Preterm birth and weight-for-gestational age for risks of autism spectrum disorder and intellectual disability: A nationwide population-based cohort study.

PURPOSE: To evaluate gestational age (GA) and small-for-gestational age (SGA) as continuums and gender on the incidences of autism spectrum disorder (ASD) and co-occurring intellectual disability (ID). METHODS: This is a population-based cohort study using the 2004-2008 Taiwan Maternal and Child Health Database. The diagnosis of ASD was determined by International Classification of Diseases, 9th Revision (ICD-9). Generalized estimating equations models were fit to evaluate associations between perinatal variables and ASD. RESULTS: This study included 916,315 individuals. A total of 9474 (1.0%) children were diagnosed with ASD, among whom 1594 (16.8%) had co-occurring ID. Lower GA carried higher odds of ASD with ID (GA < 28 weeks, aOR: 4.26, 95% CI: 2.13, 8.50; GA 28-30 weeks, aOR: 2.80, 95% CI: 1.57, 4.97; GA 31-33 weeks, aOR: 1.63, 95% CI: 1.05, 2.55; GA 34-36 weeks, aOR: 1.39, 95% CI: 1.16, 1.67) and ASD without ID (GA < 28 weeks, aOR:2.05, 95% CI: 1.25, 3.36; GA 28-30 weeks, aOR: 2.02, 95% CI: 1.46, 2.79; GA 31-33 weeks, aOR: 1.42, 95% CI: 1.13, 1.77; GA 34-36 weeks, aOR: 1.18, 95% CI: 1.08, 1.29). Male preterm infants had ASD risks negatively correlated to GA, while ASD risks were significantly increased only among female infants born late preterm. The degree of SGA showed a stepwise increased risk for ASD with and without ID in both male and female infants. CONCLUSION: Lower GA and the degree of SGA are both associated with ASD susceptibility, either with or without co-occurring ID, and remarkably increased the risk of ID.

Predicting in-hospital length of stay for very-low-birth-weight preterm infants using machine learning techniques.

BACKGROUND/PURPOSE: The in-hospital length of stay (LOS) among very-low-birth-weight (VLBW, BW < 1500 g) infants is an index for care quality and affects medical resource allocation. We aimed to analyze the LOS among VLBW infants in Taiwan, and to develop and compare the performance of different LOS prediction models using machine learning (ML) techniques. METHODS: This retrospective study illustrated LOS data from VLBW infants born between 2016 and 2018 registered in the Taiwan Neonatal Network. Among infants discharged alive, continuous variables (LOS or postmenstrual age, PMA) and categorical variables (late and non-late discharge group) were used as outcome variables to build prediction models. We used 21 early neonatal variables and six algorithms. The performance was compared using the coefficient of determination (R2) for continuous variables and area under the curve (AUC) for categorical variables. RESULTS: A total of 3519 VLBW infants were included to illustrate the profile of LOS. We found 59% of mortalities occurred within the first 7 days after birth. The median of LOS among surviving and deceased infants was 62 days and 5 days. For the ML prediction models, 2940 infants were enrolled. Prediction of LOS or PMA had R2 values less than 0.6. Among the prediction models for prolonged LOS, the logistic regression (ROC: 0.724) and random forest (ROC: 0.712) approach had better performance. CONCLUSION: We provide a benchmark of LOS among VLBW infants in each gestational age group in Taiwan. ML technique can improve the accuracy of the prediction model of prolonged LOS of VLBW.

Neurodevelopmental Outcomes after Bevacizumab Treatment for Retinopathy of Prematurity: A Meta-analysis.

