20 twin apparatus , operating at an accelerating voltage of 200 kV.b), was 10 mm/min. A minimum of five tensile specimens were tested for each reported value.An Instron 5569 tensile tester  was used for the tensile tests. Young’s modulus was measured at a crosshead speed of 1 mm/min using an extensometer. The crosshead speed for the determination of the yield stress and ductility, measured as the break strain . The notches (depth 2.54 mm and radius 0.25 mm) were machined after injection molding. A minimum of eight impact specimens were tested for each reported value.Electrical conductivity values were obtained from the measured volume resistances, according to the ASTM D4496-87 standard. A Keithley 6487 picoammeter  and a Keithley 8009 Resistivity Test Fixture at 1 V were used for the measurement. Three measurements were performed for each reported value.As explained in the experimental section, the CNTs were available both as pristine CNTs (powder) and as a PA6-based masterbatch, and therefore PA410/Pebax/CNT ternary NCs and PA410/PA6/Pebax/CNT quaternary NCs were prepared, respectively .g of the PA410 and to the superposition of the Tg of the Pebax2 and the β transition of the PA410. As can be observed, in the PA410/Pebax/CNT ternary system, the position of the high temperature peak hardly changes with the CNT content, and the decrease observed in Tg is very slight with respect to the pure PA410. Previous studies [g; on the contrary, the addition of Pebax causes slight decreases in the Tg  than in the Pebax phase. In biphasic CNT-filled systems, the CNTs can be located either in the matrix ,31,52,54With respect to the matrix-dispersed phase morphology of the systems under study, In conclusion, in both systems under study (PA410/Pebax/CNT and PA410/PA6/Pebax/CNT), the CNTs are located both in the matrix (mainly) and in the dispersed phase. This would explain the lack of effect of the CNTs on the size of the dispersed phase, in view of the fact that the usual effects caused by the location of the CNTs in the continuous phase  would offset the usual effects caused by the location of the CNTs in the dispersed phase . Finally, it must be mentioned that the geometry of the dispersed phase remains unchanged as well, even if the presence of CNTs in the dispersed phase usually causes changes in its geometry ,61. The c) values of the four systems represented in c values of the two systems containing Pebax are very similar, and lower than those of the systems without Pebax. As discussed in our previous work [c values were similar in both systems. Therefore, the lower pc values observed in the systems containing Pebax must, necessarily, be related to the presence of this component. Given the lower CNT content in the Pebax dispersed phase than in the PA continuous phase owing to the higher affinity of the CNTs for the polyamide phase (as stated in the previous section), a partial exclusion effect of the Pebax dispersed phase over the CNTs is proposed. The effective CNT concentration is higher in the continuous PA phase, and therefore the electrical percolation occurs at lower overall CNT concentrations.ous work , the disc decreased from 0.5% in the PC/CNT NCs to 0.45% after the addition of 10% of mSEBS. Bin et al. [c of the PA6/CNT binary NCs decreased from 4–5% to 3.5% after the addition of the elastomer, because, in the ternary NCs, the CNTs were located exclusively in the PA6 phase, and the percolation threshold decreased due to the volume-exclusion effect exerted by the elastomer. Dasari et al. [The volume-exclusion effect has been observed in many other thermoplastic/thermoplastic/CNT systems. For example, Gonzalez et al.  studied n et al.  studied n et al. ,64. The n et al.  studied i et al.  studied i et al. ,66.Regardless of the effect on the mechanical properties, which are discussed below, the addition of Pebax to the PA410/CNT and the PA410/PA6/CNT systems has a positive effect on the electrical performance of the corresponding systems, as the percolation threshold is achieved at lower CNT contents due to the partial exclusion effect of the Pebax phase.Regarding the deformability of the systems under study, Nevertheless, the effect of Pebax on the performance of these systems is noteworthy since even if the deformability decreases as a result of the addition of CNTs, all the compositions remain ductile, with strain at break values over 30%. It is indeed a remarkable improvement with respect to the systems without Pebax, which were brittle in the whole CNT-composition range studied  and showFinally, No clear conclusions can be drawn from the comparison between the ternary and the quaternary systems. This is because, at low CNT contents, the values of the PA410/PA6/Pebax/CNT system are slightly higher than those of the PA410/Pebax/CNT system, but, at CNT contents above 0.6%, the values of both systems are very similar, and the differences are close to the experimental error of the measurements.The combined addition of 30% Pebax and CNTs (either in the form of pristine nanotubes or PA6-based masterbatch) to the PA410 results in NCs with improved electrical properties, i.e., lower percolation thresholds, which is attributed to the lower CNT concentration needed to achieve electrical percolation due to the partial volume exclusion effect of the dispersed Pebax over the CNTs. Moreover, the mechanical properties attained are of considerable interest, since all the materials remain ductile, and the impact strength increases from three to five-fold with respect to the pure PA410, despite the usual decrease observed in low deformation mechanical properties . It is noteworthy, for instance, that, after the addition of only 0.6% CNTs, the system turns semiconductive, with an impact strength loss of only 20% with respect to the corresponding unfilled PA410/Pebax 70/30 blend but a 300% increase with respect to pure PA410. It is also remarkable that the addition method used to incorporate the CNTs to the system does not affect the final properties of the NCs, pointing to the beneficial use of the masterbatch regarding its manipulation.


