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Endometrial cancer (EC) is a prevalent gynecological malignancy with an escalating incidence and mortality rate, notably in China. Surgical options are limited, and drug resistance development poses significant challenges to EC treatment. Ketamine, a rapidacting anesthetic, exhibits anti-inflammatory, analgesic, and antidepressant properties, yet its potential in cancer therapy remains largely unexplored. Here, we investigate ketamine’s effects on EC cell growth, motility and ferroptosis, alongside its impact on the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/the mammalian target of rapamycin (mTOR) axis. Ketamine significantly inhibits EC cell growth and motility while promoting ferroptosis. Mechanistically, ketamine’s anti-tumor effects correlate with suppression of the PI3K/Akt/mTOR axis crucial for cell survival and growth. In summary, our findings highlight ketamine’s potential therapeutic application in EC treatment by impeding cell growth and motility, fostering ferroptosis, and suppressing the PI3K/Akt/mTOR axis. These insights offer novel avenues for EC therapy. [ABSTRACT FROM AUTHOR]

Various environmental factors, including H2 availability, metabolic tradeoffs, optimal growth temperature, stochasticity, and hydrology, were examined to determine if they affect microbial competition between three autotrophic thermophiles. The thiosulfate reducer Desulfurobacterium thermolithotrophum (Topt72°C) was grown in mono- and coculture separately with the methanogens Methanocaldococcus jannaschii (Topt82°C) at 72°C and Methanothermococcus thermolithotrophicus (Topt65°C) at 65°C at high and low H2 concentrations. Both methanogens showed a metabolic tra deoff shifting from high growth rate-low cell yield at high H2 concentrations to low growth rate-high cell yield at low H2 concentrations and when grown in coculture with the thiosulfate reducer. In 1:1 initial ratios, D. thermolithotrophum outcompeted both methanogens at high and low H2, no H2S was detected on low H2, and it grew with only CO2 as the electron acceptor indicating a similar metabolic tradeoff with low H2. When the initial methanogen-to-thiosulfate reducer ratio varied from 1:1 to 104:1 with high H2, D. thermolithotrophum always outcompeted M. jannaschii at 72°C. However, M. thermolithotrophicus outcompeted D. thermolithotrophum at 65°C when the ratio was 103:1. A reactive transport model that mixed pure hydrothermal fluid with cold seawater showed that hyperthermophilic methanogens dominated in systems where the residence time of the mixed fluid above 72°C was sufficiently high. With shorter residence times, thermophilic thiosulfate reducers dominated. If residence times increased with decreasing fluid temperature along the flow path, then thermophilic methanogens could dominate. Thermophilic methanogen dominance spread to previously thiosulfate-reducer-dominated conditions if the initial ratio of thermophilic methanogen-to-thiosulfate reducer increased. [ABSTRACT FROM AUTHOR]

Transfer RNAs (tRNAs) are the most highly modified cellular RNAs, both with respect to the proportion of nucleotides that are modified within the tRNA sequence and with respect to the extraordinary diversity in tRNA modification chemistry. However, the functions of many different tRNA modifications are only beginning to emerge. tRNAs have two general clusters of modifications. The first cluster is within the anticodon stem-loop including several modifications essential for protein translation. The second cluster of modifications is within the tRNA elbow, and roles for these modifications are less clear. In general, tRNA elbow modifications are typically not essential for cell growth, but nonetheless several tRNA elbow modifications have been highly conserved throughout all domains of life. In addition to forming modifications, many tRNA modifying enzymes have been demonstrated or hypothesized to also play an important role in folding tRNA acting as tRNA chaperones. In this review, we summarize the known functions of tRNA modifying enzymes throughout the lifecycle of a tRNA molecule, from transcription to degradation. Thereby, we describe how tRNA modification and folding by tRNA modifying enzymes enhance tRNA maturation, tRNA aminoacylation, and tRNA function during protein synthesis, ultimately impacting cellular phenotypes and disease. [ABSTRACT FROM AUTHOR]

To clarify the growth mechanisms of Rhodococcus in the alkane phase, we measured oxygen utilization in the alkane phase. The results showed that dissolved oxygen decreased significantly when viable cells were present in the alkane phase. The findings suggested that Rhodococcus strains can grow in alkanes and utilize the resident dissolved oxygen. [ABSTRACT FROM AUTHOR]

Melanoma, a cancer arising from melanocytes, requires a novel treatment strategy because of the ineffectiveness of conventional therapies in certain patients. Fustin is a flavanonol found in young fustic (Cotinus coggygria). However, little is known about its antimelanoma effects. Our study demonstrates that fustin suppresses the growth of B16 melanoma cells. Phalloidin staining of cytoskeletal actin revealed that fustin induced a conformational change in the actin structure of melanoma cells, accompanied by suppressed phosphorylation of myosin regulatory light chain 2 (MLC2), a regulator of actin structure. Furthermore, the protein kinase A (cAMP-dependent protein kinase) inhibitor H89 completely attenuated fustin-induced downregulation of phosphorylated myosin phosphatase targeting subunit 1, which is involved in dephosphorylation of MLC2. In a mouse model, administration of fustin suppressed tumor growth in B16 melanoma cells without adverse effects. In conclusion, our findings suggest that fustin effectively suppresses melanoma cell growth both in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Motivation Lineage tracing and trajectory inference from single-cell RNA-sequencing data hold tremendous potential for uncovering the genetic programs driving development and disease. Single cell datasets are thought to provide an unbiased view on the diverse cellular architecture of tissues. Sampling bias, however, can skew single cell datasets away from the cellular composition they are meant to represent. Results We demonstrate a novel form of sampling bias, caused by a statistical phenomenon related to repeated sampling from a growing, heterogeneous population. Relative growth rates of cells influence the probability that they will be sampled in clones observed across multiple time points. We support our probabilistic derivations with a simulation study and an analysis of a real time-course of T-cell development. We find that this bias can impact fate probability predictions, and we explore how to develop trajectory inference methods which are robust to this bias. Availability and implementation Source code for the simulated datasets and to create the figures in this manuscript is freely available in python at https://github.com/rbonhamcarter/simulate-clones. A python implementation of the extension of the LineageOT method is freely available at https://github.com/rbonhamcarter/LineageOT/tree/multi-time-clones. [ABSTRACT FROM AUTHOR]

Summary Quantification of growth parameters and extracellular uptake and production fluxes is central in systems and synthetic biology. Fluxes can be estimated using various mathematical models by fitting time–course measurements of the concentration of cells and extracellular substrates and products. A single tool is available to non-computational biologists to calculate extracellular fluxes, but it is hardly interoperable and is limited to a single hard-coded growth model. We present our open-source flux calculation software, PhysioFit, which can be used with any growth model and is interoperable by design. PhysioFit includes some of the most common growth models, and advanced users can implement additional models to calculate extracellular fluxes and other growth parameters for metabolic systems or experimental setups that follow alternative kinetics. PhysioFit can be used as a Python library and offers a graphical user interface for intuitive use by end-users and a command-line interface to streamline integration into existing pipelines. Availability and implementation PhysioFit v3 is implemented in Python 3 and was tested on Windows, Unix, and MacOS platforms. The source code and the documentation are freely distributed under GPL3 license at https://github.com/MetaSys-LISBP/PhysioFit/ and https://physiofit.readthedocs.io/. [ABSTRACT FROM AUTHOR]

