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Electronic and electric waste (e-waste) management strategies often fall short in dealing with the plastic constituents of printed circuit boards (PCB). Some plastic materials from PCB, such as epoxy resins, may release contaminants, but neither potential environmental impact has been assessed nor mitigation strategies have been put forward. This study assessed the biodegradation of microplastics (1-2 mm in size) from PCB by the fungus Penicillium brevicompactum over 28 days, thus contributing to the discussion of mitigation strategies for decreasing the environmental impact of such plastics in the environment. The capacity of P. brevicompactum to induce microplastic fragmentation and degradation has been determined by the increased the number of smaller-sized particles and microplastic mass reduction (up to 75 % within 14 days), respectively. The occurrence of chain scission and oxidation of microplastics exposed to P. brevicompactum when compared with the control conditions (which occurred only after 28 days of exposure) can be observed. Furthermore, Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy performed in dried biomass put in evidence an increase in the absorption intensities in regions that could be attributed to functional groups associated with carbohydrates. The results underline the potential role of the genus Penicillium, particularly P. brevicompactum, in the biodegradation of microplastics from PCB, thus providing the basis for further exploration of its potential for e-waste bioremediation and research on the underlying mechanisms for sustainable approaches to mitigate e-waste pollution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: In the last couple of decades, there has been a shift in use of endovascular procedures in vascular surgery. We aim to examine the impact of this endovascular shift on vascular trainees, determine whether the surgical experiences of trainees in the integrated residency and fellowship program changed over time, and identify differences between the two training paradigms., Methods: Data were extracted from the Accreditation Council for Graduate Medical Education National Data Case Logs for the vascular surgery fellowship (1999-2021) and integrated residency (2012-2021) programs. Every procedure was categorized as open or endovascular, then designated into the following subcategories: thoracic aneurysm repairs, cerebrovascular, abdominal aneurysm repairs, venous, vascular access, peripheral arterial disease, visceral, or miscellaneous. We compared the prevalence of open and endovascular cases in the fellowship and integrated residency using data from overlapping years (2012-2021). In addition, we compared the mean number of cases per trainee per year within designated time intervals. The vascular surgery fellowship was grouped into three intervals: 1999 to 2006, 2006 to 2013, and 2013 to 2021; the integrated vascular surgery residency was grouped into two intervals: 2012 to 2017 and 2017 to 2021. Data were standardized to represent the average number of cases per trainee per year., Results: Within the fellowship, we found a 362.37% increase in endovascular procedures (mean, 56.80 ± 32.57 vs 262.63 ± 9.91; P < .001), although there was only a 32.47% increase in open procedures (220.19 ± 4.55 vs 291.68 ± 8.20) between the first and last time intervals. There was a decrease in abdominal aneurysm repair (24.46 ± 7.30 vs 13.85 ± 0.58; P < .001) and visceral (6.41 ± 0.44 vs 5.80 ± 0.42; P = .039) open procedures. For the integrated residency, there was an increase in open procedures by 8.52% (352.18 ± 8.23 vs 382.20 ± 5.84; P < .001). Residents had greater total, open, and endovascular procedures per year than fellows (all P < .001). Chief residents had approximately one-half as many cases as vascular fellows per year. Fellows performed more open abdominal aneurysm repair (14.04 ± 0.80 vs 12.40 ± 1.32; P = .007) and visceral (5.83 ± 0.41 vs 4.88 ± 0.46; P > .001) procedures than residents. Overall, 52% to 53% of cases performed by trainees per year were open procedures in both the fellowship and integrated residency (288.56 ± 12.10 vs 261.27 ± 10.13, 365.52 ± 17.23 vs 319.58 ± 6.62; both P < .001). Within the subcategories, only cerebrovascular, vascular access, and miscellaneous had more open procedures performed per trainee., Conclusions: Vascular surgery training has incorporated new endovascular techniques and technologies while maintaining operative training in open procedures. Despite changes in vascular surgery training, trainees are still performing more open procedures than endovascular procedures per year. However, there are evolving deficits in specific types of procedures., Competing Interests: Disclosures None., (Copyright © 2024 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

The human head lice is a cosmopolitan ectoparasite that causes pediculosis. The main way of spreading lice is through direct head-to-head contact. It is popular knowledge that some individuals are more susceptible to contracting head lice than others. Reports of individuals who have never been affected by the disease are common, even living in the same environment and under the same conditions as people who regularly have lice infestations. Previous research has been carried out on the risk of this infection associated with different human factors like gender or age. However, studies on the influence of the individual hair characteristics are scarce. The objective of the study was to analyze the pediculosis risk using geographical location, gender, age and individual hair characteristics as variables. Pediculosis was diagnosed through the detection of living lice in the hair. This cross-sectional school-based epidemiological study was conducted in 310 schoolchildren aged 1 to 13 years of schools in 4 municipalities situated in the State of Paraná, Brazil. The prevalence of head louse infection in primary school students was 49.35 %. The Odds Ratio of presence of pediculosis (OR) was estimated using multivariate logistic regression analysis. The results obtained indicate that hair length and thickness increase the risk of infection. Furthermore, the inclusion of hair color, hair shape, kind of hair-scale as covariates increases the risk of pediculosis, indicating that these variables partly explain this susceptibility and that pediculosis is independent of gender. A smaller hair diameter may favor insect fixation to the hair in the nymphal phases. These results may explain why girls are a greater risk as they let their hair grow for cultural reasons, i.e., being of female gender is an agglutinating variable. The conclusions drawn may explain the discrepancies obtained in previous analyses., Competing Interests: Declaration of Competing Interest None declared., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Disposable facemasks