Phosphoglycerate kinase 1 acts as a cargo adaptor to promote EGFR transport to the lysosome.

The epidermal growth factor receptor (EGFR) plays important roles in multiple cellular events, including growth, differentiation, and motility. A major mechanism of downregulating EGFR function involves its endocytic transport to the lysosome. Sorting of proteins into intracellular pathways involves cargo adaptors recognizing sorting signals on cargo proteins. A dileucine-based sorting signal has been identified previously for the sorting of endosomal EGFR to the lysosome, but a cargo adaptor that recognizes this signal remains unknown. Here, we find that phosphoglycerate kinase 1 (PGK1) is recruited to endosomal membrane upon its phosphorylation, where it binds to the dileucine sorting signal in EGFR to promote the lysosomal transport of this receptor. We also elucidate two mechanisms that act in concert to promote PGK1 recruitment to endosomal membrane, a lipid-based mechanism that involves phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] and a protein-based mechanism that involves hepatocyte growth factor receptor substrate (Hrs). These findings reveal an unexpected function for a metabolic enzyme and advance the mechanistic understanding of how EGFR is transported to the lysosome.

MoodSensing: A smartphone app for digital phenotyping and assessment of bipolar disorder.

BACKGROUND: Daily life tracking has proven to be of great help in the assessment of patients with bipolar disorder. Although there are many smartphone apps for tracking bipolar disorder, most of them lack academic verification, privacy policy and long-term maintenance. METHODS: Our developed app, MoodSensing, aims to collect users' digital phenotyping for assessment of bipolar disorder. The data collection was approved by the Institutional Review Board. This study collaborated with professional clinicians to ensure that the app meets both clinical needs and user experience requirements. Based on the collected digital phenotyping, deep learning techniques were applied to forecast participants' weekly HAM-D and YMRS scale scores. RESULTS: In experiments, the data collected by our app can effectively predict the scale scores, reaching the mean absolute error of 0.84 and 0.22 on the scales. The statistical data also demonstrate the increase in user engagement. CONCLUSIONS: Our analysis reveals that the developed MoodSensing app can not only provide a good user experience, but also the recorded data have certain discriminability for clinical assessment. Our app also provides relevant policies to protect user privacy, and has been launched in the Apple Store and Google Play Store.

Reconstructing missing time-varying land subsidence data using back propagation neural network with principal component analysis.

Land subsidence, a complex geophysical phenomenon, necessitates comprehensive time-varying data to understand regional subsidence patterns over time. This article focuses on the crucial task of reconstructing missing time-varying land subsidence data in the Choshui Delta, Taiwan. We propose a novel algorithm that leverages a multi-factorial perspective to accurately reconstruct the missing time-varying land subsidence data. By considering eight influential factors, our method seeks to capture the intricate interplay among these variables in the land subsidence process. Utilizing Principal Component Analysis (PCA), we ascertain the significance of these influencing factors and their principal components in relation to land subsidence. To reconstruct the absent time-dependent land subsidence data using PCA-derived principal components, we employ the backpropagation neural network. We illustrate the approach using data from three multi-layer compaction monitoring wells from 2008 to 2021 in a highly subsiding region within the study area. The proposed model is validated, and the resulting network is used to reconstruct the missing time-varying subsidence data. The accuracy of the reconstructed data is evaluated using metrics such as root mean square error and coefficient of determination. The results demonstrate the high accuracy of the proposed neural network model, which obviates the need for a sophisticated hydrogeological numerical model involving corresponding soil compaction parameters.

Oct4 and Hypoxia Dual-Regulated Oncolytic Adenovirus Armed with shRNA-Targeting Dendritic Cell Immunoreceptor Exerts Potent Antitumor Activity against Bladder Cancer.

