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Industrial waste, including wastewater and solid waste, often contains toxic heavy metals that necessitate extraction and separation prior to safe disposal or reusing them. Sewage sludge incineration ash (SSIA), a non-incinerable waste, holds significant amounts of heavy metals such as iron, zinc, copper, nickel, chromium, and lead. Recovery and reuse of heavy metals from SSIA and further application of treated SSIA sludge remain challenging. Ethylenediaminetetraacetic acid (EDTA) is widely used for heavy metals chelation in different applications. While its chelation with heavy metals is rapid and easy to achieve, the de-chelation of the metal complexes is otherwise slow (∼3 days) and challenging due to their high stability constants. In this study, we investigate the recovery of heavy metals from SSIA through chelation using EDTA, and develop, for the first time, a method to rapidly de-chelate the EDTA-metal complexes through the facile chilling process (1 – 3 h) that accelerates the separation of EDTA and metal ions. A sequential precipitation of high-purity heavy metals from the EDTA-metal complexes was demonstrated with and without de-chelation. This novel and versatile method allows the separation of many valuable compounds from the treated SSIA, including regenerated EDTA, potassium hexafluorosilicate, iron phosphate, iron(III) hydroxide, iron silicate, titanium phosphate, calcium phosphate, copper(I) thiocyanate, nickel bis(dimethylglyoximate), lead(II) sulfate, and zinc sulfide. This approach opens doors for more sustainable waste management and the recovery of valuable resources from industrial waste.

Low-value investigations provide marginal benefit and may result in harm to the patient or disproportionate healthcare costs. The introduction of Singapore’s Choosing Wisely (CW) campaign in 2012 aims to reduce such investigations. CW was designed to encourage conversations between physicians and patients to weigh the risk-benefit ratio behind each clinical decision. A CW campaign was launched in July 2023 at Singapore General Hospital to reduce unnecessary tests and treatments.

Objective: This study aimed to construct an enzymolysis and membrane separation method to improve the production efficiency of low molecular hyaluronic acid (HA). Methods: A continuous enzymolysis and membrane separation coupling reaction system was designed, which consisted of an enzyme biological reaction tank and a flat polyethersulfone ultrafiltration membrane separation module. By evaluating membrane flux, membrane contamination, hyaluronic acid concentration, and yield, the effects of various factors on the separation of low molecular HA in the enzyme-membrane coupling system were systematically investigated. The process parameters of the enzyme-membrane coupling reaction were further optimized through orthogonal experiments. Results: The optimal process parameters for the preparation of low molecular hyaluronic acid by enzyme-membrane coupling reaction system were a stirring speed of 200 r/min, transmembrane pressure of 0.15 mPa, enzymolysis time of 4.0 h, and enzyme dosage of 5 g/100 g. In this method, two low molecular hyaluronic acids with different molecular weights (Mr) can be quickly prepared and separated, the low molecular weight hyaluronic acid (LMW-HA) with Mw between 10 000~50 000 and the HA oligosaccharides (O-HA) with Mr ＜3 000 can be prepared rapidly by one-step approach. Both of these different molecular weights hyaluronic acid could scavenge DPPH and hydroxyl free radicals. Conclusion: The enzymolysis and membrane separation coupling method provides an efficient method for simultaneous and continuous production of LMW-HA and O-HA with different molecular weights and certain antioxidant properties.

Androglobin (ADGB) is a highly conserved and recently identified member of the globin superfamily. Although previous studies revealed a link to ciliogenesis and an involvement in murine spermatogenesis, its physiological function remains mostly unknown. Apart from FOXJ1-dependent regulation, the transcriptional landscape of the ADGB gene remains unexplored. We, therefore, aimed to obtain further insights into regulatory mechanisms governing ADGB expression. To this end, changes in ADGB promoter activity were examined using luciferase reporter gene assays in the presence of a set of more than 475 different exogenous transcription factors. MYBL2 and PITX2 resulted in the most pronounced increase in ADGB promoter-dependent luciferase activity. Subsequent truncation strategies of the ADGB promoter fragment narrowed down the potential MYBL2 and PITX2 binding sites within the proximal ADGB promoter. Furthermore, MYBL2 binding sites on the ADGB promoter were further validated via a guide RNA-mediated interference strategy using reporter assays. Chromatin immunoprecipitation (ChIP)-qPCR experiments illustrated enrichment of the endogenous ADGB promoter region upon MYBL2 and PITX2 overexpression. Consistently, ectopic MYBL2 expression induced endogenous ADGB mRNA levels. Collectively, our data indicate that ADGB is strongly regulated at the transcriptional level and might have functions beyond ciliogenesis.

