Search Results (407)

Lesion mimic mutants (LMMs) represent valuable biological tools for investigating plant defense mechanisms and cell death. Although multiple genes triggering lesion mimic formation have been identified, the connection between the lesion mimic phenotype and primary nutrient biosynthesis remains poorly understood. In our study, we characterized a novel rice LMM, lmm9, which exhibited persistent reddish-brown necrotic lesions from seedling stage to maturity, coupled with compromised agronomic traits and increased mortality rates. Map-based cloning and whole-genome sequencing identified a causal insertion in the promoter of Os03g03450/OsPAT1, the sole homolog of Arabidopsis PAT1 in rice, resulting in reduced gene expression. Genetic complementation and RNAi assays confirmed that downregulation of OsPAT1 led to lesion mimic formation in lmm9. OsPAT1 could translate into two variants—the predominant OsPAT1.1 and the C-terminal variant OsPAT1.2. Structural modeling demonstrated high conservation between OsPAT1 and yeast TRP4, and OsPAT1.1 combining the plastid signal sequence of Arabidopsis PAT1 successfully complemented the trp4 mutant in yeast. Notably, OsPAT1.1 and OsPAT1.2 showed different localization patterns, with OsPAT1.1 targeted to mitochondria and OsPAT1.2 localized to chloroplasts. Transcription analysis showed significant upregulation of tryptophan biosynthesis pathway genes in lmm9, consequently increasing the relative abundance of tryptophan and associated metabolites. Our findings provided further evidence that mutations in tryptophan biosynthetic genes can induce lesion mimic phenotypes in rice and would enhance the understanding of metabolic homeostasis in plant stress responses and cell death regulation. Full article

Cyclophyllideans, which diverged from diphyllobothriideans, have evolved compact genomes to meet ecological and biological demands associated with rapid development, early maturation, and prolific asexual reproduction. This streamlining is accompanied by inactivation of transposable elements (TEs), including retrotransposons. In contrast, diphyllobothriideans retain large, retrotransposon-rich genomes, but information on their individual retrotransposons is lacking. Here, Saci2-like long terminal repeat (LTR) retrotransposons, formerly annotated as lennie in taeniid cestodes, were identified in the diphyllobothriideans Spirometra erinaceieuropaei and Sparganum proliferum, along with orthologs from Schistocephalus solidus and Ligula intestinalis. The Saci2 homologs in these genomes diversified into at least eight families, exhibiting substantial variation in LTR and primer binding site sequences, reflecting ongoing regulatory diversification. Phylogenetic and divergence analyses indicated that they maintain structural and functional integrity under purifying selection, while early signs of inactivation appeared in S. proliferum. These findings suggest that diphyllobothriideans have faced little pressure for genome compaction, permitting the retention of functional retrotransposons, whereas cyclophyllideans, particularly taeniids, underwent genome streamlining linked to shortened life cycles and high fecundity, resulting in retrotransposon degradation. This contrast underscores the reciprocal relationship between biological demands and genome remodeling with TE inactivation in metazoans. Full article

The mammalian epidermis forms a critical barrier against environmental insults and water loss. The formation of its outermost layer, the stratum corneum, involves a rapid terminal differentiation process that has traditionally been explained by the “bricks and mortar” model. Recent advances reveal a more dynamic mechanism governed by intracellular liquid–liquid phase separation (LLPS). This review proposes that the lifecycle of the granular layer is orchestrated by LLPS. Evidence is synthesized showing that keratohyalin granules (KGs) are biomolecular condensates formed by the phase separation of the intrinsically disordered protein filaggrin (FLG). The assembly, maturation, and pH-triggered dissolution of these condensates are essential for cytoplasmic remodeling and the programmed flattening of keratinocytes, a process known as corneoptosis. In parallel, an LLPS-based signaling pathway is described in which the kinase RIPK4 forms condensates that activate the Hippo pathway, promoting transcriptional reprogramming and differentiation. Together, these structural and signaling condensates drive skin barrier formation. This review further reinterprets atopic dermatitis, ichthyosis vulgaris, and Bartsocas-Papas syndrome as diseases of aberrant phase behavior, in which pathogenic mutations alter condensate formation or material properties. This integrative framework offers new insight into skin biology and suggests novel opportunities for therapeutic intervention through biophysics-informed biomaterial and regenerative design. Full article

