Haspin mediates H3.3S31 phosphorylation downstream of Aurora B in mouse embryonic stem cells.

Histone phosphorylation is instrumental in regulating diverse cellular processes across eukaryotes. Unraveling the kinases that target specific histone sites is key to deciphering the underlying mechanisms. Among the various sites on histone tails that can undergo phosphorylation, the kinase responsible for H3.3S31 phosphorylation remained elusive. Since both H3.3S31ph and H3T3ph occur specifically during mitosis, and Haspin is the known kinase for H3T3 phosphorylation, we investigated its potential role in H3.3S31 phosphorylation. We employed CRISPR/Cas9, RNA interference, and specific small molecule inhibitors to eliminate Haspin function in various cell types. Our data consistently revealed a link between Haspin and H3.3S31ph. Furthermore, in vitro kinase assays provided evidence supporting Haspin's contribution to H3.3S31ph. Loss- and gain-of-function experiments targeting Haspin and Aurora B further suggested a hierarchical relationship. Haspin acts as a downstream kinase of Aurora B, specifically orchestrating H3.3S31 phosphorylation in mESCs. This study unveils a novel role for Haspin as a kinase in regulating H3.3S31 phosphorylation during mitosis. This discovery holds promise for expanding our understanding of the functional significance of Haspin and H3.3S31ph in mammals.

Is the digital economy empowering high-quality tourism development? A theoretical and empirical research from China.

How digital economy (DE) empowers high-quality development of tourism (HQDT) has become a common concern among scholars. Given this, this study clarifies the theoretical connotation of DE enabling HQDT,and finds that: Micro, DE promotes efficiency improvements in tourism enterprises, with its economies of scale and Matthew effect reducing average costs, its economies of scope meeting diversified demand, and its long-tail effect improving supply-demand matching mechanism. Meso, DE can transform and upgrade tourism industry structure through industrial digitization and digital industrialization, and also form a new tourist industry form and value chain through cross-border integration. Macro, DE can stimulate innovation and flexibility of market players, increase new factor inputs in tourism, improve factor allocation efficiency, and advance macro regulation of the tourism market. Accordingly, the study conducts an empirical test based on panel data for 31 provinces in mainland China during 2011-2020. Results show that: ① DE positively influences HQDT, and the sub-dimensions all positively influence HQDT. ② DE has a heterogeneous impact on HQDT and shows spatial spillover effects. Finally, the study concludes with effective paths for DE promoting HQDT: "Promote digital infrastructure construction, accelerate tourism digital transformation, strengthen integration and innovation development, and overcome the challenges of tourism enterprises".

Advancing Organoid Engineering for Tissue Regeneration and Biofunctional Reconstruction.

Tissue damage and functional abnormalities in organs have become a considerable clinical challenge. Organoids are often applied as disease models and in drug discovery and screening. Indeed, several studies have shown that organoids are an important strategy for achieving tissue repair and biofunction reconstruction. In contrast to established stem cell therapies, organoids have high clinical relevance. However, conventional approaches have limited the application of organoids in clinical regenerative medicine. Engineered organoids might have the capacity to overcome these challenges. Bioengineering-a multidisciplinary field that applies engineering principles to biomedicine-has bridged the gap between engineering and medicine to promote human health. More specifically, bioengineering principles have been applied to organoids to accelerate their clinical translation. In this review, beginning with the basic concepts of organoids, we describe strategies for cultivating engineered organoids and discuss the multiple engineering modes to create conditions for breakthroughs in organoid research. Subsequently, studies on the application of engineered organoids in biofunction reconstruction and tissue repair are presented. Finally, we highlight the limitations and challenges hindering the utilization of engineered organoids in clinical applications. Future research will focus on cultivating engineered organoids using advanced bioengineering tools for personalized tissue repair and biofunction reconstruction.

Use of municipal solid waste incineration fly ash as a supplementary cementitious material: CO2 mineralization coupled with mechanochemical pretreatment.

