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Introduction: In lambs, the function of the rumen is incompletely developed at weaning, and the inclusion of yeast cultures in the diet can profoundly influence the morphological and functional development of the rumen. Methods: In this study, the effects of Saccharomyces cerevisiae and Kluyveromyces marxianus (NM) yeast co-cultures on ruminal histomorphology were assessed, and corresponding transcriptomic changes within the rumen epithelium were identified. In total, 24 lambs were grouped into four groups of six lambs including a control (C) group fed a basal diet, and N, M, and NM groups in which lambs were fed the basal diet, respectively, supplemented with Saccharomyces cerevisiae yeast cultures (30 g/d per head), Kluyveromyces marxianus yeast cultures (30 g/d per head), and co-cultures of both yeasts (30 g/d per head), the experiment lasted for 42 d. Results: In morphological analyses, lambs from the NM group presented with significant increases in papilla length, papilla width, and epithelial thickness in the rumen relative to lambs in the C group (p < 0.05). Transcriptomic analyses revealed 202 genes that were differentially expressed between samples from the C and NM groups, with the largest proportion of these genes being associated with the oxidative phosphorylation pathway. In a weighted gene coexpression network analysis, a positive correlation was observed between the MEgreen and MEpurple modules and rumen morphology. Of these modules, the MEgreen module was found to be more closely linked to fatty acid metabolism and oxidative phosphorylation, whereas the MEpurple module was linked to oxidative phosphorylation and fatty acid degradation. Ultimately, these results suggest that dietary supplementation with NM has driven the degradation of fatty acids, the induction of oxidative phosphorylation, the acceleration of lipid metabolism, the production of ATP to sustain ruminal growth, and the maintenance of intracellular NADH/NAD+ homeostasis on weaned lambs and is superior to single yeast fermentation. Discussion: These results thus offer a theoretical foundation for further studies examining the mechanisms through which NM cultures can influence ruminal development in lambs. [ABSTRACT FROM AUTHOR]

Purpose: Pancreatic Ductal Adenocarcinoma (PDAC) remains a challenging disease due to its complex biology and aggressive behavior with an urgent need for efficient therapeutic strategies. To assess therapy response, pre-clinical PDAC organoid-based models in combination with accurate real-time monitoring are required. Methods: We established stable live-imaging organoid/peripheral blood mononuclear cells (PBMCs) co-cultures and introduced OrganoIDNet, a deep-learning-based algorithm, capable of analyzing bright-field images of murine and human patient-derived PDAC organoids acquired with live-cell imaging. We investigated the response to the chemotherapy gemcitabine in PDAC organoids and the PD-L1 inhibitor Atezolizumab, cultured with or without HLA-matched PBMCs over time. Results obtained with OrganoIDNet were validated with the endpoint proliferation assay CellTiter-Glo. Results: Live cell imaging in combination with OrganoIDNet accurately detected size-specific drug responses of organoids to gemcitabine over time, showing that large organoids were more prone to cytotoxic effects. This approach also allowed distinguishing between healthy and unhealthy status and measuring eccentricity as organoids' reaction to therapy. Furthermore, imaging of a new organoids/PBMCs sandwich-based co-culture enabled longitudinal analysis of organoid responses to Atezolizumab, showing an increased potency of PBMCs tumor-killing in an organoid-individual manner when Atezolizumab was added. Conclusion: Optimized PDAC organoid imaging analyzed by OrganoIDNet represents a platform capable of accurately detecting organoid responses to standard PDAC chemotherapy over time. Moreover, organoid/immune cell co-cultures allow monitoring of organoid responses to immunotherapy, offering dynamic insights into treatment behavior within a co-culture setting with PBMCs. This setup holds promise for real-time assessment of immunotherapeutic effects in individual patient-derived PDAC organoids. [ABSTRACT FROM AUTHOR]

In this study, an ultrasonic synergistic semi-bionic method was explored to extract intracellular polysaccharides from Sanghuangporus vaninii and Pleurotus sapidus liquid co-cultures. By conducting single-factor experiments, Plackett-Burman design, and response surface design, the impact of 12 factors on co-culture intracellular polysaccharide (CC-IPS) extraction content, including extraction material-liquid ratios, extraction time, extraction temperature, and extraction solution pH were investigated. The antioxidant capacity of CC-IPS in vitro was evaluated by the DPPH and ABTS+ radical scavenging rates. Results showed that, the optimal extraction process parameters for the first, second, and third extractions, respectively, included material-to-liquid ratios of 1: 17, 1:40, and 1:30 (g/mL), extraction times of 31, 30, and 30 min, extraction temperatures of 44, 40, and 50 °C, and extraction solution pH levels of 3, 8, and 10. Under these optimal conditions, the CC-IPS extraction content was 86.4921±1.1924 mg/g, closed to the predicted value of 85.3981 mg/g. Additionally, the IC50 values of CC-IPS in the DPPH and ABTS+ free radical scavenging assays were determined to be 0.2425 and 0.1376 mg/mL, respectively. This study offers a theoretical foundation for the efficient utilization of intracellular polysaccharides in liquid fungal co-cultures. [ABSTRACT FROM AUTHOR]

Introduction and purpose Diabetes is a disease resulting from impaired action or secretion of insulin. The number of patients currently amounts to approximately 422 million, and approximately 1.5 million deaths per year are directly attributed to this disease. Diabetes significantly reduces the quality of life and, if poorly controlled, can lead to serious complications. Mesenchymal stem cells are multipotent cells capable of differentiation. They have the ability to self-renew and have a modulating function. There are reports that they can be used in the treatment of this disease. The aim of the review was to present a new method of therapy and their possible effects. Material and methods The review was based on articles obtained from PubMed scientific database in the years 2015-2023, using the following keywords: diabetes mellitus, stem cells, diabetes complications. Results Implanted stem cells are able to transform into cells that produce and secrete insulin, and also enable better glycemic control. They can alleviate chronic inflammation and reduce fibrosis. Taking into account the complications that occur during long-term diabetes, the use of stem cells may be associated with improving the function of specific organs and tissues such as the kidneys, heart, eyes and nerves. Studies also report a positive effect of these cells on the healing process of wounds and ulcers. Conclusions Stem cells are a promising object of analysis. They can control glycemia and have a positive effect on the functioning of the kidneys, heart, eyes and nerves, and accelerate wound healing, but further, extensive research is needed to assess the effectiveness and safety of this therapy. [ABSTRACT FROM AUTHOR]

Background Peyronie's disease (PD) is a fibrotic disorder affecting the penile tunica albugínea, with unclear pathophysiology despite centuries of recognition. Aim This scoping review maps the effects of interventions in basic PD research, synthesizing evidence from in vivo and in vitro studies to guide future investigation. Methods In October–November 2023, a systematic search was conducted across PubMed, Embase (Ovid), Science of Web, and Scopus, following SRYCLE's guidelines. Relevant studies were screened for data on interventions targeting PD in vivo and in vitro, with no language or time restrictions. Outcomes Primary outcomes included changes in extracellular matrix (ECM) proteins, myofibroblast activity, and plaque size. Results Of 683 articles screened, 40 studies were included. Key interventions such as phosphodiesterase inhibitors and stem cell therapies reduced ECM proteins and myofibroblast activity, particularly in early-stage PD models. However, none of the studies adhered to the ARRIVE guidelines, highlighting a gap in reporting standards. Clinical translation Findings suggest potential benefits of early and multimodal treatment strategies, but further human trials are needed to bridge the gap in clinical practice. Strengths and limitations This review systematically synthesizes animal and cellular research on PD, highlighting significant preclinical findings. However, the lack of standardized reporting and limited human studies restricts direct clinical applicability. Conclusion Further research should prioritize adherence to reporting standards, optimize treatment timing, and explore combination therapies to advance PD management. [ABSTRACT FROM AUTHOR]