represent a new form of environmental contamination worldwide. This study aimed at addressing the abundance of facemasks in an overlooked natural environment with high ecological and economic value - the wetlands (Ria de Aveiro, Portugal, as study case), evaluating their potential biodegradation using naturally occurring fungi and assessing the potential ecotoxicity of released microfibres on local bivalves. All masks collected within 6500 m 2 area of Aveiro wetland were 100 % disposable ones (PP-based, confirmed by Fourier transform infrared spectroscopy - FTIR) with an initial abundance of 0.0023 items/m 2 in Sept. 2021, which was reduced by ∼40 % in Apr. 2022 and ∼87 % in Sept. 2022, as a reflection of the government policies. Analysis of the carbonyl index (0.03 to 1.79) underlined their state of degradation, primarily due to sun exposure during low tides. In laboratory conditions, 1 mm 2 microplastics obtained from new disposable facemasks were prone to biodegradation by Penicillium brevicompactum and Zalerion maritimum inferred from microplastics mass loss (∼22 to -26 % and ∼40 to 50 %, respectively) and FTIR spectra (particularly in the hydroxyl and carbonyl groups). In addition, microfibres released from facemasks induced sublethal effects on the clam, Venerupis corrugata, mostly in their UV-aged form when compared to pristine ones, characterised by a decrease in cellular energy allocation (CEA) and an increase in aerobic energy metabolism (ETS). Concomitantly, clams exposed to 1250 items/L of UV-aged microplastics (similar to field-reported concentrations) expressed greater clearance capacity, indicating a need to compensate for the potential energy unbalance. This study provides the first baseline monitoring of facemasks in wetlands while bringing new evidence on their biodegradation and ecotoxicity, considering environmentally relevant conditions and keystone organisms in such environments. Such studies require scientific attention for rapid regulatory action against this emerging and persistent pollutant, also targeting remediation and mitigation strategies considering these items under pandemic scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Recreational activities are important source of disturbance on sandy beaches and have a negative effect on benthic communities as they alter species abundance, diversity, and/or distribution. Here, we report for the first time the effects of the recreational activities on the distribution pattern of macrobenthic communities of Amazonian sandy beaches. Sampling was performed during three consecutive months with different beach use intensity in 2017 (before, during vacation, and one month after the vacation period) on two contrasting beaches regarding disturbance (Urban: Atalaia; and Protected: Corvinas) in the intertidal zone along two equidistant transects at seven equidistant sampling stations: from the high-tide water mark to the swash zone. At each sampling station, four biological and sediment samples were randomly collected. Also, in each station, the sediment compaction was determined using a manual penetrometer. Physical sediment variables remained constant over time in both beaches, whereas differences were found in sediment compaction over the periods. Sediment compaction was considered the most important factor for the differences observed in macrobenthic community structure in the study beaches, and it was related to intensity of recreational activities. Variations in density and changes in richness between periods and sampling stations were observed in Atalaia beach. On the other hand, Corvinas beach remained the same throughout the study. Overall, three faunal zones were distinguished: upper intertidal, intermediate intertidal, and low-intertidal, whereas in the vacation period, only two faunal zones were distinguished: upper-intertidal and intermediate-low intertidal., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

As the World slowly emerged from the then-ongoing pandemic, War broke out in Europe with the invasion of Ukraine by Russia. The enduring military conflict in Ukraine has had sweeping consequences at the human, social, economic, and environmental levels, not only for the nations involved but across Europe and globally. Damaged infrastructures, severe disruption of economic activity, and forced migration have led to negative impacts on sustainability. The COVID-19 pandemic has added another layer of complexity to this already challenging situation, as the virus has further disrupted economic activity and strained healthcare systems. Herein, we examine how the intersection of war and COVID-19 affect the United Nations' 2030 Agenda for Sustainable Development. How these intersecting challenges have impacted efforts to build a more sustainable future, and how these impacts have a global reach are also assessed. The broader implications of this case for understanding the linkages between conflict, pandemics, and sustainability more generally are also considered, relating these with the United Nations' Sustainable Development Goals (SDG) Agenda for 2030., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study investigates the potential of MPs as carriers of pollutants as they can strengthen bioaccumulation of toxic metals on marine organisms. For the first time, the interaction of the metal palladium (Pd) with the widespread MPs, both with increasing concentrations in water environments from anthropogenic sources, was tested. Mytilus galloprovincialis, an important seafood product, was exposed to Pd (24 h) in two ways: water-dissolved and MPs-adsorbed, with depuration followed for 144 h. Quantification of Pd in tissues shown an accumulation 2-3 times higher (59 % of initial Pd) for mussels exposed to MPs-adsorbed Pd and higher in digestive gland than when exposed to water-dissolved Pd (25 %; higher in gills). Additionally, it was demonstrated that Pd induced oxidative stress and altered the feeding behavior of mussels. Therefore, this work support MPs as being vectors of metals (i.e. Pd) to enhance their bioaccumulation on marine organisms which highlights ecological risk of these emerging pollutants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Introduction: Simultaneous pancreas-kidney transplantation (SPKT) has demonstrated favorable impact on the progression of chronic complications in type-1 diabetes (T1D) and terminal chronic kidney disease (CKD). However, some CKD mineral and bone disorders (CKD-MBD) may persist, even after transplantation. There are only a few studies addressing the long-term progression of bone mineral density (BMD) in these patients. Our aim was to assess baseline BMD and long-term progression and consequences in patients with T1D undergoing SPKT., Methods: A retrospective cohort included patients undergoing SPKT in our tertiary center between 2000 and 2017. BMD progression was assessed on dual X-ray absorptiometry (DXA). Only patients with baseline data and a minimum follow-up of 2 years were included., Results: Seventy-three patients were included, 53.4% male, with a median age at SPKT of 35 years (interquartile range [IQR] 31; 39). At transplantation, the median T-scores for the lumbar spine (LS) and femoral neck (FN) were -1.6 (IQR -2.6; -1.1) and --2.1 (IQR -2.7; -1.6), respectively. Seventy-five percent of patients presented low BMD (osteopenia or osteoporosis) in the LS and 90% in the FN, with 33% osteoporosis in the LS and 36% in the FN. On multivariate analysis, male gender (odds ratio [OR] 10.82, 95% confidence interval (CI) 2.88-40.70) and low body-mass index (BMI) (OR 0.73, 95% CI 0.55-0.97) were significantly associated with lumbar but not femoral osteoporosis. At long-term follow-up, BMD significantly improved in the LS (ΔT-score +0.41, P<0.001) and FN (ΔT-score +0.29, P=0.01), at a median 4 years after SPKT. Twelve (16.4%) and 9 (12.3%) patients showed persistent FN and LS osteoporosis, respectively. Multivariate linear regression showed that high BMI was predictive of improvement in BMD., Conclusions: This study demonstrated severe skeletal fragility in T1D patients with terminal CKD undergoing SPKT, more than a quarter of whom showed osteoporosis. The significant improvement in BMD may result from metabolic correction by SPKT and from physiological skeleton mineralization, which continues in this age group. BMD progression was positively associated with BMI, due to improved nutritional balance after transplantation., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Metal(oid)s concentrations have been quantified in plastic pieces collected from four beaches located in the Mediterranean coast of Spain with different characteristics (i.e. anthropogenic pressure, zone). Metal(oid)s content was also related to selected plastic criteria (i.e. color, degradation status, polymer). The selected elements were quantified with mean concentrations in the sampled plastics with the following order: Fe > Mg > Zn > Mn > Pb > Sr > As > Cu > Cr > Ni > Cd > Co. Moreover, black, brown, PUR, PS, and coastal line plastics concentrated the higher metal(oid)s levels. Local of sampling (influence of mining exploitation) and severe degradation were key factors for uptake of metal(oid)s from water by plastics as modification of surfaces strengths their adsorption capacity. Determined high levels of Fe, Pb and Zn in plastics reflected the pollution degree of the marine areas. Therefore, this study is a contribution for the potential use of plastics as pollution monitors., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Juan Santos Echeandia reports financial support was provided by Spanish State Research Agency (AEI) and the Ministry of Science, Innovation and Universities (MICIU). Teresa Rocha-Santos reports financial support was provided by Portuguese Science Foundation (FCT). Armando Costa Duarte reports financial support was provided by Portuguese Science Foundation (FCT). Joana Patricio Rodrigues reports financial support was provided by Portuguese Science Foundation (FCT)., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Freshwater benthic environments are among the major sinks of microplastics (MPs, < 5 mm) sourced on inland anthropogenic activities. The ecotoxicological effects of MPs on benthic macroinvertebrates have been assessed preferably in collectors, shredders, and filter-feeders, but resulting in insufficient knowledge on the potential trophic transfer and its effects on macroinvertebrates with predator behaviour such as planarians. This work evaluated the behavioural (feeding, locomotion), physiological (regeneration) and biochemical responses (aerobic metabolism, energy reserves, oxidative damage) of the planarian Girardia tigrina after consuming contaminated live prey Chironomus riparius larvae previously exposed to microplastics of polyurethane (PU-MPs; 7-9 μm in size; 375 mg PU-MPs/kg). After the feeding period (3 h), planarians consumed 20 % more contaminated prey than uncontaminated prey, probably related to increased curling/uncurling movements of larvae (that might be more appellative to planarians). Histological analysis revealed planarians' limited intake of PU-MPs, mainly detected near the pharynx. The consumption of contaminated prey (and intake of PU-MPs) did not result in oxidative damage but slightly increased the aerobic metabolism and energy reserves which show that the consumption of more prey was sufficient to cope with the potential adverse effects of internalized MPs. Moreover, no effects were observed in the locomotion of planarians in good agreement with the hypothesis of sufficient energy acquired by the exposed planarians. Despite the previous, it seems that the energy acquired was not allocated for planarians' regeneration since a significant delay in the regeneration of the auricles was observed for planarians feeding on contaminated prey. Therefore, further studies should be performed considering the potential long-term effects (i.e., reproduction/fitness) and the effects of MPs that might result from continuous feeding on contaminated prey, representing a more realistic exposure scenario., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

During the first year of the COVID-19 pandemic, facemasks became mandatory, with a great preference for disposable ones. However, the benefits of face masks for health safety are counteracted by the environmental burden related to their improper disposal. An unprecedented influx of disposable face masks entering the environment has been reported in the last two years of the pandemic, along with their implications in natural environments in terms of their biodegradability, released contaminants and ecotoxicological effects. This critical review addresses several aspects of the current literature regarding the (bio)degradation and (eco)toxicity of face masks related contaminants, identifying uncertainties and research needs that should be addressed in future studies. While it is indisputable that face mask contamination contributes to the already alarming plastic pollution, we are still far from determining its real environmental and ecotoxicological contribution to the issue. The paucity of studies on biodegradation and ecotoxicity of face masks and related contaminants, and the uncertainties and uncontrolled variables involved during experimental procedures, are compromising eventual comparison with conventional plastic debris. Studies on the abundance and composition of face mask-released contaminants (microplastics/fibres/ chemical compounds) under pre- and post-pandemic conditions should, therefore, be encouraged, along with (bio)degradation and ecotoxicity tests considering environmentally relevant settings. To achieve this, methodological strategies should be developed to overcome technical difficulties to quantify and characterise the smallest MPs and fibres, adsorbents, and leachates to increase the environmental relevancy of the experimental conditions., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