Mining gene expression data is valuable for discovering novel biomarkers and therapeutic targets in hepatocellular carcinoma (HCC). Although emerging data mining tools are available for pan-cancer–related gene data analysis, few tools are dedicated to HCC. Moreover, tools specifically designed for HCC have restrictions such as small data scale and limited functionality. Therefore, we developed IHGA, a new interactive web server for discovering genes of interest in HCC on a large-scale and comprehensive basis. Integrative HCC Gene Analysis (IHGA) contains over 100 independent HCC patient-derived datasets (with over 10,000 tissue samples) and more than 90 cell models. IHGA allows users to conduct a series of large-scale and comprehensive analyses and data visualizations based on gene mRNA levels, including expression comparison, correlation analysis, clinical characteristics analysis, survival analysis, immune system interaction analysis, and drug sensitivity analysis. This method notably enhanced the richness of clinical data in IHGA. Additionally, IHGA integrates artificial intelligence (AI)–assisted gene screening based on natural language models. IHGA is free, user-friendly, and can effectively reduce time spent during data collection, organization, and analysis. In conclusion, IHGA is competitive in terms of data scale, data diversity, and functionality. It effectively alleviates the obstacles caused by HCC heterogeneity to data mining work and helps advance research on the molecular mechanisms of HCC.

This work is aimedto design and analyze thestaircase climbing aid, hybrid with the function of walking cane. The tremendous demand for living space increasesthe essentiality of staircase, corresponding to the escalation of fall injuries. The handrails complemented along with staircasehad provideddeficient safeguard for staircase user, particularly senior citizen and people with lower limb disability, who need staircase climbing aid to provide extra support during escalating or declining staircase.

Objective To study the molecular characterization of Cronobacter sakazakii(C. sakazakii) from infant foods. Methods The C. sakazakii isolates were characterized using antibiotic susceptibility tests and whole genome sequencing(WGS). BioNumerics software was used to analyze and assemble WGS data. Then, the multilocus sequence typing(MLST), core-genome multi-locus sequence typing(cgMLST), virulence genes profiles were performed using assembled genome sequences. The resistance genes profiles and antimicrobial susceptibility were investigated. Furthermore, STs of C. sakazakii in this study were compared with those from PubMLST database. Results In this study, nine C. sakazakii isolates were detected and subtyped into five STs. ST1 and ST64 were the predominant STs, which were also the predominant STs of C. sakazakii in PubMLST database. Three C. sakazakii isolates were found to carry the mcr-9 gene. However, all C. sakazakii isolates were sensitive to 12 antibiotics including polymyxin B and E. ST1, ST64 and ST458 carried gtrB in addition to the common virulence genes. The nine C. sakazakii isolates showed highly genomic diversity according to cgMLST results. Conclusion ST1 and ST64 were predominant STs in this study, which were also predominant STs in clinical isolates in PubMLST database. These results suggested the potential pathogenicity of C. sakazakii. Our data underscored the importance of continuous surveillance of C. sakazakii in infant foods for prevention of foodborne disease caused by C. sakazakii.

Spermatogenesis is a highly specialized differentiation process driven by a dynamic gene expression program and ending with the production of mature spermatozoa. Whereas hundreds of genes are known to be essential for male germline proliferation and differentiation, the contribution of several genes remains uncharacterized. The predominant expression of the latest globin family member, androglobin (Adgb), in mammalian testis tissue prompted us to assess its physiological function in spermatogenesis. Adgb knockout mice display male infertility, reduced testis weight, impaired maturation of elongating spermatids, abnormal sperm shape, and ultrastructural defects in microtubule and mitochondrial organization. Epididymal sperm from Adgb knockout animals display multiple flagellar malformations including coiled, bifid or shortened flagella, and erratic acrosomal development. Following immunoprecipitation and mass spectrometry, we could identify septin 10 (Sept10) as interactor of Adgb. The Sept10-Adgb interaction was confirmed both in vivo using testis lysates and in vitro by reciprocal co-immunoprecipitation experiments. Furthermore, the absence of Adgb leads to mislocalization of Sept10 in sperm, indicating defective manchette and sperm annulus formation. Finally, in vitro data suggest that Adgb contributes to Sept10 proteolysis in a calmodulin-dependent manner. Collectively, our results provide evidence that Adgb is essential for murine spermatogenesis and further suggest that Adgb is required for sperm head shaping via the manchette and proper flagellum formation.

