Search

Nicotinamide adenine dinucleotide phosphate oxidase complex 2 (NOX2) is an effector molecule expressed predominantly in neutrophils. Its deficiency is present in immune disorders, including systemic lupus erythematosus, chronic granulomatous disease, and rheumatoid arthritis. Recent reports indicated that NOX2 regulates autoimmunity and programmed cell death. However, the exact mechanism is unclear. In this study, we explored the effect of NOX2 on neutrophil apoptosis. We demonstrated that NOX2 deficiency caused neutrophil pyroptosis. Mechanistically, the NOX2 inhibitor GSK2795039 application or knockdown of NOX2 components resulted in increased mitochondrial ROS, inhibition of AMP-activated protein kinase (AMPK), and activation of Gasdermin E. AMPK activators, metformin and epigallocatechin gallate, inhibited deficient NOX2-induced pyroptosis. Together, these findings illustrate the involvement of NOX2 in regulating neutrophil death and emphasize its importance in autoimmunity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Multicolour electrochromic materials have been considered as a promising alternative to achieve dynamic full-colour tuning towards next-generation electronic display technology. However, the development of electrochromics with wide colour gamut and subtle multicolour tunability still remains challenging due to inflexible energy level structures in intrinsic active materials. Herein, the electrochromic π-conjugated polymers with rich and subtle colour tunability were designed and developed based on a fine adjustment on the energy level structures. The chromatic transition covers almost full-colour gamut, and each colour scheme has a rich variety of categories stemming from versatile hues, chromas and lightnesses. Moreover, the multicolour π-conjugated polymers also demonstrate superior overall electrochromic performance, including fast switching (∼1.0 s), high colouration efficiency (160.4 cm 2 C -1 @550 nm) and good reversibility (over 90 % retention after 10,000 cycles). As a proof of concept, ultrathin and flexible prototype devices are developed by utilizing the multicolour π-conjugated polymers as electrochromic active layer, exhibiting a wide colour gamut and highly saturated multicolour tunability. The design principles proposed in this work may also be applicable to diverse optoelectronic applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Bioaugmentation is a bioremediation approach to treat groundwater contaminated with chlorinated ethenes, but currently it faces challenges such as poor microbiome stability and effectiveness, due to blind species integration and metabolic inhibition. The objective of this study was to create a controllable and functionally stable microbial community for dichlorination application. For this, we utilized targeted screening to identify dechlorinating bacteria from contaminated groundwater, that in combination would form an anaerobic dechlorination microbial community with stabilizing metabolic interactions between the constituents. The results showed that two organohalide-respiring bacterial (OHRB) species were isolated, and these were identified as Enterobacter bugandensis X4 and Enterobacter sichuanensis C4. Upon co-cultivation with lactic acid as the carbon source, the strains demonstrated metabolic interactions and synergistic dehalogenation ability towards trichloroethene (TCE). It was further demonstrated that the functional microbiome constructed with the strains was stable over time and exhibited a robust TCE degradation rate of 80.85% at 13.13 mg/L TCE within 10 days. Additionally, the complete conversion of TCE was achieved through microbiome bioaugmentation, this augmented microbiome increased the degradation rate towards 52.55 mg/L TCE by approximately 30% within 6 days. Additionally, bioaugmentation stimulated the growth of indigenous OHRB, such as Dehalobacter and Desulfovibrio. It also promoted a positive succession of the microbial community. These findings offer valuable insights into the microbial remediation of chlorinated ethenes-contaminated groundwater and offers novel ideas for the construction of an artificial functional microbiome., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In recent decades, China's rapid development has led to significant environmental pollution from the widespread use of chemical products. Per- and polyfluoroalkyl substances (PFAS) are among the most concerning pollutants due to their persistence and bioaccumulation. This article assesses PFAS exposure levels, distribution, and health risks in Chinese blood, environment, and food. Out of 4037 papers retrieved from November 2022 to December 31, 2023, 351 articles met the criteria. Findings show perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) as the main PFAS in both Chinese populations and the environment. The highest PFOA levels in Chinese populations were in Shandong (53.868 ng/mL), while Hubei had the highest PFOS levels (43.874 ng/mL). Similarly, water samples from Sichuan (2115.204 ng/L) and Jiangsu (368.134 ng/L) had the highest PFOA and PFOS levels, respectively. Although localized areas showed high PFAS concentrations. Additionally, developed areas had higher PFAS contamination. The researches conducted in areas such as Qinghai and Hainan remain limited, underscoring the imperative for further investigation. Temporal analysis indicates declining levels of some PFAS, but emerging alternatives require more research. Limited studies on PFAS concentrations in soil, atmosphere, and food emphasize the need for comprehensive research to mitigate human exposure., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In spite of the considerable advancements achieved in enhancing the power conversion efficiency (PCE) of lead-based all-inorganic perovskite solar cells, there persists a need for materials that are both more stable and environmentally friendly. This investigation systematically explores the structural and thermodynamic stability, and electronic properties of Ge-based all-inorganic perovskite CsGeX 3 (X = Cl, Br, I) in two space groups, Pm 3̄ m and R 3 m , utilizing first-principles calculations. Introducing the novel concept of the "imaginary frequency coefficient" alongside the tolerance factor and stabilizing the chemical potential window, we collectively characterize the stability of CsGeX 3 based on the phonon spectrum and phonon density of states calculations. The findings reveal that the stable phase of the Ge-based perovskite differs from that of lead-based systems, with the R 3 m structure of CsGeX 3 being the most stable in the rhombohedral phase. Moreover, the stability of R 3 m -CsGeX 3 can be manipulated by adjusting the halide composition with a gradual increase in stability observed as halogen atoms shift from I to Cl. This comprehensive approach, integrating the phonon spectrum, innovative measurement indicators, and tolerance factor, presents an effective strategy for designing materials that are both non-toxic and stable.

Background: To evaluate the new preoperative Changgung classification (CC) system of portal vein thrombosis (PVT) in choosing suitable operative procedures to reconstruct portal veins during liver transplantation (LT) in patients with portal vein cavernous transformation (PVCT)., Methods: This retrospective observational study analyzed data from allograft LTs performed for various liver diseases., Results: The study included 22 males and 4 females with LT indications comprising cirrhosis ( n  = 9), hepatocellular carcinoma ( n  = 12), PVCT ( n  = 2), liver failure from fulminant hepatitis B ( n  = 1), dysfunction of transplanted liver ( n  = 1), and chronic rejection of transplanted liver ( n  = 1). Patients were classified according to Yerdel (21 Yerdel II and 5 Yerdel III) and CC (C1-C5). In total 16 simple operations were performed on C1-C3 cases and 9 complex operations on C4-C5 cases, with one additional simple operation. The distribution according to the Yerdel classification was 16 simple and 5 complex operations in Yerdel II cases and 1 simple and 4 complex operations in Yerdel III cases. The median follow-up time was 27.5 months with overall one-year and three-year OS rates of 88.1% and 83.9% for the cohort. Specifically, the one-year OS rates for patients classified as C1-3 vs. C4-5 were 93.3% and 80.0%, while the three-year OS rates were 86.7% and 80.0%, respectively ( p  = 0.526)., Conclusion: The CC proposed in this study shows comparable potential to the Yerdel classification in preoperatively identifying the need for complex surgical techniques in LT patients with PVCT and may also have predictive power for the survival benefits following LT.

Background: The prognostic significance of margin-to-depth ratio (MDR) in oral cavity squamous cell carcinoma (OCSCC) remains unclear, particularly in comparison to traditional margin status. We aimed to examine the association between MDR and clinical outcomes in a large Taiwanese cohort., Methods: A total of 18,324 patients with first primary OCSCC were categorized by margin status: positive (1013), <5 mm (8371), and ≥ 5 mm (8940). Disease-specific survival (DSS) and overall survival (OS) served as the main outcome measures., Results: After excluding patients with positive margins (MDR = 0), the optimal MDR cutoff value for DSS and OS was 0.6. Patients with MDR > 0.6 showed significantly better 5-year DSS and OS rates (87 %, 81 %) compared to those with MDR ≤ 0.6 (71 %, 63 %) and MDR = 0 (53 %, 43 %). Multivariable analysis identified MDR ≤ 0.6 as independently associated with both DSS and OS in the entire cohort (hazard ratio [HR] = 1.34/1.32). This finding was consistent in the subgroups with surgical margins < 5 mm (HR = 1.39 for DSS and 1.38 for OS) and margins ≥ 5 mm (HR = 1.21 for both DSS and OS). In subgroups with surgical margins < 5 mm and ≥ 5 mm, an MDR > 0.6 was associated with better survival outcomes., Conclusions: An MDR (cutoff: 0.6) is independently associated with prognosis in OCSCC, offering improved risk stratification compared to margin status alone. While MDR may guide surgical margin modification, further research is needed to determine whether MDR could serve as a postoperative indicator for adjuvant therapy in patients with close or clear margins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