Immunotherapy has emerged as a promising modality for cancer treatment. Dendritic cell immunoreceptor (DCIR), a C-type lectin receptor, is expressed mainly by dendritic cells (DCs) and mediates inhibitory intracellular signaling. Inhibition of DCIR activation may enhance antitumor activity. DCIR is encoded by CLEC4A in humans and by Clec4a2 in mice. Gene gun-mediated delivery of short hairpin RNA (shRNA) targeting Clec4a2 into mice bearing bladder tumors reduces DCIR expression in DCs, inhibiting tumor growth and inducing CD8+ T cell immune responses. Various oncolytic adenoviruses have been developed in clinical trials. Previously, we have developed Ad.LCY, an oncolytic adenovirus regulated by Oct4 and hypoxia, and demonstrated its antitumor efficacy. Here, we generated a Clec4a2 shRNA-expressing oncolytic adenovirus derived from Ad.LCY, designated Ad.shDCIR, aimed at inducing more robust antitumor immune responses. Our results show that treatment with Ad.shDCIR reduced Clec4a expression in DCs in cell culture. Furthermore, Ad.shDCIR exerted cytolytic effects solely on MBT-2 bladder cancer cells but not on normal NIH 3T3 mouse fibroblasts, confirming the tumor selectivity of Ad.shDCIR. Compared to Ad.LCY, Ad.shDCIR induced higher cytotoxic T lymphocyte (CTL) activity in MBT-2 tumor-bearing immunocompetent mice. In addition, Ad.shDCIR and Ad.LCY exhibited similar antitumor effects on inhibiting tumor growth. Notably, Ad.shDCIR was superior to Ad.LCY in prolonging the survival of tumor-bearing mice. In conclusion, Ad.shDCIR may be further explored as a combination therapy of virotherapy and immunotherapy for bladder cancer and likely other types of cancer.

A Flexible 8-in-1 Microsensor Embedded in Proton Battery Stack for Real-Time Microscopic Measurements.

According to the latest literature, it is difficult to measure the multiple important physical parameters inside a proton battery stack accurately and simultaneously. The present bottleneck is external or single measurements, and the multiple important physical parameters (oxygen, clamping pressure, hydrogen, voltage, current, temperature, flow, and humidity) are interrelated, and have a significant impact on the performance, life, and safety of the proton battery stack. Therefore, this study used micro-electro-mechanical systems (MEMS) technology to develop a micro oxygen sensor and a micro clamping pressure sensor, which were integrated into the 6-in-1 microsensor developed by this research team. In order to improve the output and operability of microsensors, an incremental mask was redesigned to integrate the back end of the microsensor in combination with a flexible printed circuit. Consequently, a flexible 8-in-1 (oxygen, clamping pressure, hydrogen, voltage, current, temperature, flow, and humidity) microsensor was developed and embedded in a proton battery stack for real-time microscopic measurement. Multiple micro-electro-mechanical systems technologies were used many times in the process of developing the flexible 8-in-1 microsensor in this study, including physical vapor deposition (PVD), lithography, lift-off, and wet etching. The substrate was a 50 µm-thick polyimide (PI) film, characterized by good tensile strength, high temperature resistance, and chemical resistance. The microsensor electrode used Au as the main electrode and Ti as the adhesion layer.

Matrix transformation of lunar regolith and its use as a feedstock for additive manufacturing.

Building a sustainable human habitat on the Moon requires advances in excavation, paving, and additive manufacturing to construct landing pads, surface transportation arteries, resilient shelters, and scientific outposts. Construction of infrastructure elements on the lunar surface necessitates exploration of the interfacial reactivity of locally sourced regolith and the adaptation of Earth-based construction techniques. Various crosslinking frameworks and sintering methods have been proposed as a means of consolidating lunar regolith into load-bearing structures but each have challenges related to incomplete understanding of reaction chemistry, excessive thermal budgets, and lack of universal applicability to different mineral components of regolith. We describe here a versatile experimental and computational study of the consolidation of a regolith simulant through formation of siloxane networks enmeshing mineral particles by surface dissolution-precipitation and polycondensation reactions. Furthermore, by tailoring the rheological properties of the formulation an additive manufacturing feedstock can be developed for the construction of lunar infrastructure.

Reactive Green 19 dye-ligand immobilized on the aminated nanofiber membranes for efficient adsorption of lysozyme: Process development and optimization in batch and flow systems.

