Uptake Pathways Research Articles

Host cell responses to Candida albicans biofilm-derived extracellular vesicles

Candida albicans is a prevalent fungal pathogen responsible for infections in humans. As described recently, nanometer-sized extracellular vesicles (EVs) produced by C. albicans play a crucial role in the pathogenesis of infection by facilitating host inflammatory responses and intercellular communication. This study investigates the functional properties of EVs released by biofilms formed by two C. albicans strains—3147 (ATCC 10231) and SC5314—in eliciting host responses. We demonstrate the capability of C. albicans EVs to trigger reactions in human epithelial and immune cells. The involvement of EVs in pathogenesis was evidenced from the initial stages of infection, specifically in adherence to epithelial cells. We further established the capacity of these EVs to induce cytokine production in the epithelial A549 cell line, THP-1 macrophage-like cells, and blood-derived monocytes differentiated into macrophages. Internalization of EVs by THP-1 macrophage-like cells was confirmed, identifying macropinocytosis and phagocytosis as the most probable mechanisms, as demonstrated using various inhibitors that target potential vesicle uptake pathways in human cells. Additionally, C. albicans EVs and their cargo were identified as chemoattractants for blood-derived neutrophils. After verification of the in vivo effect of biofilm-derived EVs on the host, using Galleria mellonella larvae as an alternative model, it was demonstrated that vesicles from C. albicans SC5314 increased mortality in the injected larvae. In conclusion, for both types of EVs a predominantly pro-inflammatory effect on host was observed, highlighting their significant role in the inflammatory response during C. albicans infection.

Exercise-Intervened Circulating Extracellular Vesicles Alleviate Oxidative Stress in Cerebral Microvascular Endothelial Cells Under Hypertensive Plus Hypoxic Conditions

Our group has recently demonstrated that exercise intervention affects the release and function of bone marrow endothelial progenitor cell-derived extracellular vesicles (EVs) in transgenic hypertensive mice. Whether such an exercise regimen can impact circulating EVs (cEVs) remains unknown. In this study, we investigated the influence of exercise on cEV level and function. Transgenic hypertensive mice (Alb1-Ren) underwent 8-week treadmill exercise (10 m/min for 1 h, 5 days per week). Age- and sex-matched sedentary Alb1-Ren mice served as controls. cEVs were isolated from the blood of exercised and sedentary mice and are denoted as ET-cEV and nET-cEV, respectively. cEVs were labeled to determine their uptake efficiency and pathways. The functions of cEVs were assessed in an Angiotensin II (Ang II) plus hypoxia-injured cerebral microvascular endothelial cell (mBMEC) injury model. Cellular migration ability and oxidative stress were evaluated. We found that treadmill exercise stimulated cEV release, and ET-cEVs were more prone to be internalized by mBMECs. The ET-cEV internalization was mediated by macropinocytosis and endocytosis pathways. Functional studies showed that ET-cEVs can improve the compromised migration capability of mBMECs challenged by Ang II plus hypoxia. Additionally, ET-cEV treatment upregulated the expression of p-Akt/Akt in mBMECs. Compared to nET-cEVs, ET-cEVs significantly reduced ROS overproduction in Ang II plus hypoxia-injured mBMECs, associated with decreased Nox2 expression. All these findings suggest that exercise-intervened cEVs can protect cerebral microvascular endothelial cells against hypertensive and hypoxic injury.

Exposure pathways (diet, dissolved or particulate substrate) of rare earth elements to aquatic organisms.

