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Plastic pollution is a consequential problem worldwide, prompting the widespread use of biodegradable plastics (BPs). However, not all BPs are completely degradable under natural conditions, but instead produce biodegradable microplastics (BMPs), release chemical additives, and absorb micropollutants, thus causing toxicity to living organisms in similar manners to conventional plastics (CPs). The new problems caused by biodegradable plastics cannot be ignored and requires a thorough comparison of the differences between conventional and biodegradable plastics and microplastics. This review comprehensively compares their environmental fates, such as biodegradation and micropollutant sorption, and ecotoxicity in soil and water environments. The results showed that it is difficult to determine the natural conditions required for the complete biodegradation of BPs. Some chemical additives in BPs differ from those in CPs and may pose new threats to ecosystems. Because of functional group differences, most BMPs had higher micropollutant sorption capacities than conventional microplastics (CMPs). The ecotoxicity comparison showed that BMPs had similar or even greater adverse effects than CMPs. This review highlights several knowledge gaps in this new field and suggests directions for future studies.

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We aimed to assess the association between medical financial hardship and psychological burden and the moderating role of social and mental health support. 2021 United States National Health Interview Survey was used. Financial hardship was defined as having financial worry, material hardship, or cost-related care nonadherence. Psychological burden was measured using perceived general health status, satisfaction with life, and serious psychological distress (SPD). Of 29,370 included adults, 49% experienced financial hardship in the last 12 months. Financial hardship was associated with a higher psychological burden (odds ratio [OR], 3.58; 95% confidence interval [CI], 2.43-5.47 for SPD). Eleven percent received counseling/therapy from mental health professionals, and 90% had experienced frequent social support. Frequent social support was associated with lower financial hardship (OR, 0.71; 95% CI, 0.63-0.80) and psychological burden (OR, 0.28; 95% CI, 0.19-0.42 for SPD). Previous mental health support was associated with higher financial hardship (OR,1.40; 95% CI, 1.28-1.54) and psychological burden (OR, 9.75; 95% CI, 6.97-13.94 for SPD). Those experiencing financial hardship had lower odds of SPD if they received mental health support in the last 12 months (OR, 0.57; 95% CI, 0.39-0.85). Future interventions should also focus on improving social support and mental health for patients as a way of mitigating medical financial hardship.

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Background: Allostatic load (AL) is the accumulation of physiological dysregulation attributed to repeated activation of the stress response over a lifetime. We assessed the utility of AL as a prognostic measure for high-risk benign breast biopsy pathology results. Method: Eligible patients were women 18 years or older, with a false-positive outpatient breast biopsy between January and December 2022 at a tertiary academic health center. AL was calculated using 12 variables representing four physiological systems: cardiovascular (pulse rate, systolic and diastolic blood pressures, total cholesterol, high-density lipoprotein, and low-density lipoprotein); metabolic (body mass index, albumin, and hemoglobin A1C); renal (creatinine and estimated glomerular filtration rate); and immune (white blood cell count). Multivariable logistic regression was used to assess the association between AL before biopsy and breast biopsy outcomes controlling for patients' sociodemographics. Results: In total, 170 women were included (mean age, 54.1 ± 12.9 years): 89.4% had benign and 10.6% had high-risk pathologies (radial scar/complex sclerosing lesion, atypical ductal or lobular hyperplasia, flat epithelial atypia, intraductal papilloma, or lobular carcinoma in-situ). A total of 56.5% were White, 24.7% Asian, and 17.1% other races. A total of 32.5% identified as Hispanic. The mean breast cancer risk score using the Tyrer-Cuzick model was 11.9 ± 7.0. In multivariable analysis, with every one unit increase in AL, the probability of high-risk pathology increased by 37% (odds ratio, 1.37; 95% confidence interval, 1.03, 1.81; p = 0.03). No significant association was seen between high-risk pathology and age, ethnicity, breast cancer risk, or area deprivation index. Conclusion: Our findings support that increased AL, a biological marker of stress, is associated with high-risk pathology among patients with false-positive breast biopsy results.