The performance of lysozyme adsorption by the aminated nanofiber membrane immobilized with Reactive Green 19 (RG19) dyes was evaluated in batch and flow systems. The physicochemical properties of the dye-immobilized nanofiber membrane were characterized. The parameters of batch-mode adsorption of lysozyme (e.g., pH, initial dye concentration, and lysozyme concentration) were optimized using the Taguchi method. In a flow process, the factors influencing the dynamic binding performance for lysozyme adsorption in the chicken egg white (CEW) solution include immobilized dye concentration, adsorption pH value, feed flow rate, and feed CEW concentration. The impact of these operating conditions on the lysozyme purification process was investigated. Under optimal conditions, the recovery yield and purification factor of lysozyme achieved from the one-step adsorption process were 98.52% and 143 folds, respectively. The dye-affinity nanofiber membrane also did not exhibit any significant loss in its binding capacity and purification performance after five consecutive uses.

Baseline Circulating miR-125b Levels Predict a High FIB-4 Index Score in Chronic Hepatitis B Patients after Nucleos(t)ide Analog Treatment.

The regulatory role of microRNAs (miRNAs) in HBV-associated HCC pathogenesis has been reported previously. This study aimed to investigate the association between serum miR-125b and liver fibrosis progression in chronic hepatitis B (CHB) patients after nucleos(t)ide analog (NA) treatment. Baseline serum miR-125b levels and other relevant laboratory data were measured for 124 patients who underwent 12-month NA therapy. Post-12-month NA therapy, serum miR-125, platelet, AST, and ALT levels were measured again for post-treatment FIB-4 index calculation. Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for a higher post-treatment FIB-4 index. Results showed that baseline miR-125b levels were inversely correlated with the post-treatment FIB-4 index (ρ = −0.2130, p = 0.0082). In logistic regression analyses, age (OR = 1.17, p < 0.0001), baseline platelet level (OR = 0.98, p = 0.0032), and ALT level (OR = 1.00, p = 0.0241) were independent predictors of FIB-index > 2.9 post-12-month treatment. The baseline miR-125b level was not significantly associated with a higher post-treatment FIB-4 index (p = 0.8992). In 59 patients receiving entecavir (ETV) monotherapy, the alternation of serum miR-125b in 12 months and age were substantially associated with a higher post-treatment FIB-4 index (>2.9), suggesting that miR-125b is a reliable biomarker for detecting early liver fibrosis under specific anti-HBV NA treatments (e.g., ETV).

Factors Associated with the Patient/Client Use of Report Cards, Physician Rating Websites, Social Media, and Google for Hospital and Physician Selection: A Nationwide Survey.

OBJECTIVE: To explore the factors associated with the different uses of report cards, physician rating websites, social media, and Google, including awareness, physician finding, and decision-making based on reviews from the patient/client perspective. METHODS: We used computer-assisted telephone interviews to conduct a nationwide representative survey in Taiwan. RESULTS: The urbanization level of the area, income, and long-term health conditions were not associated with the three kinds of usage of the websites studied. Seeking health information was an important factor in the three kinds of website use. The employment industry was associated with awareness, and education level was associated with physician seeking and actions based on reviews. CONCLUSIONS: Different factors influenced the three kinds of usage: awareness, actual use (i.e., finding an appropriate physician), and decision-making based on reviews. Seeking health information is of primary importance regardless of how the websites are used. PRACTICAL IMPLICATIONS: Policy-makers should focus on educating individuals working outside the health care sector to increase awareness of these websites and to assist individuals with low levels of education in increasing their use of these websites.

Corrigendum: Targeting the N-terminus domain of the coronavirus nucleocapsid protein induces abnormal oligomerization via allosteric modulation.

[This corrects the article DOI: 10.3389/fmolb.2022.871499.].

A collaborative study to establish the national standard for SARS-CoV-2 RNA nucleic acid amplification techniques (NAAT) in Taiwan.

The conventional PCR remains a valuable method to detect the newly emergent coronavirus rapidly and accurately. Our investigation aimed to establish the standard materials of SARS-CoV-2 for NAAT detection. We provided formalin-inactivated SARS-CoV-2 and confirmed RNA copy numbers. In addition, the virus genome was confirmed with whole-genome sequencing and identified as Wuhan/WI04/2019. Seven laboratories were invited for this collaborative study, according to the reporting data, we determined the SARS-CoV-2 with the unit of 6.35 Log10 copies/mL as the national standard. The availability of the national standard (NS) of SARS-CoV-2 will facilitate the standardization and harmonization of SARS-CoV-2 NAAT assays.