Salmonid rickettsial septicemia (SRS) is a critical sanitary problem in the Chilean aquaculture industry since it induces the highest mortality rate in salmonids among all infectious diseases. Piscirickettsia salmonis, a facultative intracellular bacterium, is the biological agent of SRS. In Chile, two genogroups of P. salmonis, designated as LF-89 and EM-90, have been identified. Previous studies suggested that their cohabitation triggers the expression of virulence effectors, which may be related to a higher pathogenicity in salmonids during co-infection with both P. salmonis genogroups. Therefore, we aimed to evaluate if the physical contact between two isolates from LF-89 and EM-90 is necessary to activate this effect. Through a spatially separated in vivo co-culture inside Atlantic salmon (Salmo salar) post smolts and RNA-seq analysis, we compared the differentially expressed genes (DEGs) with previous results from an in vivo mixed co-culture. The results showed that although the LF-89-like isolate and the EM-90-like isolate had a similar DEG profile under both co-culture conditions, important virulence factors observed during the mixed co-cultures (i.e., flagellar-related genes, CydD, and NCS2) were absent in the spatially separated co-cultures. Hence, the synergistic effect linked to increased pathogenicity to the host may be driven by the physical co-localization and contact between the P. salmonis LF-89-like and EM-90-like isolates. [ABSTRACT FROM AUTHOR]

Bioremediation, involving the strategic use of microorganisms, has proven to be a cost-effective alternative for restoring areas impacted by persistent contaminants such as polycyclic aromatic hydrocarbons (PAHs). In this context, the aim of this study was to explore hydrocarbon-degrading microbial consortia by prospecting native species from soils contaminated with blends of diesel and biodiesel (20% biodiesel/80% diesel). After enrichment in a minimal medium containing diesel oil as the sole carbon source and based on 16S rRNA, Calmodulin and β-tubulin gene sequencing, seven fungi and 12 bacteria were identified. The drop collapse test indicated that all fungal and four bacterial strains were capable of producing biosurfactants with a surface tension reduction of ≥20%. Quantitative analysis of extracellular laccase production revealed superior enzyme activity among the bacterial strains, particularly for Stenotrophomonas maltophilia P05R11. Following antagonistic testing, four compatible consortia were formulated. The degradation analysis of PAHs and TPH (C5–C40) present in diesel oil revealed a significantly higher degradation capacity for the consortia compared to isolated strains. The best results were observed for a mixed bacterial-fungal consortium, composed of Trichoderma koningiopsis P05R2, Serratia marcescens P10R19 and Burkholderia cepacia P05R9, with a degradation spectrum of ≥91% for all eleven PAHs analyzed, removing 93.61% of total PAHs, and 93.52% of TPH (C5–C40). Furthermore, this study presents the first report of T. koningiopsis as a candidate for bioremediation of petroleum hydrocarbons. [ABSTRACT FROM AUTHOR]

The December 2024 issue of Virologica Sinica features research articles on topics such as viral load dynamics in Omicron BA. 2 patients, immunological responses to genital herpes, and the discovery of severe fever with thrombocytopenia virus in Beijing. Additionally, the issue includes studies on mutations in viral proteins, the development of Nipah virus glycoprotein vaccines, and a reverse genetics system for bovine rotavirus. The issue also presents a case study on human norovirus infection and a mouse model for studying severe human adenovirus infections. The cover article discusses the development of a live attenuated hepatitis A virus vaccine and its genetic analysis for vaccine development. [Extracted from the article]

Colorectal cancer is a common malignant tumor. Revealing the interaction between tumor cells and tumor microenvironment （TME） can provide new targets for the prevention or therapy of colorectal cancer. Three-dimensional models such as cell spheroids, organoids, organ chips, and three-dimensional printing have been widely used to simulate TME in colorectal cancer, which can better reproduce the structure and heterogeneity of TME. This paper reviews the progress in the application of three-dimensional models in TME research of colorectal cancer to provide references for future research. [ABSTRACT FROM AUTHOR]

AIM: This study investigates the apoptotic and autophagic effects of atractylodin on lung cancer cells, elucidating the underlying molecular mechanisms. METHODS: Non-small cell lung cancer （NSCLC） A549 and H460 cells, in addition to non-cancerous HBE cells, were cultured in vitro. The effects of atractylodin at various concentrations on cell viability were assessed using CCK-8 assay. Apoptotic effects were evaluated through Hoechst staining and flow cytometry, while Western blot analysis was performed to detect changes in protein expressions associated with apoptosis and autophagy, including P62, beclin-1, microtubule-associated protein 1 light chain 3 （LC3）, Kelch-like epichlorohydrin（ ECH）-associated protein-1（ Keap-1）, nuclear factor E2-related factor 2（ Nrf2）, heme oxygenase-1（ HO-1）, and NAD（P）H: quinone oxidoreductase 1 （NQO1）. Autophagic flux was further analyzed using acridine orange （AO） staining, and immunofluorescence for LC3 and Nrf2. Additionally, autophagy inhibition experiments were conducted using chloroquine （CQ）, followed by analyses of autophagy and apoptosis. Reactive oxygen species （ROS） levels were quantified using DCFH-DA. RESULTS: Treatment with atractylodin significantly reduced the viability of A549 and H460 lung cancer cells, promoting apoptosis and inducing autophagy. This was evidenced by an increase in acidic autophagic vesicles, upregulation of LC3 and beclin-1, and downregulation of P62. Inhibition of autophagy by chloroquine reversed atractylodin-induced apoptosis. Moreover, atractylodin heightened ROS production, inhibited Keap-1, and stimulated the expression of Nrf2, HO-1 and NQO1. CONCLUSION: Atractylodin effectively inhibits the proliferation of lung cancer cells by inducing apoptosis and autophagy. These effects are mediated through the modulation of the ROS/Nrf2/HO-1 signaling pathway, underscoring its potential as a therapeutic agent in lung cancer treatment. [ABSTRACT FROM AUTHOR]

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Many tissues have a laminar structure, but there are limited technologies for establishing laminar co-cultures for in vitro testing. Here, we demonstrate that collagen–alginate–fibrin (CAF) hydrogel scaffolds produced using the reactive jet impingement bioprinting technique can produce osteochondral laminar co-cultures with well-defined interfaces between cell types and high cell densities to support cell–cell interaction across the interfaces. The influence of cell density and the presence of the two cell types on the production of extracellular matrix (ECM) and the emergent mechanical properties of gels is investigated using IHC, ELISA, gel mass, and the compression modulus. The results indicate that high-cell-density cultures and co-cultures with these specific cell types produce greater levels of ECM and a more biomimetic in vitro culture than low-cell-density cultures. In laminar scaffolds produced using TC28a2 chondrocytes and SaoS-2 osteoblasts, both cell density and the presence of the two cell types enhance ECM production and the mechanical properties of the cultures, presenting a promising approach for the production of more biomimetic in vitro models. [ABSTRACT FROM AUTHOR]

Introduction: Recent legislative advancements emphasize the need for alternative methods in assessing efficacy and safety while adhering to the 3Rs principles (reduce, replace, and refine) of animal experimentation. In this context, Galleria mellonella has become a well-established model in safety and efficacy research. However, existing studies predominantly focus on its use in injectable routes concerning pathogens and chemical compounds, leaving a significant gap in understanding other administration pathways, particularly topical application. To address this gap and contribute to the validation of G. mellonella as a model for cutaneous absorption studies, our investigation compares the toxicity data of gallic acid (GA) in mammalian models, alternative animal models such as G. mellonella and Caenorhabditis elegans, as well as monolayer cell cultures, providing insights valuable for future research. Methods: This was an experimental study aimed at evaluating the toxicity and permeation of gallic acid (GA) using different in vivo and in vitro alternative models, including G. mellonella, C. elegans, and monolayer cell cultures. Our study specifically evaluated the toxicity of GA in solution using human keratinocytes (HaCat), human dermal fibroblasts (HDFa), and human liver cell lines (HepG2), in addition to invertebrate animal models of C. elegans and semi-solid formulations in G. mellonella via topical administration. The results demonstrated GA's higher toxicity in C. elegans, followed by sequential susceptibility in HaCat, HepG2, and HDFa cells. Results: Surprisingly, G. mellonella displayed a notably high tolerance to GA, presenting no discernible alterations in cellular immune responses in injectable nor topical administration. Observations of the humoral immune response in G. mellonella larvae showcased melanization both administration conditions, indicating absorption following topical administration of higher GA concentrations. Conclusions: GA demonstrated low toxicity in cellular models, whereas G. mellonella revealed promise as a methodology for topical toxicity testing in cutaneous absorption. However, further research is essential to validate its efficacy. These findings demonstrated the potential to establish correlations in toxicology data between various alternative methodologies and mammals, potentially aiding in the prediction of toxicity in humans. [ABSTRACT FROM AUTHOR]

