DLL4-NOTCH1 signaling mediates juxtacrine communication between tip- and stalk-like endothelial cells during vascular sprouting, contributing to complementary aspects of endothelial function. Thus, simultaneous modulation of DLL4 and NOTCH1 may provide a strategy to broadly interfere with the endothelial functions involved in tumor angiogenesis. Accordingly, this study aimed to identify a novel natural compound capable of modulating DLL4-NOTCH1-mediated endothelial communication and to investigate how such modulation influences tumor angiogenesis and growth. We screened 66 natural compounds to identify those that suppress DLL4 and NOTCH1 signaling in endothelial cells. Using a dual-reporter system that combines a DLL4 promoter-driven reporter and a CSL luciferase reporter, we simultaneously assessed ligand-associated activity and downstream transcriptional output of the DLL4-NOTCH1 axis. We confirmed VEGF-induced target gene expression via Western blotting and quantitative real-time polymerase chain reaction. We further performed in vitro angiogenesis assays and in vivo Lewis lung carcinoma (LLC) allograft tumor experiments. Among the 66 screened natural compounds, para-coumaric acid (p-CA) dose-dependently suppressed DLL4 transcription and NOTCH1 signaling activity in endothelial cells, with IC50 values of 4.57 and 8.34 µm, respectively. Furthermore, p-CA significantly suppressed VEGF-induced endothelial cell proliferation, migration, and tube formation. In the LLC allograft model, p-CA treatment suppressed tumor growth and vascular density, increased pericyte coverage, reduced tumor hypoxia, and enhanced CD3+ T-cell infiltration and tumor cell apoptosis. In summary, p-CA is a natural compound that functionally suppresses VEGF-driven DLL4-NOTCH1 signaling in endothelial cells, exhibiting strong anti-angiogenic effects and promoting functional vascular normalization, resulting in marked tumor suppression and enhanced antitumor immunity in vivo. These findings suggest that p-CA warrants further investigation as an anti-angiogenic compound for cancer therapy. Tumors need a supply of oxygen and nutrients to grow. To obtain this, they form new blood vessels in a process called angiogenesis. This process is controlled by signals between cells that line blood vessels, known as endothelial cells. Two important molecules involved in this communication are called DLL4 and NOTCH1. These molecules help coordinate how blood vessels form and grow. In this study, we looked for natural compounds that could interfere with this process and potentially slow tumor growth. We tested 66 natural compounds and found that one compound, called para-coumaric acid (p-CA), was particularly effective. p-CA reduced the activity of DLL4 and NOTCH1 signaling in endothelial cells. We then examined how this affected blood vessel formation. In laboratory experiments, p-CA reduced the ability of endothelial cells to grow, move, and form vessel-like structures. In a mouse model of cancer, treatment with p-CA slowed tumor growth and reduced the number of blood vessels within tumors. In addition, tumors treated with p-CA showed features associated with improved blood vessel structure, including better support from surrounding cells and reduced signs of low oxygen levels. These changes were accompanied by increased presence of immune cells that can attack tumor cells. Overall, this study shows that p-CA can interfere with blood vessel formation in tumors and may help limit tumor growth.
Diffuse midline glioma (DMG) is a devastating pediatric brain tumor with an unmet need for novel therapies. Immune checkpoint inhibitors have failed to prolong survival of DMG patients. In this study, we screened for immune checkpoint molecules in DMG, evaluated immunological responses to checkpoint targeting by co-culture assays and depletion of immune cells in vivo, studied the effects of CD155 silencing by whole-transcriptome analyses and performed in vivo treatments with Thiostrepton. In human and murine DMG cells, as well as primary brain tumor samples, we identified CD155 as the most highly expressed immune checkpoint. When murine DMG cells were co-cultured with CD8+ T cells, silencing of CD155 led to a marked increase in T cell-mediated killing. Strikingly, CD155-deficient DMG cells failed to grow in immunocompetent mice, and depletion of CD8+ T cells allowed these tumors to grow. CD155 also exerted cell-autonomous effects on tumor cells: silencing of CD155 led to induction of apoptosis of DMG cells and delayed tumor growth in immunodeficient mice. Transcriptomic analyses identified FOXM1 as a key target of CD155. Notably, FOXM1 silencing also led to reduced proliferation of DMG cells in vitro and in vivo. Finally, treatment of DMG-bearing mice with Thiostrepton, a FOXM1-targeting agent, delayed tumor growth and prolonged survival. These studies demonstrate that CD155 regulates immune evasion and tumor growth in DMG, and suggest that targeting CD155 could be a valuable two-pronged therapeutic strategy for this disease. Diffuse midline glioma (DMG) is the deadliest pediatric brain tumor and novel therapies are urgently needed. Immunotherapy has failed to improve survival for these patients. We propose that one reason is the low expression of previously targeted immune-checkpoint molecules, and introduce CD155 as a highly expressed molecule across DMG and other brain tumors. Excitingly, we discover that blocking CD155 not only improves anti-tumor immune responses, but also directly kills tumor cells. Finally, we uncover FOXM1 as a target of CD155 and show that the FOXM1-targeting agent Thiostrepton prolongs survival of DMG-bearing mice, thereby introducing a novel therapeutic strategy for DMG.