PURPOSE: To evaluate neurodevelopmental outcomes after intravitreal bevacizumab (IVB) therapy in retinopathy of prematurity (ROP) infants compared with those not exposed to IVB. CLINICAL RELEVANCE: The primary concern regarding IVB treatment of ROP is the potential systemic side effects, especially the risk of causing severe neurodevelopmental impairment (sNDI). Results regarding neurodevelopmental outcomes after IVB therapy are conflicting. METHODS: We conducted a meta-analysis and searched PubMed, Embase, and Web of Science for related publications from inception through March 12, 2020. The eligibility criteria were as follows: comparative studies of ROP patients that (1) included IVB as a treatment arm, (2) included a control group without bevacizumab treatment, and (3) reported on at least 1 neurodevelopmental outcome, such as sNDI, Bayley Scales of Infant and Toddler Development, Third Edition (Bayley III), composition scores, or cerebral palsy (CP). The primary outcome was sNDI, with the odds ratio (OR) calculated. Secondary outcomes were mean differences (MDs) for cognitive, language, and motor scores (Bayley III) and OR for CP. The quality of evidence was assessed using the Grades of Recommendation, Assessment, Development, and Evaluation approach. RESULTS: Eight studies, 6 including laser-controlled ROP infants and 2 including ROP infants not requiring treatment, were included. The weighted OR for sNDI in the IVB group was 1.39 (95% confidence interval [CI], 0.98-1.97). The weighted MDs were -1.92 (95% CI, -4.73 to 0.88), -1.32 (95% CI, -4.65 to 1.99), and -3.66 (95% CI, -6.79 to -0.54) for cognitive, language, and motor scores in Bayley III, respectively. The OR for CP was 1.20 (95% CI, 0.56-2.55). No differences were observed between the preset subgroups comprising laser-controlled ROP infants and ROP infants not requiring treatment. The current quality of evidence was rated as low (sNDI and all Bayley III scores) to very low (CP). CONCLUSIONS: Risk of sNDI was not increased in ROP patients after IVB treatment. Bayley III scores were similar in the IVB and control groups, except for a minor difference in motor performance. These findings suggest that the risk of additional sNDI after IVB treatment is low. Randomized trials are warranted to provide a higher quality of evidence.

A synergistic effect of variability in estimated glomerular filtration rate with chronic kidney disease on all-cause mortality prediction in patients with type 2 diabetes: a retrospective cohort study.

BACKGROUND: The combination of diabetes mellitus (DM) and chronic kidney disease (CKD) is associated with a high risk of mortality. Annual assessment of the estimated glomerular filtration rate (eGFR) is recommended for patients with DM. We investigated the effect of variability in annual eGFR values on all-cause mortality in patients with type 2 DM. METHODS: In this retrospective cohort study, we enrolled patients with eGFR data between 01 Aug 2017 and 31 July 2018. We defined the index eGFR as the first available eGFR value within the enrollment year and collected additional annual eGFR data from the previous three years. A total of 3592 patients with type 2 DM were enrolled, including 959 patients with CKD (index eGFR < 60 mL/min/1.73 m2) and 2633 patients without CKD. We assessed eGFR variability by using the standard deviation (SD) of the three annual eGFR and index eGFR values. We divided patients into subgroups according to the median SD of their annual eGFR (7.62 mL/min/1.73 m2). The primary endpoint was all-cause mortality after the index eGFR was assessed. RESULTS: During a median follow-up of 19 months (interquartile range: 18‒20 months), 127 (3.5%) deaths occurred among all 3592 enrolled patients. The highest mortality risk was observed in the high SD with CKD group, with a hazard ratio (HR) of 2.382 [95% confidence interval (CI) 1.346‒4.215] in comparison to the low SD without CKD group after adjusting for the associated factors. In patients without CKD, a high SD was an independent risk factor for mortality (HR = 2.105, 95% CI 1.256‒3.528). According to the C-index, the mortality prediction ability was better for the index eGFR + SD model than for the index eGFR alone model (0.671 vs. 0.629, P < 0.001). CONCLUSION: There was a synergistic effect of eGFR variability with single-measured eGFR for the prediction of mortality in patients with type 2 DM. The SD of the annual eGFR values was also an independent predictor of mortality in patients with an eGFR > 60 mL/min/1.73 m2.