The gut microbiota is a key factor in the maintenance of physiological homeostasis and immunity. Correlational studies have demonstrated that alterations in microbiota composition have been associated with addiction. Moreover, animal studies have confirmed a link between reward and social processes, which may be shaped by the gut microbiota thus influencing neurodevelopment and the programming of social behaviors across diverse animal species. However, whether there is an interaction between the microbiota and social reward processes in the context of drug reward remains unclear. To this end, we explored the influence of gut microbiota in regulating behaviourally conditioned responses to different rewards (cocaine and social interactions). Depletion of the intestinal microbiota resulted in differential reward responses to both drug and social stimuli with an attenuation of the former and enhancement of the latter independent of concomitant immune changes. Moreover, the combination of depleting the gut microbiota in the presence of a positive social stimulus attenuates cocaine reward. Together these data suggest that the two-pronged approach of targeting the microbiota and enhancing social behaviour could constitute a valuable component in reducing harm in drug use by altering the salient effects of cocaine., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Purpose: Chlamydia trachomatis/Neisseria gonorrhea (CT/NG) retesting three months after diagnosis is a guideline-recommended strategy to detect re-infections. Adolescents and young adults are priority populations in the U.S. Sexually Transmitted Infections National Strategic Plan, but there is a lack of research examining CT/NG retesting among these populations. This study describes retesting following CT/NG diagnosis among adolescent and young adult patients at Title X and non-Title X clinics and measures the association of patient-level factors with CT/NG retesting., Methods: We evaluated electronic medical records from 2014 to 2020 from an academic urban-suburban primary care network. The primary outcome was retesting, defined as a diagnostic test for CT or NG ordered 8-16 weeks after index diagnosis. Mixed effects logistic regression modeling stratified by Title X funding was conducted to evaluate the association of patient-level factors with CT/NT retesting., Results: Overall, 23.5% (n = 731) of patients were retested within 8-16 weeks following index CT/NG diagnosis. A significantly greater proportion of Title X patients were retested compared to non-Title X patients. Males were significantly less likely to be retested compared to females, and the proportion of patients retested decreased significantly over the study period., Discussion: Guideline-recommended retesting following CT/NG diagnosis was low in this young primary care cohort, especially among male and non-Title X clinic patients. Decreases in CT/NG retesting over the study period may be contributing to worsening of the STI epidemic. Our results provide insights into CT/NG retesting that can inform efforts to end the STI epidemic., (Copyright © 2022 Society for Adolescent Health and Medicine. Published by Elsevier Inc. All rights reserved.)

In this work, we investigated in vitro different biological activities of alkaline lignins extracted from the species Buchenavia viridiflora, a tree from the Amazon rainforest used as a wood product. The chemical composition results for the twig and leaves were, respectively (%): cellulose (30.88 and 24. 28), hemicellulose (21.62 and 23.03), lignin (29.93 and 25.46), extractives (13.06 and 20.52), and ash (4.51 and 6.72). The yield was higher for the lignin of the branches (67.9 %) when compared to the leaves (60.2 %). Lignins are of the GSH type, low molecular weight and thermally stable. They promoted moderate to low antioxidant activity, highlighting the lignin of the branches, which presented an IC 50 of 884.56 μg/mL for the DPPH assay and an IC 50 of 14.08 μg/mL for ABTS. In the cytotoxicity assays, they showed low toxicity against macrophage cells (IC 50 28.47 and 22.58 μg/mL). In addition, they were not cytotoxic against splenocytes and erythrocytes at concentrations ranging from 100 to 6.25 μg/mL. These were able to promote splenocyte proliferation and induce the production of anti-inflammatory cytokines. And inhibit the growth of tumor cells with IC 50 ranging from 12.63 to values >100 μg/mL and microbial at a concentration of 512 μg/mL. Finally, they showed antiparasitic activity by inhibiting the growth of chloroquine-sensitive and resistant Plasmodium falciparum strains. These findings reinforce that the lignins in this study are promising for potential pharmaceutical and biomedical applications., Competing Interests: Declaration of competing interest None., (Copyright © 2022. Published by Elsevier B.V.)

Background: Peripheral artery disease (PAD) disproportionately affects nonwhite, Hispanic/Latino, and low socioeconomic status patients, who are less likely to have insurance and routine healthcare visits. Medicaid expansion (ME) has improved insurance rates and access to care, potentially benefitting these patients. We sought to assess the impact of ME on disparities in outcomes after peripheral vascular intervention (PVI) for PAD., Methods: A retrospective analysis of prospectively-collected Vascular Quality Initiative PVI procedures between 2011 and 2019 was conducted. The sample was restricted to first-record procedures in adults under the age 65 in states that expanded Medicaid on January 1, 2014 (ME group) or had not expanded before January 1, 2019 (non-expansion [NE] group). ME and NE groups were compared between pre-expansion (2011-2013) and post-expansion (2014- 2019) time periods to assess baseline demographic and operative differences. We used difference-in-differences multivariable logistic regression adjusted for patient factors and clinical center and year fixed effects. Our primary outcome was 1-year major amputation. Secondary outcomes included trends in presentation, 30-day mortality, 1-year mortality, and 1-year primary and secondary patency. Outcomes were stratified by race and ethnicity., Results: We examined 34,313 PVI procedures, including 20,378 with follow-up data. Rates of Medicaid insurance increased post-expansion in ME and NE states (ME 16.7% to 23.0%, P < 0.001; NE 10.0% to 11.9%, P = 0.013) while rates of self-pay decreased in ME states only (ME 4.6% to 1.8%, P < 0.001; NE 8.1% to 8.4%, P = 0.620). Adjusted difference-in-differences analysis revealed lower odds of urgent/emergent PVI among all patients and all nonwhite patients in ME states post-expansion compared to NE states (all: odds ratio [OR] 0.53 [95% confidence interval 0.33-0.87], P = 