The use of municipal solid waste incineration fly ash, commonly referred to as "fly ash", as a supplementary cementitious material (SCM), has been explored to mitigate the CO2 emissions resulting from cement production. Nevertheless, the incorporation of fly ash as an SCM in mortar has been shown to weaken its compressive strength and increase the risk of heavy metal leaching. In light of these challenges, this study aims to comprehensively evaluate the influence of CO2 pressure, temperature, and residual water/binder ratio on the CO2 uptake and compressive strength of mortar when combined with fly ash. Additionally, this study systematically examines the feasibility of mechanochemical pretreatment, which enhances the homogenization of fly ash and augments the density of the mortar's microstructure. The results indicate that the use of mechanochemical pretreatment leads to a notable 43.6% increase in 28-day compressive strength and diminishes the leaching of As, Ba, Ni, Pb, Se, and Zn by 17.9-77.8%. Finally, a reaction kinetics model is proposed to elucidate the CO2 sequestration process under varying conditions. These findings offer valuable guidance for incorporating fly ash as an SCM and CO2 sequestrator in mortar.

Development and validation of a novel nomogram model for predicting the survival of patients with T2-4a, N0-x, M0 bladder cancer: a retrospective cohort study.

OBJECTIVE: Recent developments in bladder cancer treatment strategies have significantly improved the prognosis of clinically curable muscle invasive bladder cancer (MIBC) patients. Here, the prognostic factors of T2-4a, N0-x, M0 MIBC patients were investigated using the Surveillance, Epidemiology, and End Results (SEER) database and a novel nomogram model was established for prognosis prediction. METHODS: The data of 7,292 patients with T2-4a, N0-x, M0 MIBC were retrieved from the SEER database (2000-2020) and randomly classified into a training set (n = 5,106) and validation set (n = 2,188). Kaplan-Meier analysis was used to calculate cancer-specific survival (CSS) and overall survival (OS) rates of patients, and differences between survival curves were analyzed using the log-rank test. Cox regression analysis was used to screen and incorporate patient prognosis-affecting independent risk factors into the nomogram model. Consistency index (C-index) values and areas under the time-dependent receiver operating characteristic curve (AUC) were used to evaluate the discriminatory ability, and the calibration curve was used to assess the calibration of the model. Its predictive performance and American Joint Committee on Cancer (AJCC) stage were compared using decision curve analysis (DCA). RESULTS: The 1-, 3-, and 5-year CSS and OS rates of patients with T2-4a, N0-x, M0 MIBC were 76.9%, 56.0%, and 49.9%, respectively, and 71.3%, 47.9%, and 39.5%, respectively. Cox regression analysis showed that age, marital status, race, pathological type, tumor size, AJCC stage, T stage, N stage, surgery of primary tumor, regional lymph node dissection, radiation, and chemotherapy were independent prognostic risk factors of both CSS and OS (P < 0.05). The C-index and AUC of the nomogram model constructed based on the training and validation sets were both > 0.7, and calibration curves for predicting the 1-, 3-, and 5-year survival were consistent with the ideal curve. The nomogram model showed a higher net benefit with DCA than AJCC stage analysis. CONCLUSION: The nomogram model could accurately predict the prognosis of patients with T2-4a, N0-x, M0 MIBC. It may help clinicians perform personalized prognosis evaluations and formulate treatment plans.

Low-temperature catalytic oxidation of PCDD/Fs over MnCeCoOx/PPS catalytic filter.

Catalytic destruction of nitrogen oxides (NOx) combined with dust removal technique has attracted much attention, yet the application in the solid waste incineration air pollution control process is still lacking due to the complex flue gas atmosphere. In this work, the Mn-Ce-Co-Ox catalyst-coated polyphenylene sulfide (PPS) filter fiber with efficient dust removal and low-temperature polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) destruction has been prepared with a redox-precipitation method. The catalyst was uniformly grown around the PPS fiber with appropriate catalyst loading. The effects of several key operating parameters (e.g., reaction temperature, catalyst loading amount, and filtration velocity) on the catalytic efficiency were comprehensively investigated. The results show that the Mn-Ce-Co-Ox/PPS has a decomposition yield of 78.0% in PCDD/Fs and 96% in nitric oxide (NO) conversion at 200 °C. The poisoned catalytic filter exhibits a removal efficiency of 88.6% for PCDD/Fs. In addition, the catalytic filter can completely reject particles smaller than 1.0 µm with a low filtration resistance. Therefore, this efficient and energy-conserving catalytic filter shows promising applications in flue gas pollution treatments.