The global extraction and use of rare earth elements (REEs) continue to rise as they are implemented in technologies that improve human and environmental livelihoods. However, the general understanding of transfer processes and fates of REEs in aquatic systems remains limited. Here, we aim to determine the REEs' main exposure pathways, e.g., particulate fraction, diet, or dissolved (ionic) fractions, to three benthic and three pelagic organisms. They were maintained under laboratory conditions and exposed to natural river water, with or without a sand substrate and an adapted diet. The organisms include northern clearwater crayfish (Faxonius propinquus), chinese mystery snail (Cipangopaludina chinensis), black sandshell mussel (Ligumia recta), striped shiner minnows (Luxilus chrysocephalus), Daphnia magna, and Euglena gracilis. The combined results of REE concentrations, fractionations, and anomalies highlighted that pelagic organisms are characterized by heavy REEs enrichment indicating they mainly uptake REEs in the dissolved form with high bioaccumulation potential, i.e., bioconcentration (BCF) > 1 and diet accumulation factors (DAF) < 1. Pelagic organisms exhibited relatively low REE concentrations in their tissues ([La] ranging from 4.6 to 57.7 µg kg-1 in minnows, 18.4 µg kg-1 in whole body D. magna, and 32.2 µg kg-1 in E. gracilis). On the other hand, snails and mussels were enriched in light REEs showing they mainly uptake REEs through their respective diets and particulate sand substrate. Relative to pelagic organisms, mussels and snails have higher DAFs (161.2 and 18.6, respectively) and REE levels in their soft tissues ([La] of 5700 µg kg -1 and 650 µg kg -1, respectively), but DAF for crayfish remains < 1. In summary, under environmental-relevant conditions, the six aquatic organisms has the potential to accumulate REEs through various uptake pathways. Nevertheless, our results confirming preferential uptake pathways of the six organisms can help select appropriate species in future studies to monitor REE exposure from vaious fractions: dissolved, particulate forms or in the food webs (i.e., diet).

An environment-responsive platform based on acid-resistant metal organic framework for efficient oral insulin delivery

Oral insulin delivery is considered a revolutionary alternative to daily subcutaneous injections in terms of compliance and convenience. However, significant challenges remain in terms of inactivation in gastrointestinal environment and limited permeation across the intestinal epithelium. Herein, we used acid-resistant metal-organic framework (PCN-222) to load insulin and modified the exterior with sodium dodecyl sulfate (SDS) to achieve efficient oral insulin delivery. The PCN-222 nanocarrier with ordered mesoporous cage structure and suitable pore size achieved a high insulin loading of 75 %. The SDS on the surface of nanocarrier reduces its hydrophilicity while reversibly altering cell morphology and increasing epithelial cell permeability, thereby promoting intestinal epithelial absorption. The constructed particle (I@P@S) was encapsulated in sodium alginate (SA) microspheres to protect it from gastric acid degradation and releases it upon entry into the intestinal tract. Through an uptake pathway dominated by clathrin-mediated endocytosis, the released I@P@S realized efficient intestinal permeability and controlled insulin release under physiological conditions due to the phosphate sensitivity of PCN-222, leading to an in vivo bioavailability of 12.9 %. This work provides a valuable reference for the design of oral insulin delivery systems.

Anionic polymer coating for enhanced delivery of Cas9 mRNA and sgRNA nanoplexes.

Polymeric carriers have long been recognized as some of the most effective and promising systems for nucleic acid delivery. In this study, we utilized an anionic di-block co-polymer, PEG-PLE, to enhance the performance of lipid-modified PEI (C14-PEI) nanoplexes for delivering Cas9 mRNA and sgRNA targeting KRAS G12S mutations in lung cancer cells. Our results demonstrated that PEG-PLE, when combined with C14-PEI at a weight-to-weight ratio of 0.2, produced nanoplexes with a size of approximately 140 nm, a polydispersity index (PDI) of 0.08, and a zeta potential of around -1 mV. The PEG-PLE/C14-PEI nanoplexes at this ratio were observed to be both non-cytotoxic and effective in encapsulating Cas9 mRNA and sgRNA. Confocal microscopy imaging revealed efficient endosomal escape and intracellular distribution of the RNAs. Uptake pathway inhibition studies indicated that the internalization of PEG-PLE/C14-PEI primarily involves scavenger receptors and clathrin-mediated endocytosis. Compared to C14-PEI formulations, PEG-PLE/C14-PEI demonstrated a significant increase in luciferase mRNA expression and gene editing efficiency, as confirmed by T7EI and ddPCR, in A549 cells. Sanger sequencing identified insertions and/or deletions around the PAM sequence, with a total of 69% indels observed. Post-transfection, the KRAS-ERK pathway was downregulated, resulting in significant increases in cell apoptosis and inhibition of cell migration. Taken together, this study reveals a new and promising formulation for CRISPR delivery as potential lung cancer treatment.