0.011; nonwhite: OR 0.41 [0.19-0.88], P = 0.023). No differences were observed for 1-year major amputation (OR 0.70 [0.43-1.14], P = 0.152), primary patency (OR 0.93 [0.63-1.38], P = 0.726), or secondary patency (OR 1.29 [0.69-2.41], P = 0.431). Odds of 1-year mortality were higher in ME states post-expansion compared to NE states (OR 2.50 [1.07-5.87], P = 0.035), although 30-day mortality was not different (OR 2.04 [0.60-6.90], P = 0.253). Notably, odds of 1-year major amputation among Hispanic/Latino patients decreased in ME states post-expansion compared to NE states (OR 0.11 [0.01-0.86], P = 0.036)., Conclusions: ME was associated with lower odds of 1-year major amputation among Hispanic/Latino patients who underwent PVI for PAD. ME was also associated with lower odds of urgent/emergent procedures among patients overall and nonwhite patients specifically. However, 1-year mortality increased in the overall cohort. Further study is needed to corroborate our findings that ME may have benefits for certain underserved populations with PAD., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Biobased and biodegradable plastic mulch films (aka, mulch biofilm) have emerged as a sustainable alternative to conventional plastic mulch films in agriculture, promising to reduce soil contamination with plastic residues through in situ biodegradation. However, current standards certifying biodegradable plastics cannot predict biodegradability in natural settings. The scarce studies considering the possible biodegradation and ecotoxicity of mulch biofilms in soil systems question the environmental friendliness of these alternative options. This study assessed the biodegradation of a commercially available mulch biofilm by the soil-dwelling fungus Penicillium brevicompactum (in solid culture media and soil for 15 and 28 days, respectively), and the ecotoxicological effects of mulch biofilm microplastics on the earthworm Eisenia andrei (pristine or UV-weathered, at 0.125-0.250-0.500 g/kg). Results (from microplastics' mass loss, microscopy, and FTIR spectroscopy) suggest that the presence of P. brevicompactum promotes mulch biofilm's biodegradation. Exposure to environmental concentrations of pristine biofilm microplastics (and its ingestion) increased earthworms' sensitivity to touch, induced physiological alterations, decreased energy reserves, and decreased their reproduction (>30%). Conversely, exposure to weathered biofilm microplastics slightly increased earthworms' sensitivity, as well as carbohydrate reserves,without affecting their reproduction. The tested mulch biofilm seems to be, at first sight, an environmentally friendly alternative as it presented susceptibility for biodegradation by a widespread fungus, and the absence of ecotoxicological chronic effects on a key macroinvertebrate species in soil ecosystems when considering environmental relevant concentrations and plastics weathered conditions. Notwithstanding, the obtained results highlight the need to revise current standards, as they often neglect the role of, and their chronic effects on, naturally occurring organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Sirtuins play an important role in female mammalian reproductive function, participating in folliculogenesis and oocyte maturation. Studies exploring the consequences of inhibition/deletion of a specific sirtuin (SIRT) have demonstrated a deleterious effect on follicular growth, oocyte maturation, fertilization rates and embryo development, suggesting that sirtuins must have a relevant role in these processes. However, the exact mechanisms behind sirtuin function are still unclear. Most of the knowledge currently available derives from mouse studies and the literature is scarce in other species. So far, there is insufficient information about the subcellular localization of sirtuins during bovine meiosis, which would contribute to understanding the role and participation of sirtuins in the process of oocyte maturation, due to the close relation between location and function. Using in vitro maturation (IVM) of bovine oocytes we comprehensively documented and illustrated the subcellular localization pattern and distribution of SIRT1, 2 and 3 during meiotic progression. Moreover, we also detailed and quantified the colocalization of those sirtuins with the meiotic spindle, from the germinal-vesicle (GV)-stage until the Metaphase-II (MII)-stage. Our study demonstrated an increase in the expression of SIRT1, 2 and 3 during in vitro oocyte maturation and, for the first time, colocalization of SIRT1, 2 and 3 with both metaphase-I and metaphase-II spindles. These findings suggest that all three sirtuins may have a role in meiotic spindle assembly and microtubule dynamics in the bovine model. In addition, we have demonstrated the nuclear presence of SIRT1 and SIRT2 in the GV-stage. The apparent perinucleolar location of SIRT2 suggests that SIRT2 may shuttle into the nucleus at the GV-stage to regulate heterochromatin. This study reinforces the value of sirtuins during in vitro bovine meiotic progression and indicates potential molecular targets to improve maturation rates and embryo development., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Published by Elsevier Inc.)

This study assessed the effects of combined supplementation with canthaxanthin (Cx) and 25-hydroxycholecalciferol (25-OH-D 3 ) on incubation performance, fertility, and chick quality in European quail breeders. A total of 240 birds were distributed in a completely randomized design with 5 diets and 8 replicates. The animals were fed a basal diet containing 50 µg of vitamin D 3 or the basal diet supplemented with 3 ppm Cx and 34.5 µg 25-OH-D 3 , 6 ppm Cx, and 69 µg 25-OH-D 3 , 9 ppm Cx and 103.5 µg 25-OH-D 3 , or 12 ppm Cx and 138 µg 25-OH-D 3 . Incubation performance was analyzed in 2 periods (32 and 38 wk). Breeders aged 32 wk produced eggs with higher hatchability (P = 0.024), hatchability of fertile eggs (P = 0.026) and lower initial plus mid embryonic mortality (P = 0.021), whereas 38-week-old breeders generated chicks with a higher length at hatching (P < 0.001) and lower final plus pipped embryonic mortality (P = 0.021). In both age groups, Cx + 25-OH-D 3 levels had a quadratic effect on egg fertility (P < 0.001), hatchability of total (P < 0.001), and fertile eggs (P < 0.001). The fertility and the number of sperm cells in the perivitelline membrane was analyzed in two periods (26 and 40 wk). A quadratic effect of diet and days after mating on both parameters (P < 0.05) was observed. Eggs from supplementing breeders showed a high fertility (P < 0.001) and sperm cell counts (P < 0.001) for up to 7 and 3 d after mating, respectively, then the control group. Moreover, the supplementation of quail breeder diets with 6 ppm Cx + 69 µg 25-OH-D 3 enhances sperm cell longevity in sperm storage tubules, hatchability of total and fertile eggs, fertility, and