Effects of everolimus on circadian gene expression and cell fate in synchronized Caco-2 cells.

Increasing temperature affects plant development, with the assumption that roots are among the tissues particularly sensitive to heat stress (HS). However, a comprehensive analysis of the impact of high temperature on the dynamics of cell cycle and mitosis in barley root cells remains limited. Here, we analyzed barley root growth across a temperature gradient from 15°C to 37°C, encompassing ambient, thermomorphogenic, and HS conditions. Root growth was stimulated by moderately elevated temperatures but arrested at approximately 35°C. HS-changed nuclear architecture parameters, including expanded nuclear area and altered circularity. Although HS led to a temporary mitosis arrest, we demonstrated that DNA replication and mitotic activity were efficiently reinitiated upon recovery at a lower temperature. Finally, we showed that moderately higher temperatures speed up mitosis. Notably, anaphase was the least affected compared to other mitotic phases. In summary, we show that germinating barley plantlets sustain active growth at high speed to temperatures above 30°C and that HS blocks cell cycle around the two critical cell cycle stages – S phase and mitosis in barley. These observations expand the knowledge of barley root growth under high temperatures and will help develop HS-resilient cereals.

Multiple myeloma (MM), characterized by uncontrolled proliferation of malignant plasma cells, remains a hard-to-treat malignancy due to the development of drug resistance, even to proteasome inhibitors such as Bortezomib. Recent studies highlight the role of tumor-derived exosomes in mediating drug resistance, immune escape, and stress adaptation in MM. Pharmacologic inhibition of exosome secretion may potentiate the cytotoxic effects of Bortezomib by impairing cellular stress responses.RPMI 8226 cells were treated with Bortezomib, Ketotifen, or their combination. Cell viability was assessed using the MTT assay. Intracellular reactive oxygen species and cell cycle progression were analyzed via flow cytometry using 2',7'-dichlorodihydrofluorescein diacetate and propidium iodide staining. Exosomes were isolated and characterized by dynamic light scattering and flow cytometry. Total exosomal protein content was quantified via BCA assay. Gene expression levels of ALIX and MUNC13-4 were measured using quantitative reverse transcriptase PCR.Ketotifen showed the least toxicity at 1 µM, and co-treatment with Bortezomib increased cell viability compared to Bortezomib alone. Ketotifen significantly reduced ALIX expression and total exosomal protein content without altering MUNC13-4 expression. It attenuated Bortezomib-induced ROS accumulation and altered cell cycle dynamics. Exosomes from Bortezomib-treated cells were more cytotoxic than those from the co-treatment group, while the supernatant of the combination induced greater cell death.Ketotifen suppresses exosome biogenesis and modulates oxidative and cell cycle responses in MM cells, potentially enhancing Bortezomib efficacy. These findings support further investigation of Ketotifen as an adjunct strategy to modulate exosome-mediated drug resistance in MM.

BackgroundDiffuse large B-cell lymphoma (DLBCL) is the most prevalent form of non-Hodgkin’s lymphoma globally. SPAG5, a mitotic spindle protein, plays a significant role in DLBCL, where its abnormal expression is often associated with tumor growth, chemotherapy resistance, local recurrence, and poor prognosis.MethodsA comprehensive analysis of SPAG5 expression across various cancer types was conducted using Timer 2.0 and Sanger Box 3.0. Subsequently, the expression levels of SPAG5 in DLBCL were investigated in comparison to normal samples. Receiver operating characteristic (ROC) curve was then generated to evaluate the diagnostic performance of SPAG5 for DLBCL. Furthermore, the functional role of SPAG5 was characterized, and its impact on the immune microenvironment of DLBCL patients was analyzed. Its potential in predicting immune checkpoint status and responses to immunotherapy was also evaluated.ResultsSPAG5 expression demonstrated significant heterogeneity across various cancer types, with a marked upregulation in DLBCL. The diagnostic efficacy of SPAG5 was moderate, yielding an area under curve (AUC) of 0.75. SPAG5 exerted a multifaceted influence on DLBCL progression by regulating critical cellular processes, including cell cycle dynamics, chromosomal segregation, and DNA homeostasis. Notably, patients with elevated SPAG5 expression had poorer survival outcomes than those with low expression. Analysis of the tumor immune microenvironment revealed a distinct pattern: high SPAG5 expression correlated with increased infiltration of resting natural killer (NK) cells, while being associated with reduced presence of regulatory T cells (Tregs) and follicular helper T cells (Tfh).ConclusionOur bioinformatics study elucidated the expression profile, diagnostic potential, and prognostic significance of SPAG5 in DLBCL, emphasizing the complex interplay between SPAG5 expression and the tumor immune landscape. Our findings suggested SPAG5 could be a candidate prognostic marker and potential therapeutic target for DLBCL.