Harnessing seaweed farming for climate mitigation in South Korea: evaluating carbon dioxide removal potential and future research directions

Seaweed farming is emerging as a scalable and effective carbon dioxide removal (CDR) strategy, offering direct sequestration benefits through carbon uptake pathways and indirect climate advantages by substituting carbon-intensive products. This paper evaluates the potential of seaweed farming within the context of South Korea, leveraging its advanced aquaculture infrastructure, extensive coastal resources, and supportive policy frameworks. By synthesizing current literature, this study examines key sequestration mechanisms, including biomass storage, dissolved organic carbon (DOC) release, and particulate organic carbon (POC) burial, while addressing uncertainties such as the stability of recalcitrant DOC and the efficiency of POC burial under site-specific conditions. The analysis highlights South Korea’s unique strengths, such as its established seaweed farming industry and innovative technological developments, alongside challenges like ecological trade-offs, nutrient competition, and the absence of robust monitoring, reporting, and verification (MRV) systems. The paper identifies opportunities to scale offshore farming, adopt integrated multi-trophic aquaculture, and enhance lifecycle climate benefits through product innovation and renewable energy integration. To guide future research and policy, the paper outlines critical gaps, including the need for precise quantification of long-term carbon sequestration pathways, development of MRV frameworks, and exploration of socio-economic impacts. By addressing these gaps, seaweed farming can become a central pillar of South Korea’s climate mitigation strategy, providing valuable insights for other regions seeking to integrate marine-based solutions into global CDR efforts.

The Effect of Copper Limitation on the Bioavailability of Cu-Organic Complexes.

We examined the bioavailability of synthetic organic Cu complexes (Cu-L) and inorganic Cu species (Cu') to Thalassiosira oceanica growing under Cu-limiting and Cu-inhibiting conditions. Copper bioavailability depended on the phytoplankton Cu nutritional state and whether Cu' diffusion to the cell surface was sufficient to meet the cellular demands for growth. Under Cu-limiting conditions when [Cu'] was less than the diffusion concentration threshold (DCT: 10-14.13 M), growth rate was a hyperbolic function of [Cu-L]. Increasing [Cu'] above the DCT caused growth rate to increase proportionally, but growth rate also increased with increasing [Cu-L] so that both Cu' and Cu-L were bioavailable. Short-term photosynthesis assays conducted under Cu-limiting conditions showed a similar response to Cu speciation. In contrast, the growth rate of Cu-inhibited cells at high [Cu'] varied inversely with [Cu2+] and was independent of [Cu-L], as previously reported. The change in Cu-L bioavailability correlated with expression levels of genes encoding the reduction-dependent, high-affinity Cu uptake pathway, which was regulated by [Cu']. Our analysis shows that at [Cu'] typical of the open sea, T. oceanica-like phytoplankton are diffusion limited and must rely on Cu-organic complexes to fulfill their Cu requirements for growth.

Quantifying Altitudinal Mercury Accumulation in Biomonitors along Himalayan Valleys Using Mercury Isotopes.

The Himalayan valleys are important transport channels of atmospheric pollutants from South Asia to the Tibetan Plateau. This study aims to demonstrate the use of biomonitors (i.e., tree foliage, bark, mosses, and lichens) in the Himalayas to understand the sources and accumulation of mercury (Hg), including the transboundary atmospheric Hg transport across the Himalayas. Results showed that the significant variability in the physiological characteristics and nutrient uptake pathways, coupled with rapid changes in topography and climate-forced precipitation, led to significant differences in concentrations and isotopic compositions among biomonitor species. Δ199Hg values (-0.32 to -0.10‰) at the lower altitudes were slightly more positive than values at upper altitudes, likely reflecting signals of transboundary transport of anthropogenic Hg from South Asia. The isotope mixing model determined atmospheric Hg0 as the main source of Hg in most biomonitors (67 ± 13% to 88 ± 13%), except for Usnea longissimas (i.e., a unique type of lichen) with 61 ± 16% contribution of atmospheric Hg2+. Additionally, the morphological structure and epiphytic environment of U. longissimas facilitate aqueous Hg secondary reactions. Our results suggest that the Hg cycling in the Himalayan valleys could mix multiple impacts from montane environments and signals of transboundary transport of anthropogenic Hg from South Asia.

