Sorting Mechanism Research Articles

Tomato Sorting Machine Design

This design is based on the existing fruit sorting mechanism, and the sorting mechanism has been improved by using a spiral roller to design a tomato sorting machine. Based on the size of tomatoes, this tomato sorter can classify tomatoes into three categories to meet the needs of different occasions for different sizes of tomatoes, improve the utilization rate of tomatoes, and also speed up the rate of tomato sorting. The threads on the outer layer of the drum push the tomatoes forward while the drum rotates, and the tomatoes fall through the corresponding drum gaps to complete this sorting stage, with the largest grade going all the way to the end of the drum for grading. And the whole machine is equipped with sponge as a cushioning material in the parts of tomatoes that are easy to be impacted to prevent tomatoes from being damaged and to facilitate storage and sale of tomatoes. The sorter is inexpensive, easy to maintain and suitable for small-scale farmers.

Experimental examination of the incentive and sorting effects of pay-for-performance on creative performance.

There is long-standing debate over whether pay-for-performance (PFP) enhances or undermines creative performance. Traditional motivation and revised creativity theories suggest that PFP and intrinsic task interest combine additively to enhance creative performance, whereas cognitive evaluation theory and self-determination theory posit that PFP undermines task interest and thus intrinsic motivation and creative performance. To help resolve these conflicting predictions and provide a more comprehensive understanding of how and when PFP influences creative performance, this study incorporates both the incentive and sorting mechanisms of PFP, varying strengths of PFP, and task autonomy as a key moderator. A novel laboratory experiment was designed to capture key elements of workplace contexts, including in the design of the creative tasks, PFP strengths based on benchmarking of U.S. companies' practices, and allowing subjects to sort into different pay conditions, consistent with the opportunity for mobility in the labor market. The results showed that, through both incentive and sorting mechanisms, high PFP intensity enhanced creative performance more so than low PFP intensity, and both were superior to fixed pay. Importantly, task autonomy positively moderated the PFP-creative performance relationship, such that creative performance under PFP increased much more under higher task autonomy. Finally, the difference in creative performance under PFP versus fixed pay was greater when subjects were allowed to sort into their preferred pay conditions than when they worked only under randomly assigned pay conditions. Theoretical and practical implications and future research directions are discussed. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Circular RNA landscape in extracellular vesicles from human biofluids

BackgroundCircular RNAs (circRNAs) have emerged as a prominent class of covalently closed single-stranded RNA molecules that exhibit tissue-specific expression and potential as biomarkers in extracellular vesicles (EVs) derived from liquid biopsies. Still, their characteristics and applications in EVs remain to be unveiled.MethodsWe performed a comprehensive analysis of EV-derived circRNAs (EV-circRNAs) using transcriptomics data obtained from 1082 human body fluids, including plasma, urine, cerebrospinal fluid (CSF), and bile. Our validation strategy utilized RT-qPCR and RNA immunoprecipitation assays, complemented by computational techniques for analyzing EV-circRNA features and RNA-binding protein interactions.ResultsWe identified 136,327 EV-circRNAs from various human body fluids. Significantly, a considerable amount of circRNAs with a high back-splicing ratio are highly enriched in EVs compared to linear RNAs. Additionally, we discovered brain-specific circRNAs enriched in plasma EVs and cancer-associated EV-circRNAs linked to clinical outcomes. Moreover, we demonstrated that EV-circRNAs have the potential to serve as biomarkers for evaluating immunotherapy efficacy in non-small cell lung cancer (NSCLC). Importantly, we identified the involvement of RBPs, particularly YBX1, in the sorting mechanism of circRNAs into EVs.ConclusionsThis study unveils the extensive repertoire of EV-circRNAs across human biofluids, offering insights into their potential as disease biomarkers and their mechanistic roles within EVs. The identification of specific circRNAs and the elucidation of RBP-mediated sorting mechanisms open new avenues for the clinical application of EV-circRNAs in disease diagnostics and therapeutics.

Waste Segregator: Waste Management Prototype

Smart Waste Dustbins utilize IoT, AI, and computer vision for efficient waste management. They feature multiple compartments, sensors, and automatic lid mechanisms for real-time waste sorting. Computer vision and AI algorithms accurately identify and segregate waste. Smart Waste Dustbins monitor fill levels and offer a user-friendly interface for public engagement. Sensors include ultrasonic, moisture, and metal sensors for proximity, moisture analysis, and metal item segregation. Smart Waste Dustbins revolutionize waste management, enhancing efficiency in segregation, recycling, and collection processes, leading to reduced environmental impact and improved resource utilization for sustainable communities.

