Controlled release microsphere drug product performance is contingent upon the structural intricacies of the microspheres, both within individual microspheres and between them. This paper describes a novel method for characterizing the structure of microsphere drug products, employing X-ray microscopy (XRM) and AI-based image analysis for efficiency and reliability. Minocycline-containing PLGA microspheres were generated in eight batches, each with uniquely calibrated production parameters, ultimately influencing their underlying microstructures and culminating in varied release performances. Employing high-resolution, non-invasive X-ray micro-radiography (XRM), a representative amount of microsphere samples from each batch was imaged. Through the application of reconstructed images and AI-based segmentation, the size distribution, intensity of the XRM signal, and intensity variation of thousands of microspheres per sample were determined. Consistent signal intensities were observed across the eight batches, irrespective of the microsphere diameter range, indicating a high level of structural similarity within each batch of spheres. Discrepancies in signal intensity across batches suggest variations in the underlying microstructures, a consequence of different manufacturing settings. High-resolution focused ion beam scanning electron microscopy (FIB-SEM) structures and in vitro release performance of the batches were found to correlate with the intensity variations. A discussion of the potential of this method for quick, on-the-spot and off-line appraisal of product quality, quality control, and quality assurance is presented.
Considering that a hypoxic microenvironment is a feature of the majority of solid tumors, a considerable investment has been made in developing approaches to address the issue of hypoxia. An investigation into ivermectin (IVM), a medication used against parasites, reveals its capability to mitigate tumor hypoxia through the inhibition of mitochondrial respiration. We examine this strategy to reinforce the effectiveness of oxygen-dependent photodynamic therapy (PDT), with chlorin e6 (Ce6) acting as the photosensitizer. Stable Pluronic F127 micelles encapsulate Ce6 and IVM, enabling a unified pharmacological response. The micelles' uniformity in size suggests their appropriateness for co-delivering Ce6 and IVM. The micelles' passive targeting action could direct drugs to tumors, enhancing their cellular penetration. A key consequence of mitochondrial dysfunction, induced by the micelles, is a decrease in oxygen consumption, lessening the hypoxic nature of the tumor. Subsequently, the augmented generation of reactive oxygen species would lead to a heightened efficacy of PDT in targeting hypoxic tumors.
Intestinal epithelial cells (IECs), though capable of expressing major histocompatibility complex class II (MHC II), particularly in circumstances of intestinal inflammation, remain indeterminate in their role of antigen presentation in driving either pro- or anti-inflammatory CD4+ T cell responses. By selectively ablating MHC II in IECs and their organoid counterparts, we explored the influence of IEC MHC II expression on CD4+ T cell responses and disease progression caused by enteric bacterial pathogens. lipid mediator We observed that colonic intestinal epithelial cells, in response to intestinal bacterial infections, demonstrated a substantial surge in the expression of MHC II processing and presentation molecules, driven by inflammatory signals. While IEC MHC II expression exhibited minimal influence on disease severity subsequent to Citrobacter rodentium or Helicobacter hepaticus infection, a colonic IEC organoid-CD4+ T cell co-culture system revealed that intestinal epithelial cells (IECs) can activate antigen-specific CD4+ T lymphocytes in an MHC II-dependent process, thereby modulating both regulatory and effector T helper cell subsets. Moreover, we evaluated adoptively transferred H. hepaticus-specific CD4+ T cells during intestinal inflammation in a live setting, and observed that enterocyte MHC II expression diminishes the activity of pro-inflammatory effector Th cells. The results of our study show that intestinal epithelial cells act as a novel type of antigen-presenting cells, with the expression of MHC class II molecules on IECs serving to delicately control the local effector CD4+ T cell response during intestinal inflammatory processes.
