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Stem Cell Research & Therapy Jun 2024There is a significant demand for intermediate-scale bioreactors in academic and industrial institutions to produce cells for various applications in drug screening...
BACKGROUND
There is a significant demand for intermediate-scale bioreactors in academic and industrial institutions to produce cells for various applications in drug screening and/or cell therapy. However, the application of these bioreactors in cultivating hiPSC-derived immune cells and other blood cells is noticeably lacking. To address this gap, we have developed a xeno-free and chemically defined intermediate-scale bioreactor platform, which allows for the generation of standardized human iPSC-derived hematopoietic organoids and subsequent continuous production of macrophages (iPSC-Mac).
METHODS
We describe a novel method for intermediate-scale immune cell manufacturing, specifically the continuous production of functionally and phenotypically relevant macrophages that are harvested on weekly basis for multiple weeks.
RESULTS
The continuous production of standardized human iPSC-derived macrophages (iPSC-Mac) from 3D hematopoietic organoids also termed hemanoids, is demonstrated. The hemanoids exhibit successive stage-specific embryonic development, recapitulating embryonic hematopoiesis. iPSC-Mac were efficiently and continuously produced from three different iPSC lines and exhibited a consistent and reproducible phenotype, as well as classical functionality and the ability to adapt towards pro- and anti-inflammatory activation stages. Single-cell transcriptomic analysis revealed high macrophage purity. Additionally, we show the ability to use the produced iPSC-Mac as a model for testing immunomodulatory drugs, exemplified by dexamethasone.
CONCLUSIONS
The novel method demonstrates an easy-to-use intermediate-scale bioreactor platform that produces prime macrophages from human iPSCs. These macrophages are functionally active and require no downstream maturation steps, rendering them highly desirable for both therapeutic and non-therapeutic applications.
Topics: Humans; Induced Pluripotent Stem Cells; Macrophages; Bioreactors; Organoids; Cell Differentiation; Cell Culture Techniques; Hematopoiesis
PubMed: 38886860
DOI: 10.1186/s13287-024-03785-2 -
Environmental Science & Technology Jun 2024Groundwater contamination by 1,2,3-trichloropropane (TCP) poses a unique challenge due to its human toxicity and recalcitrance to degradation. Previous work suggests...
Role of Nitrogenous Functional Group Identity in Accelerating 1,2,3-Trichloropropane Degradation by Pyrogenic Carbonaceous Matter (PCM) and Sulfide Using PCM-like Polymers.
Groundwater contamination by 1,2,3-trichloropropane (TCP) poses a unique challenge due to its human toxicity and recalcitrance to degradation. Previous work suggests that nitrogenous functional groups of pyrogenic carbonaceous matter (PCM), such as biochar, are important in accelerating contaminant dechlorination by sulfide. However, the reaction mechanism is unclear due, in part, to PCM's structural complexity. Herein, PCM-like polymers (PLPs) with controlled placement of nitrogenous functional groups [i.e., quaternary ammonium (QA), pyridine, and pyridinium cations (py)] were employed as model systems to investigate PCM-enhanced TCP degradation by sulfide. Our results suggest that both PLP-QA and PLP-py were highly effective in facilitating TCP dechlorination by sulfide with half-lives of 16.91 ± 1.17 and 0.98 ± 0.15 days, respectively, and the reactivity increased with surface nitrogenous group density. A two-step process was proposed for TCP dechlorination, which is initiated by reductive ß-elimination, followed by nucleophilic substitution by surface-bound sulfur nucleophiles. The TCP degradation kinetics were not significantly affected by cocontaminants (i.e., 1,1,1-trichloroethane or trichloroethylene), but were slowed by natural organic matter. Our results show that PLPs containing certain nitrogen functional groups can facilitate the rapid and complete degradation of TCP by sulfide, suggesting that similarly functionalized PCM might form the basis for a novel process for the remediation of TCP-contaminated groundwater.
Topics: Sulfides; Polymers; Groundwater; Water Pollutants, Chemical; Carbon; Propane
PubMed: 38848107
DOI: 10.1021/acs.est.3c11010 -
Phytotoxicity and hormesis in common mobile organic compounds in leachates of wood-derived biochars.Biochar 2024Although addition of pyrolyzed organic materials (biochars) to soil generally results in increased growth and physiological performance of plants, neutral and negative...
