Establishment and transcriptome analysis of single blastomere-derived cell lines from zebrafish.

Maintaining chromosome euploidy in zebrafish embryonic cells is challenging because of the degradation of genomic integrity during cell passaging. In this study, we report the derivation of zebrafish cell lines from single blastomeres. These cell lines have a stable chromosome status attributed to BMP4 and exhibit continuous proliferation in vitro. Twenty zebrafish cell lines are successfully established from single blastomeres. Single-cell transcriptome sequencing analysis confirms the fidelity of gene expression profiles throughout long-term culturing of at least 45 passages. The long-term cultured cells are specialized into epithelial cells, exhibiting similar expression patterns validated by integrative transcriptomic analysis. Overall, this work provides a protocol for establishing zebrafish cell lines from single blastomeres, which can serve as valuable tools for in vitro investigations of epithelial cell dynamics in terms of life-death balance and cell fate determination during normal homeostasis.

Improving the academic resilience of hospital nursing interns through a hybrid multi-criteria decision analysis model.

Purpose: To identify the main variables affecting the academic adaptability of hospital nursing interns and key areas for improvement in preparing for future unpredictable epidemics. Methods: The importance of academic resilience-related variables for all nursing interns was analyzed using the random forest method, and key variables were further identified. An importance-performance analysis was used to identify the key improvement gaps regarding the academic resilience of nursing interns in the case hospital. Results: The random forest showed that five items related to cooperation, motivation, confidence, communication, and difficulty with coping were the main variables impacting the academic resilience of nursing interns. Moreover, the importance-performance analysis revealed that three items regarding options examination, communication, and confidence were the key improvement areas for participating nursing interns in the case hospital. Conclusions: For the prevention and control of future unpredictable pandemics, hospital nursing departments can strengthen the link between interns, nurses, and physicians and promote their cooperation and communication during clinical practice. At the same time, an application can be created considering the results of this study and combined with machine learning methods for more in-depth research. These will improve the academic resilience of nursing interns during the routine management of pandemics within hospitals.

Recent Progress of Induced Spermatogenesis In Vitro.

Sperm, a crucial gamete for reproduction in sexual reproduction, is generated through the proliferation, differentiation, and morphological transformations of spermatogonial stem cells within the specialized microenvironment of the testes. Replicating this environment artificially presents challenges. However, interdisciplinary advancements in physics, materials science, and cell engineering have facilitated the utilization of innovative materials, technologies, and structures for inducing in vitro sperm production. This article offers a comprehensive overview of research progress on inducing in vitro sperm production by categorizing techniques into two major systems based on matrix-based and non-matrix-based approaches, respectively. Detailed discussions are provided for both types of technology systems through comparisons of their similarities and differences, as well as research advancements. The aim is to provide researchers in this field with a comprehensive panoramic view while presenting our own perspectives and prospects.

Dichloroacetate attenuates brain injury through inhibiting neuroinflammation and mitochondrial fission in a rat model of sepsis-associated encephalopathy.

Sepsis-associated encephalopathy (SAE) is a severe complication of sepsis, characterized by neuroinflammation, mitochondrial dysfunction, and oxidative stress, leading to cognitive decline and high mortality. The effectiveness of dichloroacetate (DCA) in modulating mitochondrial function provides a novel therapeutic strategy for SAE. In this study, we evaluated the neuroprotective effects of DCA in a rat model of SAE induced by cecal ligation and puncture (CLP). Rats treated with DCA exhibited significant improvements in neurological function and survival, as evidenced by less neuron loss from histopathologic analysis, restored neurologic deficit scores, improved Y-maze alternation percentages, and enhanced recognition index performance. Biochemical analyses showed that DCA administration at 25 mg/kg and 100 mg/kg reduced astrocyte and microglial activation, indicating reduced neuroinflammation. Furthermore, DCA simultaneously reduced the production of circulating and cerebral inflammatory cytokines (including TNF-α, IL-1ß, and IL-10), concomitant with mitigating oxidative stress through down-regulating expression of 8-Hydroxy-2'-deoxyguanosine (8-OHdG) and reactive oxygen species (ROS) in the brain. Mechanistically, DCA modulated mitochondrial dynamics by suppressing Drp1 and pDrp1 expression, which are indicators of mitochondrial fission. This was corroborated by transmission electron microscopy, quantification of mitochondrial area, and Western blot analyses. Furthermore, DCA treatment improved ATP levels, mitochondrial complex I activity, and NAD+/NADH ratio, indicating a significant attenuation of brain mitochondrial dysfunction. In conclusion, our findings suggest that DCA confers neuroprotection in SAE by curtailing neuroinflammation and mitochondrial fission, outlining a promising therapeutic strategy for treating SAE in critically ill patients.