chick quality, especially in older quail's breeders and reduces embryonic mortality., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Telomerase, oncogenes and tumor suppressors are closely associated with tumour occurrence, therefore these structures are being recognized as targets for the development of new anticancer drugs. The efficacy of several molecules in telomerase inhibition and regulation of genes expression, by adduct formation with G-quadruplexes (G4), has been studied by biophysical and biochemical methods with promising results. We report here the synthesis and structural characterization of a small positively charged diketopyrrolo[3,4-c]pyrrole derivative, identified as DPP(PyMe) 2 , that showed very promising results as G4 stabilizing ligand. The data obtained from UV-Vis and fluorescence experiments suggest that DPP(PyMe) 2 presents high affinity to G4 structures. Docking studies and molecular dynamics simulations unraveled the binding modes of the ligand with four G4 structures. The obtained results also allowed us to conclude that the DPP(PyMe) 2 ligand binds into the top G-tetrad or in a mixed binding mode depending on the GQ structure. A remarkable selectivity of DPP(PyMe) 2 for c-MYC and KRAS 32R in the presence of ds26 was observed by circular dichroism (CD) and fluorescence resonance energy transfer (FRET) melting experiments. CD titrations revealed a stabilization higher than 30 °C in the case of c-MYC G4 structure and, for the same sequence, DPP(PyMe) 2 showed the ability to block the activity of Taq polymerase in a dose-dependent manner. The subcellular localization obtained with confocal microscopy corroborates the results obtained by the other techniques and the obtained data suggest that DPP(PyMe) 2 is an attractive ligand for the development of G4 labelling probes., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Microplastics are ubiquitous contaminants of freshwater ecosystems. However, few ecotoxicity assays have been conducted on freshwater organisms using environmentally relevant concentrations of virgin and weathered microplastics. This work assessed the adverse effects of virgin and artificially weathered fragments of polystyrene and polypropylene on the microalga Raphidocelis subcapitata (72 h growth inhibition assay) and on embryos of the fish Danio rerio (96 h fish embryo assay) under environmentally relevant concentrations (2000-200,000 MP L -1 ) and high concentrations (12.5-100 mg L -1 ). Sizes of microplastics were measured as tens (polystyrene) to hundreds (polypropylene) of micrometers, while aging was assessed by measuring the carbonyl index. In the microalga, the tested high concentrations promoted growth, while environmentally relevant concentration induced either growth inhibition or promotion. In zebrafish embryos, environmentally relevant concentrations decreased body length and heart rates. No relevant effects were observed in organisms exposed to high concentrations for mortality, malformations, hatching rates, and swimming bladder inflation. Virgin microplastics presented slightly higher toxicity but direct comparison was hindered by the lack of a linear dose-response curve. Despite the lack of a clear pattern, adverse effects were often observed in the lowest environmentally relevant concentrations, raising concerns over the impacts of microplastics on freshwater ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The extraction of microplastics from organic-rich freshwater samples is challenging and limited information is available in the literature. This study aims at developing efficient methods for water volume reduction and organic matter removal in freshwater samples, while focusing on the reduction of the economic and environmental costs, maintaining microplastics integrity and avoiding contamination. For the water volume reduction approach, centrifuging freshwater samples (water, sediment, algae, leaves, driftwood, fish tissue) at different speeds (3500, 6000 rpm) and times (5, 10 min) showed that 3500 rpm for 5 min was efficient to settle the mineral and organic material, while preserving the polymers and showing high microplastic recovering rates (93 ± 6%). These recovery rates were significantly higher than the traditional sieving approach (77 ± 22%). The posterior minimal consumption of reagents resulting from the reduction of water volume helped to reduce the economic and environmental costs of the devised methodology, becoming more aligned with green chemistry principles. For biogenic organic matter removal, four digestion solutions were tested on freshwater samples, namely 10% potassium hydroxide, Fenton reagent (30% H 2 O 2  + Fe(II)), 7% and 10% sodium hypochlorite (NaClO), under 3 periods of time (1, 6 and 15 h), at 50 °C. Both 7% and 10% NaClO showed the highest rates of organic matter removal (86 ± 1% and 90 ± 1%, respectively), after 6 h at 50 °C. Exposure of virgin and aged polymers (polyethylene, polypropylene, polystyrene, polyvinyl chloride, nylon, polyethylene terephthalate) to NaClO showed no weight, visual, surface structure, Fourier transform infrared spectra and carbonyl index changes, except for nylon, although not to an extent that affected its identification. This method resulted in high recovery rates of polymers (92 ± 6%). Thus, 7% NaClO at 50 °C for 6 h (or overnight) may be efficiently used for microplastic analysis in organic-rich freshwater samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Marine ecosystems represent major sinks for persistent organic pollutants (POPs). Yet, while their regulations fit localized activity and emissions, POPs are mobile and can persist away from their source. The present review draws an environmental diagnostic of the organic substances studied over the past forty months, which ones accumulated the most, and where. Maximum reported concentration was used as a proxy for the accumulation of contaminants. POPs occurrences studied in the Jan 2018-April 2021 period were recorded into a database, along with (i) the geographical location of the sample and its coastal or offshore origin, (ii) the type of compartment analyzed (water vs sediment), as well as (iii) the POPs and the sample physical-chemical parameters reported. In the articles reviewed, maximum reported concentrations of POPs were in the ng/L range in seawater and in the μg/kg range in sediments. Some hotspots presented concentrations high enough to represent a hazard for sea organisms in the water columns (μg/L range) or in surficial sediments (mg/kg range). On a global scale, offshore (>1 km from the coast) maximum reported concentrations were, for the majority of the POPs, equivalent or higher than coastal ones. Finally, a POP solubility threshold (900 mg/L) was observed above which POPs would not be found in offshore waters, but only in sediments. This review highlights that studying POP accumulation away from their sources