Microbial interactions within specific environments yield diverse changes, with impacts ranging from harmful to beneficial depending on the resulting compounds. Laboratory co-culturing enables microbes to interact in controlled settings, fostering the production of beneficial compounds influenced by the type of microbial interactions. Methods like direct cellto-cell contact, shared liquid mediums, and membrane separation simulate natural microbial ecosystems, enhancing metabolic exchanges critical for drug production. Such techniques have garnered attention for their potential to activate silent metabolic pathways and stimulate cooperative interactions, leading to the discovery of novel drugs. Microbial coculturing can uncover unexpressed biosynthetic pathways, revealing new metabolites absent in monocultures. This collaborative culturing maximizes resource use, demonstrating cost-effectiveness—an advantage in large-scale drug production. However, both positive and negative outcomes from these interactions can influence medication quality and yield. To optimize the benefits and address challenges in microbial coculturing, ongoing studies aim to refine these techniques. This article explores microbial co-culturing methodologies and highlights co-culture examples where novel metabolite production can contribute to drug discovery. [ABSTRACT FROM AUTHOR]

IntroductionIn lambs, the function of the rumen is incompletely developed at weaning, and the inclusion of yeast cultures in the diet can profoundly influence the morphological and functional development of the rumen.MethodsIn this study, the effects of Saccharomyces cerevisiae and Kluyveromyces marxianus (NM) yeast co-cultures on ruminal histomorphology were assessed, and corresponding transcriptomic changes within the rumen epithelium were identified. In total, 24 lambs were grouped into four groups of six lambs including a control (C) group fed a basal diet, and N, M, and NM groups in which lambs were fed the basal diet, respectively, supplemented with Saccharomyces cerevisiae yeast cultures (30 g/d per head), Kluyveromyces marxianus yeast cultures (30 g/d per head), and co-cultures of both yeasts (30 g/d per head), the experiment lasted for 42 d.ResultsIn morphological analyses, lambs from the NM group presented with significant increases in papilla length, papilla width, and epithelial thickness in the rumen relative to lambs in the C group (p < 0.05). Transcriptomic analyses revealed 202 genes that were differentially expressed between samples from the C and NM groups, with the largest proportion of these genes being associated with the oxidative phosphorylation pathway. In a weighted gene coexpression network analysis, a positive correlation was observed between the MEgreen and MEpurple modules and rumen morphology. Of these modules, the MEgreen module was found to be more closely linked to fatty acid metabolism and oxidative phosphorylation, whereas the MEpurple module was linked to oxidative phosphorylation and fatty acid degradation. Ultimately, these results suggest that dietary supplementation with NM has driven the degradation of fatty acids, the induction of oxidative phosphorylation, the acceleration of lipid metabolism, the production of ATP to sustain ruminal growth, and the maintenance of intracellular NADH/NAD+ homeostasis on weaned lambs and is superior to single yeast fermentation.DiscussionThese results thus offer a theoretical foundation for further studies examining the mechanisms through which NM cultures can influence ruminal development in lambs.

T cell receptor-engineered T cells (TCR-Ts) therapy is promising for cancer immunotherapy. Most studies have focused on identifying tumor-specific T cell receptors (TCRs) through predicted tumor neoantigens. However, current algorithms for predicting tumor neoantigens are unreliable and many neoantigens are derived from non-coding regions. Thus, the technological platform for identifying tumor-specific TCRs using natural antigens expressed on tumor cells is urgently needed. In this study, tumor organoids-enriched tumor infiltrating lymphocytes (oeT) were obtained by repeatedly stimulation of autologous patient-derived organoids (PDO) in vitro. The oeT cells specifically responded to autologous tumor PDO by detecting CD137 expression and the secretion of IFN-γ using enzyme-linked immunospot assay. The measurement of oeT cell-mediated killing of three-dimensional organoids was conducted using a caspase3/7 flow cytometry assay kit. Subsequently, tumor-specific T cells were isolated based on CD137 expression and their TCRs were identified through single-cell RT-PCR analysis. The specificity cytotoxicity of TCRs were confirmed by transferring to primary peripheral blood T cells. The co-culture system proved highly effective in generating CD8+ tumor-specific oeT cells. These oeT cells effectively induced IFN-γ secretion and exhibited specificity in killing autologous tumor organoids, while not eliciting a cytotoxic response against normal organoids. The analysis conducted by TCRs revealed a significant expansion of T cells within a specific subset of TCRs. Subsequently, the TCRs were cloned and transferred to peripheral blood T cells generation engineered TCR-Ts, which adequately recognized and killed tumor cell in a patient-specific manner. The co-culture system provided an approach to generate tumor-specific TCRs from tumor-infiltrating lymphocytes of patients with colorectal cancer, and tumor-specific TCRs can potentially be used for personalized TCR-T therapy. [ABSTRACT FROM AUTHOR]

Triple‐negative breast cancer (TNBC) is the most invasive type of breast cancer with high risk of brain metastasis. To better understand interactions between breast tumors with the brain extracellular matrix (ECM), a 3D cell culture model is implemented using a thiolated hyaluronic acid (HA‐SH) based hydrogel. The latter is used as HA represents a major component of brain ECM. Melt‐electrowritten (MEW) scaffolds of box‐ and triangular‐shaped polycaprolactone (PCL) micro‐fibers for hydrogel reinforcement are utilized. Two different molecular weight HA‐SH materials (230 and 420 kDa) are used with elastic moduli of 148 ± 34 Pa (soft) and 1274 ± 440 Pa (stiff). Both hydrogels demonstrate similar porosities. The different molecular weight of HA‐SH, however, significantly changes mechanical properties, e.g., stiffness, nonlinearity, and hysteresis. The breast tumor cell line MDA‐MB‐231 forms mainly multicellular aggregates in both HA‐SH hydrogels but sustains high viability (75%). Supplementation of HA‐SH hydrogels with ECM components does not affect gene expression but improves cell viability and impacts cellular distribution and morphology. The presence of other brain cell types further support numerous cell–cell interactions with tumor cells. In summary, the present 3D cell culture model represents a novel tool establishing a disease cell culture model in a systematic way. [ABSTRACT FROM AUTHOR]

Biosilicate/carbon composites were fabricated in the form of highly porous foams by the polymer-derived ceramic route, and their biological response was analysed. Two different commercial silicone polymers (a poly-methyl-siloxane, MK, and a polymethyl-phenyl-silsesquioxane, H44) were considered as a silica source, mixed with active fillers yielding Na 2 O, CaO and P 2 O 5. The samples were heat treated either in air or in N 2 atmosphere to obtain products resembling the known Biosilicate® glass-ceramic, with or without free carbon. All fabricated samples exhibited acellular in-vitro bioactivity upon immersion in SBF as well as antibacterial activity against S. aureus and E. coli. Direct contact cell viability test, assessed by using a WST-8 assay, indicated that both carbon-containing and carbon-free samples were cytocompatible. [ABSTRACT FROM AUTHOR]

AIM: To investigate the effect of a disintegrin and metalloproteinase (ADAM) domain-like decy-sin 1(ADAMDEC1) knockdown on the proliferation, migration and invasion of pancreatic carcinoma cells. METHODS: Expression levels of ADAMDEC1 in pancreatic carcinoma tissues were analyzed using the GEPIA and UALCAN online databases. Western blot analysis was employed to detect the protein expression levels of ADAMDEC1 in pancreatic carcinoma cell lines (MIA PaCa-2 and PANC-1) and pancreatic ductal cell line (hTERT-HPNE). The effects of ADAMDEC1 knockdown on cell proliferation, migration and invasion were evaluated using CCK-8, colony formation, wound-healing and Transwell assays. Additionally, Western blot analysis was used to detect the effects of ADAMDEC1 knockdown on the expression levels of migration and invasion markers, as well as Wnt/β-catenin signaling pathway-related proteins in pancreatic carcinoma cells. Furthermore, a recovery experiment was conducted to assess the role of Wnt/β-catenin signaling pathway agonist CHIR-99021 in ADAMDEC1 knockdown-induced inhibition of pancreatic carcinoma cell growth and migration. RESULTS:(1) ADAMDEC1 was highly expressed in pancreatic carcinoma cells. (2) Knockdown of ADAMDEC1 led to a significant reduction in the proliferation, migration and invasion of pancreatic carcinoma cells. (3) Knockdown of ADAMDEC1 resulted in increased E-cadherin protein expression and decreased levels of matrix metalloproteinase 9, N-cadherin and vimentin proteins, alongside a reduction in the expression of Wnt/β-catenin signaling pathway-related proteins. (4) Co-treatment of pancreatic carcinoma cells with CHIR-99021 and ADAMDEC1 small interfering RNA reversed the inhibitory effects of ADAMDEC1 knockdown on cell proliferation, migration, and invasion. CONCLUSION: ADAMDEC1 is highly expressed in pancreatic carcinoma. Targeted silencing of ADAMDEC1 has the potential to inhibit the proliferation, migration and invasion of pancreatic carcinoma cells by regulating the Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