We aimed to evaluate the presence of sleep-disordered breathing (SDB) in generalized anxiety disorder (GAD) patients and its impacts after 12-week of escitalopram treatment. In this prospective observational study, GAD patients were followed up to determine whether comorbid SDB affects their treatment response. They underwent a home sleep apnea test (HSAT), and oxygen desaturation index ≥5 was considered indicative of SDB. All participants were treated with escitalopram 5 mg in the first 2 weeks and 10 mg in the remaining 10 weeks. They underwent assessments of heart rate variability and completed questionnaires for sleep, mood, attention, and daytime sleepiness on the day of enrollment, and 2 weeks, 4 weeks, and 12 weeks later. A total of 45 GAD participants (mean age, 56.2 years) were included in the study. Of these, 53.3% had comorbid SDB. Participants with GAD and SDB had higher baseline Beck Anxiety Inventory (BAI) and STOP-BANG scores. Both groups of participants with GAD showed overall improvement in anxiety, depression, and insomnia symptoms after 12 weeks of escitalopram treatment. Additionally, there were no statistically significant differences in the changes in BAI, Beck Depression Inventory, and Epworth Sleepiness Scale scores between the two groups at the Week 2, Week 4, and Week 12 assessments compared to baseline. However, the GAD group with comorbid SDB seemed to have a lower tendency for improvement in depressive symptoms than the group with non-comorbid SDB. Compared with the GAD group without comorbid SDB, the GAD group with comorbid SDB exhibited more severe anxiety symptoms at baseline and tended to show less improvement in depressive symptoms after 12 weeks of escitalopram treatment., Competing Interests: Declaration of competing interest The authors declare that this study was conducted in the absence of any commercial or financial relationships that could be construed as potential conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Introduction: When switching from premixed insulin to insulin degludec/aspart (IDegAsp), IDegAsp usually starts at the same dose as the premixed insulin according to limited clinical experience or at a dose according to clinician discretion. The dose of insulin degludec used in the real world after switching has been poorly investigated., Methods: A retrospective analysis was conducted on patients with type 2 diabetes who switched from premixed insulin to IDegAsp from October 2016 to December 2023. Repeated measures analysis of variance was used to compare changes in insulin dose, glycated hemoglobin (HbA1c), fasting blood glucose (FBG), and postprandial blood glucose (PBG) before and after switching., Results: Sixty-six patients with prior low-ratio premixed insulin and 22 with prior mid-ratio premixed insulin were included. Among the low-ratio insulin users, the total daily dose of insulin degludec (IDeg) decreased by 21.43% and 19.05% at 3 and 6 months, respectively, after switching, compared with prior basal insulin dose (both p < 0.001). Conversely, among mid-ratio insulin users, the IDeg daily dose increased by 10.71% and 32.14% at 3 and 6 months, respectively, after switching, compared with prior basal insulin dose (both p < 0.001). In all patients, HbA1c levels decreased by 0.70%, FBG decreased by 1.00 mmol/l, and PBG decreased by 1.61 mmol/l after 6 months of switching (all p < 0.05); the total daily insulin dose and injection frequency significantly decreased after switching (both p < 0.05); age and disease duration did not affect IDegAsp effects on HbA1c reduction., Conclusions: In the setting of transition to IDegAsp from premixed insulin, the dose of basal insulin in the premixed formulation can be a valuable reference for adjusting insulin degludec dose. IDegAsp is superior to premixed insulin in blood glucose control with reduced total daily dose and injection frequency. IDegAsp could be the best choice for the management of diabetes in elderly patients., Competing Interests: Declarations Conflict of Interest All authors (Yiming Wu, Junqing Zhang, and Ang Li) declare that they have no competing interests. Ethical Approval This study was approved by the Ethics Committee of the Peking University First Hospital (approval number: 2024 Yan 240-001) and registered with the Chinese Clinical Trial Registry (identifier: ChiCTR2400085570). Informed consent of the patients was waived by the Ethics Committee due to the retrospective nature of the study., (© 2024. The Author(s).)

Introduction: This retrospective study aimed to evaluate the prognostic value of [ 18 F]FDG parameters in patients with visceral and bone metastatic hormone-sensitive prostate cancer (mHSPC)., Patients and Methods: This analysis included the mHSPC patients who underwent [ 18 F]FDG PET/CT at the initial diagnosis. Baseline characteristics were analyzed, and the uptake of [ 18 F]FDG was quantified using SUV max . Kaplan-Meier and Cox proportional hazard regression analysis were employed to evaluate the correlation between SUV max and patient survival., Results: Among the 267 patients enrolled, 90 (33.7%) presented with visceral metastases and 177 (66.3%) had bone metastases. The median follow-up for the visceral metastasis group was 35.5 months (IQR 26-53.8 months). The median overall survival for patients with lung, liver, or both metastases were 30, 21 and 17 months, respectively. Patients exhibiting higher [ 18 F]FDG uptake in metastatic lesions experienced shorter overall survival (OS) in comparison to those with lower [ 18 F]FDG uptake, both in the visceral metastases group (17 vs. 31 months, p  = 0.002) and the bone metastases group (27.5 vs. 34.5 months, p  < 0.001). Cox regression analysis further revealed that increased [ 18 F]FDG uptake in metastatic lesions emerged as a significant risk factor in both OS and progression-free survival (PFS). In contrast, the variability in [ 18 F]FDG uptake in primary lesions did not provide a reliable indicator for predicting prognosis., Conclusions: In mHSPC patients, higher [ 18 F]FDG uptake in metastatic lesions indicates shorter survival and increased risk of disease progression. The [ 18 F]FDG SUV max in primary tumors did not show significant prognostic value. Our study underscores the unique prognostic potential of [ 18 F]FDG PET/CT in mHSPC patients, highlighting its importance in the management of both bone and visceral metastases.

Dissimilatory iron-reducing bacteria (DIRB) with extracellular electron transfer (EET) capabilities have shown significant potential for bioremediating halogenated hydrocarbon contaminated sites rich in iron and humic substances. However, the role and microbial molecular mechanisms of iron-humic acid (Fe-HA) complexes in the reductive dehalogenation process of DIRB remains inadequately elucidated. In this study, we developed a sustainable carbon cycling approach using Fe-HA complexes to modulate the electron flux from sawdust (SD), enabling almost complete reductive dechlorination by most DIRB (e.g., Shewanella oneidensis MR-1) that lack complex iron-sulfur molybdo enzymes. The SD-Fe-HA/MR-1 system achieved a 96.52% removal efficiency of tetrachloroethylene (PCE) at concentrations up to 250 μmol/L within 60 days. Material characterization revealed that DIRB facilitated the hydrolysis of macromolecular carbon sources by inducing the formation of amorphous ferrihydrite (FeOOH) in Fe-HA complexes. More importantly, the bioavailable FeOOH activated additional intracellular electron flow pathways, increasing the activity of potential dehalogenases. Transcriptome further highlight the innovative role of biogenic amorphous FeOOH in integrating intracellular redox metabolism with extracellular charge exchange to facilitate reductive dechlorination in DIRB. These findings provide novel insights into accelerating reductive dechlorination in-situ contaminated sites lacking obligate dehalogenating bacteria., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Leukaemia inhibitory factor (LIF), a member of the interleukin-6 (IL-6) family, is a multifunctional cytokine. The maturation-to-ovulation process of poultry follicles is determined by granulosa cell proliferation and differentiation. Granulosa cell apoptosis and degeneration lead to follicular atresia, which reduces the number of normally developing follicles and leads to a decrease in the poultry egg production rate, thus affecting the large-scale development of poultry breeding. In this study, the LIF gene overexpression vector pCDH-CMV-LIF and a siRNA that inhibits LIF gene expression were transfected into primary granulosa cells from white Muscovy duck ovaries for functional study. Compared with that in the control group, LIF gene expression was confirmed to be significantly decreased or increased in the transfection groups (P < 0.01). After LIF overexpression, the expression of the cell cycle-related genes CCND1, CDK-1 and PCNA was decreased (P < 0.05); apoptosis was promoted; the proapoptotic genes Bax and caspase-3 were significantly upregulated (P < 0.01); and the antiapoptotic gene Bcl-2 was significantly downregulated (P < 0.01). After LIF interference, the expression of the cell cycle-related genes CCND1, CCNE1, CDK-1 and PCNA and the antiapoptotic gene Bcl-2 significantly increased (P < 0.01), whereas the expression of the proapoptotic genes Bax, caspase-3 and caspase-9 significantly decreased (P < 0.01). In summary, the LIF gene is involved in regulating the biological function of ovarian granulosa cells in white Muscovy ducks. LIF gene expression promotes granulosa cell apoptosis and inhibits cell cycle progression. These experimental results provide insights into the follicular development mechanism of white Muscovy ducks., Competing Interests: Declaration of competing interest No potential conflicts of interest are reported by the authors., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Although guidelines recommend risk assessment for hospital-acquired venous thromboembolism (HA-VTE) to inform prophylaxis decisions, studies demonstrate inappropriate utilization of pharmacoprophylaxis in hospitalized medical patients. Predictors of pharmacoprophylaxis initiation in medical inpatients remain largely unknown., Objectives: To determine factors associated with HA-VTE pharmacoprophylaxis initiation in adults hospitalized on medical services., Methods: We performed a cohort study using electronic health record data from adult patients hospitalized on medical services at 4 academic medical centers between 2016 and 2019. Main measures were candidate predictors of HA-VTE pharmacoprophylaxis initiation, including known HA-VTE risk factors, predicted HA-VTE risk, and bleeding diagnoses present on admission., Results: Among 111 550 admissions not on intermediate or full-dose anticoagulation, 48 520 (43.5%) received HA-VTE pharmacoprophylaxis on the day of or the day after admission. After adjustment for age, sex, race/ethnicity, and study site, the strongest clinical predictors of HA-VTE pharmacoprophylaxis initiation were malnutrition and chronic obstructive pulmonary disease. Thrombocytopenia and history of gastrointestinal bleeding were associated with decreased odds of HA-VTE pharmacoprophylaxis initiation. Patients in the highest 2 tertiles of predicted HA-VTE risk were less likely to receive HA-VTE pharmacoprophylaxis than patients in the lowest (first) tertile (OR, 0.84; 95% CI, 0.81-0.86 for the second tertile; OR, 0.95; 95% CI, 0.92-0.98 for the third tertile)., Conclusion: Among patients not already receiving anticoagulants, HA-VTE pharmacoprophylaxis initiation during the first 2 hospital days was lower in patients with a higher predicted HA-VTE risk and those with risk factors for bleeding. Reasons for not initiating pharmacoprophylaxis in those with a higher predicted HA-VTE risk could not be assessed., Competing Interests: Declaration of competing interests There are no competing interests to disclose., (Copyright © 2024 International Society on Thrombosis and Haemostasis. All rights reserved.)