The symptoms, resourcefulness and quality of life in community-based patients with schizophrenia.

AIMS AND OBJECTIVES: To test Zauszniewski's resourcefulness and quality of life theory in community-based patients with schizophrenia. The objectives were to: (a) examine the relationship among psychotic symptoms, resourcefulness and quality of life, and (b) explore the determinants of quality of life. BACKGROUND: According to Zauszniewski's resourcefulness and quality of life theory, for patients, the skills constituting resourcefulness should centre on helping oneself or the use of resources to achieve their health. While the significant associations between resourcefulness and quality of life have been consistently delineated in published research, little is known about resourcefulness in patients with schizophrenia. DESIGN: A cross-sectional study design was used in this study. METHODS: A total of 120 patients were recruited with convenience sampling who received home care service and community rehabilitation programme. The participants were assessed with the Brief Psychiatric Rating Scale, the Resourcefulness Scale and the World Health Organization Quality of Life Scale-BREF. The psychotic symptoms were rated by the researchers with structured face-to-face interviews. Resourcefulness and quality of life were assessed through self-reporting. STROBE checklist was followed. RESULTS: The data were analysed using one-way analysis of variance, t test, Pearson's correlation coefficients and stepwise multiple regression analysis. Results showed that employment status and psychotic symptoms were correlated with resourcefulness, while employment status, psychotic symptoms and resourcefulness were correlated with quality of life. Resourcefulness and psychotic symptoms were the determinants of quality of life. CONCLUSION: Patients with employment and less psychotic symptoms had greater resourcefulness. In addition, patients with employment, less psychotic symptoms and greater resourcefulness had better quality of life. RELEVANCE TO CLINICAL PRACTICE: Community rehabilitation programmes should include supported employment, symptom management and resourcefulness skills training to enhance patients' resourcefulness and quality of life.

Hybrid lens design for ultrawide-angle imaging with a high Strehl ratio in a folded system.

When UV-curable material is used with a spherical glass lens, it is classified as a hybrid lens and can provide a new degree of freedom in designing a lens system. This method is used to explore the optical limitation of an ultrawide-angle imaging system of a folded structure, which is utilized to achieve a compact volume. The f-number and field of view of the system are 2.0 and 190°, respectively. Diffraction-limited performance is achieved within a field of view of 80° with Strehl ratios greater than approximately 80%. Even at the maximum field of view of 190°, the Strehl ratio is nearly 75%. The lens configuration is NNPNN, where N and P denote negative and positive optical power, respectively; this configuration uses only two aspherical hybrid lens surfaces. Comparisons of different optimizations are also provided.

Adjustable-focus ultracompact endoscopic lens design with ultrahigh optical performance.

An internally focusing design is developed for a compact endoscopic lens to have optical performance close to the diffraction limit. When changing the working distance from infinity to a predetermined short distance, the overall system length and the field of view remain constant. It is numerically demonstrated that the diffraction-limit-like performance is maintained over a wide range of working distances, showing a novel capability of adjustable focuses. The design concept is numerically proven to be feasible and can enable very high-resolution examination for doctors checking the positions of biological entities through the immediate capture of clear images at different working distances. The corresponding lens configuration, PNN, is shown to be useful in achieving the high-resolution performance once the number of lens elements is four, where P and N are used to denote positive and negative optical power, respectively, for the lens groups. A corresponding tolerance analysis is also presented.

[Learning Portfolio: A New Strategy in Health Education].

Health education is the teaching by healthcare professionals of healthcare-related knowledge and skills to students in order that these students learn to help patients self-manage their disease and maintain health. This article introduces a new strategy in health education known as the learning portfolio and presents the theoretical basis and function of the learning portfolio and the current application of this approach in academic and health education. The learning portfolio is a learner-centric approach that collects evidence related to an individual's learning process systematically. This approach helps educators understand learner needs and conditions, while allowing the learner to observe his / her learning process in a manner that promotes self-reflection, continual inspection, and behavioral modification throughout the learning process. The results enhance the motivation of learners and strengthen their care confidence in accomplishing learning tasks.