AIM: To investigate the effect of tea polyphenols (TP) on the sensitivity of human gastric cancer cells to oxaliplatin (L-OHP) in vitro and its mechanism. METHODS: Human gastric cancer HGC-27 and N87 cells, and human gastric mucosal GES-1 cells were used in this study. The HGC-27 and N87 cells were randomly assigned into control group, TP (5 μmol/L) group, L-OHP (5. 2 μmol/L for HGC-27 cells, 7. 7 μmol/L for N87 cells) group, and TP (5 μmol/L) combined with L-OHP (5. 2 μmol/L for HGC-27 cells, 7. 7 μmol/L for N87 cells) group, with 3 replicate wells per group. The cell viability was detected by CCK-8 assay, and the IC50 value of L-OHP was calculated. The proliferation of the cells was assessed by colony formation assay. The migration and invasion abilities of the cells were evaluated by scratch test and Transwell assay. Flow cytometry was performed to evaluate the antiapoptotic effect of the treatments. Ade-novirus infection was conducted to evaluate cell autophagy. The levels of intracellular reactive oxygen species (ROS) were assessed using a ROS assay kit. Western blot analysis was performed to evaluate the protein expression levels of microtubule-associated protein 1 light chain 3 (LC3), nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and superoxide dismutase 1 (SOD1). RESULTS: Combination of TP and L-OHP significantly reduced the viability of HGC-27 and N87 cells, and markedly inhibited cell proliferation and migration compared with L-OHP alone. Significant increases in autophagosomes and ROS levels were observed in combination group compared with L-OHP alone group. The ratio of LC3-II/LC3-I significantly increased in combination group, whereas the expression of Nrf2, HO-1 and SOD1 significantly decreased compared with L-OHP alone group (P<0. 05). CONCLUSION: Treatment with TP enhanced the sensitivity of HGC-27 and N87 cells to L-OHP by inhibiting the Nrf2 pathway, promoting the production of intracellular ROS, and inducing cell autophagy. [ABSTRACT FROM AUTHOR]

Introduction: Significant impacts of heavy work investment on employee wellbeing and organizational performance have prompted its increasing importance as a research topic. The findings about good or evil of these repercussions are nonetheless inconclusive. The intersection of Heavy Work Investment construct with gender has not been explicitly addressed by previous literature review and research. Besides, the relevance of flexibility for women, as one of the key factors for successful work-family balance management, still remains to be analyzed. Methods: A literature review on Heavy Work Investment was conducted using the SPAR-4-SLR protocol, wherein 83 articles were selected from a pool of 208 previously identified works. Bibliometric and content analysis techniques were employed, including co-word analysis, to evaluate research production, impact, and trends in the gender perspective within Heavy Work Investment. Results: As a result, a strategic diagram illustrates thematic topics, providing a clear understanding of the field's structure and evolution. Six thematic groups were identified, around work-family conflict as the central theme. Discussion: The explicit consideration of a gender perspective in literature involves nuanced differences regarding the conclusions of studies with a broader focus. First, the emerging prominence of studies on China and Japan becomes clear with gender as the specific focus of the review, aiming to clarify the experiences women face in more traditional societies with a more decisive division of roles. Second, there is a shift in interest regarding the analysis of Job Demands and Job Resources. Despite the apparent decline in interest in the former, the focus in gender literature clearly shifts toward the side of Job Resources, showing potential for the future. It could be understood that in a context of talent war and employee retention efforts, priority is given to better understanding of facilitating individual and organizational factors for work-life balance, especially for women. Future research areas are identified, including gender differences in organizational support and the impact of flexible work on the work-life balance, providing valuable insights for academia, practitioners, and organizations. The need for more comprehensive cross-cultural and gender research is also made clear. [ABSTRACT FROM AUTHOR]

Microalgae have considerable potential as a renewable feedstock for biochemical and bioethanol production that can be employed in processes associated with carbon capture. Large-scale microalgae cultivations are often non-axenic and are often cohabited by bacteria. A better understanding of the influence of cohabiting bacteria on microalgae productivity is required to develop sustainable synthetic co-culture processes at scale. Nutrient limitation is a frequently employed strategy in algal cultivations to accumulate energy reserves, such as lipids and carbohydrates. Here, a non-axenic culture of an estuarine green microalga, Chlorella vulgaris CCAP 211/21A, was studied under nutrient replete and deplete conditions to assess how changes in nutrient supply influenced the cohabiting bacterial population and its association with intracellular carbohydrate accumulations in the alga. Nutrient limitation resulted in a maximum carbohydrate yield of 47%, which was 74% higher than that in nutrient replete conditions. However, the latter condition elicited a 2-fold higher carbohydrate productivity. Three cohabiting bacterial isolates were cultivable from the three culture conditions tested. These isolates were identified using the 16S rRNA gene sequence to belong to Halomonas sp. and Muricauda sp. The composition of the bacterial population varied significantly between the growth conditions and time points. In all cases and at all time points, the dominant species was Halomonas isolates. Nutrient depletion resulted in an apparent loss of Muricauda sp. This finding demonstrates that nutrient supply can be used to control cohabiting bacterial populations in algal cultures, which will enable the development of synthetic co-culture strategies for improving algae productivity.

The marine holothurian-derived fungal strain KMM 4401 has been identified as Paragliomastix luzulae using 28S rDNA, ITS regions and the partial TEF1 gene sequences. The metabolite profile of the fungal culture was studied by UPLC-MS technique. The strain KMM 4401 is a source of various virescenoside-type isopimarane glycosides suggested as chemotaxonomic feature for this fungal species. Also Px. luzulae KMM 4401 was proposed as possible source of new bioactive secondary metabolites especially antimicrobials. Moreover, the co-cultures of Px. luzulae KMM 4401 with another marine fungus Penicillium hispanicum KMM 4689 inoculated simultaneously or after two weeks were investigated by same way. It was shown, that P. hispanicum KMM 4689 suppressed the production of most of Px. luzulae KMM 4401 metabolites. On the other hand, the co-cultivation of P. hispanicum KMM 4689 and Px. luzulae KMM 4401 resulted in increasing of production of main deoxyisoaustamide alkaloids of P. hispanicum KMM 4689 on 50–190%. [ABSTRACT FROM AUTHOR]

In the field of biomedicine, two-dimensional (2D) cell lines and animal models have played an important role in the study of cell pathways and drug targets. However, due to species differences between humans and other animals, and the lack of hierarchy, cellular diversity, and cell-cell or cell-matrix interactions, 2D cell lines could not fully reflect what cells actually look like in the human body. Organoids are three-dimensional (3D) in vitro culture models derived from autologous tissue stem cells, which make up for the defects of 2D culture and can simulate the structure and function of real human organs to a certain extent, providing new ideas for disease diagnosis and treatment. Among them, lung organoids (LO) are a typical case studying the development process of human lung and the generation principle of lung diseases. Relevant studies have provided help for the treatment of pulmonary fibrosis, lung cancer, lung injury and other diseases. This paper aims to summarize and analyze the research progress of lung organoids in recent years, and further summarize the application of LO in the diagnosis and treatment of lung diseases. [ABSTRACT FROM AUTHOR]