Fragment-based development of small molecule inhibitors targeting Mycobacterium tuberculosis cholesterol metabolism.

Mycobacterium tuberculosis ( Mtb ) is the world's most deadly infectious pathogen and new drugs are urgently required to combat the emergence of multi-(MDR) and extensively-(XDR) drug resistant strains. The bacterium specifically upregulates sterol uptake pathways in infected macrophages and the metabolism of host-derived cholesterol is essential for Mtb's long-term survival in vivo. Here, we report the development of antitubercular small molecules that inhibit the Mtb cholesterol oxidases CYP125 and CYP142, which catalyze the initial step of cholesterol metabolism. An efficient biophysical fragment screen was used to characterize the structure-activity relationships of CYP125 and CYP142, and identify a non-azole small molecule 1a that can bind to the heme cofactor of both enzymes. A structure-guided fragment-linking strategy was used to optimize the binding affinity of 1a , yielding a potent dual CYP125/142 inhibitor 5m (K D CYP125/CYP142 = 0.04/0.16 µM). Compound 5m potently inhibits the catalytic activity of CYP125 and CYP142 in vitro (K I values < 0.1 µM), and rapidly depletes Mtb intracellular ATP (IC 50 = 0.15 µM). The compound has antimicrobial activity against both drug susceptible and MDR Mtb ( MIC 99 values 0.4 - 1.5 µM ) in extracellular assays, and inhibits the growth of Mtb in human macrophages (MIC = 1.7 µM) with good selectivity over mammalian cytotoxicity (LD 50 ≥ 50 µM). The combination of small molecule inhibitors and structural data reported here provide useful tools to study the role of cholesterol metabolism in Mtb and are a promising step towards novel antibiotics targeting bioenergetic pathways, which could be used to help combat MDR-TB.

Cellular uptake of CPX-351 by scavenger receptor class B type 1-mediated nonendocytic pathway

The proper uptake of drugs in liposome formulations into target cells markedly impacts therapeutic efficacy. The protein corona (PC), formed by the adsorption of serum proteins onto the liposome surface, binds to specific surface receptors of target cells, influencing the uptake pathway. We investigated the uptake pathway into leukemia cells based on PC analysis of CPX-351, a liposome containing cytarabine and daunorubicin in a fixed 5:1 synergistic molar ratio. The PC of CPX-351 mixed with fetal bovine serum was analyzed by nanoflow liquid chromatography-tandem mass spectrometry. CPX-351 uptake in HL-60, K562, and THP-1 leukemia cell lines, was measured by flow cytometry using daunorubicin fluorescence. The major components of CPX-351 PC were apolipoproteins A-I and A-II, which bind to scavenger receptor class B type 1 (SR-BI), a nonendocytic pathway that takes up only liposome contents. SR-BI was expressed in each cell, and its expression correlated with CPX-351 uptake. The uptake was significantly decreased by the inhibition of clathrin-mediated endocytosis and macropinocytosis. Additionally, blocks lipid transport-1 (BLT-1), a selective inhibitor of SR-BI, decreased the uptake; however, high-dose BLT-1 addition significantly increased the uptake, which was more strongly inhibited by macropinocytosis suppression compared with clathrin-mediated endocytosis. BLT-1 enhances the binding of SR-BI to liposomes in a dose-dependent manner. These findings indicate that the enhancement of binding between SR-BI and CPX-351 activates different pathways, such as macropinocytosis, distinct from CPX-351 alone. SR-BI may be a biomarker for CPX-351 therapy, and the combination of CPX-351 with high-dose BLT-1 may augment therapeutic efficacy.