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Infochemicals refer to chemicals responsible for information exchange between organisms. We evaluated the effects of Daphnia magna and Daphnia galeata infochemicals on Microcystis aeruginosa for 15d. The Daphnia infochemicals were obtained from spent medium after culturing Daphnia in Elendt M4 medium for 48 h. Both Daphnia infochemicals significantly increased (p < 0.05) the intracellular reactive oxygen species level and microcystin-LR concentration in M. aeruginosa. This cellular effect increased colony formation of M. aeruginosa, thereby inhibiting the growth of M. aeruginosa. D. galeata infochemicals provoked significantly greater (p < 0.05) adverse effects on M. aeruginosa than those of D. magna infochemicals, which were further exaggerated by pre-exposure of Daphnia to M. aeruginosa. This result seems to be related to the different compositions and concentrations of Daphnia infochemicals. Several Daphnia infochemicals, such as methyl ferulate, cyclohexanone, 3, 5-dimethyl, hexanedioic acid, and bis(2-ethylhexyl) ester, showed a high correlation with M. aeruginosa cell concentration (|r | >0.6), suggesting that they may play a key role in controlling harmful cyanobacteria. Additionally, pre-exposure of D. magna and D. galeata to M. aeruginosa produced oleic acid, methyl ester, and n-hexadecanoic acid, with a highly correlation with M. aeruginosa cell concentration (|r | >0.6). p-tolyl acetate and linoleic acid were detected only in the pre-exposed D. galeata infochemicals. These findings suggest that some of Daphnia infochemicals identified in this study can be a promising tool to control M. aeruginosa growth. However, further studies are required to verify the specific actions of these infochemicals against cyanobacteria.

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OBJECTIVES: Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease globally, with an estimated prevalence exceeding 25%. Variants in the PNPLA3 and HSD17B13 genes have been a focus of investigations surrounding the etiology and progression of NAFLD and are believed to contribute to a greater burden of disease experienced by Hispanic Americans. However, little is known about socioeconomic factors influencing NAFLD progression or its increased prevalence among Hispanics. MATERIALS AND METHODS: We cross-sectionally analyzed 264 patients to assess the role of genetic and socioeconomic variables in the development of advanced liver fibrosis in individuals at risk for NAFLD. RESULTS: Adjusting for age, sex, body mass index, and PNPLA3 genotype, lacking a college degree was associated with 3.3 times higher odds of advanced fibrosis (95% confidence interval [CI]: 1.21-8.76, p = 0.019), an effect comparable to that of possessing the major PNPLA3 risk variant. Notably, the effect of PNPLA3 genotype on advanced fibrosis was attenuated to nonsignificance following adjustment for education and other socioeconomic markers. The effect of the protective HSD17B13 variant, moreover, diminished after adjustment for education (odds ratio [OR]: 0.39 [95% CI: 0.13-1.16, p = 0.092]), while lower education continued to predict advanced fibrosis following multivariable adjustment with an OR of 8.0 (95% CI: 1.91-33.86, p = 0.005). DISCUSSION: Adjusting for education attenuated the effects of genotype and Hispanic ethnicity on liver fibrosis, suggesting that social factors-rather than genes or ethnicity-may be driving disease severity within some populations. Findings reveal the importance of including socioenvironmental controls when considering the role of genetics or ethnicity in complex disease.

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BACKGROUND: This study systematically reviewed interventions mitigating financial hardship in patients with cancer and assessed effectiveness using a meta-analytic method. METHODS: PubMed, Cochrane, Scopus, CINAHL, and Web of Science were searched for articles published in English during January 2000-April 2023. Two independent reviewers selected prospective clinical trials with an intervention targeting and an outcome measuring financial hardship. Quality appraisal and data extraction were performed independently by two reviewers using a quality assessment tool. A random-effects model meta-analysis was performed. Reporting followed the preferred reporting items for systematic review and meta-analyses guidelines. RESULTS: Eleven studies (2211 participants; 55% male; mean age, 59.29 years) testing interventions including financial navigation, financial education, and cost discussion were included. Financial worry improved in only 27.3% of 11 studies. Material hardship and cost-related care nonadherence remained unchanged in the two studies measuring these outcomes. Four studies (373 participants; 37% male, mean age, 55.88 years) assessed the impact of financial navigation on financial worry using the comprehensive score of financial toxicity (COST) measure (score range, 0-44; higher score = lower financial worry) and were used for meta-analysis. There was no significant change in the mean of pooled COST score between post- and pre-intervention (1.21; 95% confidence interval, -6.54 to 8.96; p = .65). Adjusting for pre-intervention COST, mean change of COST significantly decreased by 0.88 with every 1-unit increase in pre-intervention COST (p = .02). The intervention significantly changed COST score when pre-intervention COST was ≤14.5. CONCLUSION: A variety of interventions have been tested to mitigate financial hardship. Financial navigation can mitigate financial worry among high-risk patients.