Objectives: Studies have increasingly focussed on the relationship between periodontitis (PD) and preeclampsia (PE). However, conclusions have not been consistent, and it is unclear whether any causal relationship exists between them and whether causality is bidirectional. This study employed Mendelian randomisation (MR) analysis to investigate the potential bidirectional causal relationship between PD and PE., Methods: Genetic variants strongly linked to PD (17,353 cases and 28,210 controls), chronic periodontitis (CP; 1817 cases and 2215 controls), aggressive periodontitis (AgP; 851 cases and 6580 controls), and PE (7212 cases and 194,266 controls) in the genome-wide association study (GWAS) of European ancestry were used as instrumental variables (IVs). Inverse variance weighting (IVW) served as the primary method for causal inference. MR Pleiotropy RESidual Sum and Outlier (MR-PRESSO) was utilised to analyse horizontal pleiotropy. Cochrane Q tests and leave-one-out analyses were used to assess heterogeneity and stability amongst IVs., Results: The MR analysis revealed no causal impacts of PD or its 2 subtypes-CP and AgP-on PE. Similarly, no significant causal effect of PE on PD was found in the reverse-MR analysis (IVW odds ratio, 0.97; 95% confidence interval, 0.91-1.05; P = .58). The findings from MR-Egger, weighted median, weighted mode, and the simple modelling approaches, as well as the pleiotropy and sensitivity analyses, aligned with those of the IVW method., Conclusions: The MR analysis suggests no bidirectional causal relationship between PD and PE; hence, PD and PE might not increase or prevent the risk of one other., Clinical Relevance: Genetically, periodontitis or its subtypes chronic periodontitis and aggressive periodontitis may not require specific clinical attention to prevent the development of preeclampsia., Competing Interests: Conflict of interest None disclosed., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Oocyte meiotic maturation failure and chromosome abnormality is one of the main causes of infertility, abortion, and diseases. The mono-orientation of sister chromatids during the first meiosis is important for ensuring accurate chromosome segregation in oocytes. MEIKIN is a germ cell-specific protein that can regulate the mono-orientation of sister chromatids and the protection of the centromeric cohesin complex during meiosis I. Here we found that MEIKIN is a maternal protein that was highly expressed in mouse oocytes before the metaphase I (MI) stage, but became degraded by the MII stage and dramatically reduced after fertilization. Strikingly, MEIKIN underwent phosphorylation modification after germinal vesicle breakdown (GVBD), indicating its possible function in subsequent cellular event regulation. We further showed that MEIKIN phosphorylation was mediated by PLK1 at its carboxyl terminal region and its C-terminus was its key functional domain. To clarify the biological significance of meikin degradation during later stages of oocyte maturation, exogenous expression of MEIKIN was employed, which showed that suppression of MEIKIN degradation resulted in chromosome misalignment, cyclin B1 and Securin degradation failure, and MI arrest through a spindle assembly checkpoint (SAC)-independent mechanism. Exogenous expression of MEIKIN also inhibited metaphase II (MII) exit and early embryo development. These results indicate that proper MEIKIN expression level and its C-terminal phosphorylation by PLK1 are critical for regulating the metaphase-anaphase transition in meiotic oocyte. The findings of this study are important for understanding the regulation of chromosome segregation and the prevention meiotic abnormality., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Glioblastoma (GBM), one of the most aggressive brain cancers, presents significant treatment challenges due to its complex biology and resistance to conventional therapies, necessitating the development of new, low-toxicity, and effective treatments. This study explores the antitumor potential of phlorizin, a naturally occurring dihydrochalcone, as a standalone agent and in combination with temozolomide (TMZ), the standard chemotherapeutic for GBM. Phlorizin was found to significantly inhibit cell viability and migration in vitro, with synergistic effects observed when combined with TMZ. Comprehensive analyses, including protein-protein interaction network construction, enrichment analysis, and molecular docking with AKT1, identified the PI3K/AKT/mTOR signaling pathway as a critical mediator of glioblastoma cell survival and proliferation targeted by phlorizin. Pathway enrichment analysis of 88 intersection targets further highlighted this pathway's role in phlorizin's activity. Western blot validation confirmed that phlorizin inhibits the expression of key proteins within the PI3K/AKT/mTOR pathway, providing a mechanistic basis for its antitumor effects. These findings suggest that phlorizin, particularly in combination with TMZ, holds significant potential as a therapeutic strategy for glioblastoma by targeting molecular pathways critical for cancer cell survival and proliferation., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests. Dear author, please, make sure that you can answer YES to all these questions before submission:, (Copyright © 2024. Published by Elsevier B.V.)

Aggregation of TAR-DNA binding protein-43 (TDP-43) is a pathological feature present in nearly 97 % cases of amyotrophic lateral sclerosis (ALS), making it an attractive target for pathogenic studies and drug screening. Here, we have performed a high-throughput screening of 1500 compounds from a natural product library and identified that lycorine, a naturally occurring alkaloid, significantly decreases the level of TDP-43 A315T in a cellular model. We further demonstrate that lycorine reduces the level of TDP-43 A315T both through inhibiting its synthesis and by promoting its degradation by the ubiquitin-proteasome system (UPS). Importantly, treatment with lycorine significantly attenuates TDP-43 proteinopathy and improves functional recovery in TDP-43 A315T -expressing Caenorhabditis elegans and mouse models. These findings suggest that lycorine is a promising lead compound that has therapeutic potential for ALS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Early screening of hepatocellular carcinoma (HCC) is strongly recommended for hepatitis B virus (HBV)-infected patients. We aimed to develop and validate a predictive nomogram based on HCC occurrence trajectory for screening precancerous patients with HCC., Methods: Peripheral blood mononuclear cells (PBMC) samples from 22 patients with HCC with their precancerous stage (n = 55) and 18 healthy controls were measured using HumanMethylation EPIC BeadChip assay. HCC trajectory was assessed by pseudotime based on TimeAx algorithm and chronological time. The 43 candidate CpG sites were selected from the methylation signature and measured using multiplex bisulfite sequencing in a retrospective cohort of HBV-infected patients (n = 604). A 5-CpG-classifier was built using the LASSO Cox regression model, based on the association between the methylation level of every CpG and the duration from enrollment to HCC occurrence of individual patient. We validated the risk stratification and predictive accuracy of this classifier in both the primary cohort (n = 300) and independent validation cohort (n = 304)., Results: Pseudotime and chronological time of HCC trajectory analysis revealed that the PD-1/PD-L1 pathway underwent changes in the precancerous stage. Based on the trajectory of methylation signature, we built a 5-CpG-classifier which remained powerful and independent predictive efficiency after stratified analysis by clinicopathological risk factors in both primary cohort and independent validation cohort. A predicting nomogram including the 5-CpG-classifier was constructed after multivariate analysis. One-year cumulative hazard of HCC in low- and high-risk groups of HBV-infected patients was 3.0% (0.1%-5.8%) and 17.90% (11.00%-24.3%) (P < .0001) in primary cohort, 4.5% (1.20%-7.80%) and 27.3 (18.90-34.90) (P < .0001) in the independent validation cohort., Conclusions: One-year before HCC was a critical period of transitional time when parts of the methylation profile underwent shifting toward HCC like. The nomogram could identify precancerous stage patients with HCC who should be screened for early diagnosis and intervention., (© The Author(s) 2024. Published by Oxford University Press.)