In the fission yeast Schizosaccharomyces pombe, the response to sulfur depletion has been less studied compared to the response to nitrogen depletion. Our study reveals that the fission yeast gene, SPCC417.09c, plays a significant role in the sulfur depletion response. This gene encodes a protein with a Zn2Cys6 fungal‐type DNA‐binding domain and a transcription factor domain, and we have named it sdr1+ (sulfur depletion response 1). Interestingly, while sulfur depletion typically induces autophagy akin to nitrogen depletion, we found that autophagy was not induced under sulfur depletion in the absence of sdr1+. This suggests that sdr1+ is necessary for the induction of autophagy under conditions of sulfur depletion. Although sdr1+ is not essential for the growth of fission yeast, its overexpression, driven by the nmt1 promoter, inhibits growth. This implies that Sdr1 may possess cell growth‐inhibitory capabilities. In addition, our analysis of Δsdr1 cells revealed that sdr1+ also plays a role in regulating the expression of genes associated with the phosphate depletion response. In conclusion, our study introduces Sdr1 as a novel transcription factor that contributes to an appropriate cellular nutrient starvation response. It does so by inhibiting inappropriate cell growth and inducing autophagy in response to sulfur depletion. [ABSTRACT FROM AUTHOR]

Recent advances in genetic sequencing are transforming our approach to rare-disease care. Initially identified in cancer, gain-of-function mutations of the PIK3CA gene are also detected in malformation mosaic diseases categorized as PIK3CA-related disorders (PRDs). Over the past decade, new approaches have enabled researchers to elucidate the pathophysiology of PRDs and uncover novel therapeutic options. In just a few years, owing to vigorous global research efforts, PRDs have been transformed from incurable diseases to chronic disorders accessible to targeted therapy. However, new challenges for both medical practitioners and researchers have emerged. Areas of uncertainty remain in our comprehension of PRDs, especially regarding the relationship between genotype and phenotype, the mechanisms underlying mosaicism, and the processes involved in intercellular communication. As the clinical and biological landscape of PRDs is constantly evolving, this review aims to summarize current knowledge regarding PIK3CA and its role in nonmalignant human disease, from molecular mechanisms to evidence-based treatments. [ABSTRACT FROM AUTHOR]

Background: Lung adenocarcinoma (LUAD) is a highly aggressive and rapidly fatal malignancy worldwide. Collagen XVII (COL17A1) has been implicated in various protumorigenic processes. However, the functions and mechanisms of COL17A1 in LUAD progression still remain elusive. Methods: COL17A1 and ubiquitin‐specific protease 22 (USP22) mRNA analysis was performed by quantitative PCR, and their protein levels were detected by immunoblotting and immunohistochemistry. The functional influence was evaluated by determining cell viability, proliferation, apoptosis, invasion, migration, and ferroptosis in vitro, as well as xenograft growth in vivo. Co‐immunoprecipitation (Co‐IP) and IP experiments were used to examine the USP22/COL17A1 interaction and COL17A1 deubiquitination. Cycloheximide treatment was used to analyze COL17A1 protein stability. Results: COL17A1 and USP22 were upregulated in human LUAD tissues and cell lines. Functionally, COL17A1 knockdown acted for the suppression of LUAD cell growth, invasion, and migration as well as promotion of cell apoptosis and ferroptosis in vitro. COL17A1 knockdown could diminish the tumorigenicity of LUAD cells in vivo. Mechanistically, USP22 stabilized and upregulated COL17A1 by enhancing the deubiquitination of COL17A1. Additionally, reexpression of COL17A1 could reverse USP22 silencing‐induced phenotype changes of LUAD cells in vitro. Conclusion: Our findings demonstrate that USP22‐stabilized COL17A1 possesses oncogenic activity in LUAD. We propose that USP22 and COL17A1 would be potential targets for the establishment of therapeutic approaches against LUAD. [ABSTRACT FROM AUTHOR]

The tumor suppressor p14/19ARF regulates ribosomal RNA (rRNA) synthesis by controlling the nucleolar localization of Transcription Termination Factor 1 (TTF1). However, the role played by TTF1 in regulating the rRNA genes and in potentially controlling growth has remained unclear. We now show that TTF1 expression regulates cell growth by determining the cellular complement of ribosomes. Unexpectedly, it achieves this by acting as a "roadblock" to synthesis of the noncoding LncRNA and pRNA that we show are generated from the "Spacer Promoter" duplications present upstream of the 47S pre‐rRNA promoter on the mouse and human ribosomal RNA genes. Unexpectedly, the endogenous generation of these noncoding RNAs does not induce CpG methylation or gene silencing. Rather, it acts in cis to suppress 47S preinitiation complex formation and hence de novo pre‐rRNA synthesis by a mechanism reminiscent of promoter interference or occlusion. Taken together, our data delineate a pathway from p19ARF to cell growth suppression via the regulation of ribosome biogenesis by noncoding RNAs and validate a key cellular growth law in mammalian cells. [ABSTRACT FROM AUTHOR]

Reduced nicotinamide adenine dinucleotide phosphate (NADPH) is a crucial cofactor in metabolic networks. The efficient regeneration of NADPH is one of the limiting factors for productivity in biotransformation processes. To date, many metabolic engineering tools and static regulation strategies have been developed to regulate NADPH regeneration. However, traditional static regulation methods often lead to the NADPH/NADP+ imbalance, causing disruptions in cell growth and production. These methods also fail to provide real-time monitoring of intracellular NADP(H) or NADPH/NADP+ levels. In recent years, various biosensors have been developed for the detection, monitoring, and dynamic regulate of the intracellular NADP(H) levels or the NADPH/NADP+ balance. These NADPH-related biosensors are mainly used in the cofactor engineering of bacteria, yeast, and mammalian cells. This review analyzes and summarizes the NADPH metabolic regulation strategies from both static and dynamic perspectives, highlighting current challenges and potential solutions, and discusses future directions for the advanced regulation of the NADPH/NADP+ balance. [ABSTRACT FROM AUTHOR]

The development of in vitro models is essential for a comprehensive understanding and investigation of pulmonary fibrosis (PF) at both cellular and molecular levels. This study presents a literature review and an analysis of various cellular models used in scientific studies, specifically focusing on their applications in elucidating the pathogenesis of PF. Our study highlights the importance of taking a comprehensive approach to studing PF, emphasizing the necessity of considering multiple cell types and organs and integrating diverse analytical perspectives. Notably, primary cells demonstrate remarkable cell growth characteristics and gene expression profiles; however, their limited availability, maintenance challenges, inability for continuous propagation and susceptibility to phenotypic changes over time significantly limit their utility in scientific investigation. By contrast, immortalized cell lines are easily accessible, cultured and continuously propagated, although they may have some phenotypic differences from primary cells. Furthermore, in vitro coculture models offer a more practical and precise method to explore complex interactions among cells, tissues and organs. Consequently, when developing models of PF, researchers should thoroughly assess the advantages, limitations and relevant mechanisms of different cell models to ensure their selection is consistent with the research objectives. [ABSTRACT FROM AUTHOR]