UNLABELLED
Although addition of pyrolyzed organic materials (biochars) to soil generally results in increased growth and physiological performance of plants, neutral and negative responses have also commonly been detected. Toxicity of organic compounds generated during pyrolysis, sorbed by biochars, and then released into the soil solution, has been implicated as a possible mechanism for such negative effects. Conversely, water-soluble biochar constituents have also been suggested to have "hormetic" effects (positive effects on plants at low concentrations); however, no specific compounds responsible have been identified. We investigated the relative phytotoxicity-and possible hormetic effects-of 14 organic compounds common in aqueous extracts of freshly produced lignocellulosic biochars, using seed germination bioassays. Of the compounds examined, volatile fatty acids (VFAs: acetic, propionic, butyric, valeric, caproic, and 2-ethylbutyric acids) and phenol, showed acute phytotoxicity, with germination-based ED50 values of 1-30 mmol L, and 2-ethylbutyric acid showed ED50 values of 0.1-1.0 mmol L. Other compounds (benzene, benzoic acid, butanone, methyl salicylate, toluene, and 2,4-di-tert-butylphenol) showed toxic effects only at high concentrations close to solubility limits. Although phytotoxic at high concentrations, valeric and caproic acid also showed detectable hormetic effects on seedlings, increasing radicle extension by 5-15% at concentrations of ~ 0.01-0.1 mmol L. These data support the hypothesis that VFAs are the main agents responsible for phytotoxic effects of lignocellulosic biochar leachates, but that certain VFAs also have hormetic effects at low concentrations and may contribute to positive effects of biochar leachates on early plant development in some cases.
SUPPLEMENTARY INFORMATION
The online version contains supplementary material available at 10.1007/s42773-024-00339-w.
PubMed: 38799720
DOI: 10.1007/s42773-024-00339-w -
International Journal of Molecular... May 2024Macrophages are well known for their involvement in the biocompatibility, as well as biodistribution, of nano(bio)materials. Although there are a number of rodent cell...
Macrophages are well known for their involvement in the biocompatibility, as well as biodistribution, of nano(bio)materials. Although there are a number of rodent cell lines, they may not fully recapitulate primary cell responses, particularly those of human cells. Isolation of tissue-resident macrophages from humans is difficult and may result in insufficient cells with which to determine the possible interaction with nano(bio)materials. Isolation of primary human monocytes and differentiation to monocyte-derived macrophages may provide a useful tool with which to further study these interactions. To that end, we developed a standard operating procedure for this differentiation, as part of the Regulatory Science Framework for Nano(bio)material-based Medical Products and Devices (REFINE) project, and used it to measure the secretion of bioactive molecules from M1 and M2 differentiated monocytes in response to model nano(bio)materials, following an initial assessment of pyrogenic contamination, which may confound potential observations. The SOP was deployed in two partner institutions with broadly similar results. The work presented here shows the utility of this assay but highlights the relevance of donor variability in responses to nano(bio)materials. Whilst donor variability can provide some logistical challenges to the application of such assays, this variability is much closer to the heterogeneous cells that are present in vivo, compared to homogeneous non-human cell lines.
Topics: Humans; Macrophages; Cell Differentiation; Monocytes; Biocompatible Materials; Phenotype; Cells, Cultured
PubMed: 38791527
DOI: 10.3390/ijms25105491 -
Journal of Hazardous Materials Jul 2024Polycyclic aromatic compounds (PACs), including polycyclic aromatic hydrocarbons (PAHs) and heteroatom-containing analogues, constitute an important environmental...
Polycyclic aromatic compounds (PACs), including polycyclic aromatic hydrocarbons (PAHs) and heteroatom-containing analogues, constitute an important environmental contaminant class. For decades, limited numbers of priority PAHs have been routinely targeted in pollution investigations, however, there is growing awareness for the potential occurrence of thousands of PACs in the environment. In this study, untargeted Fourier transform ion cyclotron resonance mass spectrometry was used for the molecular characterisation of PACs in a sediment core from Chiswick Ait, in the River Thames, London, UK. Using complex mixture analysis approaches, including aromaticity index calculations, the number of molecular PAC components was determined for eight core depths, extending back to the 1930s. A maximum of 1676 molecular compositions representing PACs was detected at the depth corresponding to the 1950s, and a decline in PAC numbers was observed up the core. A case linking the PACs to London's coal consumption history is presented, alongside other possible sources, with some data features indicating pyrogenic origins. The overall core profile trend in PAC components, including compounds with oxygen, sulfur, nitrogen, and chlorine atoms, is shown to broadly correspond to the 16 priority PAH concentration profile trend previously determined for this core. These findings have implications for other industry-impacted environments.