AIM: To investigate the oncolytic effect of human umbilical cord mesenchymal stem cells (hUMSCs) delivering reovirus type 3 (Reo3) on chronic myeloid leukemia (CML) K562 cells. METHODS: The expression of junctional adhesion molecule-A (JAM-A), a receptor susceptible to Reo3, on the surface of hUMSCs and K562 cells was assessed by flow cytometry. Intracellular viral inclusion body distribution 72 h after Reo3 infection in hUMSCs was observed by electron microscopy. The hUMSCs were infected with various multiplicities of infection (MOI) of Reo3 (M01=0, 1, 2 and 3) for 24, 48, 72, 96 and 120 h, and the most suitable MOI was identified by CCK-8 assay. Subsequently, hUMSCs were infected with the optimal titer of Reo3 for the same durations, and supernatants were collected. The titer of Reo3 in the supernatant from each group was measured using mouse fibroblast L929 cells combined with median tissue culture infectious dose (TCID50) method, determining the optimal infection time. The K562 cells were divided into 4 groups: control group, hUMSCs group, Reo3 group, and hUMSCs-Reo3 group. Ratios of hUMSCs to K562 cells in hUMSCs group and hUMSCs-Reo3 group were set at low, medium and high (5^1, 10^1 and 20: 1). The changes of K562 cell viability after co-cultured with hUMSCs-Reo3 for 24, 48 and 72 h were analyzed by CCK-8 assay. The apoptosis of K562 cells was evaluated by flow cytometry. The half maximal effective concentration (EC50) of anti-Reo3 monoclonal antibody was determined using L929 cells. The oncolytic effect of hUMSCs-Reo3 on K562 cells with antibody present in vitro was verified. Western blot analysis was used to detect the protein levels of Bcl-2, Bax, survivin and cleaved caspase-3 in K562 cells after treatment. A BALB/c nude mouse subcutaneous tumor model was constructed with K562 cells (n=6) to analyze the in vivo anti-tumor effect of hUMSCs-Reo3. RESULTS: The expression levels of JAM-A on the surfaces of hUMSCs and K562 cells were found to be 11. 0% and 99. 0%, respectively. Electron microscopy revealed a significant presence of viral inclusion bodies within hUMSCs 72 h following infection with Reo3. Within 120 h, no statistically significant difference was observed in the viability of hUMSCs between Reo3 (MOI=1) group and uninfected group, establishing the optimal MOI. The TCID50 results indicated that the highest virus titer in the lysate of hUMSCs in Reo3 (MOI=1) group occurred 48 h after infection, determining 48 h as the optimal infection time. The K562 cells co-cultured with hUMSCs-Reo3 for 24, 48 and 72 h showed a dose- and time-dependent inhibition of cell viability. The EC50 of the anti-Reo3 monoclonal antibody was found to be 1: 34. Even in the presence of antibodies at various concentrations (1: 34, 1 • 300 and 1: 600), hUMSCs were capable of transporting Reo3 to inhibit K562 cell viability and induce apoptosis in vitro. Compared with control group, significant down-regulation of Bcl-2 and survivin expression levels in K562 cells was noted after 48 h of co-culture with hUMSCs-Reo3 (P<0. 05), while Bax and cleaved caspase-3 expression levels were significantly up-regulated (P< 0. 05 or P<0. 01). In the BALB/c nude mouse tumor-bearing model, determination of tumor volume changes, pathological examination of tumor tissue and major organs, and assessment of cathepsin B/L activity using a small animal live imaging system confirmed the oncolytic effect of hUMSCs-Reo3 on K562 cells in vivo without adverse effects on normal tissues. CONCLUSION: The hUMSCs are effective in transporting Reo3, and this delivery system is capable of releasing sufficient quantities of Reo3 in both in vivo and in vitro settings to inhibit the malignant proliferation of K562 cells and promote apoptosis, thereby exerting an oncolytic effect. [ABSTRACT FROM AUTHOR]

To prepare bio-scaffolds with multi-stage pores, excellent mechanical and biological properties to meet the requirements of personalized bone repair, digital light processing using Pickering emulsion as paste has been proposed here firstly for porous hydroxyapatite scaffolds with complex shape and fine microstructure. It is found that dispersant content, solid loading and oil to water ratio of Pickering emulsions play vital roles on the controlling of pore morphology and pore size distribution of final hydroxyapatite ceramic foams. The multi-stage pores consist of millimeter-scale pores designed by 3D models and micro-pores below 10 µm derived from emulsion droplets. Due to the integration of hierarchical pores at different scale levels and micropores, hydroxyapatite ceramic foams possess outstanding compressive strength of 4.46–8.03 MPa at porosity of 76.59-72.96%. In vitro cell experiments illustrate that rat bone mesenchymal stem cells can spread and adhere to bio-scaffolds, promoting cell proliferation effectively and further verifying their good biocompatibility. [ABSTRACT FROM AUTHOR]

The Fusarium fungi is found in cereals and feedstuffs and may produce mycotoxins, which are secondary metabolites, such as the T-2 toxin (T-2). In this work, we explored the hepatotoxicity of T-2 using microfluidic 3D hepatic cultures. The objectives were: (i) exploring the benefits of microfluidic 3D cultures compared to conventional 3D cultures available commercially (Aggrewell plates), (ii) establishing 3D co-cultures of hepatic cells (HepG2) and stellate cells (LX2) and assessing T-2 exposure in this model, (iii) characterizing the induction of metabolizing enzymes, and (iv) evaluating inflammatory markers upon T-2 exposure in microfluidic hepatic cultures. Our results demonstrated that, in comparison to commercial (large-volume) 3D cultures, spheroids formed faster and were more functional in microfluidic devices. The viability and hepatic function decreased with increasing T-2 concentrations in both monoculture and co-cultures. The RT-PCR analysis revealed that exposure to T-2 upregulates the expression of multiple Phase I and Phase II hepatic enzymes. In addition, several pro- and anti-inflammatory proteins were increased in co-cultures after exposure to T-2. [ABSTRACT FROM AUTHOR]

This article explores the potential use of pathobiome strains as live biotherapeutic products for human health. It emphasizes the importance of evaluating these strains at a detailed level, considering factors such as functional genes, ecological niche, and drug interactions. The article acknowledges the role of the gut microbiota in human health and discusses the potential benefits and risks of using pathobiome strains. It also addresses the challenges in developing these products, particularly in terms of safety, efficacy, and quality management. The authors propose further research into gene functions, ecological influences, and the pathogenicity of specific strains, and suggest expanding the applications of live biotherapeutics beyond gastrointestinal diseases. They discuss various development methods, such as large-scale screening and gene editing technology, and highlight the need for further investigation in this field. [Extracted from the article]

The infection caused by porcine epidemic diarrhea virus (PEDV) is associated with high mortality in piglets worldwide. Host factors involved in the efficient replication of PEDV, however, remain largely unknown. Our recent proteomic study in the virus-host interaction network revealed a significant increase in the accumulation of CALML5 (EF-hand protein calmodulin-like 5) following PEDV infection. A further study unveiled a biphasic increase of CALML5 in 2 and 12 h after viral infection. Similar trends were observed in the intestines of piglets in the early and late stages of the PEDV challenge. Moreover, CALML5 depletion reduced PEDV mRNA and protein levels, leading to a one-order-of-magnitude decrease in virus titer. At the early stage of PEDV infection, CALML5 affected the endosomal trafficking pathway by regulating the expression of endosomal sorting complex related cellular proteins. CALML5 depletion also suppressed IFN-ß and IL-6 production in the PEDV-infected cells, thereby indicating its involvement in negatively regulating the innate immune response. Our study reveals the biological function of CALML5 in the virology field and offers new insights into the PEDV-host cell interaction. [ABSTRACT FROM AUTHOR]

Microalgae have considerable potential as a renewable feedstock for biochemical and bioethanol production that can be employed in processes associated with carbon capture. Large-scale microalgae cultivations are often non-axenic and are often cohabited by bacteria. A better understanding of the influence of cohabiting bacteria on microalgae productivity is required to develop sustainable synthetic co-culture processes at scale. Nutrient limitation is a frequently employed strategy in algal cultivations to accumulate energy reserves, such as lipids and carbohydrates. Here, a non-axenic culture of an estuarine green microalga, Chlorella vulgaris CCAP 211/21A, was studied under nutrient replete and deplete conditions to assess how changes in nutrient supply influenced the cohabiting bacterial population and its association with intracellular carbohydrate accumulations in the alga. Nutrient limitation resulted in a maximum carbohydrate yield of 47%, which was 74% higher than that in nutrient replete conditions. However, the latter condition elicited a 2-fold higher carbohydrate productivity. Three cohabiting bacterial isolates were cultivable from the three culture conditions tested. These isolates were identified using the 16S rRNA gene sequence to belong to Halomonas sp. and Muricauda sp. The composition of the bacterial population varied significantly between the growth conditions and time points. In all cases and at all time points, the dominant species was Halomonas isolates. Nutrient depletion resulted in an apparent loss of Muricauda sp. This finding demonstrates that nutrient supply can be used to control cohabiting bacterial populations in algal cultures, which will enable the development of synthetic co-culture strategies for improving algae productivity. [ABSTRACT FROM AUTHOR]