Flow and performance: a quantitative study of elicitation modeling in a piano performance perspective.

Introduction: Flow, defined as a heightened state of consciousness characterized by intense concentration during an activity, is influenced primarily by the perceived challenge and the dynamic equilibrium of skills. This investigation focuses on the patterns of flow state attainment and its elicitation mechanisms within the context of piano performance among Chinese music college students. Methods: Our study establishes a framework for accessing flow, utilizing quantitative data from music ontology to gauge the level of challenge and the level of music acquisition to assess skills. Additionally, we integrate external factors such as music culture heterogeneity and demographic variables to elucidate the causes and moderating effects of flow on piano performance. Results: The findings reveal a positive correlation between flow and performance, with the model of challenge and skill induction partially explaining these results. Notably, melodic Shannon Entropy emerges as a potential indicator of challenge, suggesting its relevance in future studies on flow. Discussion: This research provides multidimensional insights into the interplay between performance and flow in piano performance, guiding future investigations to explore the musical quantitative perspective more deeply.

Recommendations for the reference concentration of cadmium exposure based on a physiologically based toxicokinetic model integrated with a human respiratory tract model.

Cadmium (Cd) poses a significant threat to human health. However, chronic toxicity parameters for inhalation exposure are lacking, especially for noncritical systemic toxic effects. A physiologically based toxicokinetic (PBTK) model can be used to extrapolate toxicity parameters across various exposure routes. We combined a PBTK model with a human respiratory tract (HRT) model, which is applicable to the general population and capable of simulating the deposition and clearance processes of various airborne Cd compounds in the respiratory tract. Monte Carlo analysis was used to simulate the distribution of sensitive parameters to reflect individual variability. Validation based on datasets from general and occupational populations showed that the improved model had acceptable or better predictive performance, outperforming the original model with a 14.45 % decrease in the root mean square error (RMSE). Using this PBTK-HRT model, we extrapolated toxicity parameters from oral exposure to inhalation exposure for four systemic toxic effects with doseresponse relationships but no known inhalation toxicity parameters, and ultimately recommended reference concentrations (RfCs) for four diseases (chronic kidney disease: 0.01 µg/m3, osteoporosis: 0.01 µg/m3, stroke: 0.04 µg/m3, diabetes mellitus: 0.13 µg/m3), contributing to a comprehensive assessment of the health risks of Cd inhalation exposure. ENVIRONMENTAL IMPLICATION: Cadmium (Cd), a heavy metal, can cause lung cancer, chronic kidney disease, and osteoporosis and pose a significant threat to human health. We combined a physiologically based toxicokinetic (PBTK) model with a human respiratory tract (HRT) model to achieve better predictive performance and wider applicability; this model was subsequently employed for route-to-route extrapolation of toxicity parameters. Additionally, for the first time, we focused on multiple subchronic and chronic systemic toxic effects in addition to critical effects and derived their reference concentrations (RfCs), which can be used to assess the health risk of Cd inhalation exposure more comprehensively and accurately.

Design and Verification of Piano Playing Assisted Hand Exoskeleton Robot.

Finger technique is a crucial aspect of piano learning, and hand exoskeleton mechanisms effectively assist novice piano players in maintaining correct finger technique consistently. Addressing current issues with exoskeleton robots, such as the inability to provide continuous correction of finger technique and their considerable weight, a novel hand exoskeleton robot has been developed to enhance finger technique through continuous correction and reduced weight. Initial data are gathered using finger joint angle sensors to analyze movements during piano playing, focusing on the trajectory and angular velocity of key strikes. This analysis informs the design of a 6-bar double-closed-loop mechanism with an end equivalent sliding pair, using analytical methods to establish the relationship between motor extension and input rod rotation. Simulation studies assess the exoskeleton's motion space and dynamics, confirming its capability to meet structural and functional demands for accurate key striking. Prototype testing validates the exoskeleton's ability to maintain correct finger positioning and mimic natural strike speeds, thus improving playing technique while ensuring comfort and safety.