Cell wall anchored surface proteins are integral components of the Gram-positive bacterial cell envelope and are vital for bacterial survival in different environmental niches. The trafficking of many surface proteins carrying a YSIRK/G-S signal peptide is synchronized with cell envelope biogenesis during cell division, whereby YSIRK proteins traffic to the septal membrane and anchor to septal peptidoglycan (cross-wall). Previous work demonstrated that LtaS-mediated lipoteichoic acid (LTA) synthesis restricts YSIRK proteins septal trafficking. Here we did a comprehensive immunofluorescence microscopy screen of the entire S. aureus Nebraska Transposon Mutant Library (NTML) for additional factors regulating cross-wall trafficking of staphylococcal protein A (SpA), an archetype of YSIRK proteins. We characterized the top nine major hits that drastically diminished SpA cross-wall localization, including ypfP and ltaA (LTA glycolipid anchor synthesis genes), lcpB (LytR-CpsA-Psr family protein), mprF (lysyl-phosphatidylglycerol synthase), lytH (cell wall hydrolase), scdA (nitrite reductase), yjbH (protease adaptor protein ), cbiO (cobalt transporter) and SAUSA300_2311 (LytTR regulatory system) along with ΔtagO (wall teichoic acid synthesis). Interestingly, unlike the ltaS mutant that delocalizes SpA at both the septal membrane and peptidoglycan (PG) layer, all the hits only delocalized SpA at the PG layer, suggesting that these mutants affect the late-stage SpA trafficking. In addition, mutants of lcpB, yjbH, cbiO and 2311 exhibit both transcriptional and spatial regulation. All the hits showed defects in cell cycle, cell morphology and spatially dysregulated PG synthesis. The shared phenotypes among the mutants suggest that impaired PG homeostasis and cell cycle defects are the mechanisms underlying dysregulated SpA localization. Overall, this work not only expands our understanding of YSIRK protein cross-wall trafficking but also identifies new leads that have a broader impact on the dynamics of cell cycle and cell envelope homeostasis.

Early immune perturbations increase the risk of neurodegenerative and neurodevelopmental disorders, yet the mechanisms underlying the maturation of microglia, the resident immune cells of the brain parenchyma, remain poorly defined. Specifically, how proliferation, morphological differentiation, and phagocytosis are coordinated among microglia progenitors as they colonize the embryonic brain remains unclear. Here, we combined mathematical modeling with spatiotemporal analyses of the murine hippocampus and cerebellum from postnatal day 2 (P2) to P60 to reconstruct the trajectory of microglial development. We identified a proliferative-to-quiescent (P/Q) switch around P3/P4 that preceded the acquisition of morphological complexity and efficient phagocytosis and was accompanied by coordinated shifts in cell-cycle dynamics and metabolic state. Strikingly, this P/Q switch was recapitulated in repopulation contexts in mice and in the human fetal brain, where later stages displayed enhanced phagocytic function coupled to reduced proliferation. Perturbing the proliferative phase through pharmacological or genetic disruption of CSF1R signaling impaired subsequent microglial complexity and phagocytosis efficiency, revealing an unexpected reliance of phagocytosis on proliferation-driven colonization. Finally, we show that microglia stepwise maturation during development is associated with chromatin remodeling and driven by the epigenetic regulator Ikaros. Together, these findings uncover the sequential milestones of microglial development, revealing a potential period of early vulnerability and establishing an unexpected linkage between proliferation and phagocytosis essential to understanding how these processes are coordinated in neurodegenerative disorders.