8:2 Chlorinated polyfluoroalkyl ether sulfonate (8:2 Cl-PFESA) is a substitute for perfluorooctane sulfonate and an emerging environmental pollutant, yet its bioaccumulation and health risks are poorly understood. We established a subchronic exposure model in mice (0.04, 0.2, and 1 mg/kg/d) to evaluate its adverse effects. Our findings show extensive distribution of 8:2 Cl-PFESA in various tissues (plasma, liver, kidney, brain, heart, lung, testis, ovary, spleen, thymus, thyroid, uterus, large intestine, small intestine, muscle, and fat), with the highest accumulation in the liver, identified as the primary storage organ. Liver histopathology revealed elevated alanine aminotransferase levels, reduced triglycerides, and ballooning degeneration. Proteomics analysis indicated significant involvement of amino acid metabolism in 8:2 Cl-PFESA-induced hepatotoxicity. Metabolomics analysis further highlighted pronounced alterations in amino acid interconversion. Multi-omics integration revealed disruptions in amino acid metabolism, particularly with alanine, histidine, and tryptophan, identifying key proteins EHHADH, HAL, MAO-A, ALDH3A2, and TDO2 as crucial connectors in these metabolic processes, validated by Western blot experiments. This study provides a comprehensive analysis of 8:2 Cl-PFESA accumulation and distribution in biological systems, demonstrating that hepatotoxicity is primarily mediated through amino acid metabolism disruption, offering insights for pollution mitigation strategies and future toxicological research., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A novel buckled-layer K 1.39 Mn 3 O 6 cathode was synthesized, featuring P2-oxygen stacking and P3-K sites. The presence of two independent manganese sites significantly influences the charge distribution and diffusion pathways of potassium ions during the charge and discharge processes, resulting in exceptional electrochemical performance. A reversible capacity of 105 mA h g -1 at 100 mA g -1 and remarkable rate capability of 70 mA h g -1 at 1 A g -1 were achieved, surpassing those of most previously reported layered transition metal oxide cathodes. The findings facilitate the development of innovative cathode materials., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Dental restorative resin composites are widely used to repair tooth decay owing to attractive esthetics, adequate mechanical properties and minimally invasive tooth structure preparations. Nevertheless, dental restorative resin composites still face challenges because of their relatively high failure rate and short lifespan caused by secondary caries and bulk fracture. Thus, attempts have been carried out to explore a new generation of dental restorative resin composites with antibacterial, remineralizing, and self-healing capabilities to inhibit bacteria and lengthen the lifetime of the restorations. Such novel restorative composites can inhibit bacterial activity, reduce acid production, promote mineral regeneration and present a renewable advantage to achieve a higher performance, which are inspiring and provide support for further basic and clinical research. In this review, antibacterial dental restorative resin composites are first introduced, followed by remineralizing, self-healing, and multifunctional dental resin composites with two or more of the functions mentioned above. Meanwhile, we explain the mechanism of the corresponding dental restorative resin composites and describe their characteristics. Finally, we conclude and put forward prospects. This review will attract both researchers and clinicians in this field and help to provide innovative ideas to design new restorative resin composites for biomedical applications., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Glioblastoma (GBM) is an aggressive form of brain cancer in which treatment is associated with toxicities that can result in therapy discontinuation or death. This analysis investigated clinical and genetic markers of vascular toxicities in GBM patients during active treatment., Methods: 591 Non-Hispanic White GBM patients with clinical data were included in the analysis from NRG RTOG-0825. Genome-wide association studies (GWAS) were performed from genotyped blood samples (N=367) by occurrence of thrombosis or hypertension (grade ≥2). A clinical prediction model was produced for each vascular toxicity. Significant GWAS variants were then added to the clinical model as a single nucleotide polymorphism (SNP) -dose effect variable to produce the final genetic models., Results: Thrombosis and hypertension were experienced by 62 (11%) and 59 (10%) patients, respectively. Patients who experienced hypertension displayed improved survival over those without hypertension (median overall survival: 25.72 vs 15.47 months, p=0.002). The genetic model of thrombosis included corticosteroid use (OR: 7.13, p=0.02), absolute neutrophil count (OR: 1.008, p=0.19), body surface area (OR: 18.87, p=0.0008), and the SNP-dose effect (3 variants; OR: 3.79, p<.0001). The genetic model of hypertension included bevacizumab use (OR: 0.97, p=0.95) and the SNP-dose effect (6 variants; OR: 4.44, p<.0001)., Conclusion: In this study, germline variants were superior in predicting hypertension than clinical variables alone. Additionally, corticosteroid use was a considerable risk factor for thrombosis. Future investigations should confirm the hazard of corticosteroid use on thrombosis and the impact of bevacizumab in other malignancies after accounting for the genetic risk of hypertension., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Plesiomonas shigelloides is a gram-negative opportunistic pathogen associated with gastrointestinal and extraintestinal diseases in humans. There have been reports of specific functional genes in the study of P. shigelloides, but there are also many unknown genes that may play a role in P. shigelloides pathogenesis as global regulatory proteins or virulence factors., Results: In this study, we found a transcriptional regulator of the PadR family in P. shigelloides and named it PstR (GenBank accession number: EON87311.1), which is present in various pathogenic bacteria but whose function has rarely been reported. RNA sequencing (RNA-Seq) was used to analyze the effects of PstR on P. shigelloides, and the results indicated that PstR regulates approximately 9.83% of the transcriptome, which includes impacts on motility, virulence, and physiological metabolism. RNA-seq results showed that PstR positively regulated the expression of the flagella gene cluster, which was also confirmed by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Luminescence screening assay. Meanwhile, the ΔpstR mutant strains lacked flagella and were non-motile, as confirmed by motility assays and transmission electron microscopy (TEM). Additionally, PstR also positively regulates T3SS expression, which aids in P. shigelloides' capacity to infect Caco-2 cells. Meanwhile, we also revealed that PstR negatively regulates fatty acid degradation and metabolism, as well as the regulatory relationship between PsrA, a regulator of fatty acid degradation and metabolism, and its downstream genes in P. shigelloides., Conclusions: Overall, we revealed the effects of PstR on motility, virulence, and physiological metabolism in P. shigelloides, which will serve as a foundation for future research into the intricate regulatory network of PstR in bacteria., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

We investigated the effect of individual, joint and fluctuating exposure to air pollution (PM 2.5 , BC, NO 3 - , NH 4 + , OM, SO 4 2- , PM 10 , NO 2 , SO 2 , O 3 ) on glucose metabolisms among prediabetes, and simultaneously explored the modifying effect of lifestyle. We conducted a longitudinal study among prediabetes during 2018-2022. Exposure windows within 60-days moving averages and their variabilities were calculated. FBG, insulin, HOMA-IR, HOMA-B, triglyceride glucose index (TyG), glucose insulin ratio (GI) and allostatic load of glucose homeostasis system (AL-GHS) was included. Linear mixed-effects model and BKMR were adopted to investigate the individual and overall effects, respectively. We also explored the preventive role of lifestyle. Individual air pollutant was associated with increased FBG, insulin, HOMA-IR, HOMA-B, TyG, and decreased GI. People with FBG ≥6.1 mmol/L were more susceptible. Air pollutants mixture were only associated with increased HOMA-B, and constituents have the highest group-PIP. Air pollutants variation also exert harmful effect. We observed similar diabetic effect on AL-GHS. Finally, the diabetic effect of air pollutants disappeared if participants adopt a favorable lifestyle. Our findings highlighted the importance of comprehensively assessing multiple air pollutants and their variations, focusing on metabolic health status in the early prevention of T2D, and adopting healthy lifestyle to mitigate such harmful effect., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: Aging-related osteoporosis is considered as a serious public health concern for middle-aged and elderly people, with an intricated pathogenesis including the recently identified aging-induced immunological dysfunction and gut microbial disorder. The intervention based on dietary prebiotics is recommended to retain bone health and postpone the progression of osteoporosis., Objectives: As a well-defined prebiotic, 2'-fucosyllactose (2'-FL) has been thoroughly validated with positive effect on systemic health and was proposed in this study to unveil its intervention on aging-related osteoporosis, as well as the underlying mechanisms involving the gut microecology and innate immunity., Methods: The effects of dietary 2'-FL on osteoporosis phenotypes were identified by evaluating the severity of bone loss and microstructure damage in natural aging mice. The mechanisms relying on innate immune profile, intestinal barrier function, and gut microbial homeostasis, were analyzed to elucidate the signaling axis. The detailed molecular signaling was validated based on LPS-stimulated RAW 264.7 murine macrophages., Results: The results indicated that 12-week 2'-FL intervention retrieved bone loss and microstructure damage in natural aging mice. Also, 2'-FL alleviated aging-induced colonic inflammation, gut barrier dysfunction, and abnormal expression of intestinal tight-junction protein. The impact of 2'-FL treatment on the aging-induced gut microbial dysbiosis was validated by restoring gut microbiota diversity, recovering the abundance of Bifidobacterium, Prevotellaceae and Akkermansia, and inhibiting the growth of Stenotrophomonas. Flow cytometry analysis revealed changes in dendritic cell (DC) and macrophage subsets with age, and a decrease in M1-polarized macrophages was observed in 2'-FL-treated aged mice and RAW264.7 cells potentially through the interaction with toll-like receptor 4 (TLR4) to suppress NF-κB signaling and the secretion of proinflammatory factors., Conclusion: These findings highlight the preventive effect of 2'-FL on aging-associated osteoporosis by regulating gut microbial homeostasis and innate immune responses., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