AMPK promotes Arf6 activation in a kinase-independent manner upon glucose starvation.

AMP-activated protein kinase (AMPK) is a crucial cellular nutrient and energy sensor that maintains energy homeostasis. AMPK also governs cancer cell invasion and migration by regulating gene expression and activating multiple cellular signaling pathways. ADP-ribosylation factor 6 (Arf6) can be activated via nucleotide exchange by guanine-nucleotide-exchange factors (GEFs), and its activation also regulates tumor invasion and migration. By studying GEF-mediated Arf6 activation, we have elucidated that AMPK functions as a noncanonical GEF for Arf6 in a kinase-independent manner. Moreover, by examining the physiological role of the AMPK-Arf6 axis, we have determined that AMPK activates Arf6 upon glucose starvation and 5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside (AICAR) treatment. We have further identified the binding motif in the C-terminal regulatory domain of AMPK that is responsible for promoting Arf6 activation and, thus, inducing cell migration and invasion. These findings reveal a noncanonical role of AMPK in which its C-terminal regulatory domain serves as a GEF for Arf6 during glucose deprivation.

Operating Room Usage Time Estimation with Machine Learning Models.

Effectively handling the limited number of surgery operating rooms equipped with expensive equipment is a challenging task for hospital management such as reducing the case-time duration and reducing idle time. Improving the efficiency of operating room usage via reducing the idle time with better scheduling would rely on accurate estimation of surgery duration. Our model can achieve a good prediction result on surgery duration with a dozen of features. We have found the result of our best performing department-specific XGBoost model with the values 31.6 min, 18.71 min, 0.71, 28% and 27% for the metrics of root-mean-square error (RMSE), mean absolute error (MAE), coefficient of determination (R2), mean absolute percentage error (MAPE) and proportion of estimated result within 10% variation, respectively. We have presented each department-specific result with our estimated results between 5 and 10 min deviation would be more informative to the users in the real application. Our study shows comparable performance with previous studies, and the machine learning methods use fewer features that are better suited for universal usability.

Targeting the N-Terminus Domain of the Coronavirus Nucleocapsid Protein Induces Abnormal Oligomerization via Allosteric Modulation.

Epidemics caused by coronaviruses (CoVs), namely the severe acute respiratory syndrome (SARS) (2003), Middle East respiratory syndrome (MERS) (2012), and coronavirus disease 2019 (COVID-19) (2019), have triggered a global public health emergency. Drug development against CoVs is inherently arduous. The nucleocapsid (N) protein forms an oligomer and facilitates binding with the viral RNA genome, which is critical in the life cycle of the virus. In the current study, we found a potential allosteric site (Site 1) using PARS, an online allosteric site predictor, in the CoV N-N-terminal RNA-binding domain (NTD) to modulate the N protein conformation. We identified 5-hydroxyindole as the lead via molecular docking to target Site 1. We designed and synthesized four 5-hydroxyindole derivatives, named P4-1 to P4-4, based on the pose of 5-hydroxyindole in the docking model complex. Small-angle X-ray scattering (SAXS) data indicate that two 5-hydroxyindole compounds with higher hydrophobic R-groups mediate the binding between N-NTD and N-C-terminal dimerization domain (CTD) and elicit high-order oligomerization of the whole N protein. Furthermore, the crystal structures suggested that these two compounds act on this novel cavity and create a flat surface with higher hydrophobicity, which may mediate the interaction between N-NTD and N-CTD. Taken together, we discovered an allosteric binding pocket targeting small molecules that induces abnormal aggregation of the CoV N protein. These novel concepts will facilitate protein-protein interaction (PPI)-based drug design against various CoVs.

Galangin Reverses H2O2-Induced Dermal Fibroblast Senescence via SIRT1-PGC-1α/Nrf2 Signaling.