PubMed: 38768537
DOI: 10.1016/j.jhazmat.2024.134605 -
World Journal of Gastrointestinal... May 2024Gastrointestinal (GI) cancer is a malignancy arising in the digestive system and accounts for approximately a third of increasing global cancer-related mortality,...
Gastrointestinal (GI) cancer is a malignancy arising in the digestive system and accounts for approximately a third of increasing global cancer-related mortality, especially in the colorectum, esophagus, stomach, and liver. Interleukin-1β (IL-1β) is a leukocytic pyrogen recognized as a tumor progression-related cytokine. IL-1β secretion and maturation in inflammatory responses could be regulated by nuclear factor-kappaB-dependent expression of NLR family pyrin domain containing 3, inflammasome formation, and activation of IL-1 converting enzyme. Several studies have documented the pro-tumorigenic effects of IL-1β in tumor microenvironments, promoting proliferation and metastatic potential of cancer cells and tumorigenesis . The application of IL-1β inhibitors is also promising for targeted therapy development in some cancer types. However, as a leukocytic pro-inflammatory cytokine, IL-1β may also possess anti-tumorigenic effects and be type-specific in different cancers. This editorial discusses the up-to-date roles of IL-1β in GI cancers, including underlying mechanisms and downstream signaling pathways. Understanding and clarifying the roles of IL-1β would significantly benefit future therapeutic targeting and help improve therapeutic outcomes in patients suffering from GI cancer.
PubMed: 38764841
DOI: 10.4251/wjgo.v16.i5.1676 -
Environmental Science & Technology May 2024This study investigated the reaction pathway of 2,4-dinitroanisole (DNAN) on the pyrogenic carbonaceous matter (PCM) to assess the scope and mechanism of PCM-facilitated...
This study investigated the reaction pathway of 2,4-dinitroanisole (DNAN) on the pyrogenic carbonaceous matter (PCM) to assess the scope and mechanism of PCM-facilitated surface hydrolysis. DNAN degradation was observed at pH 11.5 and 25 °C with a model PCM, graphite, whereas no significant decay occurred without graphite. Experiments were performed at pH 11.5 due to the lack of DNAN decay at pH below 11.0, which was consistent with previous studies. Graphite exhibited a 1.78-fold enhancement toward DNAN decay at 65 °C and pH 11.5 relative to homogeneous solution by lowering the activation energy for DNAN hydrolysis by 54.3 ± 3.9%. This is supported by our results from the computational modeling using Car-Parrinello simulations by ab initio molecular dynamics/molecular mechanics (AIMD/MM) and DFT free energy simulations, which suggest that PCM effectively lowered the reaction barriers by approximately 8 kcal mol compared to a homogeneous solution. Quaternary ammonium (QA)-modified activated carbon performed the best among several PCMs by reducing DNAN half-life from 185 to 2.5 days at pH 11.5 and 25 °C while maintaining its reactivity over 10 consecutive additions of DNAN. We propose that PCM can affect the thermodynamics and kinetics of hydrolysis reactions by confining the reaction species near PCM surfaces, thus making them less accessible to solvent molecules and creating an environment with a weaker dielectric constant that favors nucleophilic substitution reactions. Nitrite formation during DNAN decay confirmed a denitration pathway, whereas demethylation, the preferred pathway in homogeneous solution, produces 2,4-dinitrophenol (DNP). Denitration catalyzed by PCM is advantageous to demethylation because nitrite is less toxic than DNAN and DNP. These findings provide critical insights for reactive adsorbent design that has broad implications for catalyst design and pollutant abatement.
Topics: Hydrolysis; Anisoles; Molecular Dynamics Simulation; Carbon
PubMed: 38739946
DOI: 10.1021/acs.est.4c01069 -
Nucleic Acids Research Jun 2024CRISPR-Cas systems serve as adaptive immune systems in bacteria and archaea, protecting against phages and other mobile genetic elements. However, phages and archaeal...