Objective To compare the species identification method of Cronobacter spp. in food, in order to establish rapid, high throughout identification method. Methods A total of 19 Cronobacter spp. in foods were identified with three method including biochemical method using VITEK 2.0 Compact, matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) and whole-genome sequencing (WGS). The advantages and disadvantages of the three identification method were analyzed. Results The result indicated that the accuracy of VITEK identification for C. sakazakii and C. malonaticus was 67% and 66% respectively. However, WGS and MALDI-TOF MS obtained the same result for 19 Cronobacter spp.. Three subspecies could not be identified by MALDI-TOF MS due to the limited database. Conclusion Biochemical method could not be used for species identification of Cronobacter spp. MALDI-TOF MS was rapid, high throughout and accurate, but still need to expand the fingerprint database for more Cronobacter spp. isolates.

Red-flowered oil-tea camellia (ROC) is an important woody oil species growing in the south, and its oil has high nutritional value. There are four main species of ROC in China, namely, Camellia chekiangoleosa (CCH), Camellia polyodonta (CPO), Camellia semiserrata (CSE) and Camellia reticulata (CRE). Reports on the comprehensive comparative analysis of ROC are limited. This study investigated the fruit characteristics and nutritional components of four ROC fruits, and the results showed that ROC had high oil content with levels of 39.13%–58.84%, especially the CCH fruit, which reached 53.6–58.84%. The contents of lipid concomitants of ROC oil were also substantial, including β-amyrin (0.87 mg/g–1.41 mg/g), squalene (0.43 mg/g–0.69 mg/g), β-sitosterin (0.47 mg/g–0.63 mg/g) and α-tocopherol (177.52 μg/g–352.27 μg/g). Moreover, the transverse diameter(TD)/longitudinal diameter (LD) of fruits showed a significant positive correlation with the oil content, and ROC fruits with thinner peels seemed to have better oil quality, which is similar to the result of the oil quality evaluation obtained by the gray correlation coefficient evaluation method. Four ROC oils were evaluated using the gray correlation coefficient method based on 11 indicators related to the nutritional value of ROC. CCH oil had the highest score of 0.8365, and YS-2 (a clone of CCH) was further evaluated as the best CCH oil. Finally, the results of heatmap analysis showed that triglycerides could be used as a characteristic substance to distinguish CCH oil from the other three ROC oils. The PLSDA (Partial least squares regression analysis) model and VIP (Variable important in projection) values further showed that P/S/O, P/O/O, P/L/L, P/L/Ln, S/S/O, S/O/O and P/S/S (these all represent abbreviations for fatty acids) could be used as characteristic differential triglycerides among the four ROC oils. This study provides a convenient way for planters to assess the nutritional quality of seed oil depending on fruit morphology and a potential way to distinguish between various ROC oils.