The purpose of this study was to determine the fate and growth kinetics of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes on the surface of whole cantaloupe and watermelon. Athena cantaloupe and mini seedless watermelon were spot inoculated (ca. 103 CFU/3.14 cm2) on the sun-side of the melons with pathogen specific cocktails of rifampicin-resistant strains of E. coli O157:H7, Salmonella or L. monocytogenes. To simulate post-harvest handling conditions, inoculated melons were stored at 4, 10, 15, 20, or 25 °C for up to 21 days. On cantaloupe surfaces, E. coli O157:H7 increased by 2.40 log CFU/3.14 cm2 at 25℃ after 3 days and L. monocytogenes increased by 2.14 log CFU/3.14 cm2 at 25℃ after 7 days. Salmonella did not grow on cantaloupes but persisted at 25℃ for 7 days before cantaloupes deteriorated. On watermelon surfaces, pathogens did not grow with the exception of L. monocytogenes at 20℃ and 25℃. Growth rates of both pathogens generally increased at higher temperatures and lag phase shortened at higher temperatures for L. monocytogenes, except at 25℃. Collectively, temperature control for storage of whole melons reduces the risk of foodborne pathogen growth and persistence on whole cantaloupes and watermelons.
The cyanobacterium Prochlorococcus MIT9312 requires helping functions from co-existing organisms for survival under stressful conditions. Here, we show that the helper strain Alteromonas macleodii EZ55 also facilitates greater exploitation of medium resources by Prochlorococcus. The presence of Alteromonas allowed Prochlorococcus to grow to greater cell densities, delay growth cessation, and avoid cell death relative to axenic cultures, phenotypes potentially associated with Alteromonas-driven degradation of autotoxic substances made by Prochlorococcus. We found that heat-labile high-molecular-weight Alteromonas exudates mediated the beneficial impacts on Prochlorococcus, although it is unclear which specific exudate-associated activities were responsible. Both the composition and activity of the secreted material changed after 500 generations of experimental evolution, suggesting genetic control. Some exudates were located within extracellular vesicles, which were capable of physically associating with Prochlorococcus cells and were responsible for at least some of the growth enhancement effects. Many of the functionalities observed in Alteromonas exudates (e.g., increasing phosphate availability, hydrogen peroxide degradation, and siderophore capabilities) are consistent with leaky Black Queen processes (i.e., services provided by one organism that benefit the entire community) and favor the evolution of interdependencies in microbial communities. We discuss the potential ramifications of such processes being packaged into vesicles as opposed to freely diffusing through the extracellular milieu.IMPORTANCEHeterotrophic bacteria can facilitate improved growth for numerous phytoplankton, including the highly abundant cyanobacterium Prochlorococcus. Here, we show that for the "helper" interaction between Alteromonas and Prochlorococcus, this facilitation is relevant across all phases of batch culture growth and is mediated by secreted products, including proteins and membrane vesicles. We explore the composition of these exudates and show evidence that they are altered during evolutionary adaptation in a changed environment.