CRISPR-Cas systems serve as adaptive immune systems in bacteria and archaea, protecting against phages and other mobile genetic elements. However, phages and archaeal viruses have developed countermeasures, employing anti-CRISPR (Acr) proteins to counteract CRISPR-Cas systems. Despite the revolutionary impact of CRISPR-Cas systems on genome editing, concerns persist regarding potential off-target effects. Therefore, understanding the structural and molecular intricacies of diverse Acrs is crucial for elucidating the fundamental mechanisms governing CRISPR-Cas regulation. In this study, we present the structure of AcrIIA28 from Streptococcus phage Javan 128 and analyze its structural and functional features to comprehend the mechanisms involved in its inhibition of Cas9. Our current study reveals that AcrIIA28 is a metalloprotein that contains Zn2+ and abolishes the cleavage activity of Cas9 only from Streptococcus pyrogen (SpyCas9) by directly interacting with the REC3 domain of SpyCas9. Furthermore, we demonstrate that the AcrIIA28 interaction prevents the target DNA from being loaded onto Cas9. These findings indicate the molecular mechanisms underlying AcrIIA28-mediated Cas9 inhibition and provide valuable insights into the ongoing evolutionary battle between bacteria and phages.
Topics: CRISPR-Associated Protein 9; CRISPR-Cas Systems; DNA; Gene Editing; Metalloproteins; Models, Molecular; Protein Binding; Protein Domains; Streptococcus; Streptococcus Phages; Viral Proteins; Zinc
PubMed: 38726868
DOI: 10.1093/nar/gkae357 -
EJNMMI Radiopharmacy and Chemistry May 2024Parkinson's disease is a neurodegenerative disorder that is characterized by a degeneration of the dopaminergic system. Dopamine transporter (DAT) positron emission...
BACKGROUND
Parkinson's disease is a neurodegenerative disorder that is characterized by a degeneration of the dopaminergic system. Dopamine transporter (DAT) positron emission tomography (PET) imaging has emerged as a powerful and non-invasive method to quantify dopaminergic function in the living brain. The PET radioligand, [F]FE-PE2I, a cocaine chemical derivative, has shown promising properties for in vivo PET imaging of DAT, including high affinity and selectivity for DAT, excellent brain permeability, and favorable metabolism. The aim of the current study was to scale up the production of [F]FE-PE2I to fulfil the increasing clinical demand for this tracer.
RESULTS
Thus, a fully automated and GMP-compliant production procedure has been developed using a commercially available radiosynthesis module GE TRACERLab FX2 N. [F]FE-PE2I was produced with a radiochemical yield of 39 ± 8% (n = 4, relative [F]F delivered to the module). The synthesis time was 70 min, and the molar activity was 925.3 ± 763 GBq/µmol (250 ± 20 Ci/µmol). The produced [F]FE-PE2I was stable over 6 h at room temperature.
CONCLUSION
The protocol reliably provides a sterile and pyrogen-free GMP-compliant product.
PubMed: 38696063
DOI: 10.1186/s41181-024-00269-9 -
Environmental Science & Technology May 2024CO is 45 to 50 times more concentrated in soil than in air, resulting in global diffusive fluxes that outpace fossil fuel combustion by an order of magnitude. Despite...
CO is 45 to 50 times more concentrated in soil than in air, resulting in global diffusive fluxes that outpace fossil fuel combustion by an order of magnitude. Despite the scale of soil CO emissions, soil-based climate change mitigation strategies are underdeveloped. Existing approaches, such as enhanced weathering and sustainable land management, show promise but continue to face deployment barriers. We introduce an alternative approach: the use of solid adsorbents to directly capture CO in soils. Biomass-derived adsorbents could exploit favorable soil CO adsorption thermodynamics while also sequestering solid carbon. Despite this potential, previous study of porous carbon CO adsorption is mostly limited to single-component measurements and conditions irrelevant to soil. Here, we probe sorption under simplified soil conditions (0.2 to 3% CO in balance air at ambient temperature and pressure) and provide physical and chemical characterization data to correlate material properties to sorption performance. We show that minimally engineered pyrogenic carbons exhibit CO sorption capacities comparable to or greater than those of advanced sorbent materials. Compared to textural features, sorbent carbon bond morphology substantially influences low-pressure CO adsorption. Our findings enhance understanding of gas adsorption on porous carbons and inform the development of effective soil-based climate change mitigation approaches.
Topics: Carbon Dioxide; Soil; Porosity; Adsorption; Carbon; Charcoal; Climate Change
PubMed: 38689207
DOI: 10.1021/acs.est.4c02015