Litchi (Litchi chinensis Sonn.) is a kind of evergreen fruit tree with good flavor and taste which has high economic value. Sufficiently low temperature in winter is essential for the successful flower formation of litchi. Therefore, in the context of global warming, litchi often experiences unstable flower formation, ultimately resulting in a decrease in litchi production. Our previous research has highlighted the pivotal role of the LcFT1 gene in regulating the flower formation of litchi and identified two AP1 homologous genes associated with LcFT1 (named LcAP1-1 and LcAP1-2) based on RNA-Seq and weight gene co-expression network analysis (WGCNA). In this study, the functions of the two AP1 homologous genes in regulating flowering time were investigated. Result showed that LcAP1-1 and LcAP1-2 were expressed in all litchi tissues. LcAP1-1 was more highly expressed in mature leaves compared to other tissues, while the LcAP1-2 has the highest expression level in flower buds. Both of them exhibited upregulation in the terminal bud of litchi under low temperature. The expression of LcAP1-1 and LcAP1-2 was highly correlated with the initiation of flower buds and the development of flower organs. They increased gradually during the floral initiation but decreased gradually during flower bud development. The transgenic tobacco of LcAP1-2 flowered about 55 days earlier than wild-type, while tobacco overexpressing the LcAP1-1 gene had no significant changes in flowering time compared to the wild-type. These results indicate that the two genes have divergent regulatory functions, and that the LcAP1-2 gene may be involved in the regulation of flower transformation and flower organ development in litchi. Our research will further reveal the molecular regulatory mechanisms of flower formation in litchi and will also provide theoretical guidance for the molecular breeding of litchi. Full article

This review presents a critical analysis of Japan’s hydrogen strategy, focusing on the broader context of its decarbonization efforts. Japan aims to achieve carbon neutrality by 2050, with intermediate targets including 3 million tons of hydrogen use by 2030 and 20 million tons by 2050. Unlike countries with abundant domestic renewables, Japan’s approach emphasizes hydrogen imports and advanced storage technologies, driven by limited local renewable capacity. This review not only synthesizes policy and project-level developments but also critically evaluates Japan’s hydrogen roadmap by examining its alignment with global trends, technology maturity, and infrastructure scalability. The review integrates recent policy updates, infrastructure developments, and pilot project results, providing insights into value chain modeling, cost reduction strategies, and demand forecasting. Three policy conclusions emerge. First, Japan’s geography justifies an import-reliant pathway, but it heightens exposure to price, standards, and supply-chain risk; diversification across LH₂ and ammonia with robust certification and offtake mechanisms is essential. Second, near-term deployment is most credible in industrial feedstocks (steel, ammonia, methanol) and the maritime sector, while refueling rollout lags materially behind plan and should be recalibrated. Third, cost competitiveness hinges less on electrolyzer CAPEX than on electricity price, liquefaction, transport; policy should prioritize bankable offtake, grid-connected renewables and transmission, and targeted CAPEX support for import terminals, bunkering, and cracking. Japan’s experience offers a pathway in the global hydrogen transition, particularly for countries facing similar geographic and energy limitations. By analyzing both the progress and the limitations of Japan’s hydrogen roadmap, this study contributes to understanding diverse national strategies in the rapidly changing state of implementation of clean energy. Full article

Automated container terminals can achieve precise matching of equipment and space, thus forming the foundation for the terminals’ efficient operation. However, the increase in container volume requires the construction of more ACTs. Existing studies lack dynamic assessment of the deep coupling between the P-shaped layout and the terminal’s system performance verification under peak operating conditions. To solve these problems, this paper aims to evaluate the system performance of the process layout in the application of ACTs through simulation methods. We have identified the differences in indicators among various schemes, thereby providing decision support for the construction of the port. In this paper, a simulation method for the configuration based on a P-shaped process layout is proposed at ACTs. The port system is constructed as a discrete event simulation model consisting of five core modules. Then two P-shaped process layout schemes and one mixed process layout scheme are proposed and the terminal models are established, respectively. Finally, by conducting numerous simulation experiments under different layout schemes, the influence of traffic organization on the efficiency of the terminal system was analyzed. The results demonstrate that on the premise of the maturity of the mixed-traffic technology at the terminal, when the proportion of cross-berth operations is low, the system efficiency of the mixed layout scheme is the highest. This article takes a new type of P-shaped process layout as the research object, reveals the correlation between its traffic organization characteristics and system performance through a customized simulation method. It provides a new theoretical perspective and quantitative tool for the optimization of automated terminal layouts. Full article