An ancient cyanobacterium has evolved through endosymbiosis to form extant chloroplasts of eukaryotic algae and higher plants. During this process, most genes have been transferred to the nuclear genomes. The chloroplast gene sets of different photosynthetic species are relatively conserved yet still exhibiting differences among different species, suggesting a dynamic process and divergence of chloroplast-to-nucleus gene transfer events. Here, based on a comparison of 15 representative green lineage species with sequenced chloroplast genomes, we selected two genes, atpB and rbcL, from a set of 41 conserved genes for chloroplast-to-nucleus gene transfer test. Using the green alga Chlamydomonas reinhardtii as a model organism, we expressed these two genes in the nuclear genome in the corresponding chloroplast mutant background. We demonstrate that transferring atpB and rbcL to the nucleus sustains photoautotrophic growth at different levels. The atpB-TN (transfer-to-nucleus) strains retained photoautotrophic growth to a substantial extent, whereas the rbcL-TN strains were able to grow photoautotrophically only under 5% CO2 and not under ambient air (∼0.04% CO2). The nucleus-encoded proteins accumulated to levels substantially lower than those of the chloroplast-expressed proteins, reaching only 7∼10% (for ATPB) and less than 6% (for RBCL) of the wild-type level. Although both two proteins were relocalized to the chloroplast and assembled into respective ATP synthase and Rubisco complexes, their low abundance appears to account for the incomplete restoration of photosynthetic capability. This study provides a framework for progressively transferring more chloroplast protein-coding genes to the nucleus towards achieving a minimal chloroplast genome in a green algal chassis.
In recent years, the production of microbial biosurfactants, increasingly used in various industries and agriculture, from low-cost renewable substrates is intensively studied. Large volumes of food waste, in particular waste cooking oil, are promising substrates for the production of biosurfactants on an industrial scale. However, studies on the ability of Rhodococcus bacteria, well-known biosurfactant producers, to grow on this substrate and the functional characteristics of synthesized biosurfactants are still scarce. In this study, the possibility of improving biosurfactant production by Rhodococcus ruber IEGM 231 grown on waste cooking oil was investigated using multifactor analysis and response surface methodology (RSM). Using standardized parameters for the concentrations of carbon and nitrogen sources in the fermentation medium, a Pareto diagram was constructed, which shows a direct dependence of the crude biosurfactant yield on the oil concentration, while the concentrations of sugar and inorganic nitrogen salts were found insignificant. Using RSM, the optimal ratio of the medium parameters was determined for the maximum yield of crude biosurfactants: oil (+ 1), sugar (0), NH4- and NO3-containing salts (- 1) in the following concentrations: 5.0 vol%, 2.5 g/L, 0.1 and 0.2 g/L. Crude biosurfactants (40 g/L) obtained from excess waste oil under nitrogen limitation contained three glycolipid fractions, including predominant monoacyl trehalose, previously identified for this strain grown on n-hexadecane, and a considerable amount of residual non-converted acylglycerides. The synthesized biosurfactants produced stable emulsions from petroleum products, food and cosmetic oils and demonstrated high antioxidant capacity in the in vitro ABTS (2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging test. Overexpression of the fbpB gene encoding mycolyltransferase, a key enzyme of the final stage of trehalolipid assembly was revealed in the cells grown on waste cooking oil, which indicated the induction of the synthesis of glycolipid biosurfactants. These findings open up the possibility of valorization of large-scale food oil waste into value-added biosurfactants using Rhodococcus.
Restoring three-dimensional electrical conduction in infarcted myocardium remains a critical challenge, as conventional conductive hydrogel patches largely remain surface-confined and prevent electrical coupling of residual cardiomyocytes within fibrotic scars. Here, we present a self-growing conductive volumetric interface (SCOVE) that transforms surface-confined biointerfaces into tissue-integrated, three-dimensional conductive networks. SCOVE is delivered as an injectable hydrogel precursor containing the tissue-permeable conductive monomer 3,4-ethylenedioxythiophene-acetic acid sodium salt (ETE), which rapidly infiltrates infarcted myocardium and undergoes endogenous glucose-triggered oxidative polymerization to self-grow a conductive polyETE network in situ. The resulting hydrogel gels within 1 min, reaches cardiac-mimetic conductivity (∼1 S m- 1) within 45 min, and preserves native myocardial mechanics without inducing tissue stiffening. In a rat myocardial infarction model, SCOVE penetrates the infarct, reduces scar resistivity by 2.54-fold compared with conventional 2D conductive patches, restores electrical coupling among residual cardiomyocytes, enhances Cx43 expression, and accelerates impulse propagation. By replacing static, surface-confined conductive patches with self-growing volumetric biointerfaces, this work establishes a generalizable strategy for reconstructing tissue electrophysiology and advancing bioelectronic therapies for myocardial infarction and other electrically dysfunctional tissues.
