Background: Chinese hamster ovary (CHO) cells are extensively used in the pharmaceutical industry for producing complex proteins, primarily because of their ability to perform human-like post-translational modifications. However, the efficiency of high-quality protein production can vary significantly for monoclonal antibody-producing cell lines, within the CHO host cell lines or by extrinsic factors. Methods: To investigate the complex cellular mechanisms underlying this variability, a phosphoproteomics analysis was performed using label-free quantitative liquid chromatography after a phosphopeptide enrichment of recombinant CHO cells producing two different antibodies and a tunicamycin treatment experiment. Using MaxQuant and Perseus for data analysis, we identified 2109 proteins and quantified 4059 phosphosites. Results: Significant phosphorylation dynamics were observed in nuclear proteins of cells producing the difficult-to-produce 2G12 mAb. It suggests that the expression of 2G12 regulates nuclear pathways based on increases and decreases in phosphorylation abundance. Furthermore, a substantial number of changes in the phosphorylation pattern related to tunicamycin treatment have been detected. TM treatment affects, among other phosphoproteins, the eukaryotic elongation factor 2 kinase (Eef2k). Conclusions: The alterations in the phosphorylation landscape of key proteins involved in cellular processes highlight the mechanisms behind stress-induced cellular responses.

• Book : 13(1)
• Pub. Date : 2025
• Page : pp.9-9
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• Pub. Date : 2025
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Ultraviolet radiation (UV) represents a significant abiotic stress, affecting green plants. Phenolic compounds have been suggested as components involved in plant photoprotective adaptation. We used a unique combination of experimental (LED lighting and leaf tagging) and analytical (unbiased, or untargeted, metabolomics) approaches to study the effects of high (approximating mid-summer) and low (approximating winter) levels of UVA on the expression of phenolic compounds. These consisted of river red gum (Eucalyptus camaldulensis ssp. camaldulensis) of five provenances. The geographically separated provenances used in our study spanned the lowest and highest latitudes of the range of this subspecies. The concentrations of gallotannins and ellagitannins (i.e., hydrolysable tannins) increased most under high levels of UVA, but responses only differed slightly among provenances. The most substantial changes in the composition of phenolic compounds were associated with leaf age. Overall, 3-month-old (herein, termed ‘young’) leaves had substantially different phenolic compositions to 6- and 12-month-old (‘old’) leaves. Hydrolysable tannins were more abundant in young leaves, whereas pedunculagin, catechin, and kaempferol galloyl glucoses were more abundant in old leaves. High levels of UVA altered the expression of phenolic compounds, but our experimental saplings were unlikely to experience photoinhibition because they were not exposed to high levels of light and low temperatures, nor were they nitrogen-limited. We expect that changes in phenolic compounds would have been more pronounced if we had induced photoinhibition.

• Book : 14(3)
• Pub. Date : 2025
• Page : pp.493-493
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Abstract The heliospheric environment is very complex and filled with diverse high-energy particles from various origins, and among these particles, cosmic rays (CRs), including anomalous and Galactic components (ACRs and GCRs), are unique as they originate from beyond the solar system. Due to their stable and long-lasting presence in the heliosphere, the study of CRs is crucial for protecting humanity and other lifeforms against the hazards of high-energy radiation. In this work, we aim to study the prolonged changes in CR nitrogen, a fundamental element that constitutes amino acids, using the measurements from the Advanced Composition Explorer (ACE) spacecraft. The comparison of solar modulation on CR nitrogen and oxygen is also made using the yearly averaged ACE observations. The ACR nitrogen has a more even spectrum than ACR oxygen while the spectrum of GCR nitrogen is more abrupt than GCR oxygen. Further model results indicate that GCR nitrogen and oxygen undergo comparable modulation processes within the heliosphere and the slight difference between the observed power-law indices of nitrogen and oxygen spectrum is due to the distinct local interstellar spectrum of the two species. Besides, an analysis is made to explain the observed more flattened ACR spectrum of nitrogen than oxygen caused by the slightly different charge-to-mass ratio Z/A.

• Book : 980(1)
• Pub. Date : 2025
• Page : pp.106-106
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This study proposes improving the process of the vertical diffusion of temperature in numerical models to enhance the accuracy of sea surface temperature (SST) simulation. SST tends to be underestimated in the coastal and tidal flat regions, such as the Yellow Sea around Korea. In particular, SST in coastal areas is highly sensitive to wet/dry treatment, implying that the sensitivity of SST increases with the slope of coastal bathymetry. Therefore, during the calculation of vertical temperature diffusion terms, the numerical model’s surface boundary condition (SBC) was modified to limit excessive temperature differences below a certain depth in the coastal regions. Under wet or dry conditions defined by the wet/dry treatment, SBC and bottom boundary condition (BBC) adjustments are stabilized within a predefined depth limit. While horizontal diffusion also plays a role in the model, SST is significantly influenced by the balance of heat advection and shortwave radiation. To demonstrate this, Heat Limit Depth (HLD) was added as an input parameter into the vertical diffusion algorithm in the model to enhance sensitivity to the SBC. If the total water depth in the tidal flat is below the HLD and less than 1.0 m, the model is changed to estimate surface sediment temperature instead of SST. The improvement in the vertical diffusion term for SST was effective primarily in tidal flat areas. In contrast, the impact was less pronounced in coastal areas with average depths exceeding 5 m. The rationale for separating SBC and BBC in the improved air–sea interaction process is twofold: SBC adjustments are suitable for reducing heat flux effects, specifically in shallow depths or tidal flats, without significantly affecting the entire model domain, while combined SBC and BBC adjustments are more appropriate for inducing coastal SST changes across the domain.