Tissue engineering has the potential to overcome the limitations of using autografts in nerve gap repair, using cellular biomaterials to bridge the gap and support neuronal regeneration. Various types of therapeutic cells could be considered for use in aligned collagen-based engineered neural tissue (EngNT), including Schwann cells and their precursors, which can be derived from human induced pluripotent stem cells (hiPSCs). Using Schwann cell precursors may have practical advantages over mature Schwann cells as they expand readily in vitro and involve a shorter differentiation period. However, the performance of each cell type needs to be tested in EngNT. By adapting established protocols, hiPSCs were differentiated into Schwann cell precursors and Schwann cells, with distinctive molecular profiles confirmed using immunocytochemistry and RT-qPCR. For the first time, both cell types were incorporated into EngNT using gel aspiration–ejection, a technique used to align and simultaneously stabilise the cellular hydrogels. Both types of cellular constructs supported and guided aligned neurite outgrowth from adult rat dorsal root ganglion neurons in vitro. Initial experiments in a rat model of nerve gap injury demonstrated the extent to which the engrafted cells survived after 2 weeks and indicated that both types of hiPSC-derived cells supported the infiltration of host neurons, Schwann cells and endothelial cells. In summary, we show that human Schwann cell precursors promote infiltrating endogenous axons in a model of peripheral nerve injury to a greater degree than their terminally differentiated Schwann cell counterparts. Full article

Background: Acute myeloid leukemia (AML) is a common and aggressive adults hematological malignancies. This study explored megakaryocyte–erythroid progenitors (MEPs) signature genes and constructed a prognostic model. Methods: Uniform manifold approximation and projection (UMAP) identified distinct cell types, with differential analysis between AML-MEP and normal MEP groups. Univariate and the least absolute shrinkage and selection operator (LASSO) Cox regression selected biomarkers to build a risk model and nomogram for 1-, 3-, and 5-year survival prediction. Results: Ten differentially expressed genes (DEGs) related to overall survival (OS), six (AHSP, MYB, VCL, PIM1, CDK6, as well as SNHG3) were retained post-LASSO. The model exhibited excellent efficiency (the area under the curve values: 0.788, 0.77, and 0.847). Pseudotime analysis of UMAP-defined subpopulations revealed that MYB and CDK6 exert stage-specific regulatory effects during MEP differentiation, with MYB involved in early commitment and CDK6 in terminal maturation. Finally, although VCL, PIM1, CDK6, and SNHG3 showed significant associations with AML survival and prognosis, they failed to exhibit pathological differential expression in quantitative real-time polymerase chain reaction (qRT-PCR) experimental validations. In contrast, the downregulation of AHSP and upregulation of MYB in AML samples were consistently validated by both qRT-PCR and Western blotting, showing the consistency between the transcriptional level changes and protein expression of these two genes (p < 0.05). Conclusions: In summary, the integration of single-cell/transcriptome analysis with targeted expression validation using clinical samples reveals that the combined AHSP-MYB signature effectively identifies high-risk MEP-AML patients, who may benefit from early intensive therapy or targeted interventions. Full article

This study investigated the effects of Bothrops moojeni (B. moojeni) venom and its high- (HMM) and low-molecular mass (LMM) fractions on human osteoclast (OC) differentiation and function in vitro, aiming to identify novel therapeutics for bone disorders. Venom preparations were applied at 5 µg/mL (crude venom and HMM) or 1 µg/mL (LMM) from day 4 of peripheral blood mononuclear cell (PBMC) differentiation through terminal OC formation, enabling evaluation across early differentiation, fusion, and maturation stages. RNA sequencing revealed 7793 genes common to all experimental groups, with unique gene expression signatures of 149 (control), 221 (HMM), 248 (crude venom), and 60 (LMM) genes, reflecting distinct molecular responses. The negative control PBMC group exhibited 1013 unique genes enriched in immune-related pathways, consistent with their undifferentiated state. Crude venom induced the broadest transcriptional modulation, upregulating key fusion (CD47) and resorption (CTSK) genes, and altering markers of OC differentiation. The HMM fraction predominantly influenced inflammatory and osteoclastogenic pathways, notably TNF and NF-κB signaling, while the LMM fraction selectively regulated fusion-related genes (e.g., CD44) and immune pathways, indicating targeted modulation of OC activity. Cytokine profiling showed that crude venom and HMM suppressed osteoclastogenic cytokines such as IL-1β and IL-6, supporting their potential use in inflammatory bone diseases. Pathway enrichment analyses confirmed these differential effects on immune response and bone resorption mechanisms. Together, these results demonstrate that B. moojeni venom and its fractions differentially impact OC biology, with crude venom exerting broad effects and HMM and LMM fractions offering more specific modulation. Future studies will isolate bioactive components and assess therapeutic efficacy in animal models of osteoporosis and rheumatoid arthritis. Full article