Objective: To investigate the impact of IGJ on the proliferation, inflammation, and motility of rheumatoid arthritis (RA) fibroblast‐like synoviocytes and elucidate the underlying mechanism. Methods: The expression of IGJ RA fibroblast‐like synoviocytes was assessed using immunoblot and qPCR. Cell growth was evaluated using CCK‐8 and FCM assays. The effects on inflammatory response were determined by ELISA and immunoblot assays. Cell motility was assessed using transwell and immunoblot assays. The mechanism was further confirmed using immunoblot assays. Results: IGJ expression was found to be elevated in fibroid synovial cells of RA. IGJ ablation inhibited the growth of MH7A cells and suppressed the inflammatory response. Knockdown of IGJ also blocked cell motility. Mechanically, the knockdown of IGJ suppressed the NF‐κB axis in MH7A cells. Conclusion: IGJ suppresses RA in fibroblast‐like synoviocytes via NF‐κB pathway. [ABSTRACT FROM AUTHOR]

Understanding the complex dynamics of telomere biology is important in the strong link between aging and cancer. Telomeres, the protective caps at the end of chromosomes, are central players in this connection. While their gradual shortening due to replication limits tumors expansion by triggering DNA repair mechanisms, it also promotes oncogenic changes within chromosomes, thus sustaining tumorigenesis. The enzyme telomerase, responsible for maintaining telomere length, emerges as a central player in this context. Its expression in cancer cells facilitates the preservation of telomeres, allowing them to circumvent the growth-limiting effects of short telomeres. Interestingly, the influence of telomerase extends beyond telomere maintenance, as evidenced by its involvement in promoting cell growth through alternative pathways. In this context, inflammation accelerates telomere shortening, resulting in telomere dysfunction, while telomere elements also play a role in modulating the inflammatory response. The recognition of this interplay has promoted the development of novel therapeutic approaches centered around telomerase inhibition. This review provides a comprehensive overview of the field, emphasizing recent progress in knowledge and the implications in understanding of cancer biology. [ABSTRACT FROM AUTHOR]

Isoamyl 4-methoxycinnamate (IMC) is widely used in various fields because of its exceptional UV-filter properties. However, due to its cytotoxicity and anti-microbial degradability, the potential eco-environmental toxicity of IMC has become a focus of attention. In this study, we propose a host–guest supramolecule approach to enhance the functionality of IMC, resulting in a more environmentally friendly and high-performance materials. Sulfobutyl-β-cyclodextrin sodium salt (SBE-β-CD) was used as the host molecule. IMC-SBE-β-CD supramolecular substances were prepared through the "saturated solution method", and their properties and biosecurity were evaluated. Meanwhile, we conducted the AOS tree evaluation system that surpasses existing evaluation approaches based on apoptosis, oxidative stress system, and signaling pathways to investigate the toxicological mechanisms of IMC-SBE-β-CD within human hepatoma SMMC-7721 cells as model organisms. The AOS tree evaluation system aims to offer the comprehensive analysis of the cytotoxic effects of IMC-SBE-β-CD. Our findings showed that IMC-SBE-β-CD had an encapsulation rate of 84.45% and optimal stability at 30 °C. Further, IMC-SBE-β-CD promoted cell growth and reproduction without compromising the integrity of mitochondria and nucleus or disrupting oxidative stress and apoptosis-related pathways. Compared to IMC, IMC-SBE-β-CD is biologically safe and has improved water solubility with the UV absorption property maintained. Our study provides the foundation for the encapsulation of hydrophobic, low-toxicity organic compounds using cyclodextrins and offers valuable insights for future research in this field. [ABSTRACT FROM AUTHOR]

Natural compounds such as thymoquinone (TQ) have recently gained increasing attention in treating glioblastoma (GBM). However, the effects of TQ in reversing drug resistance are not completely understood. Therefore, we aimed to examine TQ impacts on GBM cells with doxorubicin (DOX) resistance and the involvement of the PI3K/Akt/mTOR pathway. GBM cancer U87 and U87/DOX (resistant cells) cells were exposed to DOX and TQ, and cell proliferation was assessed by the MTT assay. ELISA was applied to evaluate cell apoptosis. The expression of apoptotic mediators such as Caspase‐3, Bax, Bcl‐2 and PI3K, Akt, mTOR, P‐gp, and PTEN was assessed via qRT‐PCR and western blot. We found that a combination of TQ and DOX suppressed dose‐dependent cell growth capacity in cells and increased the cytotoxic effects of DOX in resistant cells. In addition, TQ treatment increased DOX‐mediated apoptosis in U87/DOX cell lines via modulating the pro‐ and anti‐apoptotic markers. A combination of TQ and DOX upregulated PTEN and downregulated PI3K, Akt, and mTOR, suppressing this signal transduction in resistant cells. In conclusion, we showed TQ potentiated doxorubicin‐mediated antiproliferative and pro apoptotic function DOX‐resistant glioblastoma cells, which is mediated by targeting and suppressing PI3K/Akt/mTOR signal transduction. [ABSTRACT FROM AUTHOR]

The direct antitumor effect of bevacizumab (BEV) has long been debated. Evidence of the direct antitumor activities of drugs are mainly obtained from in vitro experiments, which are greatly affected by experimental conditions. In this study, we evaluated the effect of BEV-containing medium renewal on the results of in vitro cytotoxicity experiments in A549 and U251 cancer cells. We observed starkly different results between the experiments with and without BEV-containing medium renewal. Specifically, BEV inhibited the tumor cell growth in the timely replacement with a BEV-containing medium but promoted tumor cell growth without medium renewal. Meanwhile, compared with the control, a significant basic fibroblast growth factor (bFGF) accumulation in the supernatant was observed in the group without medium renewal but none in that with replaced medium. Furthermore, bFGF neutralization partially reversed the pro-proliferative effect of BEV in the medium non-renewed group, while exogenous bFGF attenuated the tumor cell growth inhibition of BEV in the medium-renewed group. Our data explain the controversy over the direct antitumor effect of BEV in different studies from the perspective of the compensatory autocrine cytokines in tumor cells. [ABSTRACT FROM AUTHOR]

Exosomes from cancer cells function as carriers to spread or transport specific microRNAs (miRNAs) to distant sites to exert their effects, but the mechanism of exosomal miRNA action in esophageal squamous cell carcinoma (ESCC) has not been fully explained. Therefore, in this study, we were interested in the impact of exosomal miR‐196a‐5p in ESCC progression. We found that miR‐196a‐5p was expressed enriched in clinical tissues, ESCC cells, and exosomes. Functionally, depletion of miR‐196a‐5p impeded ESCC cell growth, migration, and invasion, whereas overexpression of miR‐196a‐5p produced the opposite results. Moreover, enhancement of exosomal miR‐196a‐5p in recipient ESCC cells triggered more intense proliferation and migration. Mechanistically, we identified integral membrane protein 2B (ITM2B) as a direct target of miR‐196a‐5p. Silencing of ITM2B partially counteracted the inhibitory effect of miR‐196a‐5p inhibitors on the malignant phenotype of ESCC. Furthermore, in vivo, lower miR‐196a‐5p levels triggered by the introduction of antagomiR‐196a‐5p resulted in the generation of smaller volume and weight xenograft tumors. Thus, our results demonstrated novel mechanisms of exosomal and intracellular miR‐196a‐5p‐mediated ESCC growth and migration and identify the interaction of miR‐196a‐5p with ITM2B. These works might provide new targets and basis for the development of clinical treatment options for ESCC. [ABSTRACT FROM AUTHOR]

This study presents a short overview of the main 3D polymer composite items that are fabricated by extrusion and stereolithography printing. By coupling polymers traditionally used in additive manufacturing, such as PEGDA, and polymers of biomedical interest, such as PVA, with carbon nanostructures, such as nanodiamond and graphene nanoplates, and conductive polymers, such as PEDOT and PANI, it has been possible to produce objects with specific functional properties exploited for fabricating scaffolds for cell growth and proliferation as well as soft electrodes for developing organic compounds sensing devices and electrocardiogram monitoring systems under real‐time conditions. [ABSTRACT FROM AUTHOR]