Development of a native-locus dual reporter system for the efficient screening of the hyper-production of natural products in Streptomyces.

Streptomyces is renowned for its abundant production of bioactive secondary metabolites, but most of these natural products are produced in low yields. Traditional rational network refactoring is highly dependent on the comprehensive understanding of regulatory mechanisms and multiple manipulations of genome editing. Though random mutagenesis is fairly straightforward, it lacks a general and effective strategy for high throughput screening of the desired strains. Here in an antibiotic daptomycin producer S. roseosporus, we developed a dual-reporter system at the native locus of the daptomycin gene cluster. After elimination of three enzymes that potentially produce pigments by genome editing, a gene idgS encoding the indigoidine synthetase and a kanamycin resistant gene neo were integrated before and after the non-ribosomal peptidyl synthetase genes for daptomycin biosynthesis, respectively. After condition optimization of UV-induced mutagenesis, strains with hyper-resistance to kanamycin along with over-production of indigoidine were efficiently obtained after one round of mutagenesis and target screening based on the dual selection of the reporter system. Four mutant strains showed increased production of daptomycin from 1.4 to 6.4 folds, and significantly improved expression of the gene cluster. Our native-locus dual reporter system is efficient for targeting screening after random mutagenesis and would be widely applicable for the effective engineering of Streptomyces species and hyper-production of these invaluable natural products for pharmaceutical development.

TRIM56 coiled-coil domain structure provides insights into its E3 ligase functions.

Protein ubiquitination is a post-translation modification mediated by E3 ubiquitin ligases. The RING domain E3 ligases are the largest family of E3 ubiquitin ligases, they act as a scaffold, bringing the E2-ubiquitin complex and its substrate together to facilitate direct ubiquitin transfer. However, the quaternary structures of RING E3 ligases that perform ubiquitin transfer remain poorly understood. In this study, we solved the crystal structure of TRIM56, a member of the RING E3 ligase. The structure of the coiled-coil domain indicated that the two anti-parallel dimers bound together to form a tetramer at a small crossing angle. This tetramer structure allows two RING domains to exist on each side to form an active homodimer in supporting ubiquitin transfer from E2 to its nearby substrate recruited by the C-terminal domains on the same side. These findings suggest that the coiled-coil domain-mediated tetramer is a feasible scaffold for facilitating the recruitment and transfer of ubiquitin to accomplish E3 ligase activity.

SNHG1, a KLF4-upregulated gene, promotes glioma cell survival and tumorigenesis under endoplasmic reticulum stress by upregulating BIRC3 expression.

Increasing evidence indicates that long noncoding RNAs (lncRNAs) play crucial roles in the resistance to endoplasmic reticulum (ER) stress in many cancers. However, ER stress-regulated lncRNAs are still unknown in glioma. In the present study, we investigated the altered lncRNAs upon ER stress in glioma and found that small nucleolar RNA host gene 1 (SNHG1) was markedly increased in response to ER stress. Increased SNHG1 suppressed ER stress-induced apoptosis and promoted tumorigenesis in vitro and in vivo. Further mechanistic studies indicated that SNHG1 elevated BIRC3 mRNA stability and enhanced BIRC3 expression. We also found that KLF4 transcriptionally upregulated SNHG1 expression and contributed to the ER stress-induced SNHG1 increase. Collectively, the present findings indicated that SNHG1 is a KLF4-regulated lncRNA that suppresses ER stress-induced apoptosis and facilitates gliomagenesis by elevating BIRC3 expression.

Primary breast lymphoma in males: Incidence, demographics, prognostic factors, survival, and comparisons with females.