Comprehensive Proteomic Characterization of Intra-Golgi Trafficking Intermediates.

The Conserved Oligomeric Golgi (COG) complex is critical for efficient intra-Golgi trafficking and glycosylation. Prior research has demonstrated that COG dysfunction or RNAi-induced depletion leads to the accumulation of non-tethered COG complex-dependent (CCD) vesicles. However, the precise connection between COG deficiency, degradation of Golgi enzymes, and its impact on vesicular trafficking has not been fully elucidated. In this study, we conducted a comprehensive proteomic analysis of Golgi-derived vesicles from both wild-type and COG-depleted cells. We specifically analyzed three distinct populations of vesicles immunoisolated with antibodies targeting transmembrane proteins from the cis, medial, and trans-Golgi sub-compartments. Our findings reveal that, while the vesicle content encompasses the entire Golgi proteome, the molecular signatures of vesicles derived from wild- type cells were markedly distinct, underscoring a robust recycling mechanism for Golgi- dependent proteins. Notably, these vesicles retained various vesicular coats, and COG depletion significantly accelerated their uncoating. Furthermore, the increased overlap in molecular signatures upon COG depletion indicates that persistent defects in vesicle tethering severely compromise intra-Golgi sorting mechanisms. Crucially, our analysis highlights that the entire Golgi glycosylation machinery recycles within CCD vesicles in a COG-dependent manner, while secretory proteins and components involved in ER-Golgi and Golgi-endosome trafficking were not enriched. These results strongly support a model of multi-step intra-Golgi vesicular recycling of the glycosylation machinery, orchestrated by the COG complex in concert with a cisternae-specific array of vesicular coats, coiled-coil tethers, Rabs, and SNARE proteins.

Controlling Circular RNA Encapsulation within Extracellular Vesicles for Gene Editing and Protein Replacement.

Extracellular vesicles (EVs) are a population of vesicular bodies originating from cells, and EVs have been proven to have the potential to deliver different cargos, such as RNAs. However, conventional methods are not able to encapsulate long RNAs into EVs efficiently or may compromise the integrity of EVs. In this study, we have devised a strategy to encapsulate long circRNAs (>1000 nt) into EVs by harnessing the sorting mechanisms of cells. This strategy utilizes the inherent richness of circular RNAs in EVs and a genetic engineering method to increase the cytoplasmic concentration of target circRNAs, facilitating highly efficient RNA back-splicing to drive the circularization of RNAs. This allows target circRNAs to load into EVs with high efficiency. Furthermore, we demonstrate the practical applications of this strategy, showing that these circRNAs can be delivered by EVs to recipient cells for protein expression and to mice for gene editing.

Enumeration and gentle sorting of immune cells on chip, key to next generation advanced therapies in outpatient setting

BackgroundA thorough understanding of immune-oncology and molecular medicine has been vital in the development of cell therapeutics. At the basis of this translational research and its future implementation into a medicinal product, lies the availability of pure and viable cell populations. Currently, FACS and magnetic bead isolation are successfully used but suffer to fulfill all requirements. FACS is costly and difficult to upscale due to the limitation of shear stress, especially fragile, cells can handle. Therefore, magnetic bead isolation is often used as it is gentler, but it lacks the multiparametric aspect to isolate more complex cellular profiles. AimsWe aim to develop a versatile technology able of multi marker detection and isolation of complex cell types with high purity, viability and throughput. MethodsWe have developed a gentle sorting mechanism based on a jet flow created by micro vapor bubbles, enabling a closed microfluidic cell isolation platform capable of multiparametric sorting with high viability, purity and throughput. In this work we compared the purity, recovery and viability of sorted CD4+ CD14- cells to magnetic isolation, most often used for other cell manufacturing approaches. Futhermore, we cultured the sorted cells of both isolation strategies and compared their growth curve and expression of activation-induced IL2 and IFN-γ. ResultsWe demonstrate that this tool can achieve a pure population of CD4+ CD14- cells with high viability after sorting without compromising the recovery. On top of the viability also the growth and activation potential of sorted cells is unhampered by comparison to the benchmark gentle magnetic isolation. ConclusionsOur technology allows for the development of a compact system which sets it apart from other efforts intended to create automated cell therapeutic solutions.