• Book : 17(4)
• Pub. Date : 2025
• Page : pp.474-474
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AbstractThe role of MUTYH, a DNA repair glycosylase in the pathogenesis of acute kidney injury (AKI) is unclear. In this study, it is found that MUTYH protein levels are significantly decreased in the kidneys of cisplatin‐ or folic acid (FA)‐induced mouse AKI models and patients with AKI. MUTYH deficiency aggravates renal dysfunction and tubular injury following cisplatin and FA treatment, along with the accumulation of 7, 8‐dihydro‐8‐oxoguanine (8‐oxoG) and impairs mitochondrial function. Importantly, the overexpression of type 2 MUTYH (nuclear) significantly ameliorates cisplatin‐induced apoptosis, oxidative stress, mitochondrial dysfunction, and DNA damage in vivo and in vitro. In contrast, overexpression of type 1 MUTYH (mitochondrial) shows a marginal effect against cisplatin‐induced injury, indicating the chief role of type 2 MUTYH in antagonizing AKI. Interestingly, the results also indicate that the upregulation of the E3 ligase HUWE1 causes the ubiquitination and degradation of MUTYH in tubular epithelial cells. HUWE1 knockout or treatment with the HUWE1 inhibitor BI8622 significantly protect against cisplatin‐induced AKI. Taken together, these results suggest that the ubiquitin E3 ligase HUWE1‐mediates ubiquitination and degradation of MUTYH can aggravate DNA damage in the nucleus and mitochondria and promote AKI. Targeting the HUWE1/MUTYH pathway may be a potential strategy for AKI treatment.

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• Pub. Date : 2025
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ABSTRACT Lung cancer is a leading cause of cancer mortality, with non-small cell lung cancer (NSCLC) comprising the majority of cases. Despite the advent of immune checkpoint inhibitors (ICIs), a significant number of patients fail to achieve a durable response, highlighting the need to understand the factors influencing treatment efficacy. Saliva samples and tumor samples were collected from 20 NSCLC patients. The salivary microbiota was profiled using metagenomic next-generation sequencing, and metabolites were analyzed via liquid chromatography-mass spectrometry to identify correlations among bacteria, metabolites, and immunotherapy responses. Immunohistochemistry (IHC) analysis of tissue samples verified the result. Besides, in vitro experiments and tumor tissue microarray, including 70 NSCLC patients, were utilized to further explore the potential mechanism linking the oral microbiome and immunotherapy efficacy. The study revealed several differential species and distinct metabolite compositions between responders and non-responders to ICI therapy in NSCLC and explored correlations and mechanisms between microbiota metabolites and immunotherapy resistance. Notably, it was found that several Neisseria and Actinomyces species were significantly enriched in responders and identified lipids and lipid-like molecules associated with PD-L1 expression levels and treatment outcomes. Importantly, several differential lipid molecules were associated with differential species. Further, in vitro experiments and IHC experiments indicated that abnormal fat metabolism linked to dysbiosis is correlated with immunotherapy resistance through regulation of CD8 + T cell activity/infiltration and PD-L1 expression. Specific saliva microbiome and its associated lipids metabolites are significantly associated with the efficacy of ICI-based therapy in lung cancer. Our findings suggest that oral microbiome modulation and targeting lipid metabolism could improve immunotherapy responses, offering new avenues for personalized treatment strategies. IMPORTANCE In non-small cell lung cancer, our study links specific salivary microbiome profiles and related lipid metabolites to the efficacy of immune checkpoint inhibitor (ICI) therapies. Responders showed enrichment of certain Neisseria and Actinomyces species and distinct lipid compositions. These lipids correlate with PD-L1 expression and CD8 + T cell activity, affecting treatment outcomes. Our results imply that modulating the oral microbiome and targeting lipid metabolism may enhance ICI effectiveness, suggesting novel personalized therapeutic approaches.