Background: Primary breast lymphoma (PBL) is a rare disease condition and is mainly observed in females. No male PBL cohorts were reported previously. This study aims to investigate the incidence, clinical characteristics, prognostic factors, and survival outcomes among male PBL patients and also to perform comparisons between males and females. Methods: Patients diagnosed with PBL between 2000 and 2019 from the Surveillance, Epidemiology, and End Results (SEER) database were identified. Age-adjusted incidence rates were calculated by year and age for trend analysis. Univariate and multivariate Cox hazard proportional regression analyses were performed to identify prognostic factors. Survival comparisons were conducted using the Kaplan-Meier method and the log-rank test. The propensity matching score (PSM) method was used to balance demographics. Results: The incidence rate of 122 male PBL patients diagnosed in the period of 2000 to 2019 was 0.169 (95% CI: 0.140-2.203) per million persons, which was much lower than that of 2,543 females (1.59, 95% CI: 1.53-1.65). Clinical demographics were similar between females and males, except for lymphoma subtype distribution (P = 0.025). A higher age [hazard ratio (HR) = 1.08, 95% CI: 1.05-1.12, P < 0.001] and not receiving radiotherapy (receiving vs. not receiving: HR = 0.41, 95% CI: 0.21-0.78, P = 0.007) were significant risk factors associated with overall survival (OS) in males. Radiotherapy (OS: P = 0.023) can offer benefits in OS. Using the PSM method, we also revealed that male PBL patients had significantly worse OS and cancer-specific survival rates than females. Conclusions: This study first analyzed male patients with PBL involving incidence, clinical characteristics, and survival data. Sex disparity was also observed in the survival outcome of the disease.

Structural studies of the coiled-coil domain of TRIM75 reveal a tetramer architecture facilitating its E3 ligase complex.

Protein ubiquitination plays a vital role in controlling the degradation of intracellular proteins and in regulating cell signaling pathways. Functionally, E3 ubiquitin ligases control the transfer of ubiquitin to the target substrates. As a major family of ubiquitin E3 ligases, the structural assembly of RING E3 ligases required to exert their ubiquitin E3 ligase activity remains poorly defined. Here, we solved the crystal structure of the coiled-coil domain of TRIM75, a member of the RING E3 ligase family, which showed that two disulfide bonds stabilize two antiparallel dimers at a small crossing angle. This tetrameric conformation confers two close RING domains on the same side to form a dimer. Furthermore, this architecture allows the RING dimer to present ubiquitin to a substrate on the same side. Overall, this structure reveals a disulfide bond-mediated unique tetramer architecture and provides a tetrameric structural model through which E3 ligases exert their function.

Contrast-enhanced ultrasound manifestations of renal masses undetectable on conventional ultrasound.

This study aimed to retrospectively analyze the features of contrast-enhanced ultrasound (CEUS) of renal masses that cannot be detected by conventional ultrasound (CUS). The data of 264 patients who underwent CEUS for renal lesions from January 2016 to December 2019 were retrieved. Of these, 16 patients with renal masses which were not detected by CUS were included in the final analysis. The corresponding characteristics of CEUS were evaluated, including intensity of enhancement, homogeneity, wash-in and wash-out patterns, and perilesional rim-like enhancement. Of the 16 patients, 10 patients had clear cell renal cell carcinoma (ccRCC) and 6 patients had urothelial carcinoma of the renal pelvis (UCRP). Compared with the location on non-enhanced computed tomography (CT) scan, all tumors were detected on CEUS. Most (7/10) of the ccRCCs appeared as hyperenhancement, homogeneous enhancement, synchronous-in, and no perilesional rim-like enhancement. Most (4/6) of the UCRPs appeared as isoenhancement, slow-in, fast-out, and no perilesional rim-like enhancement. CEUS may be helpful in the diagnosis and differential diagnosis of renal tumors which were not observed on CUS, and it might be an alternative method for some patients when contrast-enhanced computed tomography (CECT) or magnetic resonance imaging (MRI) cannot be performed.

PI3K/Akt and Wnt/ß-catenin Signaling Cross-regulate NF-κB Signaling in TNF-α-induced Human Lgr5+ Intestinal Stem Cells.