Commpass explorer: An interactive platform to explore clinico-genomic data from newly diagnosed multiple myeloma patients from the commpass observational trial

Targeting CPNE8 suppresses HOXA9-dependent AML progression and overcomes chemotherapy resistance

PurposeFuchs endothelial corneal dystrophy (FECD) is characterized by corneal endothelial cell (CEnC) degeneration and excessive extracellular matrix (ECM) deposition. FECD is an age-related disorder manifested by upregulation of senescence markers. However, the association between senescence and pathological ECM accumulation remains unclear. This study investigated whether senescence mediated cell-cycle arrest drives aberrant ECM deposition and guttae formation in FECD.MethodsA chronic stress (CS) model was established by concurrently exposing immortalized human CEnCs to ultraviolet-A light (UVA; 25 J/cm2) and 4-hydroxyestradiol (4OHE2; 20 µM) on days 1, 6, and 21, followed by approximately a 60-day recovery period. In vitro guttae were characterized using brightfield microscopy, scanning electron microscopy (SEM), immunofluorescence, and compared with FECD specimens. Cell cycle dynamics, senescence markers, and ECM expression were assessed using flow-cytometry, cell sorting, immunostaining, ELISA, and RT-PCR.ResultsIn vitro guttae displayed characteristic morphology and ultrastructural similarity to FECD guttae. CS-exposed CEnCs showed significant CDKN2A upregulation (42-fold, P < 0.01) and increased p16 positivity (63 ± 12% vs. 7 ± 3% in controls, P < 0.0001), indicating senescence. Flow cytometry revealed accumulation of CS cells in G0/G1 (74 ± 6% vs. 57 ± 3%, P < 0.0001) phase. G0/G1 sorted p16+ cells exhibited upregulation of COL4A1 (188-fold, P < 0.0001) and LAMB1 (98-fold, P < 0.05). Immunostaining confirmed p16 colocalization with COL4A1 and LAMB1 in CS cells and FECD tissues. ELISA showed elevated levels of TGFBIp in the conditioned media obtained from CS-exposed cells.ConclusionsG0/G1-arrested p16+ve senescent CEnCs promote pathological ECM deposition and guttae formation. This model, integrating environmental, hormonal, and aging factors, provides a biologically relevant platform for studying FECD and testing therapeutics.

Curing the Incurable: TP53 Mutated Myeloid Neoplasms.

Heterocyclic compounds remain cornerstones of contemporary drug discovery because their ring-embedded heteroatoms confer adaptable electronics, conformational flexibility, and a broad spectrum of biological activities. The skeleton structure of 4-thiazolidinone is present in many cytotoxically active compounds and is often used in the design of new antitumor agents. This study aimed to synthesize, characterize, and evaluate the anticancer potential of fifteen new (2-imino-4-oxo-1,3-thiazolidin- 5-yl)acetic acid derivatives. Compounds were synthesized using a consistent synthetic route involving a reaction between a thiourea derivative and maleic anhydride, which formed the thiazolidin- 4-one ring through cyclization. The compounds were then categorized into three sets based on the attached heterocyclic rings (tryptamine, thiazole, and 1,2,4-triazole). The NMR and X-ray analysis followed the synthesis. Apoptotic effects, cell cycle arrest, IL-6 suppression, docking, and dynamics simulations were conducted. Preliminary cytotoxic activity was tested on metastatic colorectal cancer (SW620) and human breast adenocarcinoma (MDA-MB-231) cell lines using the MTT assay. Compounds 5, 6, and 7 demonstrated notable selectivity indexes (4.73, 2.42, 4.16, respectively) and were further investigated for their mechanisms of action, revealing pro-apoptotic properties and the ability to induce cell cycle arrest. Additionally, compound 5 inhibited IL-6 secretion by 76%. in silico studies revealed the formation of an energetically stable complex between compound 5 and the EGFR crystal structure (min/- max binding affinities of -9.4|-8.0 kcal/mol, compared to the -7.71 kcal/mol for the native ligand). This preliminary study provides compelling data on synthesized derivatives, but more advanced testing is needed to assess their therapeutic value fully. Compared with earlier reports on related thiazolidinone scaffolds, the present derivatives exhibit improved potency, clearer selectivity, and mechanistic features consistent with EGFR inhibition and cytokine modulation. These findings validate (2-imino-4-oxo-1,3-thiazolidin-5-yl)acetic acid as a privileged core for cytotoxic lead generation and indicate that strategic substitution with either a tryptamine moiety (compound 5) or a 1,2,4-triazole ring (compound 7) is particularly advantageous. These compounds are promising EGFR-targeting anticancer candidates, warranting further investigation.