Iontophoresis-driven alterations in nanoparticle uptake pathway and intracellular trafficking in carcinoma skin cancer cells.

Effective treatment of squamous cell carcinoma (SCC) poses challenges due to intrinsic drug resistance and limited drug penetration into tumor cells. Nanoparticle-based drug delivery systems have emerged as a promising approach to enhance therapeutic efficacy; however, they often face hurdles such as inadequate cellular uptake and rapid lysosomal degradation. This study explores the potential of iontophoresis to augment the efficacy of liposome and immunoliposome-based drug delivery systems for SCC treatment. The study assessed iontophoresis effects on SCC cell line (A431) viability, nanoparticle uptake dynamics, and intracellular distribution patterns. Specific inhibitors were employed to delineate cellular internalization pathways, while fluorescence microscopy and immunohistochemistry examined changes in EGFR expression and lysosomal activity. Results demonstrated that iontophoresis significantly increased cellular uptake of liposomes and immunoliposomes, achieving approximately 50 % uptake compared to 10 % with passive treatment. This enhancement correlated with modifications in endocytic pathways, favoring macropinocytosis and caveolin-mediated endocytosis for liposomes, and macropinocytosis and clathrin-mediated pathways for immunoliposomes. Moreover, iontophoresis induced alterations in EGFR distribution and triggered syncytium-like cellular clustering. It also attenuated lysosomal activity, thereby reducing nanoparticle degradation and prolonging intracellular retention of therapeutic agents. These findings underscore the role of iontophoresis in modulating nanoparticle internalization pathways, offering insights that could advance targeted drug delivery strategies and mitigate therapeutic resistance in SCC and other malignancies.

Tackling the unyielding: testing two novel approaches against NDM-producing Enterobacterales and Pseudomonas aeruginosa isolates collected in India.

Metallo-β-lactamases are therapeutically challenging due to the limited treatment options. Against such isolates, currently approved newer β-lactam/β-lactamase inhibitor combinations are ineffective. In this study, we tested siderophore cephalosporin, cefiderocol, which utilizes an unconventional iron uptake pathway for efficient cellular penetration, and cefepime/zidebactam that utilizes novel β-lactam enhancer mechanisms for overcoming diverse carbapenemases. Cefiderocol showed limited activity against Escherichia coli isolates co-harboring New Delhi metallo-β-lactamase (NDM) with PBP3 insert, dual carbapenemase (NDM with OXA-48 like)-producing Klebsiella pneumoniae, and NDM-producing Pseudomonas aeruginosa isolates, while cefepime/zidebactam potently inhibited NDM-producing Enterobacterales and P. aeruginosa isolates. NDM being a dominant carbapenemase among Enterobacterales and P. aeruginosa in India, the variable activity of cefiderocol against NDM producers is a concern. Post approval, cefepime/zidebactam could offer a promising treatment option against NDM producers.

Contribution of Cancer-Specific Protein Coronas to the Pro-Tumor Effects of Nanoplastics through Enhanced Cellular Interactions.

The potential impact of nanoplastics (NPs) on human carcinogenic processes is a matter of growing concern, particularly in light of the global plastic pollution crisis. Although the potential effects of NPs on human health have been well investigated, many uncertainties remain regarding their role in tumor behavior. Upon exposure, NPs can enter the bloodstream and are prone to interacting with plasma proteins to form a protein corona (PC), which can influence their interactions with cancer cells. However, how the PCs adsorbed on NPs affect the particle-to-tumor cell interaction and their effect on the tumor biological behavior remain unclear. To better understand the formation of PCs following NPs exposure in the bloodstream under various clinical conditions, we investigated the PC compositions of NPs derived from thyroid cancer (TC) patients and healthy volunteers. Our data revealed a significant enrichment of fibrinogen in the PCs formed on NPs derived from TC patient plasma, which in turn accelerated the endocytosis of NPs into TC cells. In addition, the uptake pathway of NPs into TC cells differed substantially between the two groups studied due to the different PC compositions in cancer patients and healthy individuals. Moreover, intriguingly alterations in the PCs induced by the clinical pathology status were also found to promote NPs engulfment by human macrophages, resulting in potent pro-inflammatory effects, in turn exerting pro-tumor effects. These findings emphasize the importance of considering the significance of a realistic biological identity on NPs and their interactions with cancer cells and also pinpoint the implications of the carcinogenesis outcomes of NPs exposure in humans.