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Micro-pollutants (specifically antibiotics and personal care products) and potential bacterial contamination pose a severe threat to human health and marine life. The study derives indigenous novel fibrous hydrophobic nanocomposite, efficient in decontaminating the micro-pollutants (tetracycline (TC) and bisphenol A (BPA)) and potential pathogens (S. pyogenes and E. coli) from aqueous wastes. A facile method synthesizes the fibrous attapulgite (ATP)- poly(4-vinylpyridine-co-styrene) (PVP) framework decorated in situ with the Ag0 nanoparticles (ATP@PVP/Ag0). A greener method using the Artocarpus heterophyllus leaf extract derives the Ag0(NPs). Various analytical methods extensively characterize the materials. A comprehensive study that includes pH, concentration, background electrolytes, and ionic strength reveals the sorptive removal insights of TC and BPA utilizing the ATP@PVP solid. The elimination of tetracycline (TC) and bisphenol A (BPA) agrees well with the pseudo-second-order kinetics. The pH 3.07 and 6.06 favor removing TC and BPA with the capacity of 10.86 mg/g and 17.36 mg/g at 25 °C. The hydrogen bonding and hydrophobic interactions predominate the sorption mechanism, and the material shows remarkable stability and reusability in repeated sorption/desorption operations. Similarly, the natural water implications and flow-bed system show fair applicability of solid in decontaminating the TC and BPA in an aqueous medium. Further, the material ATP@PVP/Ag0 exhibits very high inhibition of potential pathogens S. pyogenes and E. coli and optimizes the solid dose and solution pH.

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Fluorescent initiator tRNAs (tRNAi) play a crucial role in studying protein synthesis, yet generating highly fluorescent tRNAi complexes remains challenging. We present an optimized strategy to effectively generate highly fluorescent initiator-tRNA complexes in living cells. Our strategy allows the generation of Fluo-Met-tRNAiMet complexes. These complexes can have highly chromogenic N-terminal labeling. For generating such complexes, we use either purified fluorescent methionine (PFM) or non-purified fluorescently labeled methionine (NPFM). Furthermore, PFM promotes the active generation of endogenous tRNAi in cells, leading to highly efficient Fluo-Met-tRNAiMet complexes. Finally, PFM-tRNAiMet complexes also facilitate the visualization of native fluorescently labeled Tat binding to beads. This demonstrates the potential of our approach to advance precision protein engineering and biotechnology applications.

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The adverse effects of microplastics (MPs) on Daphnia magna have been extensively studied; however, their population-level effects are relatively unknown. This study investigated the effect of polyethylene MP fragments (33.90 ± 17.44 µm) and benzophenone-3 (BP-3), which is a widely used plastic additive (2.91 ± 0.02% w/w), on D. magna population dynamics in a 34-day microcosm experiment. In the growth phase, neither MP nor MP/BP-3 fragments changed the population size of D. magna compared with the control. However, MP/BP-3 fragments significantly reduced (p < 0.05) the population biomass compared to that of the control, whereas MP fragments did not induce a significant reduction. The MP/BP-3 group had a significantly higher (p < 0.05) neonate proportion than that in the control and MP groups. MP/BP-3 fragments upregulated usp and downregulated ecrb, ftz-f1, and hr3, altering gene expression in the ecdysone signaling pathway linked to D. magna growth and development. These findings suggested that BP-3 in MP/BP-3 fragments may disrupt neonatal growth, thereby decreasing population biomass. In the decline phase, MP fragments significantly decreased (p < 0.05) the population size and biomass of D. magna compared with the control and MP/BP-3 fragments. This study highlights the importance of plastic additives in the population-level ecotoxicity of MPs.

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Plastic products are extensively used worldwide, but inadequate management of plastic waste results in significant plastic pollution. Biodegradable plastic (BPs) offers an alternative to traditional plastics, however, not all BPs can fully degrade under natural conditions. Instead, they may deteriorate into biodegradable microplastic (BMPs) at a faster rate than conventional plastic, thereby posing an additional hazard to aquatic environments. This study provides a comprehensive overview of the fate of BPs in aquatic systems and their eco-toxicological effects on aquatic organisms such as algae, invertebrates, and fish. The findings highlight that BMPs have comparable or heightened effects compared to conventional microplastics (MPs) which physiochemical characteristic of the polymer itself or by the chemical leached from the polymeric matrix can affect aquatic organisms. While BPs is not a flawless solution to address plastic pollution, future research should prioritize investigating their production, environmental behavior, ecological impact, and whether BMPs inflict greater harm than conventional MPs.