Improving the performance of proton exchange membrane fuel cells (PEMFCs) requires deep understanding of the reactive transport processes inside the catalyst layers (CLs). In this study, a particle-overlapping model is developed for accurately describing the hierarchical structures and oxygen reactive transport processes in CLs. The analytical solutions derived from this model indicate that carbon particle overlap increases ionomer thickness, reduces specific surface areas of ionomer and carbon, and further intensifies the local oxygen transport resistance (Rother). The relationship between Rother and roughness factor predicted by the model in the range of 800-1600 s m-1 agrees well with the experiments. Then, a multiscale model is developed by coupling the particle-overlapping model with cell-scale models, which is validated by comparing with the polarization curves and local current density distribution obtained in experiments. The relative error of local current density distribution is below 15% in the ohmic polarization region. Finally, the multiscale model is employed to explore effects of CL structural parameters including Pt loading, I/C, ionomer coverage and carbon particle radius on the cell performance as well as the phase-change-induced (PCI) flow and capillary-driven (CD) flow in CL. The result demonstrates that the CL structural parameters have significant effects on the cell performance as well as the PCI and CD flows. Optimizing the CL structure can increase the current density and further enhance the heat-pipe effect within the CL, leading to overall higher PCI and CD rates. The maximum increase of PCI and CD rates can exceed 145%. Besides, the enhanced heat-pipe effect causes the reverse flow regions of PCI and CD near the CL/PEM interface, which can occupy about 30% of the CL. The multiscale model significantly contributes to a deep understanding of reactive transport and multiphase heat transfer processes inside PEMFCs. Highlights: A particle-overlapping model for reactive transport process in catalyst layers. A multiscale model coupling particle-overlapping model with cell-scale models. The model is rigorously validated from nanoscale to commercial-cell scale. Effects of catalyst layer structures on cell performance are evaluated. Phase-change-induced and capillary-driven flows in catalyst layers are studied. [ABSTRACT FROM AUTHOR]

Persistent virus infection involves modifying the host immune response and maintaining viral infection. Acute infection with Mononegavirales, such as Sendai viruses (SeVs), can give rise to viruses capable of persistent infection. SeVs establish persistent infection through generating copyback-type defective interfering (cbDI) genomes or acquiring temperature-sensitive mutations. Herein, we identify novel mutations associated with persistent infection and recombinant SeV mutants capable of persistent infection and autonomous production at physiological body temperature, independent of cbDI genomes or temperature-sensitive mutations. Diverse SeV populations were generated by passing the cDNA-recovered SeV Z strain 19 times through embryonated chicken eggs and subsequently infecting LLC-MK2 cells with the SeV populations to finally obtain SeV mutants capable of persistent infection and autonomous production in several types of cultured cells. Sequence analysis identified 4 or 5 mutations in the genome of the persistently infectious SeVs, distinguishing them from other existing strains with persistent infection. Recombinant SeVs carrying 4 or 5 mutations in the Z strain genome (designated SeV-Zpi or SeV-Zpi2, respectively) exhibited persistent infection and autonomous production in LLC-MK2, BHK-21, and Neuro2a cells at 37°C. SeV-Zpi and SeV-Zpi2 consistently produced viral particles even after long-term passages without cbDI particles or temperature-sensitive phenotypes. These results highlight the ability of acute infections of SeVs to spontaneously acquire mutations during replication, thereby endowing persistent infection and autonomous production at body temperature. The vectorization of SeV-Zpi and SeV-Zpi2 will contribute to both basic research and medical applications. [ABSTRACT FROM AUTHOR]

The balance between porosity and compressive strength, as well as the pro-osteogenic and pro-angiogenesis effects of β-tricalcium phosphate (β-TCP) are not satisfactory in the clinical application even though it has good biocompatibility, osseointegration and osteoconductivity. Herein, different contents (10, 20 and 30 wt%) of magnesium silicate (Mg 2 SiO 4, MS) and β-TCP composite bioceramic scaffolds (MS/β-TCP) were fabricated by three-dimensional fiber deposition (3DF) technology. The physicochemical, in vitro osteogenic and angiogenic properties, the early (6 weeks) cranium defect repair effects in rats and the underlying molecular mechanism on accelerating osteogenic differentiation of MS/β-TCP were systematically investigated. The results showed that MS compounding not only promoted the sintering of β-TCP and significantly enhanced the compressive strength, but also greatly improved its osteogenic and angiogenic performances, such as the viability, adhesion, and proliferation of mouse bone marrow mesenchymal stem cells (mBMSCs) and human umbilical vein endothelial cells (HUVECs), ALP activity and the expression of adhesion-related, osteogenesis-related and angiogenesis-related genes. The in vivo experiments presented that 10MS/β-TCP had more new bone formation and better angiogenesis compared to β-TCP and Blank. Analysis of the transcriptome sequencing results confirmed that the Wnt signaling pathway was involved in the regulation of osteogenic differentiation in mBMSCs by the MS/β-TCP composite bioceramic scaffold. The MS/β-TCP holds great potential in bone repair and regeneration. [ABSTRACT FROM AUTHOR]

Hydrolyzed conchiolin protein (HCP) can act as a cosmetic skin-whitening agent by inhibiting melanogenesis through effects on the enzyme tyrosinase (TYR) and reducing melanin production in B16F10 melanoma cells. However, details of the molecular mechanism underlying this effect have not been fully elucidated. miRNA is a type of ncRNA of approximately 21-25 nucleotides that exhibits binding affinity toward a specific region (r epresented by the 3' untranslated region [UTR]) of mu ltiple mRNA target molecules. We investigated the effects of HCP on the miR-26a-1-3p, which is predicted to bind the upstream regulator of TYR, microphthalmia-associated transcription factor (MITF), in B16F10 melanoma cells. We assessed whether miR-26a-1-3p regulates MITF and indirectly affects the pathways of TYR and TYR-related protein-1 (TRP-1) expression levels, thereby influencing melanogenesis. The 3' UTR of MITF was verified to be a miR-26a-1-3p t arget gene by bioinformatics prediction and a dual-luciferase reporter assay. Overexpression of miR-26a-1-3p in cultured cells decreased MITF, TYR, and TYR-related protein-1 (TRP-1) expression levels. HCP treatment increased miR-26a-1-3p levels, inhibiting MITF and leading to decreased TYR and TRP-1 levels. Our results suggest that HCP controls melanin production by increasing miR-26a-1-3p levels and inhibiting MITF, TYR, and TRP-1 expression, indicating its potential as a whitening product. [ABSTRACT FROM AUTHOR]

This research focuses on assessing the synergistic effects of Mexican oregano (Lippia graveolens) essential oil or carvacrol when combined with the antibiotic imipenem, aiming to reduce the pathogenic viability and virulence of Acinetobacter baumannii and Pseudomonas aeruginosa. The study highlighted the synergistic effect of combining L. graveolens essential oil or carvacrol with imipenem, significantly reducing the required doses for inhibiting bacterial growth. The combination treatments drastically lowered the necessary imipenem doses, highlighting a potent enhancement in efficacy against A. baumannii and P. aeruginosa. For example, the minimum inhibitory concentrations (MIC) for the essential oil/imipenem combinations were notably low, at 0.03/0.000023 mg/mL for A. baumannii and 0.0073/0.000023 mg/mL for P. aeruginosa. Similarly, the combinations significantly inhibited biofilm formation at lower concentrations than when the components were used individually, demonstrating the strategic advantage of this approach in combating antibiotic resistance. For OXA-51, imipenem showed a relatively stable interaction during 30 ns of dynamic simulation of their interaction, indicating changes (<2 nm) in ligand positioning during this period. Carvacrol exhibited similar fluctuations to imipenem, suggesting its potential inhibition efficacy, while thymol showed significant variability, particularly at >10 ns, suggesting potential instability. With IMP-1, imipenem also displayed very stable interactions during 38 ns and demonstrated notable movement and positioning changes within the active site, indicating a more dynamic interaction. In contrast, carvacrol and thymol maintained their position within the active site only ~20 and ~15 ns, respectively. These results highlight the effectiveness of combining L. graveolens essential oil and carvacrol with imipenem in tackling the difficult-to-treat pathogens A. baumannii and P. aeruginosa. [ABSTRACT FROM AUTHOR]