Unravelling the physiological adaptations that enable fish to thrive under diverse environmental stressors is crucial for predicting their resilience to climate change. However, linking molecular mechanisms to ecological contexts remains a significant challenge and such studies are greatly lacking. To close this data gap, we conducted a physiological and molecular mechanisms-integrated comparative study of representative fish species from two ecologically contrasting regions: the Indian Sundarban mangrove estuary and the Sub-Himalayan Terai-Dooars hill streams. We assessed erythropoietic activity, cell cycle dynamics, reactive oxygen species levels, and the expression of genes integral to hypoxia response, electron transport chain regulation, adaptive remodeling, and stress modulation. Our results demonstrate that fish inhabiting the Terai-Dooars region exhibit substantially higher erythropoietic efficiency, elevated reactive oxygen species levels, and significant upregulation of hypoxia-inducible and adaptive remodeling genes as compared to their Indian Sundarban counterparts. These findings suggest that the challenging environmental conditions of the Terai-Dooars—characterized by low temperatures, high water velocities, and physiological hypoxic stress—drive distinctive physiological and genetic adaptations in fish populations. Analysis of variance (ANOVA) on Principal Components (PCs) revealed species-specific cellular variability modulated by regional environmental pressures, emphasizing the interaction of intrinsic and extrinsic factors in shaping adaptive responses. This study bridges critical knowledge gaps by linking physiological traits to ecological contexts, providing insights into community-level adaptation mechanisms. The findings highlight convergent evolutionary strategies and propose molecular and cellular biomarkers for assessing resilience under future climate scenarios. Finally, these ecophysiological insights will enhance the comprehensive understanding of species responses to environmental heterogeneity, with implications for conservation and ecosystem management in the face of global environmental change.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-97749-y.

The circadian rhythms and cell cycle are closely interlinked, creating a fundamental regulatory axis vital for tissue homeostasis, which is frequently dysregulated in cancers. The circadian apparatus, which is regulated by the core clock components (BMAL1, CLOCK, PER, and CRY in mammals), establishes temporal order on cell proliferation by rhythmically regulating important cell cycle regulators such as WEE1, p21, and the oncogene MYC. This is frequently accomplished through overlapped signaling nodes that include particular kinases and ubiquitin ligases (e.g., FBXW7). Mounting evidence implicates disruption of this circadian clock-cell cycle synchrony, arising from genetic or environmental factors, as a significant contributor to tumorigenesis and progression via impacts on DNA repair fidelity, oncogene stability, and tumor suppressor pathways. This review critically evaluates the new concept of chrono-pharmacology for cancer, focusing on the substantial effects and side effects of different anticancer drugs that depend on the time-of-day efficacy. We discussed some interesting examples, like HSP90 inhibitors (ganetespib), HDAC inhibitors (quisinostat), topoisomerase inhibitors (doxorubicin), and BCL-2 family antagonists (Obatoclax, TW-37), whose therapeutic activities are tightly regulated by circadian control over their molecular targets, pharmacokinetic processes, and downstream physiological pathways. Furthermore, the circadian influence extends to the tumor microenvironment and antitumor immunity, suggesting novel chrono-immunotherapy approaches. By putting together the molecular bases of these temporal dynamics, this review underscores the significant potential of chronotherapythe timed administration of drugs to improve cancer treatment by enhancing therapeutic indices and paving the way for personalized, temporally optimized oncology strategies.