The anti‐cancer potential of Cyclopia species (honeybush) has been demonstrated in several models. The present study investigated the effects of aqueous and polyphenol‐enriched (PE) extracts of C. subternata and C. genistoides, as well as mangiferin and hesperidin, on different cell growth parameters in human liver (HepG2) and colon (HT‐29) cancer cells. Mangiferin and hesperidin were most abundant in C. genistoides and C. subternata, respectively. Cyclopia subternata extracts had the highest ferric‐reducing antioxidant capacity. Following exposure of the cells to the extracts and compounds, cell viability, proliferation, and death (apoptosis and autophagy) were determined. Cyclopia subternata extracts reduced cell viability and inhibited cell proliferation the most, associated with depletion of ATP. In HepG2 cells, the PE extracts were less effective than the aqueous extracts in reducing cell viability but more effective in inhibiting cell proliferation. Despite disrupting cell growth, none of the extracts induced apoptosis. The aqueous extracts affected autophagy in both cancer cells. Disruption of mitochondrial membrane integrity by the different extracts, presumably via polyphenol/iron interactions, is postulated to be involved; however, mangiferin and hesperidin had no effect, suggesting that other polyphenols and/or complex interactions between compounds are likely responsible for the differential cytotoxic and/or cytoprotective effects of the extracts. [ABSTRACT FROM AUTHOR]

Glycogen is a highly branched polyglucan utilized as a carbohydrate reserve in major living systems. Industrially, it is used as a prebiotic and in the nanoencapsulation of drugs and nutraceuticals. In this study, optimal fermentation conditions enabling the highest glycogen accumulation in Saccharomyces cerevisiae were experimentally evaluated for possible mass production. Production efficiency was assessed by comparing specific growth rates, specific glycogen production rates, and glycogen yields under each condition. The results demonstrated that fermentation at 30 °C with an aeration rate of 3 vvm using a medium containing 120 g/L glucose without ethanol was optimal for robust cell growth and maximum glycogen yield. Additionally, a rich medium outperformed a minimally defined medium, and a single sugar carbon source, as opposed to mixed sugars, resulted in significantly higher cell growth and glycogen yields (p < 0.05). The optimized fermentation parameters enabled a glycogen production rate of up to 0.232 ± 0.012 g-glycogen/g-cell/h and a glycogen yield of 0.603 ± 0.006 g-glycogen/g-glucose. These results provide meaningful information for future studies and/or large-scale glycogen production using S. cerevisiae. [ABSTRACT FROM AUTHOR]

This paper reports hybrid gels combining a low‐molecular‐weight gelator (LMWG) and a photoinitiated crosslinked polymer gel (PG). The presence of the PG enhanced the stiffness and strength of the gel. The gels were loaded with heparin, and in the hybrid gel, the interpenetrated LMWG and PG networks somewhat restricted its release. In terms of stem cell growth, the hybrid gel significantly improved the performance of the PG because of the presence of the LMWG, which is an excellent substrate for stem cells in its own right. Furthermore, the presence of heparin in the hybrid gels also enhanced stem cell proliferation over longer timescales. Finally, these gels were photopatterned within the well‐plates used for tissue culture, with patterning helping control stem cell proliferation. In summary, these hybrid gels combine the advantageous features of both LMWG and PG: rheological performance is endowed by the PG with stem cell compatibility provided by the LMWG. The hybrid gels also control the release of the bioactive agent heparin and have capacity to be shaped and patterned. Patterned gels such as these, capable of directing stem cell growth, have potential in regenerative medicine. [ABSTRACT FROM AUTHOR]

Cellular senescence, traditionally viewed as a consequence of proliferating and growing cells overwhelmed by extensive stresses and damage, has long been recognized as a critical cellular aging mechanism. Recent research, however, has revealed a novel pathway termed "quiescence-origin senescence", where cells directly transition into senescence from the quiescent state, bypassing cell proliferation and growth. This opinion paper presents a framework conceptualizing a continuum between quiescence and senescence with quiescence deepening as a precursor to senescence entry. We explore the triggers and controllers of this process and discuss its biological implications. Given that the majority of cells in the human body are dormant rather than proliferative, understanding quiescence-origin senescence has significant implications for tissue homeostasis, aging, cancer, and various disease processes. The new paradigm in exploring this previously overlooked senescent cell population may reshape our intervention strategies for age-related diseases and tissue regeneration. [ABSTRACT FROM AUTHOR]

Sodium butyrate (NaB) and sodium propionate (NaP) have recently garnered attention for their role in regulating inflammation and controlling signaling pathways of cell growth and apoptosis, potentially preventing cancer development. However, their therapeutic effect and the underlying mechanisms involved remain elusive in breast cancer. This study aims at investigating the anticancer role of NaB and NaP in different types of breast cancer by assessing their antiproliferative effect on MCF-7 and MDA-MB-231 cells (through an MTT assay), as well as their ability to alter the cell cycle and cyclin expression (using flow cytometry and RT-qPCR, respectively), and to promote apoptosis (using Annexin V-FITC conjugated and sub-G1 phase techniques). MDA-MB-231 cell proliferation was inhibited by NaB and NaP in a dose- and time-dependent manner with respective IC50 values of 2.56 mM and 6.49 mM. Treatment induced cell arrest in the G1 phase which was further supported by the significant reduction in cyclin A2 and cyclin B1 expressions. Finally, NaB, and less significantly NaP, induced apoptosis in a dose-dependent manner with higher concentrations required for MDA-MB-231 than MCF-7. Our findings elucidate the cyclin-dependent inhibitory effect of NaB and NaP on the progression of different breast cancer subtypes, thus highlighting their therapeutic potential in breast cancer. [ABSTRACT FROM AUTHOR]

Moxifloxacin (MOX), a widely used novel antibiotic, may pose ecological risks at its actual environmental concentrations, as has been detected in aquatic systems. However, its ecotoxicity to aquatic organisms and regulatory mechanisms of phosphorus in eutrophic aqueous environments are still limited. This study aimed to analyze its physiological and biochemical parameters, including cellular growth, chlorophyll fluorescence, photosynthetic pigments, oxidative stress biomarkers, and metabolomics to elucidate the toxicity induced by environmental concentrations of MOX in Microcystis aeruginosa at different phosphorus levels. The results revealed that the EC50 values of MOX on M. aeruginosa at different phosphorus concentrations were 8.03, 7.84, and 6.91 μg/L, respectively, indicating MOX toxicity was exacerbated with increasing phosphorus levels. High phosphorus intensified the suppression of chlorophyll fluorescence and photosynthetic pigments, while activating the antioxidant enzyme, indicating severe peroxidation damage. Metabolomic analysis showed MOX induced different discriminating metabolites under different phosphorus levels, and perturbed more biological pathways at higher phosphorus concentrations, such as starch and sucrose metabolism, pyrimidine metabolism, and glycerolipid metabolism. This indicates that phosphorus plays an important role in regulating metabolism in M. aeruginosa exposed to MOX. The findings provide valuable information on the mechanisms involved in cyanobacteria responses to antibiotic stress, and offer a theoretical basis for accurately assessing antibiotic toxicity in eutrophic aqueous environments. [ABSTRACT FROM AUTHOR]