Chile Crece Más ("Chile Grows More", CCM) is a comprehensive, multisector, equity-focussed health and social development program which provides essential support to Chilean children and families. Since its rollout in 2007 it has become the centerpiece of Chile's child development policy, but has faced challenges in its design, delivery, and functionality. This Health Reform Monitor paper identifies four key policy opportunities to improve CCM. To augment the program's impact, the Chilean government could universalize the program to cover all children, not just those insured publicly; adopt a risk-based, family-centered approach to care; modernize record-keeping practices and electronic medical record use in primary care centers; and create (over time) a nationwide, centralized medical record database for all Chilean children. These reforms will improve access, efficiency, and quality of the program. Together, they are likely to improve child development and health outcomes, and will likely be popular among program users. They are also in line with broader reforms underway in Chile, enhancing political feasibility and salience. By improving CCM, the Chilean government has an opportunity to improve child health in the country. Other countries seeking to develop or refine similar programs could utilize these principles when reforming their own policies.
For decades, discussions about the evolution of species overlooked microorganisms. Over a century ago, Neisser and Massini isolated a coliform bacterium that appeared to acquire mutations adapting it to its environment, naming it Bacterium coli mutabile to reflect this feature. With the advent of molecular biology, these widely debated experiments were subsequently forgotten. Here, we present the history of an experiment that reproduces their observations in a modern context where it has become possible to identify the nature of these mutations down to the nucleotide level. Its findings demonstrate that the transcription of gene families that ensure the long-term maintenance of the metabolism of ageing cells is a direct source of adaptive mutations: this process enables bacteria to identify previously unexploited environmental factors that can now support growth. We propose that the driving force behind this adaptation is the spontaneous dehydration/deamidation of polypeptide chains, which dictates an intrinsic lifespan for every protein. This universal mechanism of inevitable protein ageing necessitates their re-synthesis to maintain their function; however the transcription process, which involves opening the DNA double helix, is locally mutagenic. Thus, as bacteria age, the continuous re-synthesis of some of the proteins that perform the functions enabling survival triggers a local mutagenic process. This yields genetic variants, some of which may be beneficial and are therefore retained. Longtemps la réflexion sur l’évolution des espèces a ignoré les microorganismes. Il y a plus de cent ans, Neisser et Massini isolaient un colibacille qui semblait acquérir des mutations l’adaptant à son environnement et le nommaient pour cette raison Bacterium coli mutabile. Ces expériences très discutées ont été oubliées avec l’avènement de la biologie moléculaire. Nous rapportons ici l’histoire d’une expérience qui reproduit leurs observations dans un contexte moderne où il est devenu possible d’identifier au nucléotide près la nature de ces mutations. Ses résultats font apparaître que la transcription de familles de gènes assurant le maintien à long terme du métabolisme de la cellule vieillissante est source de mutations qui lui permettent de s’adapter en découvrant dans l’environnement ce qui lui permet de croître à nouveau. Le ressort de cette adaptation est la déshydratation/désamidation spontanée des chaînes polypeptidiques qui confère une durée de vie intrinsèque à chaque protéine. C’est que ce mécanisme universel du vieillissement inévitable des protéines nécessite leur resynthèse pour assurer leur fonction, alors que le processus de transcription, ouvrant la double hélice de l’ADN est localement mutagène. Ainsi, en vieillissant, les protéines qui assurent les fonctions permettant à la bactérie de survivre déclenchent un processus de mutagenèse qui en produit des variants dont certains peuvent être bénéfiques et sont alors retenus.
Individual growth is an important demographic component in population dynamics that indicates the contribution of biomass to the population, such as an increase in length and weight over time. In addition, the length-at-age provides information concerning mortality, reproduction rates, survival, and biological factors, which are essential for stock assessment and fishery management. Age and growth studies have focused on estimating average growth trajectories based on length-at-age data from only one developmental stage, assuming a constant growth throughout the life cycle. However, the individuals within a cohort grow at different rates and speeds. This causes uncertainty in the mathematical models fitted to the data, both in the growth parameters and in the mean length-at-age within a cohort. The datasets included in this article were collected from two sources: 1) Fishery-independent data of shell length frequency and age of individuals in early growth stages (0-68 days) obtained under rearing experimental conditions. 2) Fishery-dependent data of shell length-at-age of adult individuals aged 3-47 years collected from commercial landings. The straight-line distance of live specimens was measured between the anterior and posterior margin of the shell. Specimens were then dissected to extract the shells. Age was determined as the number of internal growth lines. The length and age (days) datasets comprised 797 records for larvae and juveniles, and 357 records for adult individuals. These data provide valuable information for researchers, decision makers, the government, and private and academic institutions. The dataset enables comparative analyses and further investigations. Its reuse potential extends to broader topics, such as stock assessment, population dynamics, and fishery management.