To address the need for automated sorting of synthetic diamonds based on quality in manufacturing enterprises, this study developed a dedicated dataset and an enhanced YOLOv8n model for synthetic diamonds detection and quality evaluation, named YOLOv8n-adamas. We redesigned the backbone network to improve feature extraction capabilities and introduced a dynamic detection head based on attention mechanisms to further enhance model performance. Experimental results show that on synthetic diamonds dataset, YOLOv8n-adamas achieved a 4.0% improvement in precision (P), a 2.7% increase in recall (R), and improvements of 1.5% and 1.3% in mean average precisions at 50% and 95% Intersection over Union (IoU) thresholds (mAP50 and mAP95) compared to YOLOv8. Furthermore, YOLOv8n-adamas also outperforms other commonly used, high-performing models in various metrics on this dataset, offering effective technical support for the automated quality-based sorting of synthetic diamonds., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Pancreatic β cell dysfunction is critical to the development of type 2 diabetes (T2D). We show that the complement receptor C3aR1 on β cells plays an essential role in maintaining β cell homeostasis, especially under the metabolic duress of obesity and T2D. Mice with β cell specific deletion of C3ar1 have worse glucose tolerance, lower insulin levels, and decreased β cell mass. Islets from β cell specific C3ar1 knockout (β-C3aR1 KO) mice demonstrate impaired insulin secretion. Disruption of C3ar1 on β cells ablates the insulin secretory response to C3a, establishing a signaling axis between C3a and β cell-derived C3aR1. Markers of β cell identity were decreased while stress markers were increased in β-C3aR1 KO mice. Islets from β-C3aR1 KO also exhibit increased β cell death to lipotoxicity. Finally, we show that C3AR1 is positively correlated with insulin secretion in human islets. These findings indicate that C3aR1 expression on β cells is necessary to maintain optimal β cell function and preserve β cell mass in T2D.

Lung cancer is primarily classified as NSCLC, which is distinguished by a wide range of genetic variations. This study focused on RNF144A-AS1 , a relatively unexplored lncRNA, to explore the impact of its genetic polymorphisms on the susceptibility to NSCLC. We detected RNF144A-AS1 expression and its correlation with prognosis and clinical pathological features using bioinformatics analysis. The association between RNF144A-AS1 polymorphism and NSCLC susceptibility was evaluated using case-control methods. This investigation featured a cohort of 700 NSCLC individuals and 700 healthy controls. The genotype of genetic variation was detected by PCR-RFLP and iMLDR, followed by subsequent calculation of OR and 95 % CI . Our data show that RNF144A-AS1 exhibits high expression levels in LUAD tissues and its expression is closely linked to LUAD progression and prognosis. Carrier of RNF144A-AS1 rs3806609 TT genotype increased NSCLC susceptibility compared to carrier of rs3806609 CC genotype ( OR  = 2.21, 95% CI  = 1.57-3.13). Our study identifies RNF144A-AS1 genetic variants as potential susceptibility markers in NSCLC. RNF144A-AS1 promotes cell proliferation and migration in LUAD through the IFN-γ/JAK2/STAT1 signalling pathway. Collectively, these findings pave the way for developing targeted therapies and diagnostic tools based on RNF144A-AS1 and its variants., (© 2024 The Authors.)

The same cells can die with varied immunological consequences. For the purpose of cancer therapy, stronger immunogenic death of cancer cells is considered favorable. Membrane disruptive peptides are cytotoxic agents with tunable structures capable of not just killing heterogeneous cancer cells, but also inducing immunogenic death. However, the chemo-structural principles that underlie their immunogenic cytotoxicity remain elusive. Here we investigated a series of arginine-rich amphipathic peptides with representative structures on the relationship between the mode of cell death and the immunogenic potency. Among several hydrophobic motif-appended cyclic octaarginine peptides, FC-14 was found to induce cell stress and necroptotic death, unlike apoptotic peptide RL2 and membrane-dissolving peptide RL1. Their differing abilities to release immunogenic death markers correlated well with their potential to activate innate immunity and protective vaccinal effect in a prophylactic model, with FC-14 being the most potent. FC-14 can be pre-opsonized with albumin into nanoparticles (PopAN-FC-14) using PopAN technology to improve its pharmacokinetic properties for intravenous injection. In a syngeneic mouse model of subcutaneous breast cancer, PopAN-FC-14 showed superior therapeutic effect and safety profile than the albumin formulated nanomedicine Nab-paclitaxel (Nab-PTX). Boost injections of PopAN-FC-14 significantly enhanced tumor-specific cellular and humoral immunities, acting similarly as in-situ cancer vaccine. Overall, this work demonstrates a novel focus on the immunogenic cytotoxicity of peptides and a practical approach for effective systemic therapy of cancer., Competing Interests: Declaration of competing interest The reported compounds and the PopAN methods have been patented or under application. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Lung metastasis is a leading cause of cancer-related death. mRNA-based cancer vaccines have been demonstrated to be effective at inhibiting tumor growth. Intranasal immunization has emerged as a more effective method of inducing local immune responses against cancer cells in the lungs., Methods: An innovative layered double hydroxide- and 5-OP-RU-based mRNA nanovaccine (Mg/Al LDH-5-OP-RU/mRNA) was synthesized via coprecipitation. The particle size distribution and zeta potential were measured, and the nanovaccine was observed by transmission electron microscopy. The functions and properties of the nanovaccine were evaluated via an mRNA-targeted delivery assay and measurement of dendritic cell (DC) and mucosa-associated invariant T (MAIT) cell maturation and activation. In addition, the cytotoxicity, antigen-specific T cell activation, cytokines, protective ability, and therapeutic ability of the nanovaccine were assessed in a mouse tumor model. Further, the immune cell composition was evaluated in tumors., Results: The Mg/Al LDH-5-OP-RU/mRNA nanovaccine was efficiently delivered into lung-draining mediastinal lymph nodes (MLNs), and it activated dendritic cells (DCs) and mucosa-associated invariant T (MAIT) cells after intranasal administration. Moreover, the optimized dual-activating mRNA nanovaccine efficiently transfected DC cells and expressed antigen proteins in DC cells. An HPV-associated tumor model revealed that the intranasal delivery of the Mg/Al LDH-5-OP-RU/E7 mRNA nanovaccine significantly prevented the lung metastasis of tumors and had a therapeutic effect on established metastatic tumor nodules in the lungs. Mechanistically, the enhanced activation of DC and MAIT cells induced by the Mg/Al LDH-5-OP-RU/E7 mRNA nanovaccine increased the production of immune-stimulating cytokines and decreased the secretion of immunosuppressive cytokines, which led to the expansion and activation of memory T cells targeting the E7 antigen, a reduction in the population of neutrophils, and differentiation of tumor -associated macrophages to the M1 phenotype in the lungs., Conclusion: These results highlight the potential of the innovative nasal mRNA nanovaccine for both preventing and treating tumor metastasis in the lungs., Competing Interests: The authors declare that they have no conflicts of interest., (© 2024 Li et al.)

Metal-organic frameworks (MOFs) have emerged as a highly promising platform for various sensing applications due to their tunable structures and functionalities. In the present work, a Eu 3+ -doped mixed-ligand MOF, namely, the Eu 3+ @UiO-66-IPA, exhibited excellent luminescent properties and high fluorescence stability in aqueous media, displaying dual-emission peaks under 395 and 615 nm excitation that were readily visible to the naked eye. Importantly, the presence of fluoride ions (F - ) promoted the "antenna effect" between the ligand and the Eu 3+ centers, which significantly enhanced the emission intensity of the Eu 3+ characteristic peak. In addition, the addition of F - also inhibited the quenching effect of high-energy O-H bonds existing in the aqueous environment. Notably, Eu 3+ @UiO-66-IPA demonstrated exceptional selectivity for F - over a range of competing anions, with a remarkable limit of detection as low as 0.22 μM. The developed Eu 3+ -doped mixed-ligand MOF system offers a highly promising strategy for the simple and accurate sensing of F - in practical applications.