Immune-mediated inflammatory diseases (IMIDs) are systemic conditions associated with a high social and health impact. New treatments have changed the prognosis of IMIDs and have increased patient autonomy in disease management. Mobile apps have enormous potential to improve health outcomes in patients with IMIDs. Although a large number of IMID apps are available, the app market is not regulated, and functionality and reliability remain uncertain.Our aims are to review available apps for patients with IMIDs or caregivers and to describe the main characteristics and functionalities of these apps.We performed an observational, cross-sectional, descriptive study of all apps for patients with IMIDs. Between April 5 and 14, 2021, we conducted a search of the App Store (iOS) and Play Store (Android) platforms. We used the names of the different IMIDs as search terms. The inclusion criteria were as follows: content related to IMIDs, English or Spanish language, and user population consisting of patients and health care consumers, including family and caregivers. The variables analyzed were as follows: app name, type of IMID, platform (Android or iOS), country of origin, language, category of the app, cost, date of the last update, size, downloads, author affiliation, and functionalities.We identified 713 apps in the initial search, and 243 apps met the criteria and were analyzed. Of these, 37% (n=90) were on Android, 27.2% (n=66) on iOS, and 35.8% (n=87) on both platforms. The most frequent categories were health and well-being/fitness apps  and medicine . A total of 211 (82.3%) apps were free. The mean time between the date of the analysis and the date of the most recent update was 18.5 (SD 19.3) months. Health care professionals were involved in the development of 100 (41.1%) apps. We found differences between Android and iOS in the mean time since the last update  and free apps . The functionalities were as follows: general information about lifestyles, nutrition, or exercises ; specific information about the disease or treatment ; recording of symptoms or adverse events ; agenda/calendar ; reminder medication ; and recording of patient-reported outcomes . A total of 147 (60.5%) apps had more than one functionality.IMID-related apps are heterogeneous in terms of functionality and reliability. Apps may be a useful complement to IMID care, especially inpatient education . However, more than half of the IMID apps had not been developed by health care professionals or updated in the last year. Immune-mediated inflammatory diseases (IMIDs) are systemic conditions characterized by altered immune regulation causing chronic inflammation in specific organs or systems . IMIDs iManagement and treatment of IMIDs have changed substantially in the last decade, mainly due to the emergence of biological therapies . HoweverMobile health  technologies incorporating strategies for remote self-management may offer an effective alternative to classic outpatient-based approaches . In partHealth care professionals could play an essential role not only in the review or verification of the contents of these apps but also in their prescription to the best candidates and the recommendation of the most reliable options. However, the lack of studies providing detailed IMID-related app characteristics limits health care professionals’ knowledge in this field. Available IMID apps vary substantially in terms of features, functionality, and reliability ,9-14. UpWe performed an observational, cross-sectional, descriptive study of all the smartphone apps designed for patients with IMIDs available on the iOS and Android platforms.The methodology used for the selection of the apps followed the PRISMA  system. We searched the iOS Apple Store and Android Google Play Store from April 5 to 14, 2021 using the following terms: “ankylosing spondylitis,” “Crohn's disease,” “IBD,” “inflammatory bowel disease,” “immune-mediated inflammatory diseases,” “immune-mediated inflammatory disorders,” “psoriasis,” “psoriatic arthritis,” “rheumatoid arthritis,” and “ulcerative colitis.” As the resultant app list was potentially endless (similar to a Google search), we used the approach followed in previous reviews and performed the screening process until 20 consecutive apps yielded no new potentially relevant apps ,16.Two researchers with experience in the analysis of apps, design, and development, and in the management of patients with IMIDs (authors RRJ and VEV) performed identical searches independently. The inclusion criteria were as follows: content related to IMIDs, English or Spanish language, and user population consisting of patients and health consumers, including family or caregivers. Those apps that did not target patients  were excluded.The variables analyzed were the name of the app, type of IMID , platform (Android or iOS), country of origin, language, category of the app, cost, date of