Fresh-cut produce operations often generate work-in-process (WIP) ingredients that have undergone peeling, cutting, washing, etc., and are then subjected to temporary holding, prior to packaging, creating conditions that may favor pathogen survival and growth. The objective of this study was to evaluate the survival of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica, and native microbiota on WIP fresh-cut carrot and yellow onion treated with deionized water, sodium hypochlorite (NaOCl, >10 ppm free chlorine), or peracetic acid (PAA, 80 ppm) and held at 4, 8, or 12°C for up to seven days. Pathogen behavior was strongly influenced by storage temperature and commodity matrix. At 4°C, E. coli O157:H7 and S. enterica populations generally declined or remained stable. Conversely, at 8°C and 12°C, which were evaluated to simulate potential temperature-abuse scenarios, E. coli O157:H7 and S. enterica proliferated in unwashed samples but were often suppressed by chlorine and PAA. L. monocytogenes on carrot did not grow under any treatment or temperature, likely due to intrinsic antimicrobial compounds, while it persisted on onion and grew by 0.5-1.6 logs under temperature abuse. PAA often provided the greatest pathogen reductions on onion, although chlorine was more effective against E. coli O157:H7 on carrot. Native mesophilic or psychrotrophic bacteria, and yeast and mold populations, increased rapidly across all treatments and storage conditions, with PAA-washed samples often having the greatest growth rates. Overall, while sanitizer efficacy was commodity-dependent, strict temperature and holding time management emerged as the most critical factor in limiting pathogen growth. These findings underscore the necessity of an integrated management approach for WIP produce, although further validation under dynamic industrial conditions and mechanisms for the rapid regrowth of microbiota in PAA-washed samples warrant further investigations.
Lung squamous cell carcinoma (LUSC) is a difficult cancer to treat, with few targeted therapies to improve its poor prognosis. The goal of this study was to use a drug repurposing strategy to evaluate and compare drug sensitivities using 2D adherent and 3D spheroid models of NCI-H2170 LUSC cells. Both 2D adherent and 3D spheroid models were used to grow NCI-H2170 lung squamous cell carcinoma cells and evaluate their sensitivity to a large library of food and drug administration (FDA)-approved drugs, including many not typically used as anti-cancer agents. Cell death was assessed in the 2D adherent models, and for the top drugs half maximal effective concentration (EC50) values were determined. For the 3D spheroid models, drugs reducing spheroid size after 4 days of treatment were identified. There were 263 drugs that reduced the cell viability to <20% when cells were grown in 2D in 10 µM drug. When grown in 3D the cells were generally more drug resistant, with 87 drugs capable of reducing spheroid volume when grown over 4 days in 10 µM drug. Interestingly, 60 drugs proved effective in both model systems including many drugs that typically associated with anti-cancer properties. Of these 60, four were further found to have selective effects towards metastatic NCI-H2170 cells as compared to a much less metastatic matched cell line expressing the metastasis suppressor CREB3L1, in both 2D and 3D model systems. These included the hypoxia-inducible factor 1-alpha (HIF-1α inhibitor 2-methoxyestradiol, and three anti-infection agents (cetylpyridinium chloride, chlorhexidine-2HCl, zinc pyrithione). These results suggest that the HIF-1α inhibitor 2-methoxyestradiol, and three anti-infection agents (cetylpyridinium chloride, chlorhexidine-2HCl, zinc pyrithione) may be effective for metastatic LUSCs.
Poly(acrylonitrile-butadiene-styrene) (ABS) and styrene-acrylonitrile (SAN) are extensively used in cosmetic and industrial packaging due to their mechanical strength and durability. However, their intrinsic resistance to microbial and enzymatic attack underlies their long-term environmental persistence. To date, there is limited experimental evidence regarding the microbial transformation of ABS/SAN polymers. Here, we investigated whether four previously reported polyethylene-degrading bacterial strains-Comamonas sp., Delftia sp., Stenotrophomonas sp., and Alcaligenes sp.-can grow on an ABS/SAN blend and induce measurable physicochemical modifications. Each strain was incubated for 90 days in a minimal medium containing ABS/SAN as the sole added carbon and energy source. Bacterial growth, viability, and polymer modification were assessed by fluorescence microscopy, gravimetric analysis, and attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR). All strains sustained active growth and remained viable throughout incubation. Stenotrophomonas sp. and Delftia sp. exhibited the strongest responses, with approximately 6-fold and 2.5-fold increases in viable cell counts and corresponding mass losses of 2.20% and 1.27%, respectively. ATR-FTIR profiles revealed strain-specific chemical changes on the polymer surface, including reductions in nitrile and carbonyl content consistent with partial oxidation and assimilation of ABS/SAN fragments. Comparative genomic analysis further identified genes encoding putative catabolic functions that may facilitate polymer transformation, such as nitrilases, amidases, phenylacetate and aromatic compound degradation pathways, redox-active enzymes, and efflux transporters. Together, these findings provide a novel experimental evidence that select bacterial strains can interact with and induce measurable modifications in ABS/SAN polymer blend, highlighting new perspectives for the biological degradation of recalcitrant synthetic plastics.