Staphylococcus aureus exacerbates dermal IL-33/ILC2 axis activation through evoking RIPK3/MLKL-mediated necroptosis of dry skin.

Atopic dermatitis (AD) is a persistent skin disease typified by symptoms of dry skin and recurrent eczema. Patients with AD are at heightened risk for Staphylococcus aureus infection. Group 2 innate lymphoid cells (ILC2s) are mainly activated by epithelial cell-derived cytokines IL-33 and involved in the pathogenesis of AD. However, little is known about the effect of skin delipidization on the epithelial cell-derived cytokines and dermal ILC2s in AD. In our study, we investigated the mechanism by which S. aureus infection modulates and exacerbates the pathogenesis of dry skin, leading to type 2 inflammation in the context of innate immunity. In vivo, we found that S. aureus infection aggravated delipidization-induced dermal IL-33 release and dermal ILC2 accumulation, which exacerbated skin inflammation. We also noticed that Il33fl/fl K14cre mice and Tlr2-/- mice exhibited attenuated skin inflammation. In vitro, treatment with necroptosis inhibitors reduced IL-33 release from S. aureus-infected keratinocytes. Mechanistically, we observed an increase in the necroptosis-associated kinases, MLKL and RIPK3, in S. aureus-infected mice, indicating that IL-33 release was associated with necroptotic cell death responses. Our results reveal that S. aureus infection-elicited keratinocyte necroptosis contributes to IL-33-mediated type 2 inflammation, which exacerbates the pathogenesis of dry skin.

Predicting early mortality and severe intraventricular hemorrhage in very-low birth weight preterm infants: a nationwide, multicenter study using machine learning.

Our aim was to develop a machine learning-based predictor for early mortality and severe intraventricular hemorrhage (IVH) in very-low birth weight (VLBW) preterm infants in Taiwan. We collected retrospective data from VLBW infants, dividing them into two cohorts: one for model development and internal validation (Cohort 1, 2016-2021), and another for external validation (Cohort 2, 2022). Primary outcomes included early mortality, severe IVH, and early poor outcomes (a combination of both). Data preprocessing involved 23 variables, with the top four predictors identified as gestational age, birth body weight, 5-min Apgar score, and endotracheal tube ventilation. Six machine learning algorithms were employed. Among 7471 infants analyzed, the selected predictors consistently performed well across all outcomes. Logistic regression and neural network models showed the highest predictive performance (AUC 0.81-0.90 in both internal and external validation) and were well-calibrated, confirmed by calibration plots and the lowest two mean Brier scores (0.0685 and 0.0691). We developed a robust machine learning-based outcome predictor using only four accessible variables, offering valuable prognostic information for parents and aiding healthcare providers in decision-making.

Associations between body weight trajectories and neurodevelopment outcomes at 24 months corrected age in very-low-birth-weight preterm infants: a group-based trajectory modelling study.