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Biodegradable plastics (BPs) are widely used as alternatives to non-BPs due to their inherent ability to undergo facile degradation. However, the ecotoxicological impact of biodegradable microplastics (MPs) rarely remains scientific documented especially to aquatic ecosystem and organisms compared to conventional microplastics. Therefore, this study aimed to investigate the ecotoxicity of biodegradable polylactic acid (PLA) MPs to Daphnia magna with that of conventional polyethylene (PE) MPs with and without ultraviolet (UV) treatment (4 weeks). The acute toxicity (48 h) of PLA MPs was significantly higher than that of PE MPs, potentially attributable to their elevated bioconcentration resulting from their higher density. UV treatment notably reduced the particle size of PLA MPs and induced new hydrophilic functional groups containing oxygen. Thus, the acute lethal toxicity of PLA MPs exhibited noteworthy increase, compared to before UV treatment after UV treatment, which was greater than that of UV-PE MPs. In addition, UV-PLA MPs showed markedly elevated reactive oxygen species concentration in D. magna compared to positive control. However, there was no significant increase in the level of lipid peroxidation, possibly due to successful defense by antioxidant enzymes (superoxide dismutase and catalase). These findings highlight the ecotoxicological risks of biodegradable MPs to aquatic organisms, which require comprehensive long-term studies.

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The increase in the usage of biodegradable microplastics (MPs) as an alternative to conventional plastics has necessitated comprehensive ecotoxicity assessments of biodegradable MPs alongside conventional MPs. This study aimed to assess ecotoxicity of biodegradable polylactic acid (PLA) MPs at concentration of 1 and 5 mgL-1 including a genetic analysis of Daphnia magna, and compared to effects of conventional polyethylene terephthalate (PET) MPs. The survival rate for D. magna exposed to 5 mg L-1 of PLA-MPs declined to 52.4 %, signifying a higher rate of mortality when contrasted with PET-MPs, which exhibited 85.7 % survival rate. Chronic exposure to 1 and 5 mgL-1 PLA-MPs resulted in a decrease of offspring, while increasing the sex ratio and deformed embryo. Interestingly, down-regulation of the SOD and AK genes was observed in D. magna after exposure to 5 mgL-1 of PLA-MPs, while 1 mgL-1 of PLA-MPs up-regulated. These results means that 5 mgL-1 PLA-MP could not produce energy and cope with oxidative stress, resulting in high mortality, and 1 mgL-1 of MP was maintained survival due to energy production and antioxidant action. This study contributes to our understanding of biodegradable microplastics (BMPs) toxic effects on D. magna which could be similar to conventional MPs and provide the importance of ecotoxicological data for risk assessment of BMPs in aquatic organisms.

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Microplastics and antibiotics are emerging as ubiquitous contaminants in farmland soil, harming crop quality and yield, and thus threatening global food security and human health. However, few studies have examined the individual and joint effects of degradable and/or non-degradable microplastics and antibiotics on crop plants. This study examined the individual and joint effects of polyethylene (PE) and polylactic acid (PLA) microplastics and the antibiotic oxytetracycline (OTC) on pak choi by measuring its growth, photosynthesis, antioxidant enzyme activity, and metabolite levels. Microplastics and/or oxytetracycline adversely affected root weight, photosynthesis, and antioxidant enzyme (superoxide dismutase, catalase, and ascorbate peroxidase) activities. The levels of leaf metabolites were significantly altered, causing physiological changes. Biosynthesis of plant secondary metabolites and amino acids was altered, and plant hormones pathways were disrupted. Separately and together, OTC, PE, and PLA exerted phytotoxic and antagonistic effects on pak choi. Separately and together with OTC, degradable microplastics altered the soil properties, thus causing more severe impacts on plant performance than non-degradable microplastics. This study elucidates the effects on crop plants of toxicity caused by co-exposure to degradable or non-degradable microplastic and antibiotics contamination and suggests mechanisms.