A Study on the Design of Knee Exoskeleton Rehabilitation Based on the RFPBS Model.

The gait rehabilitation knee exoskeleton is an advanced rehabilitative assistive device designed to help patients with knee joint dysfunction regain normal gait through training and activity support. This paper introduces a design framework based on the process knowledge representation method to optimize the design and control efficiency of the knee exoskeleton. This framework integrates knowledge of design objects and processes, specifically including requirements, functions, principle work areas, and the representation and multi-dimensional dynamic mapping of the Behavior-Structure (RFPBS) matrix, achieving multi-dimensional dynamic mapping of the knee exoskeleton. This method incorporates biomechanical and physiological knowledge from the rehabilitation process to more effectively simulate and support gait movements during rehabilitation. Research results indicate that the knee rehabilitation exoskeleton design, based on the RFPBS process knowledge representation model, accomplishes multi-dimensional dynamic mapping, providing a scientific basis and effective support for the rehabilitation of patients with knee joint dysfunction.

Association between flavonoids intake and dental caries in children and adolescents: a cross-sectional study from the NHANES database.

BACKGROUND: Worldwide, dental caries is a bacterial biofilm-mediated condition with a high morbidity in children and adolescents. Flavonoids are a class of active natural products with antibacterial and anti-inflammatory effect. In vivo and in vitro studies have shown that they can promote tooth mineralization and reduce inflammation. However, the association of flavonoids intake and dental caries in children and adolescents remain unclear. AIM: This study was to evaluated the association of flavonoid and its subclass intake and dental caries in children and adolescents. METHODS: Data of participants aged 2-17 years were extracted from the National Health and Nutrition Examination Survey (NHANES) database (2017-2018). Dental caries was measured via the decayed or filled surfaces in primary teeth or permanent teeth (dfs/DFS) index. The weighted univariable and multivariable logistic regression models were utilized to explore the association of flavonoids intake with dental caries in children and adolescents, with odds ratios (ORs) with 95% confidence intervals (CIs). Subgroups analyses based on age, and overweight/obesity were further assessed the association. Subgroup analysis were further performed to explore whether the association between subclasses of anthocyanidins and catechins with dental caries was robust stratified by age and individual with overweight/obesity. RESULTS: Among totally 1,818 children and adolescents, 786 (43.2%) had dental caries. High intake of anthocyanidins (OR=0.69, 95%CI: 0.52-0.92) and catechins (OR=0.64, 95%CI: 0.44-0.92) were associated with lower odds of dental caries. Similar results were discovered in individuals aged ≥6 years (anthocyanidins, OR=0.62, 95%CI: 0.43-0.90; catechins, OR=0.62, 95%CI: 0.40-0.96), and without overweight/obesity (anthocyanidins, OR=0.58, 95%CI: 0.37-0.90; catechins, OR=0.51, 95%CI: 0.31-0.84). Further investigation found that high intake of cyanidin, petunidin, malvidin, peonidin, (+)-Catechin, (-)-Epigallocatechin, and (-)-epicatechin were associated with lower odds of dental caries in children and adolescents. CONCLUSION: High intake of anthocyanidins and catechins were associated with lower odds of dental caries in children and adolescents and are a promising intervention to be further explored in children and adolescents.

Role of the hydrocarbon molecular structure in CNT growth on Fe-Al catalysts.

Upgrading plastic wastes into high-value products via the thermochemical process is one of the most attractive topics. Although carbon nanotubes (CNTs) have been successfully synthesized from plastic pyrolysis gas over Fe-, Co-, or Ni-based catalysts, a deep discussion about the reaction mechanism was seldom mentioned in the literature. Herein, this work was intended to study the growth mechanism of CNTs from hydrocarbons on Fe-Al2O3 catalysts. C5-C7 hydrocarbons were used to synthesize CNTs in a high-temperature fixed-bed reactor, and the carbon products and cracked gas were analyzed in detail. The CNT yield was in the order of cyclohexane, cyclohexene > n-hexane > n-heptane > n-pentane, 1-hexene. It was proposed that CNT growth on Fe-Al2O3 catalysts was mainly determined by the yield and structure of six-membered cyclic species, which was tailored by the carbon chain length, C-C/CC bonds, and linear/cyclic structures of C5-C7 hydrocarbons. Compared with n-hexane, the six-membered rings of cyclohexane and cyclohexene promoted six-membered cyclic species formation, increasing CNT and benzene yields; the seven-membered carbon chain of n-heptane promoted methyl-six-membered cyclic species formation, decreasing CNT and benzene yields while increasing the toluene yield; the five-membered carbon chain of n-pentane and the CC bond of 1-hexene inhibited six-membered cyclic species formation, decreasing CNT and benzene yields. This work revealed the structure-activity relationship between C5-C7 hydrocarbons and CNT growth, which may direct the process design and optimization of CNT synthesis from plastic pyrolysis gas.