the last update, size, downloads, author affiliation, and functionalities. The author’s affiliation was classified as follows: university, health administration or government, hospital/clinic, foundation, patient association, pharmaceutical company, and technology company. Concerning technology companies, we identified two categories depending on whether they were health related or not. Functionalities were further classified into the following categories: information , self-diagnosis, disease activity monitoring, symptom or patient-reported outcome (PRO) recording, medication reminders, adherence monitoring, agenda/calendar , contact with health care professionals, and social network. The app search method was developed by the authors and has not been validated. This methodology to analyze app characteristics has been used in other similar studies ,16,17.The data were collected from the description and the screenshots available at the app store websites. In cases where one of the variables was not available or there was any doubt, the app was downloaded onto an iPhone 11 (version 14.4.2) or a Xiaomi Redmi Note 7 (version 9.0). For the categorization of author affiliation, we conducted Google searches using the name or the website provided by the app store.t test or Mann-Whitney test to compare quantitative variables. A P value <.05 was considered statistically significant. Cohen kappa (κ) test was performed to determine the reliability of the data analyzed by these two independent researchers (RRJ and VEV).The researchers (RRJ and VEV) evaluated each app independently. Data were analyzed using SPSS Statistics for Windows, Version 21.0 . The homogeneity of the groups was analyzed using a univariate analysis by applying the chi-square test to compare qualitative variables and the The two researchers jointly analyzed those apps again in which the information collected varied between them. Discrepancies were discussed and an agreement was reached.We identified 713 apps in the initial search in the two app stores; of these, 470 were excluded . We finaMost of the apps belonged to the health and well-being/fitness and medicine categories. The mean time since the last update was 18.5 (SD 19.3) months, and the mean app size was 28.6 (SD 46.9) Mb. A total of 43 (17.7%) apps required payment for use, with a mean price of US $10.30 (SD US $12.00). Concerning the language, 86.8% (n=211) of the apps analyzed were in only one language, and 15 (6.2%) were in three or more languages. A total of 234 (96%) apps were in English, and 11.4% (29/243) were in Spanish. P<.001) between the Android and iOS apps, as follows: time since the last update, 16.2 (SD 14.7) months vs 30.3 (SD 25.7) months; apps with cost, 14.4% vs. 24.2%; and size, 11.2 (SD 13.9) Mb vs 50.2 (SD 77.9) Mb.We found statistically significant differences  apps were specifically aimed at one IMID, and 21 (8.6%) were aimed at all IMIDs .P<.001). No statistically significant differences were found between the rest of the recorded app characteristics and the type of IMIDs.Concerning author affiliation, we found statistically significant differences between specific ankylosing spondylitis–related apps and specific psoriasis-related apps. These differences were found in the number of apps developed by a health-related technology company (50% in psoriasis apps and 6.7% in ankylosing spondylitis), nonhealth technology company (20% in psoriasis apps and 40% in ankylosing spondylitis), and in those apps where this information was not available  had more than one functionality. A total of 135 apps offered information about lifestyle, nutrition, and physical exercise, and 102 apps offered information about the disease or its treatment . We founThe mean number of functionalities per app was 2.1 (range 1-7). No statistically significant differences were found between the number of functionalities and the remaining variables, except for author affiliation. Apps developed by health-related technology companies presented a higher average number of functionalities.We provide a comprehensive and unique review of IMID-related apps available for patients in the Play Store and Apple Store. The marketplace offers hundreds of solutions, thus making it difficult for the user to filter . Health We analyzed 243 apps for the most important IMIDs and identified features that may contribute to our understanding of the current landscape and the development of future solutions. Most of the apps targeted rheumatoid arthritis and inflammatory bowel diseases (IBDs). This distribution is proportional to the prevalence of the IMIDs in real life . Among tThe use of IMID-related apps is still in its infancy ,18. The In addition to the limited number of IMID apps, some of the negative characteristics such as a lack of robust scientific evidence prevent clinicians from using health-related apps or recommending them to patients ,13,23. ICurrently, there