The eukaryotic cell cycle comprises several processes that must be carefully orchestrated and completed in a timely manner. Alterations in cell cycle dynamics have been linked to the onset of various diseases, underscoring the need for quantitative methods to analyze cell cycle progression. Here we develop RepliFlow, a model-based approach to infer cell cycle dynamics from flow cytometry data of DNA content in asynchronous cell populations. We show that RepliFlow captures not only changes in the length of each cell cycle phase but also alterations in the underlying DNA replication dynamics. RepliFlow is species-agnostic and recapitulates results from more sophisticated analyses based on nucleotide incorporation. Finally, we propose a minimal DNA replication model that enables the derivation of microscopic observables from population-wide DNA content measurements. Our work presents a scalable framework for inferring cell cycle dynamics from flow cytometry data, enabling the characterization of replication programme alterations.

UBE2C promotes cell cycle progression and suppresses DNA damage-induced apoptosis in triple-negative breast cancer.

About one-third of renal cell carcinoma (RCC) patients present with metastatic disease upon diagnosis because of the retroperitoneal location of the kidneys, which causes many tumors to stay asymptomatic. Besides, shortly after 5years following successful curative surgery, nearly 30% of individuals develop distant cancer metastases and recurrences. This is mostly attributable to the complex and diverse characteristics of the tumor microenvironment. Although targeted treatments and immunotherapies can extend the survival of patients, they are linked to the swift emergence of resistance, constraining the therapeutic alternatives for RCC patients and drawing attention to the critical requirement for improved targeted treatments. Along the same vein, there is an urgent demand for novel biomarkers capable of detecting early RCC with significant sensitivity and specificity. Additionally, prognostic indicators are required for the stratification of RCC patients. MicroRNAs (miRNAs) are crucial regulators of mRNA and subsequent protein production in both healthy and malignant tissues. The malignant pathophysiology of RCC has been associated with miRNA dysregulation, which impacts numerous cellular processes and has been found to increase the likelihood of proliferative and invasive processes, promote angiogenesis, alter cell cycle dynamics, evade cell death, facilitate metastasis, and make cancer cells less responsive to certain treatments. Therefore, in this review, we will go over the latest findings regarding the functions of oncogenic and tumor suppressor miRNAs in RCC, how they could be used as diagnostic and prognostic indicators for RCC, and the role they play in the development of RCC and its resistance to cancer-fighting therapies.

Forkhead homologue 1 (Fkh1) is a yeast transcription factor that plays essential roles in cell-cycle dynamics. Here, we report the co-crystal structure of the DNA-binding domain (DBD) of the yeast Fkh1 protein in complex with a 19-base pair oligonucleotide containing the core binding site and flanking regions. The three-dimensional structure of the Fkh1-DBD reveals a previously unknown protein fold among all known Forkhead proteins. The winged-helix fold forms base-specific contacts of α-helix H3 with the major groove of the core binding site. Wing 1 and Wing 2 form DNA shape-mediated contacts with the minor groove of the binding site flanking regions. The conformation of Wing 2 is distinct from all known Forkhead proteins, with α-helices H5 and H6 wrapping back onto the protein core, creating a stable Wing 2 loop. Backbone interactions with β-strands S1 and S2 reveal a structural mechanism for previously observed flanking region preferences in SELEX-seq experiments. In vivo yeast experiments on Fkh1 mutants demonstrate that wing residues interacting with flanking regions are important for Fkh1 function. Molecular dynamics simulations relate Fkh1 function to conformational flexibility of wing residues. The novel Forkhead fold enables Fkh1 function with implications, such as structure-based protein design, for other DNA-binding proteins.

Cell-cycle dynamics of gene expression are fundamental to life, yet their origin remains unclear. A prevailing model, derived from cancer cells, posits that transcription occurs in one cell cycle phase, while mature RNA accumulates in subsequent phases, suggesting temporal segregation of transcriptional regulation and RNA abundance. Whether this paradigm applies to normal human cells is unclear. Here, we co-profiled nascent transcription and mature RNAs in cycling human fibroblasts. The two dynamics were strongly concordant, with no evidence of a lag extending cell-cycle phases. Our data suggest a widespread transcription-to-maturation lag is not a general feature of human cells.

155P Establishment of a clinical translational model for personalized chemotherapy of lung cancer based on cell cycle dynamics

Hydrophobic collagen peptides from grass carp scales attenuate dextran sulfate sodium (DSS)-induced ulcerative colitis in mice via dual modulation of cell cycle dynamics and nuclear factor-kappa B signaling pathway