Abstract Background Mesenchymal stem cells are heterogenous populations with hematopoietic supporting and immunomodulating capacities. Enormous studies have focused on their preclinical or clinical therapeutic effects, yet the systematic study of continuous in vitro passages on signatures and functions of UC-MSCs at both the cellular and molecular levels is still lacking. Methods In this study, to systematically evaluate the biological properties of MSCs at various passages, we analyzed biomarker expression, cell proliferation and apoptosis, chromosome karyotype, and tri-lineage differentiation potential. Subsequently, we took advantage of whole-exome sequencing to compare the somatic hypermutation of hUC-MSCs at P3, P6, and P15 including SNV and INDEL mutations. In addition, to explore the safety of the abovementioned hUC-MSCs, we performed metabolic pathway enrichment analysis and in vivo transplantation analysis. Furthermore, we cocultured the abovementioned hUC-MSCs with UCB-CD34+ HSCs to evaluate their hematopoietic supporting capacity in vitro. Finally, we transplanted the cells into acute graft-versus-host disease (aGVHD) mice to further evaluate their therapeutic effect in vivo. Results The hUC-MSCs at P3, P6, and P15 showed similar morphology, biomarker expression, and cytokine secretion. hUC-MSCs at P15 had advantages on adipogenic differentiation and some cytokine secretion such as IL-6 and VEGF, with disadvantages on cell proliferation, apoptosis, and osteogenic and chondrogenic differentiation potential. Based on the SNP data of 334,378 exons and bioinformatic analyses, we found the somatic point mutations could be divided into 96 subsets and formed 30 kinds of signatures but did not show correlation with risk of tumorigenesis, which was confirmed by the in vivo transplantation experiments. However, hUC-MSCs at P15 showed impaired hematologic supporting effect in vitro and declined therapeutic effect on aGVHD in vivo. Conclusions In this study, we systematically evaluated the biological and genetic properties of hUC-MSCs at various passages. Our findings have provided new references for safety and effectiveness assessments, which will provide overwhelming evidence for the safety of hUC-MSCs after continuous in vitro passages both at the cellular and molecular levels for the first time. Taken together, our studies could help understand the controversial effects of disease treatment and benefit the clinical research of UC-MSCs.

Adoptive cell therapy using TCR-engineered T cells (TCR-T cells) represents a promising strategy for treating relapsed and metastatic cancers. We previously established methods to identify neoantigen-specific TCRs based on patients’ PBMCs. However, in clinical practice isolation of PBMCs from advanced-stage cancer patients proves to be difficult. In this study, we substituted blood-derived T cells for tumor-infiltrating lymphocytes (TILs) and used an HLA-matched cell line of antigen-presenting cells (APCs) to replace autologous dendritic cells. Somatic mutations were determined in head and neck squamous cell carcinoma resected from two patients. HLA-A*02:01-restricted neoantigen libraries were constructed and transferred into HLA-matched APCs for stimulation of patient TILs. TCRs were isolated from reactive TIL cultures and functionality was tested using TCR- T cells in vitro and in vivo. To exemplify the screening approach, we identified the targeted neoantigen leading to recognition of the minigene construct that stimulated the strongest TIL response. Neoantigen peptides were used to load MHC-tetramers for T cell isolation and a TCR was identified targeting the KIAA1429D1358E mutation. TCR-T cells were activated, exhibited cytotoxicity, and secreted cytokines in a dose-dependent manner, and only when stimulated with the mutant peptide. Furthermore, comparable to a neoantigen-specific TCR that was isolated from the patient’s PBMCs, KIAA1429D1358E-specific TCR T cells destroyed human tumors in mice. The established protocol provides the required flexibility to methods striving to identify neoantigen-specific TCRs. By using an MHC-matched APC cell line and neoantigen-encoding minigene libraries, autologous TILs can be stimulated and screened when patient PBMCs and/or tumor material are not available anymore. Abbreviations: Head and neck squamous cell carcinoma (HNSCC); adoptive T cell therapy (ACT); T cell receptor (TCR); tumor-infiltrating lymphocytes (TIL); cytotoxic T lymphocyte (CTL); peripheral blood mononuclear cell (PBMC); dendritic cell (DC); antigen-presenting cells (APC)

Vertebrate hemoglobin (Hb) and myoglobin (Mb) were among the first proteins whose structures and sequences were determined over 50 years ago. In the subsequent pregenomic period, numerous related proteins came to light in plants, invertebrates and bacteria, that shared the myoglobin fold, a signature sequence motif characteristic of a 3-on-3 α-helical sandwich. Concomitantly, eukaryote and bacterial globins with a truncated 2-on-2 α-helical fold were discovered. Genomic information over the last 20 years has dramatically expanded the list of known globins, demonstrating their existence in a limited number of archaeal genomes, a majority of bacterial genomes and an overwhelming majority of eukaryote genomes. In vertebrates, 6 additional globin types were identified, namely neuroglobin (Ngb), cytoglobin (Cygb), globin E (GbE), globin X (GbX), globin Y (GbY) and androglobin (Adgb). Furthermore, functions beyond the familiar oxygen transport and storage have been discovered within the vertebrate globin family, including NO metabolism, peroxidase activity, scavenging of free radicals, and signaling functions. The extension of the knowledge on globin functions suggests that the original roles of bacterial globins must have been enzymatic, involved in defense against NO toxicity, and perhaps also as sensors of O2, regulating taxis away or towards high O2 concentrations. In this review, we aimed to discuss the evolution and remarkable functional diversity of vertebrate globins with particular focus on the variety of non-canonical expression sites of mammalian globins and their according impressive variability of atypical functions.