is no specific legislation on the use of the apps and standards or official guidelines on app development. Similarly, there are no official repositories, mainly due to the fast-moving pace of mHealth ,24. The The level of the update is also a potential cause for concern. Given the rapid advance in IMID care, information must be constantly updated. In our study, we found that more than half of the apps had not been updated in the previous year, while in 25.1% (n=61), the last update had been over 2 years previously. In a review of 20 rheumatoid arthritis apps, only 12 had been updated in the previous year . ConcernOn the other hand, one of the advantages of these tools is that most health-related apps are free ,17,31,33Many of the health-related apps emerged as tools for consulting information, and this is still the most frequent functionality, although 60.5% n=147) of the apps analyzed had more than one functionality. Our results are consistent with those of other studies and highlight that most apps offer passive functionalities such as information about the disease, treatments, lifestyle, nutrition, and physical exercise. Medication information was the feature most desired by a cohort of 193 patients with rheumatic disease . Mollard7 of the Several studies have shown the usefulness of apps for improving adherence ,34. ThisEncouraging enthusiastic patients to engage with apps regularly is another barrier . For exaA possible limitation is that we did not assess the privacy and confidentiality of apps since this information is not available in most cases. The relationship between the use of health apps and privacy risk is understudied . Most reThis review provides health professionals with an overview of available IMID-related apps and their characteristics. IMID-related apps continue to be heterogeneous in terms of their functionalities and reliability. Apps may act as a useful complement to IMID care, especially in patient education, as this is their most frequent functionality. However, more than half of the IMID apps had not been developed by health care professionals or updated in the previous year.


Since the quality and safety of food highly depend on its preservation and protection, the use of food packaging materials increases the risk of chemical contamination of the packaged food by migration. Herein, we focused on antioxidants, photoinitiators, UV absorbers and plasticizers which are extensive additives used in food packaging materials. In the present study, a rapid, simple, green and reliable method was developed and validated for the determination of twelve chemical additives in edible vegetable oils using SFC together with a modified QuEChERS procedure. Under the optimum conditions, twelve additives were separated within 10 min, and the consumption of the organic solvent was significantly reduced, which improved the environmentally friendliness. The performance of the developed method was evaluated. Good linearity (r > 0.999) was obtained in the range of 0.20–20.0 µg/mL and 0.50–20.0 µg/mL, respectively. The limits of detection and limits of quantification of the twelve additives in vegetable oils were 0.05–0.15 µg/mL and 0.15–0.50 µg/mL, respectively. Recoveries of all the chemical additives for the spiked samples were between 60.9% and 106.4%, with relative standard deviations (RSD) lower than 9.9%. The results demonstrated that the proposed method was efficient, reliable and robust for the routine analysis of additives in edible vegetable oils and can be an alternative to the multi-residue analysis of chemical additives for other packaged foods. Since the quality and safety of food highly depend on its preservation and protection, the use of food packaging materials increases the risk of chemical contamination of the packaged food by migration . During Herein, we focused on antioxidants, photoinitiators, UV absorbers and plasticizers which are extensive additives used in food packaging materials. Adding antioxidants can improve durability  and prevEdible vegetable oils, which are abundant in unsaturated fatty acids and other nutrients, are widely used in daily diets. Packaging materials contaminated with harmful chemical constituents and the migration of such chemical additives from packaging materials are the main reasons for the occurrence of contaminants in edible vegetable oils . Therefore, developing sensitive and reliable methods to analyze these chemical additives is necessary for the quality control of vegetable oils. However, sample pretreatment is a tricky subject, since target compounds are fat-soluble and tend to remain in the fatty food sample matrix. To date, using acetonitrile to extract analytes from the oil matrix is one of the most reported methods as the first step of preprocessing ,19,20. UMost of the present analytical methods for the additives used in packaging materials are based on gas chromatography (GC) , liquid The objective of this study was to develop a simple, rapid and reliable analytical method combining QuEChERS with SFC for the simultaneous analysis of twelve chemical additives  in spiked blank oil samples (10 µg/g). Other pretreatment conditions were as described in After determining the type of d-SPE sorbent, the dosage also needed to be investigated. Herein, the various amounts  of PSA were determined. When the dosage of PSA increased to 120 mg, recovery was lower (59.2–91.5%), since excess PSA can adsorb some target compounds, as shown in 4 is often used as a dehydrating agent, because it has a high affinity for water and the salting-out effect. Dehydrating agents are usually added twice in conventional methods: the first is in the solvent extraction, and the other is together with the d-SPE sorbents. In this study, since there was very little water in the edible oil, the first step of dehydration was omitted. In other words, anhydrous MgSO4 was only added to the acetonitrile extract together with 40 mg PSA. As can be seen from 4 had no obvious impact on recoveries, probably because very little water existed in the acetonitrile extract itself. To be clear, five additives including TBHQ, BHA, 4-MBP, UV-9 and UV-P achieved the highest recoveries in 800 mg MgSO4 conditions.In the QuEChERS process, anhydrous MgSOTM AcclaimTM 120 C18  column was chosen. The wavelength of the UV detector was kept at 220 nm.In order to improve the separation effect of twelve target additives, several SFC conditions, including the modifier, flow rate, column temperature, backpressure and gradient eluting procedure, were discussed, respectively. A Thermo Scientific2 eluent with low polarity. Therefore, the addition of a polar organic co-solvent, always called a modifier, is essential to adjust the eluent strength and enhance the solubility of the target analytes. Organic modifiers, including methanol, isopropanol and acetonitrile, were introduced into the mobile phase to improve the separation of the twelve target compounds on the C18 column. As shown in The application of SFC is limited by a supercritical COThe proportion of the modifier is the main factor in changing the polarity of the mobile phase, leading to a different retention and separation behavior. In this research, a low content of the organic modifier (methanol from 4% to 8%) was added to acquire a better separation efficiency. As depicted in Column temperature can affect retention behavior and separation efficiency via influencing the density and viscosity of the supercritical fluid. In this study, different temperatures  were investigated. As shown in 2. Different backpressures  were tested to get better separation in this study. From In SFC, backpressure (BPR) also has a significant impact on retention and separation behavior. Increasing BPR usually shortens the retention time and influences the separation effect attributed to the higher density and greater eluting strength of supercritical COFlow rate is one of the optimization parameters in SFC. Apparently, a high flow rate can shorten the analysis time due to the density of the mobile phase increases. Compared with liquid chromatography, SFC can afford a higher flow rate due to the unique properties of supercritical fluid. The results of the different flow rates  were shown in 3OH (95% CO2); 5–10 min, 3% CH3OH (97% CO2). As mentioned in 3OH. Hence, 1.4 mL/min was selected to shorten the analysis time. However, UV-531 (peak 11) and TOTM (peak 12) were not perfectly separated under such conditions. This problem was solved by changing the methanol concentration from 5% to 3% in the fifth minute, because the eluent strength of the mobile phase weakened with a lower modifier concentration. The optimal elution gradient conditions and other SFC conditions were presented in According to the above comprehensive analysis of the modifier, column temperature, backpressure and flow rate, the gradient elution program was further explored to obtain the optimal separation effect. In this research, both the flow rate gradient and concentration gradient were optimized. The results of four different gradient elution programs were shown in A series of mixed matrix matched standard solutions  were prepared to establish the matrix-matched calibration curve. Satisfactory linearity was obtained for the twelve additives, with all correlation coefficient (r) values higher than 0.999. Spiked samples were prepared by spiking standards into blank natural samples and were used to determine the limit of detection  and limit of quantification  values. The LODs and LOQs values of the twelve additives were calculated to range from 0.05–0.15 μg/mL and 0.15–0.50 μg/mL, respectively. All the results were summarized in Compared to conventional pretreatment methods such as solid phase extraction, QuEChERS is much greener and simpler. Nevertheless, some impurities may