Evolutionary constraints governing flagellar number in bacterial pathogens remain poorly understood. While related Pseudomonas species are hyperflagellated, Pseudomonas aeruginosa maintains strict monoflagellation through the FleQ-FleN regulatory circuit. Here, we demonstrate that FleN dosage is essential for maintaining monoflagellation and fitness. Confirming earlier reports, wild-type P. aeruginosa displayed unipolar monoflagellation, whereas ∆fleN mutants developed multiple flagella, frequently more than four, in unipolar or bipolar arrangements. These hyperflagellated cells showed marked fitness defects, including reduced growth, attenuated virulence in a nematode infection model, and a competitive disadvantage in co-culture. The virulence defect reflected hyperflagellation rather than the loss of motility, since a non-flagellated ∆fliC mutant retained full pathogenicity. Notably, ∆fleN cells rapidly evolved suppressor mutations in fleQ that partially restored growth and motility without always restoring monoflagellation. Five independent suppressor alleles mapped to key FleQ domains, four in the AAA+ ATPase domain, and one in the DNA-binding domain, consistent with reduced FleQ activity that rebalances the circuit. Single-cell holographic tracking showed that suppressor strains swim with heterogeneous dynamics, including subpopulations that reach wild-type (WT) speeds, despite carrying multiple flagella. Quantitative proteomics indicated that the fitness burden extends beyond flagellar components, with protein-abundance changes across metabolism, stress responses, and signaling. Conversely, selection under high-viscosity conditions drove wild-type cells to acquire enhanced spreading through a recurrent fleN mutation, fleNV178G; multiflagellation is therefore accessible. Together, these findings indicate that the FleQ-FleN circuit balances the flagellar number against fitness, holding P. aeruginosa at a single flagellum while retaining the latent capacity to reach multiflagellated states through single mutations.IMPORTANCEBacterial flagella are extracellular appendages that rotate to propel the cell and enable swimming motility. While some bacteria have multiple flagella, many pathogenic species like Pseudomonas aeruginosa have just one. Surprisingly, mutants of P. aeruginosa with multiple flagella performed worse, that is, they grew more slowly, were less infectious in laboratory animals, and were outcompeted by wild-type bacteria. Even when some mutant bacteria evolved compensatory changes, they still struggled compared to single-flagellum bacteria. This reveals an important evolutionary trade-off: while multiple flagella might seem advantageous for movement, having just one flagellum allows the bacteria to grow faster and cause more severe infections. This plasticity likely explains why P. aeruginosa is so successful both in the environment and as a human pathogen.
Alternative polyadenylation (APA) generates mRNA isoforms with distinct 3' untranslated regions (3'UTRs), thereby influencing transcript stability and translation. In cancer, 3'UTR shortening can activate oncogenes by escaping microRNA (miRNA)-mediated repression, but its role in hepatocellular carcinoma (HCC) remains poorly defined. Here, we profiled mRNA length alterations in multistage human HCC transcriptome datasets and investigated their functional consequences. Approximately 77% of mRNAs with altered length exhibited 3'UTR shortening. Glypican-3 (GPC3) was the most prominently upregulated shortened transcript, and high GPC3 expression was associated with poor prognosis in HCC. GPC3 knockdown reduced proliferation and induced apoptosis, whereas GPC3 overexpression promoted cell growth. Among APA regulators, Cleavage Stimulation Factor 2 (CSTF2) was upregulated in HCC, correlated positively with GPC3 expression, and predicted adverse clinical outcomes. Modulation of CSTF2 expression altered GPC3 3'UTR length, with CSTF2 overexpression promoting GPC3 3'UTR shortening, increasing GPC3 protein expression, enhancing proliferation, and suppressing apoptosis. Further analysis revealed that GPC3 3'UTR shortening removed binding sites for miR-96-5p and miR-140-5p, relieving miRNA-mediated translational repression. These findings identify CSTF2-driven APA as a mechanism of oncogenic GPC3 activation in HCC and suggest the CSTF2-GPC3 axis as a potential therapeutic target. Liver cancer is one of the leading causes of cancer-related death worldwide. Glypican-3 (GPC3) is often highly increased in liver cancer and is being studied as a marker and treatment target, but the reason for its increase is not fully understood. In this study, we analyzed patient datasets, liver cancer cells, and tumor samples to investigate how GPC3 is controlled. We found that liver cancer cells often produce a shortened form of GPC3 RNA. This shorter RNA form avoids regulation by small RNA molecules that normally help keep GPC3 levels low. We also identified CSTF2 as an important factor that promotes this shortening process. As a result, GPC3 becomes more stable and more highly expressed, helping cancer cells grow and survive. These findings reveal a new way that liver cancer cells increase GPC3 and may support future strategies to diagnose or treat liver cancer.