Microfluidics have revolutionized cell culture by allowing for precise physical and chemical environmental control. Coupled with electrodes, microfluidic cell culture can be activated or have its changes sensed in real-time. We used our previously developed reliable and stable microfluidic device for cell growth and monitoring to design, fabricate, and characterize a whole-channel impedance-based sensor and used it to systematically assess the electrical and electrochemical influences of microfluidic channel boundaries coupled with varying electrode sizes, distances, coatings, and cell coverage. Our investigation includes both theoretical and experimental approaches to investigate how design parameters and insulating boundary conditions change impedance characteristics. We examined the system with various solutions using a frequency range of 0.5 Hz to 1 MHz and a modulation voltage of 50 mV. The results show that impedance is directly proportional to electrode distance and inversely proportional to electrode coating, area, and channel size. We also demonstrate that electrode spacing is a dominant factor contributing to impedance. In the end, we summarize all the relationships found and comment on the appropriateness of using this system to investigate barrier cells in blood vessel models and organ-on-a-chip devices. This fundamental study can help in the careful design of microfluidic culture constructs and models that require channel geometries and impedance-based biosensing. [ABSTRACT FROM AUTHOR]

The green tea catechin epigallocatechin gallate (EGCg) has antimicrobial effects on many bacteria. In this study, we investigated the inhibitory effects of EGCg on Bacillus anthracis spores and vegetative cells. The B. anthracis spores were insensitive to EGCg, but the growth of vegetative cells derived from germinated spores was inhibited by EGCg. Moreover, EGCg decreased the minimum inhibitory concentration of penicillin and meropenem for penicillin-resistant B. anthracis. In the penicillin-resistant B. anthracis strain, the transcription levels of the beta-lactamase genes (bla1 and bla2) decreased significantly following the treatment with 50 µg/mL EGCg. These results suggest that the appropriate application of EGCg may effectively control the penicillin-resistant B. anthracis growth and beta-lactamase production. [ABSTRACT FROM AUTHOR]

Conventional methods have failed to show the ability of articular cartilage to completely restore. Decellularized extracellular matrix has achieved interest as a potential biomaterial for cartilage tissue engineering approaches. The decellularized cartilage-derived matrix (CDM) scaffolds retain the native composition of cartilage tissue, providing a conducive microenvironment for cellular growth and differentiation, while exhibit limited mechanical support. Our investigation involved the fabrication of a new hybrid CDM- polyvinyl alcohol (PVA) hydrogel scaffold with four different concentrations (5%, 10%, 15%, and 20%), followed by a comprehensive characterization of the construct's physicochemical, mechanical, and biological properties to elucidate their potential in cartilage regeneration applications. Our results demonstrated that hybridization of the CDM with PVA enhanced the mechanical properties besides ensuring biocompatibility. FTIR and DSC results also confirmed the mechanical improvements in hybrid scaffolds. The hybrid CDM/PVA (15%/5%, 15%/10%, 15%/15%, and 15%/20%) scaffolds showed significantly different compressive strengths (p < 0.0001, p < 0.0004, p < 0.0001, p < 0.012 respectively). Moreover, resazurin test showed cell attachment and growth on all four types of hybrid scaffolds during seven days of three dimensional culture. The cross-linked CDM15%/PVA5% group, demonstrated acceptable mechanical strength, pore size, physicochemical properties, swelling behavior, and cell growth and attachment. Our data indicate that our hybrid CDM/PVA scaffold possess bioactive properties suitable for a promising candidate for cartilage tissue engineering studies. [ABSTRACT FROM AUTHOR]

Abstract Benign prostatic hyperplasia (BPH) is a prevalent condition affecting the male urinary system, with its molecular mechanisms of pathogenesis remaining unclear. Y-27632, a non-isoform-selective Rho kinase inhibitor, has shown therapeutic potential in various diseases but its effects on static factors and fibrosis in BPH remain unexplored. This study investigated human prostate tissues, human prostate cell lines, and BPH rat model using immunofluorescence, flow cytometry, quantitative reverse transcription polymerase chain reaction, western blotting, and cell counting kit-8. ROCK1 and ROCK2 were significantly up-regulated in BPH tissues, correlating with clinical parameters. Y-27632 targeted the inhibition of ROCK1 & ROCK2 expression and inhibited cell proliferation, fibrosis, epithelial-mesenchymal transition (EMT), while induced cell apoptosis in a dose-dependent manner. Moreover, knockdown of either ROCK isoform inhibited fibrosis and EMT, induced apoptosis, while ROCK overexpression had the opposite effects. ROCK downregulation inhibited the β-catenin signaling pathway (such as C-MYC, Snail and Survivin) and decreased β-catenin protein stability, while inhibiting TGF-β/Smad2/3 signaling. At the in vivo level, Y-27632 reversed prostatic hyperplasia and fibrosis in BPH model rats to some extent. Our study sheds light on the therapeutic potential of Y-27632 in regulating prostate cell growth, fibrosis and EMT, and demonstrates for the first time the regulatory effect of ROCK isoforms on prostate cells, providing the basis for future research of ROCK isoform-selective inhibitors.

Objective To investigate the effect of calcium ions on the expression of kallikrein-4 (KLK4) and cell growth of ameloblast, and to provide an experimental basis for calcium ion promoting normal mineralization of enamel. Methods ALC cells were treated with 0, 2.0, 2.5, 3.0, and 3.5 mmol/L CaCl2 for 24 and 48 h. KLK4 expression was analyzed by qRT-PCR and Western blot analysis. The viability of ALC cells was determined by using CCK-8. AnnexinV-FITC/PI dual staining combined with flow cytometry and Hoechst 33342 staining were used to detect the ALC cell cycle and cell apoptosis. The protein expression level of glucose-regulated protein 78 (GRP78) was measured by Western blot analysis. Results After 24 h of treatment with 2.5, 3.0, and 3.5 mmol/L CaCl2, the expression of KLK4 mRNA was increased (P2, the expression of KLK4 protein was increased (P2, the expression of KLK4 mRNA and protein was increased (P2 (P2. Hoechst 33342 staining results showed that 3.0 mmol/L and 3.5 mmol/L CaCl2 may promote apoptosis in ALC cells. Flow cytometry showed that the proportion of G2/M phase cells and the apoptosis rate increased after 3.5 mmol /L CaCl2 treatment for 24 h (P2 groups. After 24 h of treatment with 3.0 mmol/L and 3.5 mmol/L CaCl2, the expression of GRP78 protein was reduced (P2, the expression of GRP78 protein was reduced (P

This study explored the effects of β-1,3/1,6-glucan derived from Saccharomyces cerevisiae cell walls on the growth, survival, and physiological responses of post-larvae (PL) of M. rosenbergii. Over a 3-week period, larvae were fed a formulated egg custard diet containing varying amounts of β-glucan. The findings revealed that incorporating β-glucan into the diet had a substantial positive impact. The inclusion of β-glucans significantly enhanced survival (59.14% for the PL fed 0.2% β-glucan versus 46.58% for the control; p < 0.05), promoted growth (13.58 mg wet weight for the PL fed 0.2% β-glucan versus 9.53 mg for the PL fed the control diet; p < 0.05), and accelerated the time to metamorphosis (26.67 days for the PL fed 0.2% β-glucan versus 28.0 days for the PL fed the control diet). As the amount of β-glucan in the diet increased, larval growth performance consistently improved. The group receiving 0.2% β-glucan exhibited the highest performance in terms of wet and dry weight, total length, and mean production of PL. Furthermore, the study assessed the influence of β-glucan supplementation on larval tolerance to hypersaline stress. Although the differences were not statistically significant, the addition of β-glucan resulted in incremental improvements in the ability of larvae to withstand hypersaline conditions. In conclusion, the dietary supplement containing 0.2% β-glucan exhibited the highest performance among the inclusion levels tested. Further investigation is recommended to determine the nutritional and physiological effects of β-glucan supplementation under salinity stress conditions. [ABSTRACT FROM AUTHOR]