BACKGROUND/AIM: Intestinal stem cells (ISCs) are responsible for intestinal proliferation, differentiation, and neoplasia, and also play a crucial role in inflammation. Thus, it is important to investigate the effect of TNF-α on the activities of NF-κB, PI3K/Akt, and Wnt/ß-catenin signaling pathways. MATERIALS AND METHODS: The Lgr5+ intestinal cells were isolated using fluorescence-activated cell sorting from NCM460 spheroid cells, and the potential molecular mechanisms were investigated via short hairpin RNA (shRNA) transfection or the use of an inhibitor. RESULTS: The Lgr5+ cells were termed ISCs because of the higher expression of stem cell genes, including Sox2, Nanog, Oct4, Lgr5, and CD133. The Lgr5+ ISCs had a higher proliferation capacity, invasive ability, and drug resistance to 5-fluorouracil, as well as higher expression levels of anti-apoptotic proteins but lower expression levels of pro-apoptotic proteins, compared with Lgr5~ cells. The PI3K/Akt and Wnt/ß-catenin pathways were triggered by the TNF-α-induced activation of NF-κB signaling. Notably, when p65 expression was knocked-down via shRNA transfection in Lgr5+ ISCs, the TNF-α-induced activation of the NF-κB, PI3K/Akt, and Wnt/ß-catenin pathways were reversed. The same effect was observed with regards to ß-catenin shRNA transfection. Moreover, the Akt inhibitor MK2206 inhibited the TNF-α-induced activation of the PI3K/Akt pathway, as well as the NF-κB and Wnt/ß-catenin pathways. CONCLUSION: PI3K/Akt and Wnt/ß-catenin signaling cross-regulate NF-κB signaling in TNF-α-induced human Lgr5+ ISCs.

ZFP281-BRCA2 prevents R-loop accumulation during DNA replication.

R-loops are prevalent in mammalian genomes and involved in many fundamental cellular processes. Depletion of BRCA2 leads to aberrant R-loop accumulation, contributing to genome instability. Here, we show that ZFP281 cooperates with BRCA2 in preventing R-loop accumulation to facilitate DNA replication in embryonic stem cells. ZFP281 depletion reduces PCNA levels on chromatin and impairs DNA replication. Mechanistically, we demonstrate that ZFP281 can interact with BRCA2, and that BRCA2 is enriched at G/C-rich promoters and requires both ZFP281 and PRC2 for its proper recruitment to the bivalent chromatin at the genome-wide scale. Furthermore, depletion of ZFP281 or BRCA2 leads to accumulation of R-loops over the bivalent regions, and compromises activation of the developmental genes by retinoic acid during stem cell differentiation. In summary, our results reveal that ZFP281 recruits BRCA2 to the bivalent chromatin regions to ensure proper progression of DNA replication through preventing persistent R-loops.

Comparative ecological network pattern analysis: a case of Nanchang.

Urban-ecological landscape connectivity and pattern optimization can significantly enhance biodiversity and sustainable development capacity, which play an important role in continued ecosystem functioning. Previous studies identified ecological sources based on the area threshold method or combination with morphological spatial pattern analysis and the landscape connectivity index (CMSPACI) method, but few studies have compared the advantages, disadvantages, and applicability of the two methods. In this paper, taking Nanchang as the study area, we address the ecological sources via area threshold and the CMSPACI method. Then, the minimum cost distance method is used to generate potential corridors of different methods, and the differences in ecological networks are analyzed. Finally, the circuit theory is used to identify barriers, and we provide targeted recommendations for ecological network pattern optimization in the study area. The results show that (1) the ecological sources extracted by different methods are different. The ecological sources extracted by the area threshold are far away from the surrounding sources, and the landscape connectivity is low. The ecological sources identified by the CMSPACI method are closely related to the surrounding sources, and the landscape connectivity is high. (2) Compared with the area threshold method, the habitat quality of corridors under the CMSPACI method is better, and the interaction intensity between patches is larger. (3) There is little difference in the number of ecological barriers under different methods; all of them are located between patches or on the edge of patches, and most of them are roads or construction land. Overall, the area threshold method is simpler. Ecological sources can be effectively addressed through the CMSPACI method, and the landscape connectivity of the ecological network will be better. This study provides an important reference for the selection of ecological sources in the construction of ecological networks.