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The use of data-based species distribution models (SDMs) has increased significantly in recent years. However, studies of determining the minimum requirements of occurrence sites from ecological monitoring datasets used in species distribution modelling remain insufficient. Therefore, this study proposed a framework to determine the threshold of minimum occurrence sites for SDMs by assessing compliance with Benford's law. The compliance test verified that the national-scale freshwater fish monitoring dataset was natural and reliable. Results derived from true skill statistics (TSS) determined the minimum number of occurrence sites for reliable species distribution modelling was 20 with a TSS value of 0.793 and an overall accuracy of 0.804. The Benford compliance test has shown to be a useful tool for swift and efficient evaluation of the reliability of species occurrence datasets, or the determination of the threshold of occurrence sites before species distribution modelling. Further studies regarding the evaluation of this method's transferability to other species and validation using SDM performance are required. Overall, the framework proposed in this study demonstrates that Benford compliance test applied to species monitoring datasets can be used to derive a universal and model-independent minimum occurrence threshold for SDMs.

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Healthcare price transparency is an effort to inform patient decision-making, but also to decrease prices and their variation across healthcare systems for equivalent medical services. The initiative is meaningful only for medical services that are shoppable-such as imaging examinations-for which patients incur out-of-pocket costs. Therefore, several countries in which patients commonly share a portion of their healthcare costs have been implementing mandates to improve healthcare price transparency. However, the provisional implementation has many issues, especially in the United States, including provider non-compliance and limited accessibility of price transparency tools by the general public. Many of the existing tools are not user-friendly, are difficult to navigate, focus on charges and health plan negotiated rates rather than patients' out-of-pocket costs, and disclose prices on the service level instead of per episode of care. As such, the disclosed amounts are often not reliable. Many price transparency tools also lack valid and measurable quality metrics, which can result in a selection of high-cost care as a proxy for high-value care, as well as an increase in healthcare prices when providers want to imply they offer high-quality care. Nevertheless, the impact of the initiatives on patients' decision-making and healthcare costs remains unclear. While transparency initiatives are patient-centric, efforts should be made to increase patient engagement, provide accurate patient-specific out-of-pocket cost information, compare available treatment and provider alternatives, and couple price information with quality metrics to enable making fully informed decisions.

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INTRODUCTION: Retrospective studies report that angiotensin-converting enzyme inhibitors (ACEIs) may reduce the severity of COVID-19, but prospective data on de novo treatment with ACEIs are limited. The RAMIC trial was a randomized, multicenter, placebo-controlled, double-blind, allocation-concealed clinical trial to examine the efficacy of de novo ramipril versus placebo for the treatment of COVID-19. METHODS: Eligible participants were aged 18 years and older with a confirmed diagnosis of SARS-CoV-2 infection, recruited from urgent care clinics, emergency departments, and hospital inpatient wards at eight sites in the USA. Participants were randomly assigned to daily ramipril 2.5 mg or placebo orally in a 2:1 ratio, using permuted block randomization. Analyses were conducted on an intention-to-treat basis. The primary outcome was a composite of mortality, intensive care unit (ICU) admission, or invasive mechanical ventilation by day 14. RESULTS: Between 27 May 2020 and 19 April 2021, a total of 114 participants (51% female) were randomized to ramipril (n = 79) or placebo (n = 35). The overall mean (± SD) age and BMI were 45 (± 15) years and 33 (± 8) kg/m2. Two participants in the ramipril group required ICU admission and one died, compared with none in the placebo group. There were no significant differences between ramipril and placebo in the primary endpoint (ICU admission, mechanical ventilation, or death) (3% versus 0%, p = 1.00) or adverse events (27% versus 29%, p = 0.82). The study was terminated early because of a low event rate and subsequent Emergency Use Authorization of therapies for COVID-19. CONCLUSION: De novo ramipril was not different compared with placebo in improving or worsening clinical outcomes from COVID-19 but appeared safe in non-critically ill patients with COVID-19. TRIAL REGISTRATION: Clinicaltrials.gov NCT04366050.