Essential Fluidics for a Flow Cytometer.

Flow cytometry is an inherently fluidic process that flows particles on a one-by-one basis through a sensing region to discretely measure their optical and physical properties. It can be used to analyze particles ranging in size from nanoparticles to whole organisms (e.g., zebrafish). It has particular value for blood analysis, and thus most instruments are fluidically optimized for particles that are comparable in size to a typical blood cell. The principles of fluid dynamics allow for particles of such size to be precisely positioned in flow as they pass through sensing regions that are tens of microns in length at linear velocities of meters per second. Such fluidic systems enable discrete analysis of cell-sized particles at rates approaching 100 kHz. For larger particles, the principles of fluidics greatly reduce the achievable rates, but such high rates of data acquisition for cell-sized particles allow rapid collection of information on many thousands to millions of cells and provides for research and clinical measurements of both rare and common cell populations with a high degree of statistical confidence. Additionally, flow cytometers can accurately count particles via the use of volumetric sample delivery and can be coupled with high-throughput sampling technologies to greatly increase the rate at which independent samples can be delivered to the system. Due to the combination of high analysis rates, sensitive multiparameter measurements, high-throughput sampling, and accurate counting, flow cytometry analysis is the gold standard for many critical applications in clinical, research, pharmaceutical, and environmental areas. Beyond the power of flow cytometry as an analytical technique, the fluidic pathway can be coupled with a sorting mechanism to collect particles based on desired properties. We present an overview of fluidic systems that enable flow cytometry-based analysis and sorting. We introduce historical approaches, explanations of commonly implemented fluidics, and brief discussions of potential future fluidics where appropriate. © 2024 Wiley Periodicals LLC.

Detection of walnut internal quality via improved YOLOv5 and x‐ray imaging

AbstractThis study proposes a walnut target detection algorithm based on improved YOLOv5 and x‐ray imaging to meet the demand for internal quality detection and removal in the Xinjiang walnut industry. By replacing the C3 module in the backbone layer with the C2f module and the couple‐head in the head layer with the decouple‐head, the algorithm reduces computational complexity, enhances robustness and generalizability, and retains more spatial information, thereby improving the performance of multicategory small target detection. In addition, this paper replaces the original CIOU loss function with the EIOU loss function to improve the convergence speed of the algorithm's accuracy and boundary aspect ratio. Compared with the original model, the improved model, improved YOLOv5, maintains the same average precision for normal walnuts while increasing the average precision for shriveled walnuts and empty‐shell walnuts by 8.2% and 0.4%, respectively. Compared with other mainstream models, such as VGG16, ResNet50, YOLOv5s, YOLOv7, YOLOv8s, YOLOv9s, and YOLOv10s, this model achieves the highest detection accuracy and good detection performance, with a single‐image detection time of 11.9 ms, meeting the requirements for real‐time detection. This work lays a foundation for automatic robot detection of the internal quality and removal of walnuts, showing practical application potential.Practical applicationsXinjiang stands as a prominent producer of walnuts; however, due to the decentralized nature of its cultivation and management processes, a notable prevalence of internal empty shells and shrinkage is observed, which substantially diminishes their commercial value. The integration of YOLOv5 with x‐ray imaging technology promises to enhance the precision of internal quality assessments of walnuts. The improved YOLOv5 model, as delineated in this study, exhibits the highest detection accuracy when benchmarked against a multitude of other models. It achieves a detection latency of merely 11.9 ms per image, thereby satisfying the stringent demands for real‐time detection applications. This model is designed for integration into x‐ray systems with an adjunctive sorting mechanism, which facilitates the inspection and exclusion of defective walnuts on conveyor belts.