Essential trace minerals play vital roles in maintaining human and animal health. However, an overdose of the existing inorganic trace minerals is prone to induce detrimental effects that outweigh positive benefits. In this study, an extracellular polymeric substances (EPS)-producing bacterium, identified as Bacillus licheniformis CCTCC M2020298, was isolated from marine using glutinous rice processing wastewater as enrichment medium. The EPS yield of Bacillus licheniformis CCTCC M2020298 could reach 8.62 g/L by using glutinous rice-processing wastewater containing medium. Furthermore, the potential of the EPS as a carrier for synthesizing EPS-iron (Fe) and EPS-copper (Cu) complex was explored. The results showed that the optimum condition for the synthesis EPS-Fe were the reaction temperature 70°C, pH 8.5-9.0 and mass ratio of EPS to trisodium citrate 2:1. The iron content of EPS-Fe reached 77.4 mg/g. Under the same condition, the copper content of EPS-Cu reached 90.7 mg/g. The elemental composition, functional groups and valence state of the mineral elements of EPS-Fe and EPS-Cu were well characterized. The EPSFe and EPS-Cu exhibited antioxidant activity in scavenging ·OH, DPPH and ·O2-free radicals, thereby leading to reduced oxidative stress and apoptosis levels in human colonic epithelial cells in vitro. They also inhibited the proliferation of mouse hepatocellular carcinoma H22 and the growth of intestinal pathogens in vitro. This study provided an effective avenue for EPS production from glutinous rice processing wastewater and proved the potential of EPS-Fe and EPS-Cu complexes as a new-type comprehensive essential trace mineral supplement. [ABSTRACT FROM AUTHOR]

Mesenchymal stem cells (MSCs) are an integral part of the tumor microenvironment (TME); however, their role is somewhat controversial: conflicting reports suggest that, depending on the stage of tumor development, MSCs can either support or suppress tumor growth and spread. Additionally, the influence of MSCs on drug resistance is also ambiguous. Previously, we showed that, despite MSCs proliferating significantly more slowly than cancer cells, there are chemotherapeutic drugs which proved to be similarly toxic to both cell types. Here we established 2D co-cultures and 3D co-culture spheroids from different ratios of GFP-expressing, adipose tissue-derived MSCs and A431 epidermoid carcinoma cells tagged with mCherry to investigate the effect of MSCs on cancer cell growth, survival, and drug sensitivity. We examined the cytokine secretion profile of mono- and co-cultures, explored the inner structure of the spheroids, applied MSC-(nutlin-3) and cancer cell-targeting (cisplatin) treatments separately, monitored the response with live-cell imaging and identified a new, double-fluorescent cell type emerging from these cultures. In 2D co-cultures, no effect on proliferation or drug sensitivity was observed, regardless of the changes in cytokine secretion induced by the co-culture. Conversely, 3D spheroids developed a unique internal structure consisting of MSCs, which significantly improved cancer cell survival and resilience to treatment, suggesting that physical proximity and cell–cell connections are required for MSCs to considerably affect nearby cancer cells. Our results shed light on MSC–cancer cell interactions and could help design new, better treatment options for tumors. [ABSTRACT FROM AUTHOR]

Introduction: This study is an introduction to the empirical and impact evaluation of radiofrequency electromagnetic field (RF-EMF) radiation exposure in a healthcare environment, focusing on an indoor microenvironment. It explores the expression of various genes associated with cellular responses, cell proliferation, senescence, and apoptotic cell death. The assessment analyzes current personal mobile communications (2G-5G FR1), providing a clear understanding of RF-EMF exposure and compliance with regulatory limits. Methods: The signals from different wireless communication systems at Hospital Universitario de Canarias (HUC) in Tenerife, Canary Islands, Spain, were examined in 11 locations. Four measurement campaigns were performed with frequencyselective exposimeters (PEMs) and an EME Spy 200 MVG, and experimental electric field values were compared as a long-term exposition. The frequency with the highest contribution (2.174 V/m) observed (1840 MHz) in UMTS was selected for biological effects evaluation. Results: The study focuses on four locations with the highest exposure to communication systems (downlinks), analyzing the results to verify compliance with regulations that ensure the safety of patients, the general public, and healthcare workers. LTE B20 (DL), GSM+UMTS 900 (DL), GSM 1800 (DL), UMTS 2100 (DL), and LTE B7 (DL) exhibited relatively higher E/m values throughout the campaigns, and these values consistently remained below the ICNIRP reference levels, signifying a consistently low level of exposure. In addition, this work presents the biological effects on neural stem cells (NSCs) using 3D brain organoids (BOs) exposed to RF signals in a validated and commercial experimental setting: the Gigahertz Transverse Electromagnetic cell (GTEM). The GTEM allows for the creation of homogeneous field electromagnetic fields in a small, enclosed setting and guarantees exposure conditions in a wide range of frequencies. BOs are an in vitro 3D cell-culture technology that reproduces the cellular composition and structure of the developing brain. Analyzing the expression of several genes associated with cellular responses, cell proliferation, senescence, and apoptotic cell death, we found that exposure of BOs at 1840MHzdid not affectmRNAexpression in brain genes related to apoptosis or senescence. However, a decrease in gene expression for cell proliferation and cell activity markers was observed during the differentiation stage of BOs. Discussion: The discussion emphasizes the coexistence and evolution of various heterogeneous networks and services throughout the four measurement campaigns. Across all measured results, the levels of the obtained E-field were consistently well below the exposure limits set by internationally accepted standards and guidelines. These obtained values have been established in order to consider their potential effects on cell proliferation and cell activity, especially in differentiating biological organisms. Consequently, the results obtained and the methodology presented could serve as a foundational framework for establishing the basis of RF-EMF assessment in future heterogeneous 5G developments, particularly in the millimeter wave (mmWave) frequency range, where the forecast is for massive high-node density networks. [ABSTRACT FROM AUTHOR]

Background: Tissue engineering is a multidisciplinary and interdisciplinary topic that involves the development of biological implants for tissue regeneration intending to improve or enhance tissue or organ function. Objectives: This study aimed to evaluate the mechanical and histological properties of decellularized rat pancreas scaffolds, as well as to investigate the viability of adipose mesenchymal stem cells (MSCs) on the said scaffold for use in regenerative medicine and tissue engineering. Methods: This is an experimental study that was performed in the research laboratory of Mohaghegh Ardabili University. To prepare the scaffold, male Wistar rats were anesthetized with carbon dioxide. After dissecting the mice, their pancreases were isolated and immediately transferred to a phosphate-buffered saline (PBS) solution to prepare them for decellularization. The decellularized scaffolds were evaluated histologically and mechanically. After decellularization, lipid MSCs were injected into de-cell scaffolds in the third passage. Results: Examination of the results of histological evaluations showed that scaffolding was completely decellularized. These results were confirmed by Mason trichrome and Dapi staining (coloring). Specialized tissue assessments by electron microscopy showed that the collagen and elastin strands were relatively conserved in the extracellular matrix (ECM). Conclusions: In general, the result of this research demonstrates the successful decellularization of pancreatic tissue, effective preservation of the ECM of the desired tissue, and the viability of the MSCs on the scaffold resulting from the decellularization of the tissue. [ABSTRACT FROM AUTHOR]

