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Journal of Separation Science Jul 2024Glycosylation and phosphorylation rank as paramount post-translational modifications, and their analysis heavily relies on enrichment techniques. In this work, a facile...
One-step enrichment and stepwise elution of glycoproteins and phosphoproteins by hydrophilic Ti-immobilized dendrimer poly(glycidyl methacrylate) microparticles functionalized with polyethylenimine and phytic acid.
Glycosylation and phosphorylation rank as paramount post-translational modifications, and their analysis heavily relies on enrichment techniques. In this work, a facile approach was developed for the one-step simultaneous enrichment and stepwise elution of glycoproteins and phosphoproteins. The core of this approach was the application of the novel titanium (IV) ion immobilized poly(glycidyl methacrylate) microparticles functionalized with dendrimer polyethylenimine and phytic acid. The microparticles possessed dual enrichment capabilities due to their abundant titanium ions and hydroxyl groups on the surface. They demonstrate rapid adsorption equilibrium (within 30 min) and exceptional adsorption capacity for β-casein (1107.7 mg/g) and horseradish peroxidase (438.6 mg/g), surpassing that of bovine serum albumin (91.7 mg/g). Furthermore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis was conducted to validate the enrichment capability. Experimental results across various biological samples, including standard protein mixtures, non-fat milk, and human serum, demonstrated the remarkable ability of these microparticles to enrich low-abundance glycoproteins and phosphoproteins from biological samples.
Topics: Glycoproteins; Phosphoproteins; Polyethyleneimine; Dendrimers; Humans; Titanium; Polymethacrylic Acids; Hydrophobic and Hydrophilic Interactions; Surface Properties; Animals; Particle Size; Adsorption; Cattle
PubMed: 38948935
DOI: 10.1002/jssc.202400154 -
BioRxiv : the Preprint Server For... Jun 2024Sickle cell disease is caused by a mutation in the beta subunit of hemoglobin (HbSS) that drives Hb fiber formation when the protein is in the deoxygenated (tense, T)...
UNLABELLED
Sickle cell disease is caused by a mutation in the beta subunit of hemoglobin (HbSS) that drives Hb fiber formation when the protein is in the deoxygenated (tense, T) state. The drug voxelotor was recently approved to treat sickle cell disease by preventing HbSS fiber formation. Voxelotor acts as an allosteric inhibitor of polymerization by maintaining the HbSS protein in the relaxed (R) conformation, limiting polymerization of T-state fibers. Normal blood cells contain small amounts of natural Hb fibers and a few percent of the Fe ferric form, metHb, incapable of binding oxygen. Although the drug Voxelotor is now in use, the effect of the drug on the oxidized metHb state has not been reported. Here we assessed the influence of voxelotor on normal human metHb. We compared the aggregation of metHb at two pH values (5.5 and 7.1). MetHb is known to form organized fiber structures at or below pH 5.5. We find that voxelotor significantly enhances fiber formation of metHb R-state at pH 5.5, consistent with the mode of action for this drug in maintaining the Hb R conformation. The opposite effect is observed at physiological pH values. Voxelotor significantly decreases the rate of metHb aggregate formation at pH 7.1 but did not affect protein stability. Notably, drug binding drives metHb into novel spherical particles with a morphology never seen before for Hb. The formation of these particles should be considered in patients being treated for sickle cell disease with voxelotor.
WHY IT MATTERS
Voxelotor is an FDA-approved drug for sickle cell anemia, known to prevent hemoglobin fiber formation. Here, we investigate its effect on methemoglobin, the form of hemoglobin in which iron takes on the ferric Fe state. Our study examines voxelotor's impact on methemoglobin aggregation and stability. At pH 7.1, we found voxelotor to have an effect on methemoglobin solubility as evidenced by the formation of novel methemoglobin spherical structures. We observe that voxelotor significantly increases methemoglobin fiber formation at pH 5.5 but, notably, reduces methemoglobin aggregation at physiological pH levels. Minimal impact on methemoglobin thermodynamic stability is noted. These findings suggest voxelotor's potential therapeutic efficacy for various hemoglobinopathies, including conditions characterized by Heinz body formation.