Screening of Reference Genes for Quantitative Real-Time PCR Analysis in Tissues and during Testis Development, and Application to Analyze the Expression of kifc1 in Hemibarbus labeo (Teleostei, Cypriniformes, Cyprinidae).

The selection of proper reference genes is vital for ensuring precise quantitative real-time PCR (qPCR) assays. This study evaluates the stability of the expression of nine candidate reference genes in different tissues and during testicular development in H. labeo. The results show that eef1a is recommended as a reference gene for qPCR analysis in tissues and during testicular development. Furthermore, we evaluated the optimal number of reference genes needed when calculating gene expression levels using the geomean method, revealing that two reference genes are sufficient. Specifically, eef1a and rps27 are recommended for analysis of gene expression in tissues, whereas eef1a and actb are advised for evaluating gene expression during testicular development. In addition, we examined the expression pattern of kifc1, a kinesin involved in the reshaping of spermatids. We detected peak expression levels of kifc1 in testes, with its expression initially increasing before decreasing throughout testicular development. The highest expression of kifc1 was observed in stage IV testes, the active period of spermiogenesis, suggesting a possible role for kifc1 in the regulation of the reshaping of spermatids and hence testicular development. This study represents the first investigation of reference genes for H. labeo, providing a foundation for studying gene expression patterns and investigating gene expression regulation during testicular development.

Beyond Shell-Filling: Strong Enhancement of Electron Affinity of Metal Clusters through a Noninvasive Oriented External Electric Field.

Traditional electron counting rules, like the Jellium model, have long been successfully utilized in designing superhalogens by modifying clusters to have one electron less than a filled electronic shell. However, this shell-filling approach, which involves altering the intrinsic properties of the clusters, can be complex and challenging to control, especially in experiments. In this letter, we theoretically establish that the oriented external electric field (OEEF) can substantially enhance the electron affinity (EA) of diverse aluminum-based metal clusters with varying valence electron configurations, leading to the creation of superhalogen species without altering their shell arrangements. This OEEF approach offers a noninvasive alternative to traditional superatom design strategies, as it does not disrupt the clusters' geometrical structures and superatomic states. These findings contribute a vital piece to the puzzle of constructing superalkalis and superhalogens, extending beyond conventional shell-filling strategies and potentially expanding the range of applications for functional clusters.

Two new cucurbitane-type triterpenoid saponins from the fruit of Citrullus colocynthis.

Two new cucurbitane-type triterpenoid saponins, 2,20ß,22ß-trihydroxy-16α,23(R)-epoxycucurbita-1,5,24-triene-3,11-dione 2-O-ß-D-glucopyranoside (1), 2,20ß,22α-trihydroxy-16α,23(S)-epoxycucurbita-1,5,11,24-tetraene-3-one 2-O-ß-D-glucopyranoside (2) were isolated from the fruit of Citrullus colocynthis (L.) Schrad. Their structures were elucidated by mass spectrometry, IR, 1D, and 2D NMR spectroscopy, etc. Besides, both of the compounds showed significant hepatoprotective activities at 10 µM against paracetamol-induced HepG2 cell damage.

GATA2 promotes castration-resistant prostate cancer development by suppressing IFN-ß axis-mediated antitumor immunity.