The use of co-culture aims to enhance the quality of beverages by providing a diverse range of volatile compound profiles and sensory characteristics. In this study, the fermentative, sensory characteristics and volatile compounds of meads produced from S. cerevisiae strains JP14 and IM8, both in pure cultures and co-cultures, were evaluated. A slower sugar consumption rate was observed when IM8 was inoculated solely or at a 1:100. Higher ethanol concentrations were achieved in the meads produced with pure JP14 or in 1:1 co-cultivation. Meads exclusively produced with IM8 exhibited higher concentrations of ethyl acetate and acetaldehyde, contributing fruity and nutty aromas. Conversely, meads produced in a 1:1 co-culture showed higher concentrations of isoamyl alcohol, imparting banana, pear, and marzipan aromas. All meads were well-accepted, receiving ratings between "lightly liked" and "moderately liked". However, meads produced with IM8 received higher scores in flavor, indicating a stronger preference compared to meads produced exclusively with JP14. In conclusion, the use of different inocula influenced both the fermentative and sensory characteristics of mead. The appropriate selection of strains can be explored to produce meads with specific aromatic profiles, allowing for the creation of high-quality beverages with diverse sensory characteristics. [ABSTRACT FROM AUTHOR]

To investigate the degradation effect of bovine trypsin on multispecies biofilm of caries-related bacteria and provide an experimental foundation for the prevention of dental caries. Standard strains of S. mutans, S. sanguis, S. gordonii, and L. acidophilus were co-cultured to form 24 h, 48 h, and 72 h biofilms. The experimental groups were treated with bovine trypsin for 30 s, 1 min, and 3 min. Morphological observation and quantitative analysis of extracellular polymeric substances (EPS), live bacteria, and dead bacteria were conducted using the confocal laser scanning microscope (CLSM). The morphological changes of EPS and bacteria were also observed using a scanning electron microscope (SEM). When biofilm was treated for 1 min, the minimal inhibitory concentration (MIC) of bovine trypsin to reduce EPS was 0.5 mg/mL in 24 h and 48 h biofilms, and the MIC of bovine trypsin was 2.5 mg/mL in 72 h biofilms (P < 0.05). When biofilm was treated for 3 min, the MIC of bovine trypsin to reduce EPS was 0.25 mg/mL in 24 h and 48 h biofilms, the MIC of bovine trypsin was 1 mg/mL in 72 h biofilm (P < 0.05). The ratio of live-to-dead bacteria in the treatment group was significantly lower than blank group in 24 h, 48 h, and 72 h multispecies biofilms (P < 0.05). Bovine trypsin can destroy multispecies biofilm structure, disperse biofilm and bacteria flora, and reduce the EPS and bacterial biomass in vitro, which are positively correlated with the application time and concentration. [ABSTRACT FROM AUTHOR]

This in vitro study assessed peak temperature and temperature increase (ΔT) within the pulp chamber during different extended photoactivation techniques (EPT—applying similar radiant exposure values) to resin-based composites (RBCs) placed in a Class I cavity preparation in an extracted human lower third molar. A T-type thermocouple was placed in the pulp chamber and connected to a temperature analysis device (Thermes, Physitemp). The tooth was attached to an assembly simulating the in vivo environment (controlled baseline pulp chamber temperature and fluid flow). The real-time pulp chamber temperature was evaluated throughout the photoactivation (Bluephase N, Ivoclar Vivadent) of two bulk-fill RBCs: Tetric N Ceram Bulk Fill (TBF; shade: IVA; Ivoclar Vivadent); Surefill SDR flow + (SDR, shade: Universal; Dentsply Sirona), which were exposed to different curing techniques: 40 s-occlusal surface; 20 s-occlusal + 10 s-buccal + 10 s-lingual surfaces; 10 s-buccal + 10 s + lingual + 20 s-occlusal surfaces. Each EPT delivered 42.4 J/cm2. Vickers hardness (VHN) was measured on the removed, sectioned RBC restorations at the top and bottom middle areas after curing. ΔT and VHN data were analyzed using 2-way ANOVA followed by Bonferroni post-hoc test (α = 0.05). Peak temperature data were analyzed using one-way ANOVA and Dunnett's post-hoc test (α = 0.05). SDR showed higher ΔT values than TBF (p = 0.008) in some EPTs. Neither technique resulted in ΔT values greater than 5.5 °C. Both composites had acceptable bottom/top hardness ratios (greater than 80%), regardless of the photoactivation technique. The evaluated EPTs may be considered safe as a low-temperature increase was noticed within the pulp chamber. [ABSTRACT FROM AUTHOR]

To evaluate the osteogenic potential of platelet-rich fibrin (PRF) and low-level laser therapy (LLLT) on human stem cells from the apical papilla (SCAP) we isolated, characterized, and then cultured in an osteogenic medium cells with PRF and/or LLLT (660 nm, 6 J/m2-irradiation). Osteogenic differentiation was assessed by bone nodule formation and expression of bone morphogenetic proteins (BMP-2 and BMP-4), whereas the molecular mechanisms were achieved by qRT-PCR and RNA-seq analysis. Statistical analysis was performed by ANOVA and Tukey's post hoc tests (p < 0.05* and p < 0.01**). Although PRF and LLLT increased bone nodule formation after 7 days and peaked at 21 days, the combination of PRF + LLLT led to the uppermost nodule formation. This was supported by increased levels of BMP-2 and -4 osteogenic proteins (p < 0.005). Furthermore, the PRF + LLLT relative expression of specific genes involved in osteogenesis, such as osteocalcin, was 2.4- (p = 0.03) and 28.3- (p = 0.001) fold higher compared to the PRF and LLLT groups, and osteopontin was 22.9- and 1.23-fold higher, respectively (p < 0.05), after 7 days of interaction. The transcriptomic profile revealed that the combination of PRF + LLLT induces MSX1, TGFB1, and SMAD1 expression, after 21 days of osteogenic differentiation conditions exposition. More studies are required to understand the complete cellular and molecular mechanisms of PRF plus LLLT on stem cells. Overall, we demonstrated for the first time that the combination of PRF and LLLT would be an excellent therapeutic tool that can be employed for dental, oral, and craniofacial repair and other tissue engineering applications. [ABSTRACT FROM AUTHOR]

We have recently shown that multiple tRNA synthetase inhibitors can greatly increase lifespan in multiple models by acting through the conserved transcription factor ATF4. Here, we show that these compounds, and several others of the same class, can greatly upregulate mammalian ATF4 in cells in vitro, in a dose dependent manner. Further, RNASeq analysis of these cells pointed toward changes in protein turnover. In subsequent experiments here we show that multiple tRNA synthetase inhibitors can greatly upregulate activity of the ubiquitin proteasome system (UPS) in cells in an ATF4-dependent manner. The UPS plays an important role in the turnover of many damaged or dysfunctional proteins in an organism. Increasing UPS activity has been shown to enhance the survival of Huntington's disease cell models, but there are few known pharmacological enhancers of the UPS. Additionally, we see separate ATF4 dependent upregulation of macroautophagy upon treatment with tRNA synthetase inhibitors. Protein degradation is an essential cellular process linked to many important human diseases of aging such as Alzheimer's disease and Huntington's disease. These drugs' ability to enhance proteostasis more broadly could have wide-ranging implications in the treatment of important age-related neurodegenerative diseases. [ABSTRACT FROM AUTHOR]

Bioremediation, involving the strategic use of microorganisms, has proven to be a cost-effective alternative for restoring areas impacted by persistent contaminants such as polycyclic aromatic hydrocarbons (PAHs). In this context, the aim of this study was to explore hydrocarbon-degrading microbial consortia by prospecting native species from soils contaminated with blends of diesel and biodiesel (20% biodiesel/80% diesel). After enrichment in a minimal medium containing diesel oil as the sole carbon source and based on 16S rRNA, Calmodulin and β-tubulin gene sequencing, seven fungi and 12 bacteria were identified. The drop collapse test indicated that all fungal and four bacterial strains were capable of producing biosurfactants with a surface tension reduction of ≥20%. Quantitative analysis of extracellular laccase production revealed superior enzyme activity among the bacterial strains, particularly for Stenotrophomonas maltophilia P05R11. Following antagonistic testing, four compatible consortia were formulated. The degradation analysis of PAHs and TPH (C5–C40) present in diesel oil revealed a significantly higher degradation capacity for the consortia compared to isolated strains. The best results were observed for a mixed bacterial-fungal consortium, composed of Trichoderma koningiopsis P05R2, Serratia marcescens P10R19 and Burkholderia cepacia P05R9, with a degradation spectrum of ≥91% for all eleven PAHs analyzed, removing 93.61% of total PAHs, and 93.52% of TPH (C5–C40). Furthermore, this study presents the first report of T. koningiopsis as a candidate for bioremediation of petroleum hydrocarbons.