UV radiation and H2O2 are the primary factors that cause skin aging. Both trigger oxidative stress and cellular aging. It has been reported that deacetylase silent information regulator 1 (SIRT1), a longevity gene, enhances activation of NF-E2-related factor-2 (Nrf2), as well as its downstream key antioxidant gene hemeoxygenase-1 (HO-1), to protect cells against oxidative damage by deacetylating the transcription coactivator PPARγ coactivator-1α (PGC-1α). Galangin, a flavonoid, possesses anti-oxidative and anti-inflammatory potential. In the present study, we applied Ultraviolet B/H2O2-induced human dermal fibroblast damage as an in vitro model and UVB-induced photoaging of C57BL/6J nude mice as an in vivo model to investigate the underlying dermo-protective mechanisms of galangin. Our results indicated that galangin treatment attenuates H2O2/UVB-induced cell viability reduction, dermal aging, and SIRT1/PGC-1α/Nrf2 signaling activation. Furthermore, galangin treatment enhanced Nrf2 activation and nuclear accumulation, in addition to inhibiting Nrf2 degradation. Interestingly, upregulation of antioxidant response element luciferase activity following galangin treatment indicated the transcriptional activation of Nrf2. However, knockdown of SIRT1, PGC-1α, or Nrf2 by siRNA reversed the antioxidant and anti-aging effects of galangin. In vivo evidence further showed that galangin treatment, at doses of 12 and 24 mg/kg on the dorsal skin cells of nude mice resulted in considerably reduced UVB-induced epidermal hyperplasia and skin senescence, and promoted SIRT1/PGC-1α/Nrf2 signaling. Furthermore, enhanced nuclear localization of Nrf2 was observed in galangin-treated mice following UVB irradiation. In conclusion, our data indicated that galangin exerts anti-photoaging and antioxidant effects by promoting SIRT1/PGC-1α/Nrf2 signaling. Therefore, galangin is a potentially promising agent for cosmetic skin care products against UV-induced skin aging.

Potentially conjugative plasmids harboring Tn6636, a multidrug-resistant and composite mobile element, in Staphylococcus aureus.

OBJECTIVES: This study aimed to provide detailed genetic characterization of Tn6636, a multidrug-resistant and composite mobile element, in clinical isolates of Staphylococcus aureus. METHODS: A total of 112 ermB-positive methicillin-susceptible S. aureus (MSSA) and 224 ermB-positive methicillin-resistant S. aureus (MRSA) isolates collected from 2000 to 2015 were tested for the presence of Tn6636. Detection of the plasmids harboring Tn6636 was performed by S1 nuclease digestion pulsed-field gel electrophoresis (PFGE) analysis, conjugation test, and whole genome sequencing (WGS). RESULTS: Prevalence of Tn6636 in MSSA is higher than that in MRSA. Ten MSSA isolates and 10 MRSA isolates carried Tn6636. The 10 MSSA isolates belonged to three sequence types (ST), including ST7 (n = 6), ST5 (n = 3), and ST59 (n = 1). The 10 MRSA isolates belonged to ST188 (n = 8) and ST965 (n = 2). Analysis of plasmid sequences revealed that Tn6636 was harbored by six different mosaic plasmids. In addition to resistance genes, some plasmids also harbored toxin genes. CONCLUSION: The presence of multi-resistant Tn6636 in plasmids of both MSSA and MRSA with various STs suggests its broad dissemination. Results indicate that Tn6636 has existed for at least 16 years in Taiwan. The mosaic plasmids harboring Tn6636 can be transferred by conjugation. Ongoing surveillance of Tn6636 is essential to avoid continued spreading of resistant plasmids.

Design of an Integrated Microfluidic Paper-Based Chip and Inspection Machine for the Detection of Mercury in Food with Silver Nanoparticles.

For most of the fast screening test papers for detecting Hg2+, the obtained results are qualitative. This study developed an operation for the µPAD and combined it with the chemical colorimetric method. Silver nanoparticle (AgNP) colloids were adopted as the reactive color reagent to combine and react with the Hg standards on the paper-based chip. Then, the RGB values for the color change were used to establish the standard curve (R2 > 0.99). Subsequently, this detection system was employed for the detection tests of actual samples, and the detected RGB values of the samples were substituted back to the formula to calculate the Hg2+ contents in the food. In this study, the Hg2+ content and recovery rate in commercially available packaged water and edible salts were measured. The research results indicate that a swift, economical, and simple detection method for Hg2+ content in food has been successfully developed.

Process Optimization of Silver Nanoparticle Synthesis and Its Application in Mercury Detection.