Accelerating research advancing health equity for the entire inheritable bleeding disorders community requires a new approach. It must be firmly rooted in health equity, diversity, and inclusion (HEDI) and center the knowledge of people living with inheritable bleeding disorders, the Lived Experience Experts (LEEs). The National Bleeding Disorders Foundation charged seven multidisciplinary working groups (WGs) with developing a National Research Blueprint (NRB) for this Bleeding Disorders Research Collaborative (BDRC). The Infrastructure and Workforce WGs, in collaboration with the HEDI and LEE WGs, met virtually to develop recommendations for BDRC operationalization. A progressive network of elements and processes capacitating diverse community-prioritized research ideas into successfully completed BDRC projects is proposed. Essential components for launch, iterative evolution, effective conduct, and accountability are described. Sharing resources and expertise and embedding research in inheritable bleeding disorders care will create synergistic efficiencies. Education and training to grow and empower interdisciplinary research teams, including LEEs and HEDI champions as valued members, are detailed. Shared leadership integrating LEE and HEDI expertise throughout will provide dynamic governance. BDRC infrastructure and workforce development must start small and grow iteratively in partnership with the many organizations that share its vision of health justice. The National Research Blueprint is a proposal for a new Bleeding Disorders Research Collaborative (BDRC) doing the research people with bleeding disorders need and want. The people who live with a disorder, and their close family members affected by it, are Lived Experience Experts. The new collaborative will place Lived Experience Experts at the center of research. What research is done, how it is done, and how the results are used and shared must be decided in partnership with lived experience expertise. The collaborative must also advance health equity for all. Every initiative and project must improve diversity, inclusion, and belonging.This paper proposes infrastructure and workforce development processes for the new research collaborative. Recommendations were developed by groups of clinical, research, lived experience, and health equity experts. The groups made sure everyone was able to contribute meaningfully and confidently. Every voice was heard and valued. This is also how the collaborative must operate, with shared leadership and teams that are trained to work well together. Education and processes are proposed to develop a diverse, inclusive interdisciplinary workforce, reflecting the community it serves and integrating lived experience expertise throughout. The proposed infrastructure is a network of expertise, resources, facilities, and processes, all connected by a platform. It is designed to start small, with just enough of each component to support a few simple projects. All projects will be evaluated to learn what works well and what can be improved. The whole collaborative will improve with learnings from each success and shortcoming.
Increasing atmospheric carbon dioxide (CO2) is transforming the climate space in which plants grow, severely affecting crop physiology and crop productivity. Elevated CO2 enhances photosynthesis and biomass; however, it can nitrogen (N) metabolism, inhibiting the nutritional value and the yield capacity of crops. The most important central N-regulated protein machineries are: transporters, nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, glutamate dehydrogenase, and urease which control N assimilation, distribution and remobilization in crop plants. With high CO2, these molecular components exhibit altered expression and activities mostly due to the reduction in N concentration. Complex systemic plant responses under N control like adaptation of photosynthetic capacity, flowering time, reproductive development and seed nutrient profiles further support the complexity of the interactions between C and N signaling. The high CO2 environment requires a more holistic analysis of the regulatory networks of N metabolism and anticipative crop improvement strategies. Future breeding and crop improvement strategies should focus on enhancing the resilience of N assimilation by optimizing the N transporter function and maintaining C-N stoichiometry, thereby sustaining crop performance and nutritive quality under changing climatic conditions. The present review identifies the molecular processes that regulate N responses in crops grown under elevated CO2, highlighting the differences between legume versus non-legume and C3 versus C4 plant responses and providing details that can ensure mitigation against negative impacts and outline future perspectives on crop improvement.