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BACKGROUND AND PURPOSE: Cognitive and behavioral changes are common in amyotrophic lateral sclerosis (ALS), with about 15% of patients presenting with overt frontotemporal dementia and 30%-50% with varying degrees of impairments. We aimed to develop and validate the Korean version of the Edinburgh Cognitive and Behavioral ALS Screen (ECAS-K), a brief multidomain assessment tool developed for ALS patients with physical disability. METHODS: We developed the ECAS-K according to the translation guidelines, and administered it to 38 patients with ALS and 26 age- and education-level-matched controls. We also administered the Montreal Cognitive Assessment (MoCA) and Frontal Assessment Battery (FAB) to investigate convergent validity, and the Center for Neurologic Study-Liability Scale to assess the association between pseudobulbar affect and cognitive/behavioral changes. RESULTS: Internal consistency among the ECAS-K test items was found to be high, with a Cronbach's alpha of 0.87. Significant differences were found between patients with ALS and the controls in language, fluency, and memory functions (p<0.05). Abnormal performance based on the ECAS total score was noted in 39.4% of patients, and 66.6% presented behavioral changes in at least one domain. Significant correlations were observed between the scores of the ECAS-K and those of other cognitive screening tools (MoCA and FAB, with correlation coefficients of 0.69 and 0.55, respectively; p<0.01). CONCLUSIONS: We developed and validated the ECAS-K which could be used as an effective tool to screen the cognitive and behavioral impairments in Korean patients with ALS.

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Microplastic (MP) pollution poses a growing concern in freshwater ecosystems, which are further threatened by global warming. Thus, this study investigated the effect of elevated temperature (25 °C) on acute toxicity of polyethylene MP fragments to Daphnia magna over a 48 h period. At the reference temperature (20 °C), MP fragments (41.88 ± 5.71 µm) induced over 70 times higher lethal toxicity than that induced by MP beads (44.50 ± 2.50 µm), with median effective concentrations (EC50) of 3.89 and 275.89 mg L-1, respectively. Elevated temperature significantly increased (p < 0.05) the lethal (EC50 = 1.88 mg L-1) and sublethal (lipid peroxidation and total antioxidant capacity) toxicity in D. magna exposed to MP fragments compared to those at the reference temperature. Additionally, the elevated temperature led to a significant increase (p < 0.05) in the bioconcentration of MP fragments in D. magna. Overall, the present study increases understanding for the ecological risk assessment of microplastics under global warming, highlights that elevated temperature can be seriously increased bioconcentration of MP fragments, leading to increased acute toxicity in D. magna.

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BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) and type 2 diabetes mellitus (T2DM) are independent risk factors for cardiovascular disease (CVD). AIMS: To examine the clinical utility of liver fat quantification for determining CVD risk among a well-phenotyped cohort of patients with T2DM. METHODS: This was a cross-sectional analysis of a prospective cohort of adults aged ≥50 with T2DM. Liver fat was quantified with magnetic resonance imaging proton-density-fat-fraction (MRI-PDFF), an advanced imaging-based biomarker. Patients were stratified into a higher liver fat group (MRI-PDFF ≥ 14.6%), and a lower liver fat group (MRI-PDFF < 14.6%). The co-primary outcomes were CVD risk determined by Framingham and Atherosclerotic Cardiovascular Disease (ASCVD) risk scores. High CVD risk was defined by risk scores ≥20%. RESULTS: Of the 391 adults (66% female) in this study, the mean (±SD) age was 64 (±8) years and BMI 30.8 (±5.2) kg/m2 , respectively. In multivariable analysis, adjusted for age, gender, race, and BMI, patients in the higher liver fat group had higher CVD risk [OR = 4.04 (95% CI: 2.07-7.88, p < 0.0001)] and ASCVD risk score [OR = 2.85 (95% CI: 1.19-6.83, p = 0.018)], respectively. CONCLUSION: Higher liver fat content increases CVD risk independently of age, gender, ethnicity and BMI. These findings raise the question whether liver fat quantification should be incorporated into risk calculators to further stratify those with higher CVD risk.

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Chemical conjugation of purified proteins of interest (POIs) in Escherichia coli cells is effective for high expression but has limitations for highly chromogenic dual labeling of intrinsically disordered native proteins (IDNPs). Our probes can tag IDNPs using chemical conjugation during protein synthesis and folding while preserving biologically active structures in mammalian cells. We fluorescently labeled IDNPs in mammalian cells using pure fluorescent methionine and ATTO 565-biotin at the N-or C-terminus, respectively. The dual-labeled Tat protein was used as a model for IDNPs in HeLa cells and detected using Ni-NTA beads to estimate its highly chromogenic concentration. We also demonstrated highly chromogenic double labeling of genetically encoded fluorescent-Tat expression in eukaryotic cells using a single fluorescent dye pair with Förster resonance energy transfer (FRET) ratio and two-color correlation analysis. This study aims to solve native POI processing and achieve ultra-sensitive protein folding for biological and ecological applications at the nanoscale.

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