Divergent Proteomic Profiles and Uptake Mechanisms of Exosomes Derived from Human Dental Pulp Stem Cells, Endothelial Cells, and Fibroblasts.

Effective intercellular communication is crucial for tissue repair and regeneration, with exosomes playing a key role in mediating these processes by transferring proteins, lipids, and nucleic acids between cells. This study explored the mechanisms underlying the uptake of exosomes derived from human dental pulp stem cells (hDPSCs), human umbilical vein endothelial cells (HUVECs), and human fibroblasts (HFBs). Our findings revealed that hDPSCs exhibited the greatest capacity for exosome uptake across all three cell types. Moreover, exosomes originating from hDPSCs were also taken up in the highest amounts by all three cell types. Proteomic analysis uncovered significant differences in protein expression among exosomes from these different cell types, particularly in proteins related to endocytosis. Clathrin-dependent endocytosis emerged as the primary pathway for exosome uptake in hDPSCs and HUVECs, while HFBs appeared to use a different mechanism. Additionally, proteins such as fibronectin and tetraspanins were found to be highly expressed in hDPSC-derived exosomes, suggesting their potential involvement in exosome-cell interactions. This study offers new insights into exosome uptake mechanisms and highlights the potential of exosomes in advancing tissue engineering and regenerative medicine.

The microbial phosphorus cycle in aquatic ecosystems.

Phosphorus is an essential element for life, and phosphorus cycling is crucial to planetary habitability. In aquatic environments, microorganisms are a major component of phosphorus cycling and rapidly transform the diverse chemical forms of phosphorus through various uptake, assimilation and release pathways. Recent discoveries have revealed a more dynamic and complex aquatic microbial phosphorus cycle than previously understood. Some microorganisms have been shown to use and produce new phosphorus compounds, including those in reduced forms. New findings have also raised numerous unanswered questions that warrant further investigation. There is an expanding influence of human activity on aquatic ecosystems. Advancements in understanding the phosphorus biogeochemistry of evolving aquatic environments offer a unique opportunity to comprehend, anticipate and mitigate the effect of human activities. In this Review, I discuss the wealth of new aquatic phosphorus cycle research, spanning disciplines from omics and physiology to global biogeochemical modelling, with a focus on the current comprehension of how aquatic microorganisms sense, transport, assimilate, store, produce and release phosphorus. Of note, I delve into cellular phosphorus allocation, an underexplored topic with wide-ranging implications for energy and element flux in aquatic ecosystems.

Alterations in store-operated calcium entry in neurodegenerative pathologies: history, facts, perspectives

Neurodegenerative disorders, along with cardiovascular and oncological pathologies, are one of the most actual issues facing modern medicine. Therefore, the study of the molecular mechanisms of their pathogenesis and the search for new drug targets is highly demanded. Neuronal calcium signalling has attracted close attention, as altered calcium homeostasis has been demonstrated in the pathogenesis of various neurodegenerative diseases. In this review, we focus on one of the most ubiquitous and important pathways for calcium uptake: store-operated calcium entry. Here we describe studies demonstrating disturbances in store-operated calcium entry in various neurodegenerative pathologies, including Alzheimer’s, Parkinson’s and Huntington’s diseases. Also, we analyse the molecular determinants underlying these disturbances and propose ways for pharmacological correction of altered calcium signaling. The information summarized in the review will allow us to consider store-operated calcium channels as promising targets for the drug development in order to treat neurodegenerative pathologies and outline further promising directions for the investigation.