Lightweight Deep Learning Algorithm for Sorting Medical Waste Embedded System

The recent COVID-19 pandemic has led to an increased number of hospitalized patients, and in some countries, the overwhelming problem has even caused the collapse of the healthcare system. Thus, the associated waste generated and inadequate waste disposal may have deleterious impacts on public health and the surrounding environment. This paper aims to design and implement an automatic medical waste identification and sorting mechanism that can distinguish the items disposed of via a series of image processing procedures. Specifically, the system can classify, sort, and calculate the disposed items based on their visual appearance. To establish this, both software and hardware systems have been devised. Particularly, the vision system for object detection and classification is constructed based on several popular pre-trained convolutional neural networks. Upon verifying the type of waste (i.e., general infection, dangerous infection, and general garbage), an automatic sorting mechanism will be triggered to dispose of the object in the corresponding garbage bin. The experimentation has been validated on a total of self-collected 2025 images from 3 categories, and the best accuracy attained is 99.34% when adopting GoogLeNet as the backbone architecture.

Tropomyosin Isoforms Segregate into Distinct Clusters on Single Actin Filaments.

Tropomyosins (Tpms) are rod-shaped proteins that interact head-to-tail to form a continuous polymer along both sides of most cellular actin filaments. Head-to-tail interaction between adjacent Tpm molecules and the formation of an overlap complex between them leads to the assembly of actin filaments with one type of Tpm isoform in time and space. Variations in the affinity of tropomyosin isoforms for different actin structures are proposed as a potential sorting mechanism. However, the detailed mechanisms of the spatio-temporal sorting of Tpms remain elusive. In this study, we investigated the early intermediates during actin-tropomyosin filament assembly, using a skeletal/cardiac Tpm isoform (Tpm1.1) and a cytoskeletal isoform (Tpm1.6) that differ only in the last 27 amino acids. We investigated how the muscle isoform Tpm1.1 and the cytoskeletal isoform Tpm1.6 nucleate domains on the actin filament, and tested whether (1) recruitment is affected by the actin isoform (muscle vs. cytoskeletal) and (2) whether there is specificity in recruiting the same isoform to a domain at these early stages. To address these questions, actin filaments were exposed to low concentrations of fluorescent tropomyosins in solution. The filaments were immobilized onto glass coverslips and the pattern of decoration was visualized by TIRF microscopy. We show that at the early assembly stage, tropomyosins formed multiple distinct fluorescent domains (here termed "cluster") on the actin filaments. An automated image analysis algorithm was developed and validated to identify clusters and estimate the number of tropomyosins in each cluster. The analysis showed that tropomyosin isoform sorting onto an actin filament is unlikely to be driven by a preference for nucleating on the corresponding muscle or cytoskeletal actin isoforms, but rather is facilitated by a higher probability of incorporating the same tropomyosin isoforms into an early assembly intermediate. We showed that the 27 amino acids at the end of each tropomyosin seem to provide enough molecular information for the attachment of the same tropomyosin isoforms adjacent to each other on an actin filament. This results in the formation of homogeneous clusters composed of the same isoform rather than clusters with mixed isoforms.

Plasma extracellular vesicle: a novel biomarker for neurodegenerative disease diagnosis

Extracellular vesicles (EVs) are membrane-bound structures that carry proteins, lipids, RNA, and DNA, playing key roles in cell communication and material transport. Recent research highlights their potential as disease biomarkers due to their stability in bodily fluids. This study explores using tau and TDP-43 proteins in plasma EVs as diagnostic biomarkers for frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). Analyzing plasma EVs from clinical cohorts, the study found that the 3R/4R tau ratio and TDP-43 levels effectively differentiate between diagnostic groups with high accuracy. Notably, plasma EV biomarkers demonstrate higher diagnostic accuracy and stability compared to direct plasma testing, providing new insights and approaches for future research and clinical practice. Further research is needed to validate these biomarkers in diverse populations and to establish standardized protocols. Future studies should continue to explore the potential of EV biomarkers in a broader range of neurodegenerative diseases and delve deeper into the mechanisms of EV secretion and sorting to enhance their diagnostic utility.