is fundamental for the diagnostic of long-lasting marine POPs contaminations. Further, POPs lipophilicity is a good predictor for offshore transport, and an indicator of interest for predicting sediment accumulation. Although POPs fate and transport in oceans is complex and require a finer analysis that this review could provide, the present work is a step forward identifying the hotspots in which POPs could be of particular concern, along with chemical indicators to predict for POPs accumulation in marine reservoirs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Microplastics have been found in fish, but most studies have focused on the digestive system without considering additional organs. Herein, the objective was to assess the presence of microplastics in internal organs (gills, guts, kidney, heart) of the Atlantic horse mackerel (Trachurus trachurus) captured of the coast of Portugal (Northeast Atlantic Ocean). Suspected microplastics were present in all organs, with particles of larger size (i.e., equivalent diameter) found in the gut and those of lower size in the heart and its luminal blood. Suspected microplastics of 1-10 μm were the most abundant (65.4%), more likely to translocate, owing to their minute size, but more difficult to properly characterize. These results highlight the need to expand the analytical work on organs and tissues for assessing microplastics in organisms, but also emphasize the actual need for developing analytical methods that allow for an accurate isolation, identification, and characterization of microplastics in biota., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This paper presents a convenient and efficient method to predict the mechanical solutions of a laminated Liquid Crystal Elastomers (LCEs) system subjected to combined thermo-mechanical load, based on a back propagation (BP) neural network which is trained by machine learning from a database established by analytical solutions. Firstly, the general solutions of temperature, displacement, and stress of any single layer in the LCEs system are obtained by solving the two-dimensional (2D) governing equations of both heat conduction and thermoelasticity. Then, the unknown coefficients in above general solutions are determined by a transfer-matrix method based on the continuity condition at the interface of adjacent layers and the combined thermo-mechanical loads condition at the surface of the LCEs system. The formula derivation and calculator program are verified through convergence studies and comparisons with FEM results. Finally, a database with displacements of LCEs system in a temperature field subjected to 561 sets of mechanical loads is established based on the presented analytical model. The BP neural network based on above database is further applied to establish the relationship between deformation and mechanical load to predict the elastic deformation of the LCEs system in a temperature field subjected to a mechanical load. Moreover, the BP network can also inverse the coefficients of mechanical load which induces the specific deformation in a temperature field. The numerical examples show that: (1) The deformation of a laminated LCEs system due to thermal load is limited within the range of human temperature changes from 36 °C to 40 °C. (2) The thickness of the LCE is a sensitive parameter on the deformation at the bottom surface of the system. (3) The accuracy of predicted displacements induced by the thermo-mechanical load and the inversed mechanical load based on deformation of the LCEs system in a temperature field using BP neural network reaches 99.6% and 98.5% respectively., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Landfilling and illegal waste disposal have risen to deal with the COVID-19 potentially infectious waste, particularly in developing countries, which aggravates plastic pollution and inherent environmental threats to human and animal health. It is estimated that 3.5 million metric tonnes of masks (equivalent to 601 TIR containers) have been landfilled worldwide in the first year, with the potential to increase global plastic municipal solid waste by 3.5%, alter biogas composition, and release 2.3 × 10 21 microplastics to leachates or adjacent environments, in the coming years. This paper reviews the challenges raised in the pandemic scenario on landfills and discusses the potential environmental and health implications that might drive us apart from the 2030 U.N. sustainable goals. Also, it highlights some innovative technologies to improve waste management (from collection to disposal, waste reduction, sterilization) and mitigates plastic leakage (emission control approaches, application of biotechnological and monitoring/computational tools) that can pave the way to environmental recovery. COVID-19 will eventually subside, but if no action is taken in the short-term towards effective plastic policies, replacement of plastics for sustainable alternatives (e.g., biobased plastics), improvement of waste management streams (prioritising flexible and decentralized approaches), and a greater awareness and responsibility of the general public, stakeholders, industries; we will soon reach a tipping-point in natural environments worldwide., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 Elsevier B.V. All rights reserved.)

Objective: To investigate evidence for the treatment of childhood colic by supplementing Lactobacillus reuteri in infants breastfed with breast milk., Methods: The study was conducted according to the PRISMA protocol. The databases used for acquiring data were PubMed and Web of Science, applying MeSH terms and free terms. Meta-analysis was conducted using Stata ™ 12.0. The risk of bias was evaluated by the Review Manager (RevMan) 5.3 tool, and the strength of evidence was assessed by the Grading of Recommendations Assessment, Development and Evaluation (GRADE)., Results: Ten clinical trials were included in the review. The administration of L. reuteri (DSM 17938 or ATCC55730) was tested in infants (n = 248) versus the control/placebo group (n = 229). Eight articles were included in the meta-analysis. There was a significant response in reducing crying time (minutes/day) and treatment effectiveness (reduction ≥ 50% in average daily crying time) in the first week (p = 0.001 and p = 0.003, respectively). These results were similar in the second, third weeks (p < 0.001 for both outcomes) and fourth weeks (p<0.001 and p = 0.002, respectively). The risk of bias was low for the majority of the studies. Confidence in evidence was considered very low for crying time and low for effectiveness treatment., Conclusions: The evidence shows that the administration of Lactobacillus reuteri to babies fed with breast milk reduces the crying time in babies diagnosed with colic. But our confidence in the effect estimate is limited., (Copyright © 2021. Published by Elsevier Ltd.)