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• Pub. Date : 2025
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Abstract Computed tomography (CT) is an instrument that is still widely used to diagnose and evaluate patient conditions from images produced by the CT scan process. CT scan images are formed from X-ray beams that penetrate the patient’s body and are captured by a detector and then displayed on a monitor screen. A fairly high dose of X-ray radiation is required to produce images with high resolution, good contrast, and low noise. However, this is contrary to the principles of ALARA, as low as reasonably achievable, and patient safety by using high-dose radiation. Various efforts to reduce the dose of X-ray radiation have been made. One of which is to use a low dose by operating the X-ray CT-scan machine at low voltage. Still, the consequence is that the resulting CT scan image has greater noise and lower spatial resolution compared to the image produced from the full radiation dose. This research measured spatial resolution by its modulation transfer function (MTF) and noise on open-source images taken at 25% dose by operating a 50-kV x-ray CT-scan machine on ACR phantom images using the IndoQCT application. The results are the noise of 25% dose images has an average of 86.71% greater and 41.96% lower spatial resolution than full-dose images. These results show that there are problems in using low-dose CT scans and open up opportunities to solve this problem by image reconstruction or image processing.

• Book : 2945(1)
• Pub. Date : 2025
• Page : pp.012013-012013
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Abstract Purpose Multidrug resistance-associated proteins (MRPs) have a widespread tissue distribution. They play an important role in drug disposition and drug-drug interactions (DDIs) and have been associated with various diseases. PET with 6-bromo-7-[11C]methylpurine ([11C]BMP) has been used to assess MRP1 function in the brain and lungs of mice. [11C]BMP crosses cellular membranes by passive diffusion followed by intracellular conjugation with glutathione and MRP1-mediated efflux of the radiolabelled glutathione-conjugate. In this study, we assessed the effect of the prototypical organic anion transporter inhibitor probenecid on the whole-body disposition of [11C]BMP to examine its suitability for measuring the function of MRP1 and possibly other MRP subtypes across multiple tissues. Methods Seven healthy volunteers (3 women, 4 men) underwent two dynamic whole-body PET scans on a long axial field-of-view (LAFOV) PET/CT system after intravenous injection of [11C]BMP, without and with pre-treatment with a single oral dose of probenecid. Volumes of interest were outlined for several MRP-expressing tissues (cerebral cortex, cerebellum, choroid plexus, retina, lungs, myocardium, skeletal muscle, kidneys, and liver). Tissue time-activity curves were corrected for the contribution of vascular radioactivity and the elimination rate constant (k E, h− 1) was calculated as a parameter for tissue MRP function. Results Radioactivity was primarily excreted into the urinary bladder and urinary clearance was significantly decreased after probenecid administration (− 50 ± 16%). Following probenecid administration, k E was significantly decreased in the kidneys (− 43 ± 20%), liver (− 18 ± 15%), myocardium (− 16 ± 12%), skeletal muscle (− 51 ± 34%), and retina (− 57 ± 29%, non-blood-corrected). Conclusion Our study highlights the great potential of LAFOV PET/CT to assess drug disposition and transporter-mediated DDIs in humans at a whole-body, multi-tissue level. Due to the slow elimination of [11C]BMP-derived radioactivity from the human brain, [11C]BMP appears unsuitable to measure cerebral MRP1 function in humans, but it may be used to assess the function of MRP1 and possibly other MRP subtypes in various peripheral tissues. Trial registration EudraCT 2021-006348-29. Registered 15 December 2021.

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• Pub. Date : 2025
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AbstractHerpes simplex virus 1 (HSV-1) infects about 50-80% of the entire human population and persists in the neurons of affected individuals. A fraction of affected individuals suffer from recurrent cold sores caused by reactivating virus, in rare but severe cases the virus can cause encephalitis.During lytic infection, the virus relies on host factors such as RNA polymerase II and accessory proteins involved in transcription to express its genes and ensure successful replication. In general, RNA molecules in cells are bound by RNA-binding proteins (RBPs) during their entire lifecycle. Importantly, RBPs are increasingly described to also regulate transcription, an aspect long time outside the scope of investigations, especially during viral infections. Here, we examined the impact of five nuclear proteins (FUBP1, SLBP, SFPQ, SPT5 and SAF-B) with known RNA-binding activities on HSV-1 transcription. Additionally, we evaluated their importance for human adenovirus C5 (HAdV) growth to assess whether these host factors are specific to HSV-1 infections or might have broader relevance for the general transcription of dsDNA viruses.We show that the transcriptional elongation factor SPT5 coded by SUPT5H accumulates on HSV-1 genomes early during the infection and is required for the transcription of the immediate-early gene UL54. Its depletion affects also HAdV replication, indicating a general role in transcription of viruses that depend on the host transcriptional machinery.In contrast, depletion of the transcriptional repressor and paraspeckle protein SFPQ reduces UL54 RNA levels in HSV-1 infection, but does not cause significant changes in HAdV growth. Since SFPQ does not co-localize with HSV-1 genomes, this suggests a function not directly associated to viral DNA.

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• Pub. Date : 2025
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