Introduction: This study aimed to explore the relationship between the trajectories of body weight (BW) z-scores at birth, discharge, and 6 months corrected age (CA) and neurodevelopmental outcomes at 24 months CA. Methods: Conducted as a population-based retrospective cohort study across 21 hospitals in Taiwan, we recruited 3,334 very-low-birth-weight (VLBW) infants born between 2012 and 2017 at 23-32 weeks of gestation. Neurodevelopmental outcomes were assessed at 24 months CA. Instances of neurodevelopmental impairment (NDI) were defined by the presence of at least one of the following criteria: cerebral palsy, severe hearing loss, profound vision impairment, or cognitive impairment. Group-based trajectory modeling was employed to identify distinct BW z-score trajectory groups. Multivariable logistic regression was used to assess the associations between these trajectories, postnatal comorbidity, and neurodevelopmental impairments. Results: The analysis identified three distinct trajectory groups: high-climbing, mid-declining, and low-declining. Significant associations were found between neurodevelopmental impairments and both cystic periventricular leukomalacia (cPVL) [with an adjusted odds ratio (aOR) of 3.59; p < 0.001] and belonging to the low-declining group (aOR: 2.59; p < 0.001). Discussion: The study demonstrated that a low-declining pattern in body weight trajectory from birth to 6 months CA, along with cPVL, was associated with neurodevelopmental impairments at 24 months CA. These findings highlight the importance of early weight trajectory and specific health conditions in predicting later neurodevelopmental outcomes in VLBW infants.

Relationships between environmental stressors and stress biobehavioral responses of preterm infants in NICU.

Although research has demonstrated that the neonatal intensive care unit (NICU) is a stressful environment for preterm and high-risk infants, little research validates the suspected relationships between infant biobehavioral responses and environmental stress in the NICU. This exploratory study examined the relationship between environmental stress and biobehavioral responses of preterm infants. The study used a repeated-measures research design to examine research variables in one group of preterm infants. Measurements of research variables were recorded every 2 minutes during two 60-minute observation periods for each research day (1 hour in the morning and 1 hour in the afternoon) and conducted over 2 days. A convenience sample of 37 preterm infants was recruited from 2 medical centers in Taiwan. A total of 4164 observations were made and recorded during the study. There was a statistically significant (P < .05) relationship between environmental stressors and changes in physiological signals. There were also statistically significant (P < .05) relationships between environmental stress and some specific stress behaviors. This research is applicable to neonatal clinical practice because it demonstrates the importance of recognizing the preterm infant's biological stress responses to environmental stressors, allowing for early interventions to reduce the possibility of more serious physiological or pathological changes in the status of the preterm infant.

The Treatment Experience of Anorexia Nervosa in Adolescents from Healthcare Professionals' Perspective: A Qualitative Study.

Anorexia nervosa (AN) is a serious psychiatric illness with a high mortality rate and a poor outcome. AN in adolescents can be difficult to treat. The prognosis of patients with AN depends highly on how early specialized AN treatment starts. Therefore, the purpose of this study was to explore the experiences of healthcare professionals in treating adolescents with AN. A qualitative study approach was conducted. Through semi-structured interviews, 16 healthcare professionals, including 10 nurses, 3 dieticians and 3 physicians from the paediatric ward at a university-affiliated medical centre in central Taiwan, shared their experiences. Recorded interviews were transcribed and analyzed by content analysis. Five themes and ten subthemes were identified: 1. Building a trusting relationship first: (a) spending time to build trust with the client and (b) establishing a relationship with the client's parents; 2. The key to treatment success: (a) Clients' awareness of the illness and (b) parents' support for clients; 3. Consistency of team treatment goals: (a) maintaining stable vital signs and (b) achieving caloric intake; 4. Empowerment with knowledge about anorexia: (a) continuing education for healthcare professionals and (b) interdisciplinary collaborative care; and 5. Using different interaction strategies: (a) the hard approach and (b) the soft approach. In conclusion, the findings will provide important information for healthcare professionals to apply in monitoring the psychological and emotional states of adolescents with AN. The findings indicate that healthcare professionals should invite parents to participate in the treatment, support and guide them in their adolescent care, develop scales of family stress and support for AN in adolescents, develop interventions, and establish an early therapeutic alliance.

A possible systematic culinary approach for spent duck meat: sous-vide cuisine and its optimal cooking condition.