The utilization of solar-driven technologies for the direct conversion of methanol (CH3OH) into two or multi-carbon compounds through controlled carbon-carbon (C-C) coupling is an appealing yet challenging objective. In this study, we successfully demonstrate the photocatalytic CH3OH coupling to ethylene (C2H4), a valuable chemical raw material, by employing a carbon nitride-based catalyst. Specifically, we modify the layered polymer carbon nitride (PCN) photocatalyst through the incorporation of Au single atoms (Au1/PCN) using a chemical-scissors method. The synergistic effect between the PCN substrate and the Au single atoms reduces the potential barrier associated with C-C coupling, thereby enhancing the efficiency of CH3OH reforming to C2H4. This investigation not only reveals a novel pathway for C2H4 production via CH3OH reforming but also provides fresh insights into the possibilities of C-C coupling., (© 2024 Wiley‐VCH GmbH.)

Photon-based entanglement sources, crucial for obtaining entangled states, are typically generated via spontaneous parametric down-conversion. However, birefringence in nonlinear crystals causes spatial or temporal walk-off, reducing entangled photon quality. The theoretical analysis attributes birefringent walk-off to dispersive materials curbed by pre- and post-compensation. We experimentally validate this method, enhancing polarization-entangled photon quality. We measured the Bell state along with a violation of CHSH-Bell's inequality by ∼366 standard deviations (S = 2.623 ± 0.0017). Our approach, simple and stable, does not complicate setups. Its applicability extends to optimizing various entanglement source schemes, aiding optical quantum technologies such as computation, sensing, and communication.

To improve the accuracy of photogrammetric joint roughness coefficient (JRC) estimation, this study proposes two optimization models based on ground sample distance (GSD), point density, and the root mean square error (RMSE) of checkpoints. First, an algorithm that automatically generates spatial positions for equipment based on the convergence strategy was developed, using principles of Structure from Motion and Multi-View Stereo (SfM-MVS) and the shooting parameter selection algorithm (SPSA). Second, a portable positioning plate containing ground control points and checkpoints was designed based on optical principles, and a moving camera capture strategy guided by SPSA was proposed. Combining SPSA, portable positioning plate, and moving camera capture strategy, a photogrammetric experiment for small-scale rock samples in the field was conducted, collecting 48 datasets with different shooting parameters. Subsequently, a dataset incorporating GSD, point density, RMSE, and three JRC estimation metrics was established, revealing their correlations and sensitivities. Using seven machine learning algorithms, optimization models for photogrammetric JRC accuracy were developed, with Linear Multidimensional Regression and Gaussian Process Regression models improving JRC accuracy by an average of 85.73%. Finally, the applicability and limitations of the newly proposed method were further discussed., (© 2024. The Author(s).)

While the direct impact of oyster calcification and respiration on the seawater inorganic carbon system is well-acknowledged, their indirect effect through filter feeding activities remains unclear. Here we studied the impact of large-scale oyster farming on the removal of dissolved inorganic carbon (DIC) from seawater. Field investigations showed that the DIC level in the oyster farming area in Sanggou Bay, China were significantly lower than that in the non-farming area. In-lab incubation showed that regardless of whether incubated in high or low-transparent environments, the DIC removal rate of seawater from the oyster farming area was significantly higher than that of the non-farming area. These results indicate that cultivated oysters facilitate the removal of seawater DIC in the farming area. To reveal the indirect effect of filter feeding activities on DIC removal, we used 6-m 3 ponds to simulate the oyster-farming environment. Results showed that the average DIC level of the oyster-cultivating groups was 105.83 μmol/kg lower than that of the control groups (without of oyster) after a six-day cultivation. Surprisingly, the average concentration of Chl-a in oyster-cultivating groups was significantly higher than that of the control group at the end of the experiment. Similarly, DIC level declined faster while Chl-a concentration increased faster in seawater that previously experienced 12～20 h of oyster cultivation than that in the control seawater. It was noticed that the transparency of seawater within 6-m 3 ponds increased significantly just after hours of oyster cultivation. This enhanced transparency created a favorable light environment that supported phytoplankton photosynthesis and simultaneously accelerated the DIC removal rate. Overall, oysters not only remove the inorganic carbon in the seawater through calcification but also create a suitable environment for phytoplankton photosynthesis through their filtering activity, and subsequently accelerating the removal of inorganic carbon in the seawater of the oyster farming area., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Autophagy plays an important role in determining stem-cell differentiation. During the osteogenic differentiation of mesenchymal stem cells (MSCs), autophagosome formation is upregulated but the reason is unknown. A long-standing quest in the autophagy field is to find the membrane origin of autophagosomes. In this study, cytoplasmic coat protein complex II (COPII) vesicles, endoplasmic reticulum-derived vesicles responsible for the transport of storage proteins to the Golgi, are demonstrated to be a critical source of osteoblastic autophagosomal membrane. A significant correlation between the number of COPII vesicle and the autophagy level is identified in the rat bone tissues. Disruption of COPII vesicles restrained osteogenesis and decreased the number and size of autophagosomes. SEC31a (an outer coat protein of COPII vesicle) is found to be vital to regulate COPII vesicle-dependent autophagosome formation via interacting with ATG9a of autophagosomal seed vesicles. The interference of Sec31a inhibited autophagosome formation and osteogenesis in vitro and in vivo. These results identified a novel mechanism of autophagosome formation in osteogenic differentiation of stem cells and identified SEC31a as a critical protein that mediates the interplay between COPII and ATG9a vesicles. These findings broaden the understanding of the regulatory mechanism in the osteogenic differentiation of MSCs., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Signal-molecule-mediated strategies are proposed for aerobic granular sludge (AGS), but the regulatory mechanisms behind AGS formation are largely unexplored. In this study, two sequence batch reactors (SBRs) were operated to investigate the regulation of diffusible signal factor (DSF) in AGS formation. DSF secretion in Reactor 2 (R2: 10 °C→25 °C) decreased by 15 % compared to Reactor 1 (R1: 25 °C→10 °C), correlating with a 26 % increase in extracellular polymeric substance (EPS) concentration, resulting in a 63 % acceleration of the granulation process. After temperature shocks in R2, DSF concentration increased by 70 %, while EPS concentration decreased by 47 %. Batch tests confirmed that DSF inhibited EPS secretion. Combined 16S rRNA analysis and machine learning identified key bacteria responsible for secreting EPS and signal molecule. The decrease in the abundances of these bacteria reduced EPS production. These findings on DSF regulation of EPS secretion provide an in-depth understanding of enhanced AGS granulation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The properties of materials are strongly correlated with their atomic scale structures. Achieving a comprehensive understanding of the atomic-scale structure-property relationship requires advancements of imaging and spectroscopy techniques. Aberration-corrected scanning transmission electron microscopy (STEM) has seen rapid development over the past decades and is now routinely employed for atomic-scale characterization. However, quantitative STEM imaging and spectroscopy analysis at the single-atom level is challenging due to the extremely weak signals generated from individual atom, thus imposing stringent requirements for analysis sensitivity. This review discusses the development and application of low-voltage STEM techniques with single-atom sensitivity, primarily based on recent research presented on an invited talk at the 5th 2D23 SALVE Symposium, including annular dark-field (ADF) imaging, functional imaging and electron energy-loss spectroscopy (EELS) analysis. Carbon-based nanomaterials were chosen as model systems for demonstrating the capabilities of single-atom STEM imaging and EELS analysis, due to their structural stability under low accelerating voltages and their rich physical and chemical properties. Moreover, this review summarizes recent advancements and applications of low-voltage single-atom STEM imaging and spectroscopy in the study of functional materials and discusses prospects for future developments., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Despite the advantages of high tissue penetration depth, selectivity, and non-invasiveness of photothermal therapy for cancer treatment, developing NIR-II photothermal agents with desirable photothermal performance and advanced theranostics ability remains a key challenge. Herein, a universal surface modification strategy is proposed to effectively improve the photothermal performance of vanadium carbide MXene nanosheets (L-V 2 C) with the removal of surface impurity ions and generation of mesopores. Subsequently, MnO x coating capable of T1-weighted magnetic resonance imaging can be in situ formed through surface redox reaction on L-V 2 C, and then, stable nanoplatforms (LVM-PEG) under physiological conditions can be obtained after further PEGylation. In the tumor microenvironment irradiated by NIR-II laser, multivalent Mn ions released from LVM-PEG, as a reversible electronic station, can consume the overexpression of glutathione and catalyze a Fenton-like reaction to produce ·OH, resulting in synchronous cellular oxidative damage. Efficient synergistic therapy promotes immunogenic cell death, improving tumor-related immune microenvironment and immunomodulation, and thus, LVM-PEG can demonstrate high accuracy and excellent anticancer efficiency guided by multimodal imaging. As a result, this study provides a new approach for the customization of 2D surface strategies and the study of synergistic therapy mechanisms, highlighting the application of MXene-based materials in the biomedical field., (© 2024 Wiley‐VCH GmbH.)

