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<jats:sec> <jats:title>Abstract</jats:title> <jats:p>Globally colorectal cancer ranks as the third most widespread disease and the third leading cause of cancer-associated mortality. Immunotherapy treatments like PD-L1 blockade have been used to inhibit the PD-L1 legend, which boosts the activity of cytotoxic T lymphocytes. Recently, studies suggest that some probiotics could potentially enhance the effectiveness of immunotherapy treatments for cancer patients. We found that in Caco-2 and HT-29 cells, the live <jats:italic>Leuconostoc mesenteroides</jats:italic> treatment resulted an increase in the PD-L1 expression and this treatment stimulated interferon-gamma (IFN-γ) production in Jurkat T-cells. Due to the well-established ability of IFN-γ to enhance PD-L1 expression, the combination of IFN-γ and <jats:italic>L. mesenteroides</jats:italic> was used in colon cancer cell lines and a resulting remarkable increase of over tenfold in PD-L1 expression was obtained. Interestingly, when <jats:italic>L. mesenteroides</jats:italic> and IFN-γ are present, the blockage of PD-L1 using PD-L1 antibodies not only improved the viability of Jurkat T-cells but also significantly boosted the levels of IFN-γ and IL-2, the T-cells activation marker cytokines. In addition to upregulating PD-L1, <jats:italic>L. mesenteroides</jats:italic> also activated Toll-like receptors (TLRs) and NOD-like receptors (NODs) pathways, specifically through TLR2 and NOD2, while also exerting a suppressive effect on autophagy in colon cancer cell lines. In conclusion, our findings demonstrate a significant upregulation of PD-L1 expression in colon cancer cells upon co-culturing with <jats:italic>L. mesenteroides</jats:italic>. Moreover, the presence of PD-L1 antibodies during co-culturing activates Jurkat T cells. The observed enhancement in PD-L1 expression may be attributed to the inhibition of the Autophagy pathway or activation of the hippo pathway.</jats:p> </jats:sec><jats:sec> <jats:title>Graphical abstract text</jats:title> <jats:p>The administration of Live <jats:italic>Lactobacillus mesenteroides</jats:italic> on colon cancer cells leads to the elevation of PD-L1, with a further increase observed in the presence of IFN-γ. Co-cultivation of Live <jats:italic>L. mesenteroides</jats:italic> with colon cancer cells in conjunction with anti-PD-L1 blockade antibody results in the enhanced viability of T cells.</jats:p> </jats:sec><jats:sec> <jats:title>Key Points</jats:title> <jats:p><jats:list list-type="bullet"> <jats:list-item> <jats:p>Co-culturing <jats:italic>L. mesenteroides</jats:italic> increases PD-L1 gene and protein transaction in colon cancer.</jats:p> </jats:list-item> <jats:list-item> <jats:p><jats:italic>L. mesenteroides</jats:italic> existing enhances T cells viability and activity.</jats:p> </jats:list-item> <jats:list-item> <jats:p>GPCR41/42 is a possible link between <jats:italic>L. mesenteroides</jats:italic>, YAP-1 and PD-L1.</jats:p> </jats:list-item> </jats:list></jats:p> </jats:sec>

<jats:title>Abstract</jats:title><jats:p>The viable but non-culturable (VBNC) state is considered a survival strategy employed by bacteria to endure stressful conditions, allowing them to stay alive. Bacteria in this state remain unnoticed in live cell counts as they cannot proliferate in standard culture media. VBNC cells pose a significant health risk because they retain their virulence and can revive when conditions normalize. Hence, it is crucial to develop fast, reliable, and cost-effective methods to detect bacteria in the VBNC state, particularly in the context of public health, food safety, and microbial control assessments. This research examined the biomolecular changes in <jats:italic>Escherichia coli</jats:italic> W3110 induced into the VBNC state in artificial seawater under three different stress conditions (temperature, metal, and antibiotic). Initially, confirmation of VBNC cells under various stresses was done using fluorescence microscopy and plate counts. Subsequently, lipid peroxidation was assessed through the TBARS assay, revealing a notable increase in peroxidation end-products in VBNC cells compared to controls. ATR-FTIR spectroscopy and chemomometrics were employed to analyze biomolecular changes, uncovering significant spectral differences in RNA, protein, and nucleic acid concentrations in VBNC cells compared to controls. Notably, RNA levels increased, while protein and nucleic acid amounts decreased. ROC analyses identified the 995 cm<jats:sup>− 1</jats:sup> RNA band as a consistent marker across all studied stress conditions, suggesting its potential as a robust biomarker for detecting cells induced into the VBNC state under various stressors.</jats:p>

<jats:title>Abstract</jats:title><jats:p>Recombinant protein production technology is widely applied to the manufacture of biologics used as drug substances and industrial proteins such as recombinant enzymes and bioactive proteins. Various heterologous protein production systems have been developed using prokaryotic and eukaryotic hosts. Especially methylotrophic yeast in eukaryotic hosts is suggested to be particularly valuable because such systems have the following advantages: protein secretion into culture broth, eukaryotic quality control systems, a post-translational modification system, rapid growth, and established recombinant DNA tools and technologies such as strong promoters, effective selection markers, and gene knock-in and -out systems. Many methylotrophic yeasts such as the genera <jats:italic>Candida</jats:italic>, <jats:italic>Ogataea</jats:italic>, and <jats:italic>Komagataella</jats:italic> have been studied since methylotrophic yeast was first isolated in 1969. The methanol-consumption-related genes in methylotrophic yeast are strongly and strictly regulated under methanol-containing conditions. The well-regulated gene expression systems under the methanol-inducible gene promoter lead to the potential application of heterologous protein production in methylotrophic yeast. In this review, we describe the recent progress of heterologous protein production technology in methylotrophic yeast and introduce <jats:italic>Ogataea minuta</jats:italic> as an alternative production host as a substitute for <jats:italic>K. phaffii</jats:italic> and <jats:italic>O. polymorpha</jats:italic>.</jats:p>