Differential Effects of Milk, Yogurt, and Cheese on Insulin Sensitivity, Hepatic Function, and Gut Microbiota in Diet-Induced Obese Mice

The prevalence of obesity and associated metabolic disorders, such as insulin resistance (IR) and fatty liver disease (steatosis), is often linked to a high-fat Western diet. Recent meta-analyses indicate that dairy consumption may mitigate these effects, particularly favoring low-fat dairy products. However, the underlying molecular mechanisms remain elusive. Dairy products vary in nutrient composition (calcium, fat) and structure, impacting digestion and nutrient absorption. Fermentation processes in cheese and yogurt introduce bacterial cultures with potentially positive metabolic effects. This study investigates the distinct effects of different dairy subgroups on insulin sensitivity, liver function, and gut microbiota in diet-induced obese mice, exploring potential mechanisms for mitigating Western diet-induced metabolic dysfunction.C57BL/6 mice (n = 16/group) were fed a high-fat diet (HFD, 45% fat) mimicking a Western diet rich in saturated fatty acids (SFA), or HFD supplemented with: fat-free milk (MILK), fat-free yogurt (YOG), or reduced-fat cheese (CHE, 19% fat) for 8 weeks. These supplements were administered daily at 10% of total caloric intake, reflecting typical U.S. dietary patterns. We assessed insulin resistance using the insulin tolerance test (ITT) and the homeostasis model assessment (HOMAIR). Weekly monitoring of food and water intake and body weight was conducted. After euthanasia, liver tissue was collected for Western blot analysis of key proteins in insulin signalling, gluconeogenesis, lipogenesis, lipid oxidation, uptake, and release pathways. Comprehensive lipidomics profiling of the liver and serum was performed using LC-MS, and fecal samples were analyzed for microbial diversity via 16S rRNA sequencing. Statistical analyses included QIIME2DEseq2, MetaboAnalyst 5.0, and MicrobiomeAnalyst. One-way and two-way ANOVA with Tukey’s post-hoc test were performed.The MILK and YOG interventions significantly reduced body weight, fat mass, and epididymal fat pad weight (p &lt; 0.05) compared to HFD alone. Enhancements in glucose clearance and insulin sensitivity were notable in the MILK and YOG groups, with associated upregulations in glycogen synthase (GS), phosphoenolpyruvate carboxykinase (PEPCK), and protein kinase B (AKT) signaling. Regarding hepatic steatosis, MILK and YOG increased protein levels involved in fatty acid oxidation (ACSL, CPT1) and lowered PPARγ, suggesting reduced fat storage. Additionally, higher AMPK, PPARα, ATGL, and HSL protein abundances supported enhanced fatty acid breakdown. Gut microbiota analyses revealed a higher bacterial species richness in the MILK and YOG groups, with distinct fecal microbiota compositions across all dairy interventions. Lipidomics identified unique and shared metabolic markers among the groups, implicating glycerolipid, glycerophospholipid, sphingomyelin metabolism, and inflammatory pathways.This study demonstrates that different dairy products uniquely modulate metabolic health markers in diet-induced obese mice. Milk and yogurt particularly promote improved insulin sensitivity and hepatic functions, potentially mediated through alterations in metabolic pathways and gut microbiota composition.

A novel approach to regulate glucose uptake in an anaplastic thyroid cancer cell line.