Recurrent spontaneous abortions (RSA) affect reproductive health and increase the risk of subsequent abortions. to investigate the role of KISS-1/GPR-54 signaling in RSA progression. Villus tissue was collected from Rsa patients, and human trophoblastic HTR-8/SVneo cells were used. KISS-1 and GRP54 levels were detected using RT-qPCR and immunohistochemistry. Western blotting was performed to analyze ZO-1 and ZEB1 levels. cell proliferation was determined via ccK-8 and cell clone formation assays. transwell assays were performed to assess cell migration and invasion abilities. KISS-1 was down-regulated in the villus tissues of RSA patients. KISS-1 overexpression dramatically inhibited trophoblast proliferation, migration, and invasion. Mechanistically, ZEB1 expression was down-regulated, whereas ZO-1 expression was up-regulated, after Kiss-1 overexpression. GPR54 silencing neutralized the effect of KISS-1 in HTR-8/SVneo cells. additionally, Kiss-1 overexpression inactivated the PI3K/AKT signaling pathway through GRP54. the KISS-1/GPR-54 signaling axis regulates RSA progression by regulating the PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]

Maximizing production potential of recombinant proteins such as monoclonal antibodies (mAbs) in Chinese Hamster Ovary (CHO) cells is a key enabler of reducing cost of goods of biologics. In this study, we explored various strategies to utilize adenosine mediated effects in biologics manufacturing processes. Results show that supplementation of adenosine increases specific productivity by up to two-fold while also arresting cell growth. Introducing adenosine in intensified perfusion processes in a biphasic manner significantly enhanced overall productivity. Interestingly, adenosine effect was observed to be dependent on the cell growth state. Using specific receptor antagonists and inhibitors, we identified that ENTs (primarily Slc29a1) mediate the uptake of adenosine in CHO cell cultures. Transcriptomics data showed an inverse correlation between Slc29a1 expression levels and peak viable cell densities. Data suggests that in fed-batch cultures, adenosine can be produced extracellularly. Blocking Slc29a1 using ENT inhibitors such as DZD and DP alone or in combination with CD73 inhibitor, PSB12379, resulted in a twofold increase in peak viable cell densities as well as productivities in fed batch – a novel strategy that can be applied to biologics manufacturing processes. This is the first study that suggests that adenosine production/accumulation in CHO cell cultures can potentially regulate the transition of CHO cells from exponential to stationary phase. We also demonstrate strategies to leverage this regulatory mechanism to maximize the productivity potential of biologics manufacturing processes. • Supplementation of adenosine in exponentially growing cells increases specific productivity by two-fold. • Adenosine supplementation boosts overall permeate titer in perfusion cultures. • CHO cells produce extracellular adenosine, which can trigger the shift from exponential to stationary phase in fed batch cultures. • Inhibiting extracellular adenosine uptake in fed batch cultures can lead to 2-fold increase in peak cell density and titer. [ABSTRACT FROM AUTHOR]

Regulatory authorities in biopharmaceutical industry emphasize process design by process understanding but applicable tools that are easy to implement are still missing. Soft sensors are a promising tool for the implementation of the Quality by Design (QbD) approach and Process Analytical Technology (PAT). In particular, the correlation between viable cell counting and oxygen consumption was investigated, but problems remained: Either the process had to be modified for excluding CO 2 in pH control, or complex k L a models had to be set up for specific processes. In this work, a non-invasive soft sensor for simplified on-line cell counting based on dynamic oxygen uptake rate was developed with no need of special equipment. The dynamic oxygen uptake rates were determined by automated and periodic interruptions of gas supply in DASGIP® bioreactor systems, realized by a programmed Visual Basic script in the DASware® control software. With off-line cell counting, the two parameters were correlated based on linear regression and led to a robust model with a correlation coefficient of 0.92. Avoidance of oxygen starvation was achieved by gas flow reactivation at a certain minimum dissolved oxygen concentration. The soft sensor model was established in the exponential growth phase of a Chinese Hamster Ovary fed-batch process. Control studies showed no impact on cell growth by the discontinuous gas supply. This soft sensor is the first to be presented that does not require any specialized additional equipment as the methodology relies solely on the direct measurement of oxygen consumed by the cells in the bioreactor. • Non-invasive soft sensor method for simplified on-line cell counting. • Implemented in exponential growth phase of CHO cells. • Dynamic oxygen uptake rate as only predictor resulted in a correlation coefficient of 0.92. • No negative influences on cell growth as oxygen starvation was prevented. • 5-k cross validation showed RMSE of 0.7 × 106 cells/mL. [ABSTRACT FROM AUTHOR]

Strontium is a potent osteo-stimulative element with a wide safe dosage range. In this work, calcium-strontium phosphate (Ca 2 Sr(PO 4) 2 ; abbreviation: CSP) was synthesized, and 3D-printed CSP-based composite (CSP-T) ceramic scaffolds were prepared by introduction of β-tricalcium phosphate (TCP). When sintering the CSP-T ceramic scaffolds, no new phase was produced, but the strontium from CSP was substituted for calcium in TCP; the TCP additives promoted densification of scaffold filaments, resulting in lower porosity of the CSP-T ceramic scaffolds. The porosity (25.6%–77.1 %) and compressive strength (1.5–44.5 MPa) of CSP-T ceramic scaffolds could be adjusted by altering the amount of TCP additives and the sintering temperature. The CSP-T ceramic scaffolds gradually released ions of calcium, phosphorus and strontium, which were in favor of cell growth and osteogenic differentiation. The CSP-T ceramic scaffolds modified by 25 wt% TCP possessed the greatest performances on enhancing mechanical strength and cellular response, which let them serve as promising bone grafts to treat various sites of bone defects. [ABSTRACT FROM AUTHOR]

Different contributions of dopants based on Yttrium oxide (Y 2 O 3)/titanium(IV) oxide (TiO 2)/carbon nanotube (CNT)/have been incorporated into the polymeric matrix of Polystyrene (PS). These films were fabricated in films due to their feasibility and low cost. The contact angle revealed that the dopants supported the hydrophilicity of the films that Y 2 O 3 /TiO 2 /CNT/PS represents the second highest wettability with (56.15 ± 6.35°) films, while Y 2 O 3 /PS film displays rough surface with the highest wettability with (32.5 ± 0.9°). we examined the impact of Y 2 O 3 /TiO 2 /CNT/PS composite on the viability of a human lung cell-line to better understand its biological behavior. Specifically, we measured cell viability as a means to assess the effect of the composite. The results revealed that at a concentration of 5000 μg/ml, the cell viability was measured to be 31.8 %. However, at a lower concentration of 2.4 μg/ml, the cell viability increased significantly to 117.9 %. These findings highlight a concentration-dependent relationship between the composite and the viability of the human lung cell-line. The observed cell viability exceeding 100 % indicates that the Y 2 O 3 /TiO 2 /CNT/PS composite scaffold had a positive effect on cell proliferation and generation. This suggests that the modified PS scaffold is conducive to cellular growth and can be considered safe for human use. The results imply that the composite scaffold promotes a favorable cellular environment, potentially supporting tissue regeneration and indicating its potential suitability for biomedical applications. The surface morphology shows that the topography of the films was changed clearly with the dopants of Yttrium oxide, titanium(IV) oxide and CNT components. The TiO 2 granules implanted in the PS composition cause an increase in pore size to an average of 2.2 μm. Pore diameters ranging from 100 to 500 μm may enhance cell proliferation, while lower sizes lead to more stable implantable discs. [ABSTRACT FROM AUTHOR]