The risk of asthma, encompassing treatment-resistant severe forms, is linked to the unfolded protein response (UPR). Airway structural cells have been shown in recent studies to be impacted pathologically by the activating transcription factor 6a (ATF6a or ATF6), a critical UPR sensor. However, its contribution to the activity of T helper (TH) cells has not been adequately studied. The current study found that ATF6 was selectively induced by signal transducer and activator of transcription 6 (STAT6) in TH2 cells and by signal transducer and activator of transcription 3 (STAT3) in TH17 cells. UPR genes, upregulated by ATF6, facilitated the differentiation and cytokine secretion of TH2 and TH17 cells. In vitro and in vivo studies showed that the lack of Atf6 in T cells suppressed TH2 and TH17 responses, ultimately diminishing the manifestation of mixed granulocytic experimental asthma. The ATF6 inhibitor Ceapin A7 effectively dampened the expression of ATF6 target genes and Th cell cytokines in both murine and human memory CD4+ T cell populations. Ceapin A7's administration at the chronic asthma stage decreased TH2 and TH17 responses, thereby leading to a decrease in airway neutrophilia and eosinophilia inflammation. In conclusion, our data demonstrate a vital function of ATF6 in TH2 and TH17 cell-induced mixed granulocytic airway disease, indicating a potential new therapeutic approach for steroid-resistant mixed and even T2-low asthma endotypes by targeting ATF6.
For over eighty-five years, ferritin's primary function has been recognized as an iron storage protein, since its initial discovery. Although its primary role is iron storage, new functions are being discovered. Ferritin, encompassing processes like ferritinophagy and ferroptosis, and its function as a cellular iron transporter, broadens our understanding of its multifaceted roles and presents possibilities for cancer pathway targeting. Our review investigates the efficacy of ferritin modulation as a potential cancer treatment approach. Plant symbioses In cancers, we scrutinized the novel functions and processes attributed to this protein. This review considers not only the cellular modulation of ferritin's function in cancers but also its potential use as a 'Trojan horse' delivery system in cancer therapies. The newly discovered functions of ferritin, as elaborated upon herein, reveal its complex roles within cellular biology, offering potential therapeutic opportunities and stimulating future research.
The global push for decarbonization, environmental sustainability, and the increasing interest in renewable resources, including biomass, have catalyzed the development and utilization of bio-based chemicals and fuels. Based on these developments, the biodiesel industry is expected to flourish, as the transportation sector is pursuing various strategies to accomplish carbon-neutral mobility. However, the inevitable consequence of this industry is the generation of an abundant amount of glycerol as a waste by-product. Though glycerol acts as a renewable organic carbon source, assimilated by a multitude of prokaryotes, the full-scale implementation of a glycerol-based biorefinery is currently not a practical reality. β-Aminopropionitrile cell line From a range of platform chemicals like ethanol, lactic acid, succinic acid, 2,3-butanediol, and more, 1,3-propanediol (1,3-PDO) uniquely originates via fermentation, with glycerol as its source material. Metabolic Explorer's recent commercialization of glycerol-based 1,3-PDO in France has reawakened research interest in the development of alternative, cost-effective, scalable, and marketable biological procedures. A survey of natural glycerol-assimilating microbes and their 1,3-PDO synthesis is presented, including details of their metabolic pathways and associated genes. Eventually, technical limitations related to the direct utilization of industrial glycerol as a feedstock, along with the genetic and metabolic challenges concerning microbial application, are examined with care. A comprehensive review of biotechnological interventions—such as microbial bioprospecting, mutagenesis, metabolic engineering, evolutionary engineering, bioprocess engineering, and their combinations—is presented, highlighting their successful application in the past five years to effectively overcome such challenges. Concluding thoughts revolve around the emerging and promising discoveries within microbial cell factories and/or bioprocesses, resulting in innovative, effective, and resilient systems for glycerol-based 1,3-PDO production.
Sesamol, a crucial element in the composition of sesame seeds, is well-regarded for its contribution to a healthy lifestyle. Nevertheless, the impact of this on bone metabolic processes has yet to be investigated. The current research seeks to explore the impact of sesamol on bone tissue in growing, adult, and osteoporotic individuals, and elucidate the underlying mechanism driving its effect. Sesamol, at varying dosages, was administered orally to developing rats, both ovariectomized and with intact ovaries. Utilizing micro-CT and histological studies, bone parameter alterations were scrutinized. The procedure involved Western blotting and mRNA expression analysis of long bones. We investigated the impact of sesamol on osteoblast and osteoclast function, as well as its mechanism of action, within a cellular environment. Analysis of these data revealed that sesamol promoted the maximum bone mass in developing rats. Yet, in ovariectomized rats, sesamol showed the opposite effect, leading to a clear deterioration in the organization and structure of the trabecular and cortical microarchitecture. At the same time, bone density in adult rats was increased. In vitro experiments uncovered a link between sesamol and enhanced bone formation, with the mechanism involving stimulation of osteoblast differentiation through MAPK, AKT, and BMP-2 signaling.