This study offered a possible systematic culinary approach to spent-laying ducks. Breast meat is suitable for processing due to its amount and completeness. Sous-vide cooking resulted in lower cooking loss than poaching, pan-frying (P < 0.05), and roasting. The sous-vide duck breast had higher gumminess, chewiness, and resilience than other culinary techniques (P < 0.05). Sous-vide cooking at 65°C had a lower cooking loss than 70°C (P < 0.05), and less than 1.5-h sous-vide could keep a lower cooking loss and WB shear value (P < 0.05) as the cooking period extended, the smaller (P < 0.05) quantity of myosin heavy chain and the destroyed sarcomere arrangement were observed. A condition at 65°C for 1.5 h could be the optimal sous-vide cuisine for spent-laying duck breast. These sous-vide products stored at 4°C were still safe for consumption due to no detectible microorganisms and unchangeable physicochemical properties within 7 d.

Machine learning to predict late respiratory support in preterm infants: a retrospective cohort study.

Bronchopulmonary dysplasia (BPD) has been a critical morbidity in preterm infants. To improve our definition and prediction of BPD is challenging yet indispensable. We aimed to apply machine learning (ML) to investigate effective models by using the recently-proposed and data-driven definition to predict late respiratory support modalities at 36 weeks' post menstrual age (PMA). We collected data on very-low-birth-weight infants born between 2016 and 2019 from the Taiwan Neonatal Network database. Twenty-four attributes associated with their early life and seven ML algorithms were used in our analysis. The target outcomes were overall mortality, death before 36 weeks' PMA, and severity of BPD under the new definition, which served as a proxy for respiratory support modalities. Of the 4103 infants initially considered, 3200 were deemed eligible. The logistic regression algorithm yielded the highest area under the receiver operating characteristic curve (AUROC). After attribute selection, the AUROC of the simplified models remain favorable (e.g., 0.801 when predicting no BPD, 0.850 when predicting grade 3 BPD or death before 36 weeks' PMA, and 0.881 when predicting overall mortality). By using ML, we developed models to predict late respiratory support. Estimators were developed for clinical application after being simplified through attribute selection.

Postnatal corticosteroid treatment as a risk factor for false positivity in severe combined immunodeficiency newborn screening.

From 2011, 37 children were referred to a hospital due to low levels of T cell receptor excision circles (TRECs) from newborn screening. Among them, three children were immunologically characterized and followed up to show that postnatal corticosteroid usage may be among the causes of false positivity in TRECs screening.

Formation of Supported Thylakoid Membrane Bioanodes for Effective Electron Transfer and Stable Photocurrent.

The light-dependent reactions of photosynthesis use light energy to generate photoelectrons traveling through the thylakoid membranes (TMs). Extracting the photoelectrons from the TMs to form bioanodes can have various applications. Most studies focus on modifying the electrode materials to increase the collected photocurrent. Seldom studies have investigated how the orientation of the TMs influences photocurrent collection. In addition, the formation of reactive oxygen species (ROS) during photosynthesis is a challenge for stable photocurrent generation. Here, we enhanced the photoelectron transfer from the TMs to electrodes by depositing expanded thylakoids as planar supported membranes onto an electrode. The high contact area between the external electrodes and TMs per unit mass of thylakoid allows the thylakoid to more effectively transfer electrons to the electrodes, thereby reducing the free electrons available for the ROS generation. We expanded the naturally stacked thylakoids into liposomes through osmotic pressure and dropcasted them onto an Au electrode. The electrochemical impedance measurement showed that the supported membrane bioanode formed by the expanded liposomes had a lower photoelectron transfer resistance. Additionally, we observed that the expanded TM bioanode provided a higher photocurrent and was more durable to air/water interfacial tension. These results suggest that the effective contact between the expanded TM and electrodes can lead to more efficient electron transfer and increase the system robustness. The photo fuel cell (PFC) made by the expanded TM bioanode had a higher open-circuit voltage than the one made by the stacked TM bioanode. Interestingly, we found that PFCs made of high-load TM bioanodes had fast photocurrent decay under continuous operation at high cell voltages. The poor contact of large numbers of TMs with the electrodes at the high-load TM bioanodes could cause more ROS accumulation and therefore decreased the operational stability, supporting the importance of effective contact between TMs and the electrodes.