Abstract A075: Establishing the interaction mechanism of oncoprotein, Mucin1, and extracellular vesicle marker protein, Alix, in pancreatic cancer

Abstract Pancreatic ductal adenocarcinoma (PDAC) remains an aggressive disease with devastating mortality rates largely due to a poor understanding of its progression mechanism. Small extracellular vesicles (sEVs) are used by cancer cells to aid in their progression, and tumor-derived EVs (TEVs) show a distinct protein cargo composition potentially contributing to this rapid progression and metastasis of PDAC. To investigate this phenomenon, we isolated sEVs from conditioned media of pancreatic cancer cell lines using ultracentrifugation and characterized them by transmission electron microscopy (TEM), nanoparticle tracking analysis, and immunoblotting. These sEVs were then used in several studies to investigate critical TEV cargo sorting machinery. For instance, PDAC-derived sEVs have selective enrichment of the oncogenic transmembrane protein Mucin1 (MUC1). Preliminary MUC1KO studies from our lab revealed that MUC1 potentially regulates sEV formation, where we saw a clear impact on sEV cargo in the absence of MUC1. Additional studies found that the cytoplasmic adaptor protein ALG-2-Interacting Protein X (Alix), a protein known to play a critical role during TEV formation, regulates MUC1’s presence in sEVs. From this discovery, we then identified a novel MUC1-Alix interaction, which was confirmed using several techniques, including co-immunoprecipitation (CoIP). To define the Alix-interacting domain of MUC1, we utilized in silico modeling and localized surface plasmon resonance (LSPR) of MUC1.CT and Alix to quantify the interaction. This revealed the cytoplasmic tail of MUC1 (MUC1.CT) as the Alix-interacting domain. Moving forward, our central hypothesis for this study is that this MUC1.CT-Alix interaction utilizes a conserved binding motif that targets Alix to the endosomal membrane and contributes to the formation of MUC1+ sEVs. While a distinct sEV biogenesis pathway employed by cancer cells has not yet been defined, our studies will provide insight into the potential sEV sorting mechanism of MUC1 and other cancer biomarkers and advance the understanding of PDAC progression. Citation Format: Kristine V Hoagstrom, Ying Huang, Gloria E Borgstahl, Michael A Hollingsworth. Establishing the interaction mechanism of oncoprotein, Mucin1, and extracellular vesicle marker protein, Alix, in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A075.

Agonistic freedoms: enabling disaffected working-class students to turn-back-around to mainstream school

ABSTRACT The sifting and sorting mechanisms of Australia’s education system continue to work to the detriment of groups put at a disadvantage. For mainstream schools serving working-class communities in particular, the rejection of the offers and advantages of schooling continue to result in differential class consequences and inequalities for working-class young people. In response, this paper pursues a more hopeful project of schooling by considering how schools can work to circumvent the alienation of working-class students from the mainstream secondary school system. Foucault’s conceptualisations of ‘power relations’, ‘agonistic freedom’ and ‘technologies of the self’ are used alongside Kamler and Comber’s notion of ‘turn-around pedagogies’ to draw attention to the complex relationships between students’ subjectivities, their shifting dispositions towards school and eventual enticement to turn-back-around to mainstream school.

Immunomodulatory Effects of Probiotic-Derived Extracellular Vesicles: Opportunities and Challenges.

Probiotics are known to modulate host immune responses in the course of many diseases. Recently, bacterial extracellular vesicles (EVs), which contain bioactive proteins, lipids, nucleic acids, and metabolites released by bacteria, have been identified as potentially important mediators of bacteria-bacterium and bacteria-host interactions. With the deepening of research, it has been found that probiotic-derived EVs play a significant role in regulating host immune function and, thus, exerting health-promoting effects. Nevertheless, current research is in its early stages, and there remains a long way to go to bridge the gap between basic research and clinical practice. In this review, we describe the fundamental aspects of probiotic-derived EVs, including their biogenesis, cargo sorting mechanism, and transport capabilities. We further discussed the potential mechanisms of probiotic-derived EVs in regulating the host's gut microbiota and immune responses. Finally, we speculate about the potential of probiotic-derived EVs as new postbiotics for applications in functional food, disease treatment substitutes, and immune regulatory adjuvants.

Non-canonical RNA-binding protein ANXA11 regulates microRNA resorting into small extracellular vesicles to mediate cisplatin resistance.