Background: The present study aimed to identify for the first time sex differences in the development of CPP induced by intragastric alcohol administration in mice., Methods: Male and female adult Swiss mice were submitted to 16 days of conditioning with alcohol (0.5-3.0 g/kg, N = 8/dose/sex), with 2 post-conditioning tests (after 8 and 16 sessions) during the protocol., Results: 8 days of conditioning (4 alcohol sessions, 4 saline sessions) with intragastric alcohol administration were sufficient to induce CPP in male mice at the doses of 1.0, 1.5 and 2.0 g/kg. However, only higher doses (2.0, 2.5 and 3.0 g/kg) induced CPP in female mice using an 8-day conditioning protocol, while a 16-day conditioning protocol was necessary for the development of intragastric alcohol-induced CPP at the doses of 1.0 and 1.5 g/kg. Regardless of the conditioning protocol, higher doses or alcohol that had rewarding effects in females (2.5 and 3.0 g/kg) did not induce CPP in males, with a significant difference between males and females at those doses. Analysis of the potency (EC 50 ) and efficacy (E max ) of alcohol in inducing CPP when administered intragastrically in male and female mice showed significant sex differences with 8 conditioning sessions., Conclusions: Our data show a clear protocol (8 vs 16 days) and dose difference between male and female Swiss mice regarding the development of CPP induced by intragastric alcohol administration. Intragastric alcohol administration is closer to human drinking, and our protocol provides a more translational approach to studying the rewarding effects of alcohol in mice., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Chagas Disease (CD) is a neglected illness whose immunopathological mechanisms have not yet been plainly elucidated. The asymptomatic (indeterminate) form of CD is a long-term condition and approximately 20% to 35% of the individuals with this form evolve into one of the three chronic symptomatic clinical forms of CD, namely: cardiac, digestive or cardio-digestive (mixed). A variant of blood monocytes characterized by low expression of the HLA-DR antigen (CD14 + /HLA-DR low/‒ ) constitutes a subtype of myeloid-derived suppressor cells (MDSCs) whose main function is to regulate exacerbated inflammatory processes. The development of the symptomatic forms of CD can be related to the interaction between the host's immune system and the CD14 + /HLA-DR low/‒ immunosuppressive monocytes. Here, we evaluated, by flow cytometry, the absolute number and the HLA-DR antigenic density of this population of MDSCs in 57 patients with the diagnosis of CD: 34 with the symptomatic clinical forms (26 cardiac and 8 mixed) and 23 with the asymptomatic (indeterminate) form. The asymptomatic form exhibited a greater number of CD14 + /HLA-DR low/‒ monocytes and, accordingly, a low HLA-DR antigenic density, when compared to the symptomatic forms. It is possible to speculate that the predominance of CD14 + /HLA-DR low/- monocytes in the patients with the asymptomatic (indeterminate) form might have been a factor that could delay or even prevent the evolution of the asymptomatic form to the symptomatic forms of Chagas Disease., (Copyright © 2021. Published by Elsevier B.V.)

The use of disposable face masks became essential to fight against the COVID-19 pandemic, resulting in an unprecedented rise in their production and, unfortunately, to a new form of environmental contamination due to improper disposal. Recent publications reported the abundance of COVID-19-related litter in several environments, wildlife interaction with such items, and the contaminants that can be released from such protective equipment that has the potential to induce ecotoxicological effects. This paper provides a critical review of COVID-19 face mask occurrence in diverse environments and their adverse physiological and ecotoxicological effects on wildlife. It also outlines potential remediation strategies to mitigate the environmental challenge impose by COVID-19-related litter., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper., (Copyright © 2021. Published by Elsevier B.V.)

The ubiquity of microplastics raises issues regarding contamination control measures and laboratory practices. The objective was to adapt the use of counting chambers and plastic microplates on the ecotoxicity evaluation of microplastics. Counting chambers, originally used to quantify cells, can also be used to count high concentrations of microplastics (<100 μm) used in laboratory assays. By decontaminating the chamber and mixing the test solution with Nile Red (1:1), fluorescent particles can be easily counted under optical microscopy. Microplate wells, due to their composition, can be contaminated or release microplastics to the test medium, which can interfere with the results of ecotoxicity assays or spectroscopy readings. A cleaning method based on ethanol was developed, which effectively removed particles by 91% without interfering with microalgae yield. Besides providing practical applications that can improve ecotoxicity assays, this work intends to raise awareness on the need to adapt laboratory practices when working with microplastics., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Nile Red staining enables visual identification and quantification of fluorescent particles as a proxy to microplastics at low cost and high throughput, including those of small sizes (≥2 μm), when preceded by proper natural organic matter removal, but providing no chemical characterization. On the other hand, micro-spectroscopy methods allow chemical characterization of particles based on their spectra, essential for polymer identification, but are costly and time-consuming. This work addresses the combination of both Nile Red staining with micro-Raman spectroscopy for the identification of microplastics. Besides being useful for quantification, Nile Red staining can be advantageously used as an objective criterion for pre-selection of particles for micro-Raman spectroscopy, producing little interference. The use of the 442 nm laser in micro-Raman spectroscopy induces Nile Red luminescence thus allowing to target the specific suspected microplastics when using an orange filter, reducing the number of particles subjected to identification and improving sample throughput. Staining dyes could also be used for mapping suspected microplastics before targeted analysis by micro-Raman spectroscopy. Thus, coupling Nile Red with micro-Raman spectroscopy can be useful to improve time efficiency while using this equipment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Many methods have been used to isolate and identify microplastics from biological matrices. In biological samples, Nile Red can stain undigested residues, such as fats, soaps, and gels formed during organic matter removal, hindering the identification of fluorescent microplastics (≥2 μm). Thus, adjustments on sample preparation (e.g., fat removal) are required for the accurate identification of Nile Red stained microplastics. Multiples tests allowed to identify that digestion with 10% KOH at 60 °C for 24 h, followed by treatments with boiling water, acetone, and staining, produced good results in fourteen biological samples, including vertebrates and invertebrates. Digestion efficiencies were 94-100%, except for feces, which were 87%. Recovery rates of spiked microplastics were 97-100%, and few effects were observed in the infrared spectra and carbonyl index of seven polymers, with only the occasional yellowing suggesting surface changes. Filtration rates were improved by reducing the amount of sample. Small fluorescent microplastics could be identified in all samples under the microscope. Overall, the proposed method was efficient in removing natural organic matter from biological samples for Nile Red staining, requiring minimal sample handling, improving sample throughput, and allowing quantification of fluorescent microplastics in biological samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)