Castration-resistant prostate cancer (CRPC) nearly inevitably develops after long-term treatment with androgen deprivation therapy (ADT), leading to significant mortality. Investigating the mechanisms driving CRPC development is imperative. Here, we determined that the pioneer transcription factor GATA2, which is frequently amplified in CRPC patients, inhibits interferon (IFN)-ß-mediated antitumor immunity, thereby promoting CRPC progression. Employing a genetically engineered mouse model (GEMM), we demonstrated that GATA2 overexpression hindered castration-induced cell apoptosis and tumor shrinkage, facilitating tumor metastasis and CRPC development. Notably, GATA2 drives castration resistance predominantly via repressing castration-induced activation of IFN-ß signaling and CD8+ T-cell infiltration. This finding aligns with the negative correlation between GATA2 expression and IFNB1 expression, as well as CD8+ T-cell infiltration in CRPC patients. Mechanistically, GATA2 recruited PIAS1 as corepressor, and reprogramed the cistrome of IRF3, a key transcription factor of the IFN-ß axis, in an androgen-independent manner. Furthermore, we identified a novel silencer element that facilitated the function of GATA2 and PIAS1 through looping to the IFNB1 promoter. Importantly, depletion of GATA2 augmented antitumor immunity and attenuated CRPC development. Consequently, our findings elucidate a novel mechanism wherein GATA2 promotes CRPC progression by suppressing IFN-ß axis-mediated antitumor immunity, underscoring GATA2 as a promising therapeutic target for CRPC.

Research hotspots and development trends of K-12 engineering education research: Bibliometric analysis based on CiteSpace.

To comprehensively assess the impact of K-12 engineering education research (K-12 EER) and provide insights to stakeholders and policymakers, this study uses the scientometric analysis software CiteSpace to evaluate recent advances in K-12 EER. A search of 885 articles from the Web of Science Core Collection was conducted. This study analyzed publication trends, authors, countries, and research institutions to determine the trajectory of the K-12 EER. In addition, keyword co-occurrence and clustering maps were generated to identify major hotspots, and keyword burst detection was used to demonstrate development trends. The analysis reveals that global interest in the K-12 EER is growing, and the results are accumulating rapidly. However, cooperation between universities, institutions, experts, and scholars must be strengthened. Current significant topics in K-12 EER focus on the practical form of engineering education, its impact on learners, the centrality of engineering design, and teachers' professional development. Research frontiers primarily revolve around the nature of engineering. Future research efforts should promote the systematic integration of K-12 engineering education through the learning process, develop comprehensive measurement tools to assess its impact on learners, and expand research on teachers' professional development in K-12 engineering education.

A foundation model for clinical-grade computational pathology and rare cancers detection.

The analysis of histopathology images with artificial intelligence aims to enable clinical decision support systems and precision medicine. The success of such applications depends on the ability to model the diverse patterns observed in pathology images. To this end, we present Virchow, the largest foundation model for computational pathology to date. In addition to the evaluation of biomarker prediction and cell identification, we demonstrate that a large foundation model enables pan-cancer detection, achieving 0.95 specimen-level area under the (receiver operating characteristic) curve across nine common and seven rare cancers. Furthermore, we show that with less training data, the pan-cancer detector built on Virchow can achieve similar performance to tissue-specific clinical-grade models in production and outperform them on some rare variants of cancer. Virchow's performance gains highlight the value of a foundation model and open possibilities for many high-impact applications with limited amounts of labeled training data.

Integration of metagenomics and metaproteomics in the intestinal lavage fluids benefits construction of discriminative model and discovery of biomarkers for HBV liver diseases.

Intestinal lavage fluid (IVF) containing the mucosa-associated microbiota instead of fecal samples was used to study the gut microbiota using different omics approaches. Focusing on the 63 IVF samples collected from healthy and hepatitis B virus-liver disease (HBV-LD), a question is prompted whether omics features could be extracted to distinguish these samples. The IVF-related microbiota derived from the omics data was classified into two enterotype sets, whereas the genomics-based enterotypes were poorly overlapped with the proteomics-based one in either distribution of microbiota or of IVFs. There is lack of molecular features in these enterotypes to specifically recognize healthy or HBV-LD. Running machine learning against the omics data sought the appropriate models to discriminate the healthy and HBV-LD IVFs based on selected genes or proteins. Although a single omics dataset is basically workable in such discrimination, integration of the two datasets enhances discrimination efficiency. The protein features with higher frequencies in the models are further compared between healthy and HBV-LD based on their abundance, bringing about three potential protein biomarkers. This study highlights that integration of metaomics data is beneficial for a molecular discriminator of healthy and HBV-LD, and reveals the IVF samples are valuable for microbiome in a small cohort.