Circular RNAs (circRNAs) are implicated in the progression of breast cancer (BC). However, the explorations on circRNA IQ motif containing H (circIQCH) in BC progression remain limited. Functional experiments were conducted using in vitro and murine xenograft model assays, respectively. Dual‐luciferase reporter assay and RIP assay detected the associations among circIQCH, miR‐139‐5p, and nuclear factor IB (NFIB). CircIQCH was upregulated in BC, and the silencing of circIQCH repressed BC cell growth, metastasis, and autophagy, arrested cell cycle, promoted cell apoptosis in vitro, and blocked tumor growth in vivo. CircIQCH positively modulated NFIB expression by sponging miR‐139‐5p. Moreover, the deletion of miR‐139‐5p abated the action of circIQCH deficiency on BC cell malignant behaviors. Overexpression of miR‐139‐5p repressed the malignant characteristics of BC cells, while these impacts were abolished by elevating NFIB. Collectively, CircIQCH functioned as an oncogene in BC through upregulating NFIB expression by sponging miR‐139‐5p. [ABSTRACT FROM AUTHOR]

Yeast is one of the co-products of ethanol plants, which can be used as a nutritional supplement in animal feed due to its high protein content. Given the importance of yeast contribution to the nutritional properties of DDG (dried distillers’ grains), the aim of this study was to assess how different levels of aeration affect the biomass production and the quality of yeast providing new insights into yeast production, offering an alternative source of income for the corn ethanol industry. For this purpose, yeasts were grown in a fed-batch process, and different concentrations of aeration in the medium were tested, namely 0.5, 1.0, and 1.5 volume of air per volume of wort per minute (v v−1 min−1). At the end of the cellular biomass production process, yeasts grown with 0.5 (v v−1 min−1) aeration in the reactor showed higher biomass formation (19.86 g L−1), cellular yield (g g−1), and a lower formation of succinic acid (0.70 g L−1) and acetic acid (0.11 g L−1). Aeration influenced an increase of 1.0% in the protein content in yeast. In conclusion, lower levels of aeration in the yeast production process enables more efficient sugar utilization for biomass formation and is a potential strategy to increase the protein content and the commercial value of DDG.

Abstract Background Increased level of serum cholic acid (CA) is often accompanied with decreased CYP2E1 expression in hepatocellular carcinoma (HCC) patients. However, the roles of CA and CYP2E1 in hepatocarcinogenesis have not been elucidated. This study aimed to investigate the roles and the underlying mechanisms of CYP2E1 and CA in HCC cell growth. Methods The proteomic analysis of liver tumors from DEN-induced male SD rats with CA administration was used to reveal the changes of protein expression in the CA treated group. The growth of CA-treated HCC cells was examined by colony formation assays. Autophagic flux was assessed with immunofluorescence and confocal microscopy. Western blot analysis was used to examine the expression of CYP2E1, mTOR, AKT, p62, and LC3II/I. A xenograft tumor model in nude mice was used to examine the role of CYP2E1 in CA-induced hepatocellular carcinogenesis. The samples from HCC patients were used to evaluate the clinical value of CYP2E1 expression. Results CA treatment significantly increased the growth of HCC cells and promoted xenograft tumors accompanied by a decrease of CYP2E1 expression. Further studies revealed that both in vitro and in vivo, upregulated CYP2E1 expression inhibited the growth of HCC cells, blocked autophagic flux, decreased AKT phosphorylation, and increased mTOR phosphorylation. CYP2E1 was involved in CA-activated autophagy through the AKT/mTOR signaling. Finally, decreased CYP2E1 expression was observed in the tumor tissues of HCC patients and its expression level in tumors was negatively correlated with the serum level of total bile acids (TBA) and gamma-glutamyltransferase (GGT). Conclusions CYP2E1 downregulation contributes to CA-induced HCC development presumably through autophagy regulation. Thus, CYP2E1 may serve as a potential target for HCC drug development.

Gluconobacter oxydans is an important Gram-negative industrial microorganism that produces vitamin C and other products due to its efficient membrane-bound dehydrogenase system. Its incomplete oxidation system has many crucial industrial applications. However, it also leads to slow growth and low biomass, requiring further metabolic modification for balancing the cell growth and incomplete oxidation process. As a non-model strain, G. oxydans lacks efficient genome editing tools and cannot perform rapid multi-gene editing and complex metabolic network regulation. In the last 15 years, our laboratory attempted to deploy multiple CRISPR/Cas systems in different G. oxydans strains and found none of them as functional. In this study, Cpf1-based or dCpf1-based CRISPRi was constructed to explore the targeted binding ability of Cpf1, while Cpf1–FokI was deployed to study its nuclease activity. A study on Cpf1 found that the CRISPR/Cpf1 system could locate the target genes in G. oxydans but lacked the nuclease cleavage activity. Therefore, the CRISPR/Cpf1–FokI system based on FokI nuclease was constructed. Single-gene knockout with efficiency up to 100% and double-gene iterative editing were achieved in G. oxydans. Using this system, AcrVA6, the anti-CRISPR protein of G. oxydans was discovered for the first time, and efficient genome editing was realized.

(1) The study aims to utilize a reported approach for culturing mesophilic bacteria from a plastic waste environment; (2) The work revived mesophilic microbial population from an aged PET recycling site using a culture-based approach, and determined the purified isolates in genus level in 16S identification; (3) A total of 59 bacterial isolates were obtained, in which microbial species, including Pseudomonas spp, Rhodococcus spp, and Burkholderia spp were identified as abundance. It was observed that the surviving microbes favoured sodium propionate as a short-chain carbon source for growth, rather than the intended plastic substrate, PET. The preference of sodium propionate utilization by several bacterial isolates, including 5601W (detected as Rhodococcus spp.), 5601Y, 7801, and 7802 (detected as Burkholderia spp.), was confirmed through growth curve analysis and cell enumeration conducted in a medium where sodium propionate served as the sole carbon source.; (4) The microbial demonstration revealed the metabolic complex of microbial communities in the environment and indicated the challenges associated with bacterial isolation from environments with accumulated plastic waste.