be co-extracted during the QuEChERS process to cause matrix interference and finally affect the accuracy of results. Considering the mentioned conditions, the matrix effects (ME) were studied to investigate the influence of the substrate. The equation was as shown below and the values were listed in n = 6) of all recovery experiments were lower than 9.9%, showing the satisfactory precision of this method. Herein, the recoveries of the two target analytes were below 80%, which may be attributed to the higher molecular weight and lower polarity.The recovery experiments and relative standard deviation (RSD) were investigated by spiking blank natural samples in three levels  with six replicates to evaluate the accuracy and precision, respectively. As presented in Considering that the polymer additives used in packaging materials may migrate into foodstuff, strict quality control and testing is essential to ensure the health of consumers. To further verify the applicability, the proposed method was applied to various natural samples, including two extra virgin olive oil samples, two peanut oil samples, two blend oil samples and one sunflower oil sample. Each sample was analyzed in triplicate and the results are summarized in 4) was obtained from Chronchem . The ultrapure water used in all experiments was obtained from a Millipore Simplicity purification system .Tert-butylhydroquinone , butylated hydroxyanisole , methyl 2-benzoylbenzoate , 2,2-dimethoxy-2-phenylacetophenone , 4-methylbenzophenone , benzyl butyl phthalate , 2-hydroxy-4-methoxybenzophenone , 2,2′-dihydroxy-4-methoxybenzo phenone , octabenzone , 2-(2-hydroxy-5-methylphenyl) benzotriazole , bis (2-ethylhexyl) phthalate  and trioctyl trimellitate  of analytical grade were purchased from Aladdin Chemical Co. Ltd. . Methanol, acetonitrile and isopropanol of HPLC grade were purchased from Tedia Company . Primary secondary amine , graphitized carbon black  and octadecylsilyl  were purchased from Agela . Analytical grade anhydrous magnesium sulfate  of twelve target compounds were dissolved in methanol. The mixed stock standard solutions and working solutions were prepared before use by taking certain amounts of each individual standard solutions and diluting with HPLC grade methanol. All solutions were stored at 4 °C in amber glass vessels.Chromatographic analysis and separation were carried out by using Nexera UC SFC system  with an SPD-20A UV detector. Furthermore, the system was equipped with a column manager, a convergence manager controlling the pressure of supercritical carbon dioxide, an auto-sampler (5 μL loop for injection), an online degasser and a backpressure regulator (BPR). With an LC-20ADXR pump, the modifier could be delivered and then mixed with supercritical carbon dioxide. Lab Solutions Ver5.8 was used for instrument control, data collection and processing.TM AcclaimTM 120 C18  column was used to separate twelve target analytes. The mobile phase comprised of supercritical CO2 (A) and methanol (B). The gradient elution program was set as follows: 0–5 min, 5% B; 5–10 min, 3% B. The flow rate was maintained at 1.4 mL/min. The column temperature was set at 40 °C and the backpressure was 10 MPa. The injection volume was 5 μL. The UV detection wavelength was 220 nm.The final SFC conditions were as follows: A Thermo Scientific4 and 40 mg PSA, vortexed for 3 min, followed by centrifugation at 8000 rpm for 5 min. Finally, the resulting supernatant was filtered through a 0.22 μm organic membrane into a glass auto-sampler vial for SFC analysis. The use of plastic products was avoided to prevent extra interference during the experiment.All edible vegetable oil samples  used in this study were purchased randomly from local supermarkets  and stored at room temperature. The general QuEChERS methodology was shown in 4, modifier selection, column temperature, flow rate and backpressure were investigated to obtain the optimum experimental conditions. Under the optimal conditions, twelve chemical additives were separated within 10 min, and the consumption of organic solvent was minimized. Satisfactory method validation results were achieved and demonstrated that the proposed method can be regarded as a useful alternative and a complementary method for the routine analysis of additives in edible vegetable oils. Furthermore, the QuEChERS-SFC method is more time-saving, simplified and environmentally friendly. In general, this method provides an efficient procedure for reducing the costs and work involved in the quality control of edible vegetable oils and may have a potential application in other packaged food samples.In the present study, a rapid, simple, green and reliable method was developed and validated for simultaneous determination of twelve chemical additives in edible vegetable oils using a modified QuEChERS procedure followed by SFC. Various parameters such as sorbents, dosage of MgSO