Silver nanoparticles (AgNPs) have stable reactivity and excellent optical absorption properties. They can be applied in various industries, such as environmental protection, biochemical engineering, and analyte monitoring. However, synthesizing AgNPs and determining their appropriate dosage as a coloring substance are difficult tasks. In this study, to optimize the process of AgNP synthesis and obtain a simple detection method for trace mercury in the environment, we evaluate several factors-including the reagent addition sequence, reaction temperature, reaction time, the pH of the solution, and reagent concentration-considering the color intensity and purity of AgNPs as the reaction optimization criteria. The optimal process for AgNP synthesis is as follows: Mix 10 mM of silver nitrate with trisodium citrate in a hot water bath for 10 min; then, add 10 mM of sodium borohydride to produce the AgNPs and keep stirring for 2 h; finally, adjust the pH to 12 to obtain the most stable products. For AgNP-based mercury detection, the calibration curve of mercury over the concentration range of 0.1-2 ppb exhibits good linearity (R2 > 0.99). This study provides a stable and excellent AgNP synthesis technique that can improve various applications involving AgNP-mediated reactions and has the potential to be developed as an alternative to using expensive detection equipment and to be applied for the detection of mercury in food.

Breast cancer recurrence prediction with ensemble methods and cost-sensitive learning.

Breast cancer is one of the most common cancers in women all over the world. Due to the improvement of medical treatments, most of the breast cancer patients would be in remission. However, the patients have to face the next challenge, the recurrence of breast cancer which may cause more severe effects, and even death. The prediction of breast cancer recurrence is crucial for reducing mortality. This paper proposes a prediction model for the recurrence of breast cancer based on clinical nominal and numeric features. In this study, our data consist of 1,061 patients from Breast Cancer Registry from Shin Kong Wu Ho-Su Memorial Hospital between 2011 and 2016, in which 37 records are denoted as breast cancer recurrence. Each record has 85 features. Our approach consists of three stages. First, we perform data preprocessing and feature selection techniques to consolidate the dataset. Among all features, six features are identified for further processing in the following stages. Next, we apply resampling techniques to resolve the issue of class imbalance. Finally, we construct two classifiers, AdaBoost and cost-sensitive learning, to predict the risk of recurrence and carry out the performance evaluation in three-fold cross-validation. By applying the AdaBoost method, we achieve accuracy of 0.973 and sensitivity of 0.675. By combining the AdaBoost and cost-sensitive method of our model, we achieve a reasonable accuracy of 0.468 and substantially high sensitivity of 0.947 which guarantee almost no false dismissal. Our model can be used as a supporting tool in the setting and evaluation of the follow-up visit for early intervention and more advanced treatments to lower cancer mortality.

ArfGAP1 acts as a GTPase-activating protein for human ADP-ribosylation factor-like 1 protein.

ADP-ribosylation factors (Arfs) and Arf-like (Arl) GTPases are key regulators of intracellular vesicle trafficking and Golgi structure. Both Arf and Arl proteins cycle between active GTP-bound and inactive GDP-bound forms, where guanine nucleotide exchange factors (GEFs) regulate the exchange of GDP for GTP, whereas GTPase-activating proteins (GAPs) promote the hydrolysis of bound GTP. Human Arl1 is located at the trans-Golgi network (TGN) and regulates the function and structure of the Golgi complex. However, neither GEFs nor GAPs for human Arl1 have been identified. Here, we report that ArfGAP1, an Arf1 GAP, can promote GTP hydrolysis of Arl1. We show that ArfGAP1 directly interacts with GTP-bound Arl1 and exhibits GAP activity toward Arl1 in vitro. Exogenous expression of ArfGAP1, but not ArfGAP2 and ArfGAP3, causes dissociation of endogenous Arl1 from the TGN. In addition, GAP activity-deficient ArfGAP1 fails to regulate the Golgi localization of Arl1. Using an activity pull-down assay, we demonstrated that ArfGAP1 regulates the levels of Arl1-GTP in cells expressing ArfGAP1-myc or with ArfGAP1 knockdown. Finally, we observed that, similar to expression of putative active Arl1 (Arl1QL), ArfGAP1 knockdown impairs endosome-to-TGN retrograde transport of the Shiga toxin B-subunit. Thus, our findings support the idea that ArfGAP1 acts as an Arl1 GAP to regulate the function of Arl1 in vesicle trafficking at the TGN.