The sensitivity of laryngeal squamous cell carcinoma (LSCC) to chemotherapy shows large heterogeneity. The role of miRNA in small extracellular vesicles (sEV) in chemotherapy resistance is under investigation. However, the regulation and sorting mechanism of sEV miRNAs remains unclear. In this study, small RNA sequencing was used to explore miRNA expression profiles in sEV of LSCC after cisplatin stimulation; RNA pull-down, mass spectrometry, and EMSA were used to clarify the binding of candidate RNA-binding protein (RBP) and candidate miRNA. Immunostaining and microRNA fluorescence insitu hybridization were performed to identify how candidate RBP affects miRNA stability and nuclear/cytoplasmic distribution. Invivo experiments were performed to verify the biological functions and response to cisplatin of candidate RBP. We found that cisplatin stimulation induced increased expression of miR-148a-3p and sEV sorting. ANXA11 binds to miR-148a-3p in a sequence-specific manner. ANXA11 inhibits tumor cell proliferation and drug resistance by binding to and retaining miR-148a-3p. Cisplatin stimulation reduced ANXA11 expression and promoted miR-148a-3p efflux through sEV pathways. ANXA11 overexpression reduced invivo tumor proliferation and cisplatin-resistance. Taken together, ANXA11 mediates cisplatin resistance through sEV miRNA resorting. Mechanically, ANXA11 binds to miR-148a-3p in a sequence-specific manner to regulate its resorting and thus influences tumor proliferation and chemoresistance.

Design and Preliminary Evaluation of Automated Sweetpotato Sorting Mechanisms

Design and Preliminary Evaluation of Automated Sweetpotato Sorting Mechanisms

Interactive design and development of an intelligent vision-driven 3D printed precision sorting mechanism for silk cocoons

Interactive design and development of an intelligent vision-driven 3D printed precision sorting mechanism for silk cocoons

Contribution of intraflagellar transport to compartmentalization and maintenance of the photoreceptor cell.

The first steps of vision take place in the ciliary outer segment compartment of photoreceptor cells. The protein composition of outer segments is uniquely suited to perform this function. The most abundant among these proteins is the visual pigment, rhodopsin, whose outer segment trafficking involves intraflagellar transport (IFT). Here, we report three major findings from the analysis of mice in which ciliary transport was acutely impaired by conditional knockouts of IFT-B subunits. First, we demonstrate the existence of a sorting mechanism whereby mislocalized rhodopsin is recruited to and concentrated in extracellular vesicles prior to their release, presumably to protect the cell from adverse effects of protein mislocalization. Second, reducing rhodopsin expression significantly delays photoreceptor degeneration caused by IFT disruption, suggesting that controlling rhodopsin levels may be an effective therapy for some cases of retinal degenerative disease. Last, the loss of IFT-B subunits does not recapitulate a phenotype observed in mutants of the BBSome (another ciliary transport protein complex relying on IFT) in which non-ciliary proteins accumulate in the outer segment. Whereas it is widely thought that the role of the BBSome is to primarily participate in ciliary transport, our data suggest that the BBSome has another major function independent of IFT and possibly related to maintaining the diffusion barrier of the ciliary transition zone.

The NADPH oxidases DUOX1 and DUOX2 are sorted to the apical plasma membrane in epithelial cells via their respective maturation factors DUOXA1 and DUOXA2.

The membrane-integrated NADPH oxidases DUOX1 and DUOX2 are recruited to the apical plasma membrane in epithelial cells to release hydrogen peroxide, thereby playing crucial roles in various functions such as thyroid hormone synthesis and host defense. However, it has remained unknown about the molecular mechanism for apical sorting of DUOX1 and DUOX2. Here we show that DUOX1 and DUOX2 are correctly sorted to the apical membrane via the membrane-spanning DUOX maturation proteins DUOXA1 and DUOXA2, respectively, when co-expressed in MDCK epithelial cells. Impairment of N-glycosylation of DUOXA1 results in mistargeting of DUOX1 to the basolateral membrane. Similar to DUOX1 complexed with the glycosylation-defective DUOXA1, the naturally non-glycosylated oxidase NOX5, which forms a homo-oligomer, is targeted basolaterally. On the other hand, a mutant DUOXA2 deficient in N-glycosylation is less stable than the wild-type protein but still capable of recruiting DUOX2 to the apical membrane, whereas DUOX2 is missorted to the basolateral membrane when paired with DUOXA1. These findings indicate that DUOXA2 is crucial but its N-glycosylation is dispensable for DUOX2 apical recruitment; instead, its C-terminal region seems to be involved. Thus, apical sorting of DUOX1 and DUOX2 is likely regulated in a distinct manner by their respective partners DUOXA1 and DUOXA2.