Intracerebral hemorrhage (ICH) is a subtype of stroke that is followed by primary and secondary brain injury. As a result of the injury, cell metabolism is disrupted and a series of stress responses are activated, such as endoplasmic reticulum (ER) stress and the unfolded protein response (UPR), leading to the re-establishment of cell homeostasis or cell death. As an important mechanism of cell homeostasis, autophagy has been widely studied, and the associations between autophagy, ER stress, and the UPR have also been demonstrated. Whether these mechanisms are beneficial or detrimental remains a matter of controversy, but there is no doubt as to their vital functions. An understanding of the mechanisms of injury and recovery after ICH is crucial to develop therapeutic strategies. In this review, we summarize the related studies and highlight the roles of autophagy, ER stress, and the UPR in disease, especially in ICH. We also provide an overview of therapeutic approaches that target autophagy, and we discuss the prospects for modulating autophagy, ER stress, and UPR mechanisms in ICH therapy.

In the electricity market, the real-time balance of electricity generation and consumption is a main task. In view of this, power providers usually sign contracts with their critical consumers (i.e., usually large-scale industrial companies) for managing their capacity demands. On the other hand, aggregators group commercial and residential consumers, and integrate their demands to negotiate with power providers. With a proper grouping of numerous electricity consumers, aggregators help to ensure stable electric supply, and reduce the burden of managing many consumers. In this work, we thus propose a novel data clustering approach to group complementary consumers based on their usage patterns (i.e., daily electricity consumption curves.) Furthermore, we incorporate the technique of discrete wavelet transform to speed up the clustering process. Specifically, approximations reconstructed from only a few wavelet coefficients may precisely capture the shape of original usage patterns. Experimental results based on a real dataset show that our approach is promising in practical applications.

The administration of exogenous β-hydroxybutyrate (β-HB), as well as fasting and caloric restriction, is a condition associated with β-HB abundance and decreased appetite in animals. Increased β-HB and decreased appetite exist simultaneously in some diseases, such as bovine left displaced abomasums (LDA) and human chronic gastritis. However, the effects of β-HB on stomach injuries have not been explored. To elucidate the possible effects of exogenous β-HB on the stomach, mice were injected intraperitoneally with β-HB, and bovine abomasum smooth muscle cells (BSMCs) were treated with different concentrations of β-HB. We found that β-HB induced BSMCs endoplasmic reticulum- and mitochondria-mediated apoptotic cell death. β-HB promoted Bax expression and caspase-12, -9, and -3 activation while blocking Bcl-2 expression. β-HB also promoted AIF, EndoG release and p53 expression. β-HB acted on key molecules in the apoptotic cell death pathway and increased p38 and c-June NH2-terminal kinase phosphorylation while inhibiting ERK phosphorylation and PCNA expression. β-HB upregulated P27 and P21 mRNA levels while downregulating cyclin and CDK mRNA levels, arresting the cell cycle. These results suggest that BSMCs treated with β-HB can induce oxidative stress, which can be prevented by intracellular calcium chelators BAPTA/AM but not antioxidant NAC. Additionally, these results suggest that β-HB causes ROS generation through a Ca2+-dependent mechanism and that intracellular Ca2+ levels play a critical role in β-HB -induced apoptotic cell death. The impact of β-HB on programmed cell death and oxidative stress in vivo was confirmed in murine experiments. For the first time, we show oxidative stress effects of β-HB on smooth muscle. We propose that β-HB is a possible cause of some stomach diseases, including bovine LDA.