Exploring Perceived Stress in Mothers with Singleton and Multiple Preterm Infants: A Cross-Sectional Study in Taiwan.

OBJECTIVE: The aim of this study was to explore mothers' perceived level of stress one month after hospital discharge following the birth of singleton and multiple preterm infants. DESIGN: A cross-sectional design was used to compare mother's perceived stress in two groups of postpartum mothers and the relationship of the theoretical antecedents and these variables. SETTING: A neonatal intensive care unit in a medical center in Taiwan. PARTICIPANTS: Mothers of 52 singletons and 38 multiple premature infants were recruited. One month after the infant was discharged, the participants completed a self-reported questionnaire that included demographic data about the mother and infant, the 21-item Social Support Scale, and the 15-item Perceived Stress Scale. This was returned by email or completed at the outpatient unit. ANALYSIS: Descriptive and inferential analysis. RESULTS: The mean social support scores were 76.6 and 76.5 (out of 105) for mothers with singleton and multiple birth infants, respectively. The most important supporter was the husband. The mean perceived stress scores of 25.8 and 31.0 for mothers with singleton and multiple birth infants, respectively, were significantly different (p = 0.02). Sleep deprivation and social support were predictive indicators of perceived stress in mothers with preterm infants. CONCLUSIONS: We suggest that the differences in stress and needs of mothers with singleton and multiple births should be recognized and addressed in clinics. The findings of this study serve as a reference for promoting better preterm infant care.

Exploring the Determinants of Nascent Social Entrepreneurial Behaviour.

Following the emergence of social, environmental, and public health issues, this study analysed the determinants of nascent social entrepreneurial behaviour. This research investigated the effects of empathy and prior experience with social problems on nascent social entrepreneurial behaviour through social entrepreneurial self-efficacy, outcome expectations of social entrepreneurship, and perceived social support. Through utilising the quantitative survey method, this study gathered a total of 560 valid responses, comprising 258 Chinese respondents and 302 Taiwanese respondents. Outcome expectations and perceived social support positively influenced nascent social entrepreneurial behaviour for the Chinese sample, whereas social entrepreneurial self-efficacy and perceived social support positively influenced nascent social entrepreneurial behaviour for the Taiwanese sample. This research enriches the existing literature by taking account of social entrepreneurial behaviour, instead of merely analysing social entrepreneurial intentions.

Extraction method development for nanoplastics from oyster and fish tissues.

Nanoplastics are now found in some environmental media and consumer products. However, very limited data on nanoplastics are available for one of the main human consumption sources of microplastics: seafood. Unlike microplastics, a method for extracting nanoplastics from seafood is still lacking. Herein, a combination of common extraction techniques including enzymatic digestion, sequential membrane filtration, centrifugal concentration, and purification (dialysis and sodium dodecylsulfate (SDS) incubation), was developed to extract nanoplastics from oyster and fish tissues. Corolase with subsequent lipase treatment achieved the highest digestion efficiencies (88- 89%) for non-homogenized tissues compared to other proteases and additional cellulase or H2O2 treatment. With the exception of polyethylene terephthalate (PET), enzymatic digestion did not change the morphology or structure of polyvinyl chloride (PVC), polyethylene (PE), or polystyrene (PS) nanoplastic particles, and the subsequent extraction procedures had good recoveries of 71- 110% for fluorescence-labeled 76-nm PVC and 100- and 750-nm PS, as validated by a Nanoparticle Tracking Analysis (NTA). Few of the 1011 digested residual particles of 150- 300 nm in diameter per oyster or per serving of fish tissue were left in the method blank. Consequently, this efficient approach could be used as a pretreatment protocol for current potential nanoplastic detection methods.