Curcumin's function in affecting cancer metabolic reprogramming remains poorly understood. Herein, we aimed to elucidate a novel link between Curcumin and the glucose uptake metabolism and glucose transporters (GLUTs) status in SW1736 cell line derived from anaplastic thyroid cancer. TheMTT test and flow cytometry was employed to test cell viability and cell death. For glucose uptake detection, ''GOD-PAP'' enzymatic colorimetric assay was applied to measure the direct glucose levels inside of the cells. Determination of GLUT1 and GLUT3 mRNA and protein expression in SW1736 cells was performed by qRT-PCR and western blotting. Also, the scratch wound healing assay was conducted for cell migration. The data indicated that Curcumin-induced cell death is independent of apoptosis in this type of thyroid cancer cell line. Furthermore, significantly reduced GLUT1 and GLUT3 expression was observed after treatment with Curcumin, resulting in the inhibition of glucose uptake (p < 0.05). Scratch assay indicated the inhibition of cell migration in SW1736 cells treated by Curcumin (p < 0.05). It can be concluded that GLUTs as metabolic targets can be blocked specifically by Curcumin for thyroid cancer prevention. Curcumin, as a promising anti-cancer agent, inhibits the growth of SW1736 anaplastic thyroid cancer cell line by regulating glucose uptake pathway and cell death. Altogether, these results suggest that the glucose pathway may be an important target for therapeutic intervention to sensitize tumor cells to cell death process by inhibition of glucose transporters.

Effects of different carbapenemase and siderophore production on cefiderocol susceptibility in Klebsiella pneumoniae.

The resistance mechanism of Gram-negative bacteria to the siderophore antibiotic cefiderocol is primarily attributed to carbapenemase and siderophore uptake pathways; however, specific factors and their relationships remain to be fully elucidated. Here, we constructed cefiderocol-resistant Klebsiella pneumoniae (CRKP) strains carrying different carbapenemases and knocked out siderophore genes to investigate the roles of various carbapenemases and siderophores in the development of cefiderocol resistance. Antimicrobial susceptibility testing revealed that both blaNDM and blaKPC significantly increased the minimum inhibitory concentration (MIC) of Klebsiella pneumoniae (KP) to cefiderocol, while blaOXA-48 showed a modest increase. Notably, KP expressing NDM exhibited a higher cefiderocol MIC compared to KP expressing KPC, although expression of NDM alone did not induce cefiderocol resistance. Laboratory evolutionary experiments demonstrated that combining pNDM with mutations in the siderophore uptake receptor gene cirA and pKPC with a mutation in the two-component system gene envZ led to KP reaching a high level of cefiderocol resistance. Although combining pOXA with mutations in the two-component system gene baeS did not induce cefiderocol resistance, it significantly reduced susceptibility. Moreover, siderophores could influence the development of cefiderocol resistance. Strains deficient in enterobactin exhibited increased susceptibility to cefiderocol, while deficiencies in yersiniabactin and salmochelin showed no significant alterations. In conclusion, carbapenemase gene expression facilitates cefiderocol resistance, but its presence alone is insufficient. Cefiderocol resistance in CRKP typically involves abnormal expression of certain genes and other factors, such as mutations in siderophore uptake receptor genes and two-component system genes. The enterobactin siderophore synthesis gene entB may also contribute to resistance.

Toxoplasma gondii sustains survival by regulating cholesterol biosynthesis and uptake via SREBP2 activation

Toxoplasma gondii (T. gondii) is an obligate intracellular parasite that cannot biosynthesize cholesterol via the mevalonate pathway, it sources this lipid from its host. We discovered that T. gondii infection upregulated the expression of host cholesterol synthesis-related genes HMG-CoA reductase(HMGCR), squalene epoxidase (SQLE), and dehydrocholesterol reductase-7 (DHCR7), and increased the uptake pathway gene low-density lipoprotein receptor (LDLR). We found a protein, sterol regulatory element binding protein 2 (SREBP2), which is the key protein regulating the host cholesterol synthesis and uptake during T. gondii infection. T. gondii induced a dose-dependent nuclear translocation of SREBP2. Knockdown SREBP2 reduced T. gondii-induced cholesterol biosynthesis and uptake. Consequently, the parasite's ability to acquire cholesterol was significantly diminished, impairing its invasion, replication, and bradyzoites development. Interfering cholesterol metabolism using AY9944 effectively inhibited T. gondii replication. In summary, SREBP2 played an important role in T. gondii infection in vitro, serving as a potential target for regulating T. gondii-induced cholesterol metabolism, offering insights into the prevention and treatment of toxoplasmosis.