Purpose: To provide relative citation ratio (RCR) benchmark data for the field of glaucoma., Design: Cross-sectional bibliometric analysis., Subjects: Fellowship-trained glaucoma faculty at Accreditation Council for Graduate Medical Education-accredited institutions., Methods: Glaucoma faculty were individually indexed using the National Institutes of Health (NIH) iCite website. Publication count, mean RCR score, and weighted RCR score were collected for each author between May and August 2023 and included PubMed-listed articles from 1980 to 2023. Data were compared by sex, career duration, academic rank, and acquisition of a Doctor of Philosophy (PhD)., Main Outcome Measures: Total number of publications, mean RCR value, and weighted RCR value., Results: Five hundred twenty-six academic glaucoma specialists from 113 institutions were indexed. These physicians produced highly impactful research with a median publication count of 13 (interquartile range [IQR] 4-38), median RCR of 1.41 (IQR 0.97-1.98), and median weighted RCR of 16.89 (4.80-63.39). Academic rank, career duration, and having a PhD were associated with increased publication count, mean RCR, and weighted RCR. Publication count and weighted RCR differed significantly by sex; however, no difference was observed with mean RCR., Conclusions: Current academic glaucoma specialists have high mean RCR values relative to the NIH standard RCR value of 1. This benchmark data serve as a more accurate gauge of research impact within the glaucoma community and can be used to inform self, institutional, and departmental evaluations. Additionally, the mean RCR may provide an accurate metric for quantifying research productivity among historically underrepresented groups that are disadvantaged by time-dependent factors such as number of publications., Financial Disclosure(s): Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article., (Copyright © 2024 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Sorption is one of the most efficient methods to remediate the increasing oil spill incidents, but the currently available absorbents are inadequate to tackle such a global threat. Recently, numerous researchers have attempted to develop sustainable oil sorbents. Cellulose aerogels and foams, a type of lightweight porous material with excellent sorption performance, are one of the most promising candidates. Significant progress has been made in the past decade towards the development of cellulose porous materials as effective oil sorbents, with improvements in their oil sorption capacity, reusability, and enhanced multifunctionality, indicating their potential for oil spill remediation. This article reviews recent reports and provides a comprehensive overview of the preparation and modification strategies for cellulose porous materials, with a specific emphasis on their oil sorption performance and structure control. We also focus on the burgeoning 3D printing technology within this field, summarizing the latest advances with a discussion of the potential for using 3D printing to customize and optimize the structure of cellulose porous materials. Lastly, this review addresses current limitations and outlines future directions for development., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Background: Chronic total occlusions (CTO) occur in about 20% of patients referred for coronary angiography, and right coronary artery (RCA) CTO has been reported in 38-50% of the entire CTO population. Limited data on angiographic and procedural characteristics of RCA-CTO and the risk of adverse cardiac events asks for a detailed study., Methods: From 2010 to 2013, patients with attempted revascularization of at least one CTO lesion were included and followed up to 5 years after PCI. Eligible patients are assigned to RCA-CTO and non-RCA-CTO groups based on their target vessels. The primary endpoint was major adverse cardiovascular events (MACEs; a composite of all-cause death, myocardial infarction (MI) or rehospitalization for heart failure), and secondary endpoints were cardiac death, target lesion revascularization (TLR) and target vessel revascularization (TVR)., Results: The present study included 2659 eligible patients, among which 1285 patients were assigned to the RCA-CTO group, whereas 1374 patients were assigned to the non-RCA-CTO group. Lesions in RCA had longer lesion length, higher J-CTO score, higher rates of severe vessel tortuosity, a higher percentage of Rentrop grade 2-3, and more likely to be re-try lesion than those in LAD or LCX (all P < 0.01). CTO lesions in RCA reached less successful recanalization and post-procedural TIMI 3 flow (all <0.01). Multivariate Cox analysis revealed that RCA-CTO was not associated with primary outcome MACEs. Besides MACEs, RCA-CTO was also not associated with cardiac death, but was significantly associated with TLR and TVR (adjusted HR: 1.37 [95% CI:1.07-1.76], P = 0.01; adjusted HR: 1.43 [95% CI:1.13-1.82], P = 0.003)., Conclusion: RCA-CTO lesions, which had more complex angiographic features, independently contributed to TLR and TVR but not to MACEs or cardiac death in the 5 years of follow-up., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Blastocyst morphology has been demonstrated to be associated with ploidy status. Existing artificial intelligence models use manual grading or 2D images as the input for euploidy prediction, which suffer from subjectivity from observers and information loss due to incomplete features from 2D images. Here we aim to predict euploidy in human blastocysts using quantitative morphological parameters obtained by 3D morphology measurement., Methods: Multi-view images of 226 blastocysts on Day 6 were captured by manually rotating blastocysts during the preparation stage of trophectoderm biopsy. Quantitative morphological parameters were obtained by 3D morphology measurement. Six machine learning models were trained using 3D morphological parameters as the input and PGT-A results as the ground truth outcome. Model performance, including sensitivity, specificity, precision, accuracy and AUC, was evaluated on an additional test dataset. Model interpretation was conducted on the best-performing model., Results: All the 3D morphological parameters were significantly different between euploid and non-euploid blastocysts. Multivariate analysis revealed that three of the five parameters including trophectoderm cell number, trophectoderm cell size variance and inner cell mass area maintained statistical significance (P < 0.001, aOR = 1.054, 95% CI 1.034-1.073; P = 0.003, aOR = 0.994, 95% CI 0.991-0.998; P = 0.010, aOR = 1.003, 95% CI 1.001-1.006). The accuracy of euploidy prediction by the six machine learning models ranged from 80 to 95.6%, and the AUCs ranged from 0.881 to 0.984. Particularly, the decision tree model achieved the highest accuracy of 95.6% (95% CI 84.9-99.5%) with the AUC of 0.978 (95% CI 0.882-0.999), and the extreme gradient boosting model achieved the highest AUC of 0.984 (95% CI 0.892-1.000) with the accuracy of 93.3% (95% CI 81.7-98.6%). No significant difference was found between different age groups using either decision tree or extreme gradient boosting to predict euploid blastocysts. The quantitative criteria extracted from the decision tree imply that euploid blastocysts have a higher number of trophectoderm cells, larger inner cell mass area, and smaller trophectoderm cell size variance compared to non-euploid blastocysts., Conclusions: Using quantitative morphological parameters obtained by 3D morphology measurement, the decision tree-based machine learning model achieved an accuracy of 95.6% and AUC of 0.978 for predicting euploidy in Day 6 human blastocysts., Trial Registration: N/A., (© 2024. The Author(s).)

Background: The evidence for adjuvant chemoradiotherapy (CRT) of oral cavity squamous cell carcinoma (OCSCC) with extra-nodal extension (ENE) in National Comprehensive Cancer Network (NCCN) guidelines is derived from patients with head and neck cancer. The guidelines further suggest a radiation dose ranging from 6000 to 6600 cGy. In this nationwide study, we sought to evaluate the prognostic significance of adjuvant therapy and the specific radiation dosage in Taiwanese patients with pure OCSCC and ENE., Methods: A retrospective analysis of 1577 OCSCC patients with ENE who underwent resection and received adjuvant CRT or radiotherapy (RT) between January 2011 and December 2020 was conducted., Results: Multivariable analysis revealed that adjuvant RT, more than four pathologically positive nodes, and radiation dosage below 6000 cGy were independent risk factors for unfavorable 5-year disease-specific survival (DSS) and overall survival (OS). Comparing patients who received CRT (n = 1453) to those treated with RT (n = 124) before and after propensity score (PS) matching, the 5-year outcomes were as follows: before PS matching, DSS (54% versus 30%, p < 0.0001), OS (42% versus 18%, p < 0.0001); after PS matching (n = 111 in each group), DSS (52% versus 30%, p = 0.0016), OS (38% versus 21%, p = 0.0019). For patients who underwent CRT, the 5-year outcomes for different radiation dose groups (6600 - 7000 cGy, n = 1155 versus 6000 - 6500 cGy, n = 199) were as follows: before PS matching, DSS (52% versus 54%, p = 0.1904), OS (43% versus 46%, p = 0.1610); after PS matching (n = 199 in each group), DSS (55% versus 54%, p = 0.8374), OS (46.5% versus 46.3%, p = 0.7578)., Conclusions: For OCSCC patients with ENE, our study shows CRT improved survivals than RT alone, underscoring the clinical significance of chemotherapy. Patients undergoing CRT with irradiation doses ranging from 6000 to 6500 cGy exhibited comparable survival outcomes to those receiving doses of 6600-7000 cGy. This observation suggests that irradiation doses exceeding the 6600 cGy may not confer the survival advantage in these patients. Further research is needed to confirm our results and explore the optimal irradiation dose for managing these patients., (© 2024. The Author(s).)