The Conjugated/Non-Conjugated Linked Dimer Acceptors Enable Efficient and Stable Flexible Organic Solar Cells.

The fabrication of the flexible devices with excellent photovoltaic performance and stability is critical for the commercialization of organic solar cells (OSCs). Herein, the conjugated dimer acceptor DY-TVCl and the non-conjugated dimer acceptor DY-3T based on the monomer MY-BO are synthesized to regulate the molecular glass transition temperatures (Tg) for improving the morphology stability of active layer films. And the crack onset strain values for the blend films based on dimer acceptors are superior than that of small molecule, which are beneficial for the preparation of flexible devices. Accordingly, the binary device based on PM6:DY-TVCl achieves a maximum power conversion efficiency (PCE) of 18.01%. Meanwhile, the extrapolated T80 (time to reach 80% of initial PCE) lifetimes of the PM6:DY-TVCl-based device and PM6:DY-3T-based device are 3091 and 2227 h under 1-sun illumination, respectively, which are better than that of the PM6:MY-BO-based device (809 h). Furthermore, the flexible devices based on DY-TVCl and DY-3T exhibit the efficiencies of 15.23% and 14.34%, respectively. This work affords a valid approach to improve the stability and mechanical robustness of OSCs, as well as ensuring the reproducibility of organic semiconductors during mass production.

EDIL3/Del-1 prevents aortic dissection through enhancing internalization and degradation of apoptotic vascular smooth muscle cells.

Thoracic aortic dissection (TAD) is a severe disease, characterized by numerous apoptotic vascular smooth muscle cells (VSMCs). EDIL3/Del-1 is a secreted protein involved in macrophage efferocytosis in acute inflammation. Here, we aimed to investigate whether EDIL3 promoted the internalization and degradation of apoptotic VSMCs during TAD. The levels of EDIL3 were decreased in the serum and aortic tissue from TAD mice. Global edil3 knockout (edil3-/-) mice and edil3-/- bone marrow chimeric mice exhibited a considerable exacerbation in ß-aminopropionitrile monofumarate (BAPN)-induced TAD, accompanied with increased apoptotic VSMCs accumulating in the damaged aortic tissue. Two types of phagocytes, RAW264.7 cells and bone marrow-derived macrophages (BMDMs) were used for in vitro efferocytosis assay. edil3-deficient phagocytes exhibited inefficient internalization and degradation of apoptotic VSMCs. Instead, EDIL3 promoted the internalization phase through interacting with phosphatidylserine (PtdSer) on apoptotic VSMCs and binding to the macrophage ITGAV/αv-ITGB3/ß3 integrin. In addition, EDIL3 accelerated the degradation phase through activating LC3-associated phagocytosis (LAP). Mechanically, following the engulfment, EDIL3 enhanced the activity of SMPD1/acid sphingomyelinase in the phagosome through blocking ITGAV-ITGB3 integrin, which facilitates phagosomal reactive oxygen species (ROS) production by NAPDH oxidase CYBB/NOX2. Furthermore, exogenous EDIL3 supplementation alleviated BAPN-induced TAD and promoted apoptotic cell clearance. EDIL3 may be a novel factor for the prevention and treatment of TAD.Abbreviations: BAPN: ß-aminopropionitrile monofumarate; BMDM: bone marrow-derived macrophage; C12FDG: 5-dodecanoylaminofluorescein-di-ß-D-galactopyranoside; CTRL: control; CYBB/NOX2: cytochrome b-245, beta polypeptide; DCFH-DA: 2',7'-dichlorofluorescin diacetate; EDIL3/Del-1: EGF-like repeats and discoidin I-like domains 3; EdU: 5-ethynyl-2'-deoxyuridine; EVG: elastic van Gieson; H&E: hematoxylin and eosin; IL: interleukin; LAP: LC3-associated phagocytosis; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; NAC: N-acetylcysteine; PtdSer: phosphatidylserine; rEDIL3: recombinant EDIL3; ROS: reactive oxygen species; SMPD1: sphingomyelin phosphodiesterase 1; TAD: thoracic aortic dissection; TEM: transmission electron microscopy; VSMC: vascular smooth muscle cell; WT: wild-type.