This study investigated the influence of cane diameter on vegetative, productive, and physiological behaviors in Actinidia chinensis, cv. Gold 3. Conducted over two years (2021–2022), the experiment compared canes with larger (HD) and smaller (LD) proximal diameters. This research focused on parameters such as shoot morphology, leaf gas exchange, fruit quality, and hydraulic resistance. The results revealed that HD canes promoted more vigorous growth, with a higher proportion of long and medium shoots, whereas LD canes resulted in shorter shoots. Additionally, the HD canes demonstrated a higher leaf area and more extensive leaf coverage, contributing to enhanced photosynthetic activity, as evidenced by enhanced gas exchange, stomatal conductance, and transpiration rates. This higher photosynthetic efficiency in HD canes resulted in more rapid fruit growth, with a larger fruit size and weight, particularly in fruits from non-terminate shoots. By contrast, fruits on LD canes exhibited slower growth, particularly in terms of fresh weight and dry matter accumulation. Despite these differences, maturation indices, including soluble solids and acidity levels, were not significantly affected by cane type. The findings suggest that selecting HD canes during winter pruning could lead to earlier harvests, with improved fruit quality and productivity, making this practice beneficial for optimizing vineyard management in Actinidia chinensis. Full article

The sodium–citrate cotransporter (NaCT) plays a crucial role in citrate transport during amelogenesis. Mutations in the SLC13A5 gene, which encodes the NaCT, cause early infantile epileptic encephalopathy 25 and amelogenesis imperfecta. We analyzed developing pig molars and determined that the citrate concentrations in secretory- and maturation-stage enamel are both 5.3 µmol/g, with about 95% of the citrate being bound to mineral. To better understand how citrate might enter developing enamel, we developed Slc13a5^Flag reporter mice that express NaCT with a C-terminal Flag-tag (DYKDDDDK) that can be specifically and accurately recognized by commercially available anti-Flag antibodies. The 24-base Flag coding sequence was located immediately upstream of the natural translation termination codon (TAG) and was validated by Sanger sequencing. The general development, physical activities, and reproductive outcomes of this mouse strain were comparable to those of the C57BL/6 mice. No differences were detected between the Slc13a5^Flag and wild-type mice. Tooth development was extensively characterized using dissection microscopy, bSEM, light microscopy, in situ hybridization, and immunohistochemistry. Tooth formation was not altered in any detectable way by the introduction of the Flag. The Slc13a5^Flag citrate transporter was observed on all outer membranes of secretory ameloblasts (distal, lateral, and proximal), with the strongest signal on the Tomes process, and was detectable in all but the distal membrane of maturation-stage ameloblasts. The papillary layer also showed positive immunostaining for Flag. The outer membrane of odontoblasts stained stronger than ameloblasts, except for the odontoblastic processes, which did not immunostain. As NaCT is thought to only facilitate citrate entry into the cell, we performed in situ hybridization that showed Ank is not expressed by secretory- or maturation-stage ameloblasts, ruling out that ANK can transport citrate into enamel. In conclusion, we developed Slc13a5^Flag reporter mice that provide specific and sensitive localization of a fully functional NaCT-Flag protein. The localization of the Slc13a5^Flag citrate transporter throughout the ameloblast membrane suggests that either citrate enters enamel by a paracellular route or NaCT can transport citrate bidirectionally (into or out of ameloblasts) depending upon local conditions. Full article

The yield of common bean (Phaseolus vulgaris L.) is limited by abiotic stresses such as drought and high temperatures, which frequently occur simultaneously under field conditions. This study examined 100 bean genotypes under three environmental conditions, namely, the rainy season (optimal conditions), full irrigation in the dry season (high-temperature stress), and terminal drought in the dry season (combined stress), via a 10 × 10 triple-lattice design. Agronomic parameters evaluated included days to flowering (DF), days to physiological maturity (DM), plant height (PH), aerial biomass (BIO), grain yield (YLD), and 100-seed weight (100SW). The natural temperature exceeded 35 °C during the reproductive stage of the dry season. Combined stress revealed differential adaptive mechanisms in the tested germplasms, indicating that the response to multiple stresses is more complex than the sum of individual stress responses. The average yield under optimal conditions was 1344 kg/ha, decreasing to 889 kg/ha (66.1%) under irrigation with high temperatures and to 317 kg/ha (23.6%) under terminal drought with high temperatures. Under terminal drought with high temperatures, the number of days to maturity decreased by 5%, and the seed weight decreased by 20%. The G69-33-PT and G-19158 genotypes presented high yields under high-temperature stress, with yields above 1800 kg/ha, suggesting specific physiological mechanisms for tolerance to elevated temperatures. Under combined stress, genotypes G69-Sel25, Pinto Mestizo, and Dalia presented yields above 680 kg/ha, indicating adaptations in terms of water use efficiency and tolerance to high temperature. The identification of genotypes with differential stress tolerance provides valuable genetic resources for breeding programs. The diverse origins of superior germplasms (bred lines, landraces, and commercial cultivars) highlight the importance of exploring various germplasms in the search for sources of abiotic stress tolerance for breeding projects aimed at developing cultivars adapted to climate change scenarios where drought and high temperatures occur simultaneously. Full article