Spinal cord injury (SCI) triggers a cascade of intricate molecular and cellular changes that determine the outcome. In this study, we resolve the spatiotemporal organization of the injured mouse spinal cord and quantitatively assess in situ cell-cell communication following SCI. By analyzing existing single-cell RNA sequencing datasets alongside our spatial data, we delineate a subpopulation of Igfbp2-expressing astrocytes that migrate from the white matter (WM) to gray matter (GM) and become reactive upon SCI, termed Astro-GMii. Further, Igfbp2 upregulation promotes astrocyte migration, proliferation, and reactivity, and the secreted IGFBP2 protein fosters neurite outgrowth. Finally, we show that IGFBP2 significantly reduces neuronal loss and remarkably improves the functional recovery in a mouse model of SCI in vivo. Together, this study not only provides a comprehensive molecular atlas of SCI but also exemplifies how this rich resource can be applied to endow cells and genes with functional insight and therapeutic potential., Competing Interests: Declaration of interests Y.F., Z.W., and T.L., from the Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, have filed a Patent Cooperation Treaty (PCT) application (PCT/CN2024/092024) on the use of the IGFBP2 protein and bio-material composition for treating neural injury and neurodegeneration., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The canonical G406R mutation that increases Ca 2+ influx through the CACNA1C-encoded Ca V 1.2 Ca 2+ channel underlies the multisystem disorder Timothy syndrome (TS), characterized by life-threatening arrhythmias. Severe episodic hypoglycemia is among the poorly characterized non-cardiac TS pathologies. While hypothesized from increased Ca 2+ influx in pancreatic beta cells and consequent hyperinsulinism, this hypoglycemia mechanism is undemonstrated because of limited clinical data and lack of animal models. We generated a Ca V 1.2 G406R knockin mouse model that recapitulates key TS features, including hypoglycemia. Unexpectedly, these mice do not show hyperactive beta cells or hyperinsulinism in the setting of normal intrinsic beta cell function, suggesting dysregulated glucose homeostasis. Patient data confirm the absence of hyperinsulinism. We discover multiple alternative contributors, including perturbed counterregulatory hormone responses with defects in glucagon secretion and abnormal hypothalamic control of glucose homeostasis. These data provide new insights into contributions of Ca V 1.2 channels and reveal integrated consequences of the mutant channels driving life-threatening events in TS., (© 2024. The Author(s).)

The detection of gravitational waves has ushered in a new era of observing the universe. Quantum resource advantages offer significant enhancements to the sensitivity of gravitational wave observatories. While squeezed states for ground-based gravitational wave detection have received marked attention, the generation of squeezed states suitable for mid-to-low-frequency detection has remained unexplored. To address the gap in squeezed state optical fields at ultra-low frequencies, we report on the first direct observation of a squeezed vacuum field until Fourier frequency of 4 millihertz with the quantum noise reduction of up to 8.0 dB, by the employment of a multiple noise suppression scheme. Our work provides quantum resources for future gravitational wave observatories, facilitating the development of quantum precision measurement., (© 2024. The Author(s).)

The nematode Caenorhabditis elegans is widely employed as a model organism to study basic biological mechanisms. However, transgenic C. elegans are generated by manual injection, which remains low-throughput and labor-intensive, limiting the scope of approaches benefitting from large-scale transgenesis. Here, we report a robotic microinjection system, integrating a microfluidic device capable of reliable worm immobilization, transfer, and rotation, for high-speed injection of C. elegans. The robotic system provides an injection speed 2-3 times faster than that of experts with 7-22 years of experience while maintaining comparable injection quality and only limited trials needed by users to become proficient. We further employ our system in a large-scale reverse genetic screen using multiplexed alternative splicing reporters, and find that the TDP-1 RNA-binding protein regulates alternative splicing of zoo-1 mRNA, which encodes variants of the zonula occludens tight junction proteins. With its high speed, high accuracy, and high efficiency in worm injection, this robotic system shows great potential for high-throughput transgenic studies of C. elegans., (© 2024. The Author(s).)

Competing Interests: Declaration of interests PM reports grants from National Institute for Health and Care Research (NIHR), the Medical Research Council (MRC), Bristol & Weston Hospitals Charity, and The Cassell Hospital Charitable Trust; and salary support from the NIHR Applied Research Collaboration Southwest, the NIHR Biomedical Research Centre at University Hospitals Bristol and Weston NHS Foundation, and Avon and Wiltshire Mental Health Partnership NHS Trust. NK reports grants from NIHR, Health Quality Improvement Partnership, and the Department of Health and Social Care; has received salary support from the Greater Manchester NIHR Patient Safety Research Collaboration, Mersey Care NHS Foundation Trust, and the University of Manchester; has chaired and contributed to committees for the National Institute for Health and Clinical Excellence (NICE) guidelines, including those on the management of self-harm; and is a member of the National Suicide Prevention Strategy Advisory Group (England). RCO is a trustee and science council member of MQ Mental Health Research, president of the International Association for Suicide Prevention, co-chair of the academic advisory group to the Scottish Government's National Suicide Prevention Leadership Group, and a board member of the International Academy of Suicide Research; was a member of the NICE guideline group for the management of self-harm; and reports grants from Medical Research Foundation, the Mindstep Foundation, Chief Scientist Office, MRC, Public Health Scotland, Scottish Government, NIHR, Shout 85258, Scottish Association for Mental Health, Zoetis Foundation, Jonathan's Voice, ADHD UK, and Barfil Charitable Trust. HC reports grants from the National Health and Medical Research Council (NHMRC), the Medical Research Future Fund, Paul Ramsay Foundation, and the Australian Government; is Scientia Professor at the University of New South Wales, supported by an NHMRC Elizabeth Blackburn Research Fellowship, and chief investigator on the NHMRC Centre for Research Excellence in Suicide Prevention; sits on the Million Minds Committee; and is a director of the Black Dog Institute Board and the Ramsay Health Care Research Foundation. MS declares salary support through academic scholar awards from Sunnybrook Health Sciences Centre and the University of Toronto. OJK is currently supported by a Research Foundation Flanders Senior Postdoctoral Fellowship; reports grants from Research Foundation Flanders and the King Baudouin Foundation; is co-chair of the International Association for Suicide Prevention Early Career Group, for which she receives complimentary student membership; is a former member of the Samaritans research ethics board; and has previously received travel grants and waived registration to present at conferences of the International Academy of Suicide Research. JP reports grants from NHMRC, the Medical Research Future Fund, Department of Health of the Australian Government, New South Wales Health, and the National Suicide Prevention Office. LFC is the vice president of the International Association for Suicide Prevention and is a permanent member of the Malaysian technical working group for suicide prevention; reports grants from the Centre of Pesticide Suicide Prevention at the University of Edinburgh; has received honorarium from Johnson & Johnson as a consultant and speaker; and, through her institution, has received access to the industry-sponsored medication sampling programme (compassionate patient programme) for clinical use for medication samples of esketamine (Johnson & Johnson), brexpiprazole (Lundbeck), Abilify Maintena (Lundbeck), and Trinza (Johnson & Johnson). DK is funded through the Elizabeth Blackwell Institute for Health Research at the University of Bristol, which is supported by the Wellcome Trust; has received grants from the Centre for Pesticide Suicide Prevention and the American Foundation for Suicide Prevention; and is a steering group member of the UK's National Suicide Prevention Alliance, and the Migration Health and Development Research Initiative. VP has consulted with Google and Modern Health (unrelated to the scope of this project). AC reports grants from Wellcome Trust, Leverhulme Trust, Economic and Social Research Council; has received funded consultancy from the Scottish Government and Alcohol Change UK; and is a member of the academic advisory group to the Scottish Government's National Suicide Prevention Leadership Group. SPe reports a 2021–2024 Vanier Canada Graduate Scholarship from the Canadian Institutes of Health Research. NS is a lived experience advisor to the Wellcome Trust, consumer academic at the University of Melbourne, lived experience lead at the Royal Children's Hospital, member of the Lived Experience Air Academy for University of Wollongong's Project Air, lived experience director for the Australian BPD Foundation, and associate at yLab (a division of the Foundation for Young Australians); reports previous employment with Orygen, and previous roles with the Youth Affairs Council of Victoria, Victorian Department of Families, Fairness, and Housing, Victorian Department of Premier and Cabinet, and Moonee Valley City Council; funded travel through Black Dog Institute and International Association of Suicide Prevention; and a current research tie to Orygen. FSt reports a grant from the Burdett Trust for Nursing and support from Abertay University. PSFY is a member of the advisory committee on mental health for the government of the Hong Kong Special Administrative Region. All other authors declare no competing interests.