Background: While pluripotent stem cell (PSC) therapies move toward clinical and commercial applications at a rapid rate, manufacturing reproducibility and robustness are notable bottlenecks in regulatory approval. Therapeutic applications of PSCs require large cell quantities to be generated under highly robust, well-defined, and economically viable conditions. Small-scale and short-term process optimization, however, is often performed in a linear fashion that does not account for time needed to verify the bioprocess protocols and analysis methods used. Design of a reproducible and robust bioprocess should be dynamic and include a continuous effort to understand how the process will respond over time and to different stresses before transitioning into large-scale production where stresses will be amplified. Methods: This study utilizes a baseline protocol, developed for the short-term culture of PSC aggregates in Vertical-Wheel® bioreactors, to evaluate key process attributes through long-term (serial passage) suspension culture. This was done to access overall process robustness when performed with various commercially available media and cell lines. Process output variables including growth kinetics, aggregate morphology, harvest efficiency, genomic stability, and functional pluripotency were assessed through short and long-term culture. Results: The robust nature of the expansion protocol was demonstrated over a six-day culture period where spherical aggregate formation and expansion were observed with high-fold expansions for all five commercial media tested. Profound differences in cell growth and quality were revealed only through long-term serial expansion and in-vessel dissociation operations. Some commercial media formulations tested demonstrated maintenance of cell growth rates, aggregate morphology, and high harvest recovery efficiencies through three bioreactor serial passages using multiple PSC lines. Exceptional bioprocess robustness was even demonstrated with sustained growth and quality maintenance over 10 serial bioreactor passages. However, some commercial media tested proved less equipped for serial passage cultures in bioreactors as cultures led to cell lysis during dissociation, reduction in growth rates, and a loss of aggregate morphology. Conclusions: This study demonstrates the importance of systematic selection and testing of bioprocess input variables, with multiple bioprocess output variables through serial passages to create a truly reproducible and robust protocol for clinical and commercial PSC production using scalable bioreactor systems. [ABSTRACT FROM AUTHOR]

Objective Animal Studied Procedures Results Conclusions The application of canine amniotic membrane (cAM) for corneal reconstruction is widely used in the veterinary field. However, the information on biological properties and alternative forms of cAM for corneal wound healing is limited. This study aimed to investigate the proteomic profiles and corneal wound healing properties of cAM, cAM extract (cAME), and lyophilized cAM extract (cAMX).A total number of 14 cAMs were sterilely harvested from healthy full‐term puppies and randomly divided into three different forms: cAM (n = 14), cAME (n = 14), and cAMX (n = 14).Each form of cAMs was subjected to proteomic analysis using label‐free liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS), followed by bioinformatic analysis. The proteins were classified into properties by comparing them with the literature search on human amniotic membrane (hAM) properties and the effect on corneal wound healing when given topically.The analyses identified 8136 proteins in cAM, 8211 proteins in cAME, and 7093 proteins in cAMX. A total number of 100 proteins were matched with proteins in hAM properties and were classified into anti‐inflammatory, anti‐fibrotic, anti‐microbial, anti‐angiogenic, promotion of epithelialization, analgesic, and support cell adhesion and growth properties. Furthermore, proteins with corneal wound healing effects were identified in cAME and cAMX.cAM and its extracts contain numerous proteins, including proteins related to corneal wound healing properties. Additionally, cAME and cAMX showed proteins involved in corneal wound healing and their potential benefits for topical use in ophthalmology. [ABSTRACT FROM AUTHOR]

This study investigates differences in focal adhesion (FA) morphology and Talin cleavage levels between transformed and non-transformed cell lines. Utilizing fluorescently tagged wild-type Talin and Talin mutants with calpain cleavage site mutations, FA structures were visualized. Mutations in different Talin cleavage sites showed varying impacts on FA morphology and distribution across melanoma cell lines (Meljuso, A375P, A2058) and a non-transformed cell line (HFF). Western blot analysis, ratiometric fluorescence intensity-based measurements, and FRAP experiments revealed higher Talin cleavage levels within FAs of transformed cell lines compared to non-transformed cells. Additionally, growth assays indicated that reducing calpain cleavage levels attenuated transformed cell growth. These findings suggest that Talin cleavage level is crucial for FA morphology and assembly, with higher levels observed in transformed cells, influencing their growth dynamics. [ABSTRACT FROM AUTHOR]

Ovarian cancer (OVC) is one of the most common causes of cancer-related deaths in women worldwide. Despite advancements in detection and therapy, the prognosis of OVC remains poor due to late diagnosis and the lack of effective therapeutic options at advanced stages. Therefore, a better understanding of the biology underlying OVC is essential for the development of effective strategies for early detection and targeted therapies. Nuclear receptors (NRs) are a superfamily of 48 transcription factors that, upon binding to their specific ligand, play a vital role in regulating various cellular processes such as growth, development, metabolism, and homeostasis. Accumulating evidence from several studies has shown that their aberrant expression is associated with multiple human diseases. Numerous NRs have shown significant effects in the development of various cancers, including OVC. This review summarizes the recent findings on the role of NRs inOVC, as well as their potential as prognostic and therapeutic markers. Further, the basic structure and signaling mechanism of NRs have also been discussed briefly. Moreover, this review highlights their cellular and molecular mechanisms in chemoresistance and chemosensitization. Further, the clinical trials targeting NRs for the treatment of OVC have also been discussed. [ABSTRACT FROM AUTHOR]

The extraction and recovery of valuable metals from various spent catalysts via bioleaching represents a green, low-carbon and eco-friendly process. However, the pulp density of spent catalysts is usually 1.0% or lower owing to their toxicity, denoting low process capacity and poor practical potential. In this study, an intensified bioleaching strategy was used for the first time to promote the release efficiencies of both Co and Mo from a spent Co–Mo catalyst at a high pulp density of 10% by supplementing extracellular polymeric substances (EPSs). The results showed that the addition of 0.6 g L−1 EPSs harvested a maximum release of 73.6% for Co and 72.5% for Mo after 9 days of contact, with an evident elevation of 22.6% for Co and 24.4% for Mo, in contrast to no addition, respectively. The added EPS not only promoted the growth of plankton cells to produce more active molecules but also boosted the adhesion of leaching cells to the spent catalyst to form stable aggregates. Moreover, the resulting aggregates allowed for the gathering and confinement of the active small molecules, including Fe3+ and Fe2+, inside the micro-areas between the spent catalysts and the cells for quick electronic transfer as an interface oxidation/reduction reaction to free both Co and Mo from the spent catalyst. [ABSTRACT FROM AUTHOR]

Objectives: The development of effective treatments for non-small cell lung cancer (NSCLC), particularly targeting the KRASG12C mutation, remains a challenge. In this study, we investigated the therapeutic potential of VT204, a small molecule inhibitor of KRASG12C, in NSCLC. Methods: To achieve the objectives, we conducted a comprehensive set of experimental methods. In vitro experiments involved the investigation of VT204 on proliferation, apoptosis, cell cycle dynamics, migration, invasion, and on the RAF/MEK/ERK signaling pathway in NCI-H358 cells. In addition, in vivo experiments were performed to evaluate the influence of VT204 on tumor growth. Results: We demonstrated that VT204 effectively suppressed cell proliferation in NCI-H358 cells, with significant inhibition observed at a concentration of 8 μM. Colony formation assays further supported the inhibitory effect of VT204 on NCI-H358 cell growth. Moreover, VT204 exhibited notable effects on suppressing migration and invasion capacities of NCI-H358 cells, indicating its potential as a metastasis-inhibiting agent. Mechanistic investigations revealed that VT204 induced apoptosis and G2M-phase cell cycle arrest in NCI-H358 cells. Additionally, VT204 modulated the RAF/MEK/ERK signaling pathway, leading to reduced phosphorylation of ERK. In vivo studies using xenograft models confirmed the inhibitory effect of VT204 on NCI-H358 tumor growth. Conclusion: These findings highlight VT204 as a promising therapeutic candidate for NSCLC targeting the KRASG12C mutation. [ABSTRACT FROM AUTHOR]