Cell-based immunotherapy is a promising treatment strategy for cancer. Particularly in the case of solid tumors, however, this strategy only benefits a minority of patients. A critical limitation to immunotherapy is T cell exhaustion, a terminal differentiation state characterized by loss of self-renewal and cytotoxic capacity. For over a decade, regenerative immunology approaches to overcome exhaustion and restore stem-like features of T cells have been pursued. The reprogramming of tumor-specific T cells back to a less-differentiated, stem-like state using induced pluripotent stem cell (iPSC) technology has been viewed as a powerful and highly appealing strategy to overcome the limitations imposed by exhaustion. However, clinical translation of these approaches has been stymied by the requirement for subsequent iPSC-to-T cell re-maturation strategies, vanishingly low efficiencies, and resource-intensive cell culture protocols. In this review, we discuss the emergence of transcription factor reprogramming to iPSCs, contemporary techniques for T cell reprogramming, as well as techniques for re-differentiation into mature T cells. We discuss the potential clinical utility of T cell reprogramming and re-maturation strategies alongside progress and major roadblocks toward clinical translation. If these challenges can be addressed, transcription factor reprogramming of T cells into iPSCs and subsequent re-maturation into tumor-specific stem-like T cells may represent an incredibly efficacious approach to cancer immunotherapy. Full article

Erythropoiesis, the process driving the differentiation of hematopoietic stem and progenitor cells to mature erythrocytes, unfolds through tightly orchestrated developmental stages, each defined by profound epigenetic remodeling. From the initial commitment of hematopoietic progenitors to the terminal enucleation of erythrocytes, dynamic changes in chromatin accessibility, transcription factor occupancy, and three-dimensional genome architecture govern lineage specification and stage-specific gene expression. Advances in our understanding of the regulatory genome have uncovered how non-coding elements, including enhancers, silencers, and insulators, shape the transcriptional landscape of erythroid cells. These elements work in concert with lineage-determining transcription factors to establish and maintain erythroid identity. Disruption of these epigenetic programs—whether by inherited mutations, somatic alterations, or environmental stress—can lead to a wide range of hematologic disorders. Importantly, this growing knowledge base has opened new therapeutic avenues, enabling the development of precision tools that target regulatory circuits to correct gene expression. These include epigenetic drugs, enhancer-targeted genome editing, and lineage-restricted gene therapies that leverage endogenous regulatory logic. As our understanding of erythroid epigenomics deepens, so too does our ability to design rational, cell-type-specific interventions for red blood cell disorders. Full article

This research assessed 42 lentil genotypes developed by ICARDA along with a local variety over three growing seasons (2019–2022) in Southeastern Türkiye. Phenological, morphological, and yield attributes were determined to observe earliness, yield stability, and adaptation properties. Genotype G3771 showed outstanding performance in grain yield (2579 kg ha⁻¹), 1000-seed weight (54.9 g), and harvest index (37.3%), although it had lower stability under more severe drought conditions. Early-maturing genotypes like G3744, G3715, and G3716 consistently flowered and matured sooner, making them better suited for escaping terminal drought stress areas. The highest yields were recorded during the 2019–2020 season, which experienced favorable rainfall and soil nutrient levels, while the lowest yields occurred due to changing climatic conditions in the 2020–2021 season, highlighting the crop’s sensitivity to climate. Principal component analysis, hierarchical clustering, the Modified Multi-Trait Stability Index (MTSI), and the Multi-Trait Genotype-Ideotype Distance Index (MGIDI) aided in effective genotype classification. Although G3771 was the most productive, genotypes G3687, G3715, and G3689 proved to be the most stable and early maturing based on MGIDI scores. Strong relationships between grain yield, biological yield, and seed size identified these as key selection criteria. This study underscores the value of multi-trait selection tools like MGIDI and MTSI in consistently pinpointing lentil genotypes that balance earliness, productivity, and adaptability, laying a strong foundation for developing climate-resilient varieties suited to semi-arid climates. Full article