Heterophylly, characterized by distinct leaf morphologies, is an adaptive strategy evolved by some aquatic plants to cope with the contrasting environments in which they grow. It is shaped by environmental factors and by hormones, particularly abscisic and gibberellic acids. We conducted a comparative multi-omics analysis of submerged and floating leaves of Potamogeton nodosus, a heterophyllous species of the Alismatales order. Genome sequencing has confirmed its tetraploid status. A phylogenomic analysis has identified genes associated with aquatic adaptation, notably genes possibly involved in aurone biosynthesis, which are antioxidant flavonoids. Transcriptomic and metabolomic analyses have revealed distinct gene expression and metabolite patterns depending on the leaf position in the water column. The floating leaves exhibited upregulated genes associated with photosynthetic and metabolic activities, and with the accumulation of UV-protective metabolites reflecting light capture and photoprotection optimization. Conversely, submerged leaves showed upregulated genes associated with homeostatic processes, defense responses, and cell wall remodeling, reflecting tissue maintenance. We have also identified genes involved in the abscisic and gibberellic acids pathways. The developmental analyses highlighted active growth processes in young leaves. This multi-omic study provides an integrated understanding of the complex molecular mechanisms underlying the adaptation of P. nodosus to a dual lifestyle.
<b>Background and Objective:</b> Changes in rhizosphere microbial populations have been reported in response to drought, temperature fluctuations, CO<sub>2</sub> levels and other environmental factors. However, the structure of the root-associated microbes in local North Sulawesi rice using a metagenomic approach has not yet been investigated. This study examined the microbial community structure in local North Sulawesi rice (cv. Superwin) under drought (water deficit) conditions compared to well-watered conditions at the vegetative phase. <b>Materials and Methods:</b> Rice plants were grown in polybags filled with a 5:1:1 mixture of garden soil, compost and rice husks and were allowed to grow until the four-fully-expanded leaf stage. They were then subjected to two treatments for 14 days: well-watered conditions (irrigated to 100% field capacity) and water deficit conditions (0% field capacity). Root samples were collected for next-generation sequencing analysis to assess molecular response of Superwin rice to water deficit. <b>Results:</b> During drought, several root-associated microbes were more prevalent, including <i>Nitrospirota</i> at the phylum level, <i>Rubrobacteria</i> at the class level, <i>Micrococcales</i> at the order level, Gaiellaceae at the family level, <i>Gaiella</i> at the genus level and <i>Gaiella occulta</i> at the species level. <b>Conclusion:</b> Root-associated microbes, including taxa <i>Nitrospirota</i>, <i>Rubrobacteria</i>, Micrococcales, Gaiellaceae, <i>Gaiella</i> and <i>Gaiella occulta</i>, have a higher relative abundance in rice plants under water deficit. <i>Gaiella occulta</i> serves as sensitive indicator of water deficit in North Sulawesi local rice, i.e. Superwin.
Genuine accumulation of metals/metalloids in bryophytes is limited and highly susceptible to surficial contamination with soil particles. Washing with an apolar solvent removes most surficial contamination prior to elemental analysis. Bryophytes are often the first colonisers of soils toxic from metals/metalloids derived from natural mineralisation or mining wastes. They are ostensibly highly tolerant to the prevailing high concentrations of metals and metalloids in the substrate, but little is known about their ability to (hyper)accumulate these metals/metalloids. Terrestrially growing bryophytes were collected from arsenic-thallium mineralised soils at the Allchar site in North Macedonia. Samples were analysed for elemental concentrations using monochromatic X-ray fluorescence analysis (MXRF) after stringent washing with an apolar solvent and subjected to synchrotron micro-X-ray fluorescence (µXRF) elemental imaging. Scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) were additionally used to assess extraneous contamination and test the efficiency of the washing procedure. The results show that surficial contamination with soil particles is a major challenge for assessing metal and metalloid concentrations in (terrestrial) bryophytes from metalliferous soils. Washing with an apolar solvent (hexane) removes most surficial contamination prior to elemental analysis, indicating some potential for elemental accumulation, as found in Rhynchostegium megapolitanum for thallium. Genuine accumulation of arsenic and thallium is relatively low despite the ability to grow on extremely arsenic-thallium enriched soils. Measured elemental concentrations in bryophyte samples are strongly affected by the washing procedure, highlighting the importance of appropiate sample preparation.