PubMed: 38948767
DOI: 10.1101/2024.06.16.599216 -
Analytical Science Advances Jun 2024Detecting foodborne contamination is a critical challenge in ensuring food safety and preventing human suffering and economic losses. Contaminated food, comprising... (Review)
Review
Detecting foodborne contamination is a critical challenge in ensuring food safety and preventing human suffering and economic losses. Contaminated food, comprising biological agents (e.g. bacteria, viruses and fungi) and chemicals (e.g. toxins, allergens, antibiotics and heavy metals), poses significant risks to public health. Microfluidic technology has emerged as a transformative solution, revolutionizing the detection of contaminants with precise and efficient methodologies. By manipulating minute volumes of fluid on miniaturized systems, microfluidics enables the creation of portable chips for biosensing applications. Advancements from early glass and silicon devices to modern polymers and cellulose-based chips have significantly enhanced microfluidic technology, offering adaptability, flexibility, cost-effectiveness and biocompatibility. Microfluidic systems integrate seamlessly with various biosensing reactions, facilitating nucleic acid amplification, target analyte recognition and accurate signal readouts. As research progresses, microfluidic technology is poised to play a pivotal role in addressing evolving challenges in the detection of foodborne contaminants. In this short review, we delve into various manufacturing materials for state-of-the-art microfluidic devices, including inorganics, elastomers, thermoplastics and paper. Additionally, we examine several applications where microfluidic technology offers unique advantages in the detection of food contaminants, including bacteria, viruses, fungi, allergens and more. This review underscores the significant advancement of microfluidic technology and its pivotal role in advancing the detection and mitigation of foodborne contaminants.
PubMed: 38948318
DOI: 10.1002/ansa.202400003 -
PeerJ 2024Plastic pollution is a widespread and growing concern due to its transformation into microplastics (MPs), which can harm organisms and ecosystems. This study, aimed to...
Plastic pollution is a widespread and growing concern due to its transformation into microplastics (MPs), which can harm organisms and ecosystems. This study, aimed to identify plastic pollution in the feces of terrestrial vertebrates using convenience sampling both inside and outside protected areas in Western Thailand. We hypothesized that MPs are likely to be detectable in the feces of all vertebrate species, primarily in the form of small black fragments. We predicted varying quantities of MPs in the feces of the same species across different protected areas. Furthermore, we expected that factors indicating human presence, landscape characteristics, scat weight, and the MP abundance in water, soils, and sediments would influence the presence of plastics in feces. Among 12 terrestrial species studied, potential MPs were found in 41.11% of 90 samples, totaling 83 pieces across eight species including the Asian elephant (), Eld's deer (), Dhole (), Gaur (), Sambar deer (), Wild boar (), Northern red muntjac (), and Butterfly lizard (). Specifically, 3.61% of all potential MPs (three pieces) were macroplastics, and the remaining 96.39% were considered potential MPs with the abundance of 0.92 ± 1.89 items.scat or 8.69 ± 32.56 items.100 g dw. There was an association between the numbers of feces with and without potential plastics and species (χ = 20.88, = 0.012). Most potential plastics were fibers (95.18%), predominantly black (56.63%) or blue (26.51%), with 74.70% smaller than two millimeters. Although there were no significant associations between species and plastic morphologies, colors, and sizes, the abundance classified by these characteristics varied significantly. FTIR identified 52.38% as natural fibers, 38.10% as synthetic fibers (rayon, polyurethane (PUR), polyethylene terephthalate (PET), polypropylene (PP), and PUR blended with cotton), and 9.52% as fragments of PET and Polyvinyl Chloride (PVC). Human-related factors were linked to the occurrence of potential plastics found in the feces of land-dwelling wildlife. This study enhances the understanding of plastic pollution in tropical protected areas, revealing the widespread of MPs even in small numbers from the areas distant from human settlements. Monitoring plastics in feces offers a non-invasive method for assessing plastic pollution in threatened species, as it allows for easy collection and taxonomic identification without harming live animals. However, stringent measures to assure the quality are necessitated to prevent exogenous MP contamination. These findings underscore the importance of raising awareness about plastic pollution in terrestrial ecosystems, especially regarding plastic products from clothing and plastic materials used in agriculture and irrigation systems.
Topics: Animals; Feces; Thailand; Environmental Monitoring; Plastics; Microplastics; Environmental Pollution; Vertebrates; Environmental Pollutants; Humans
PubMed: 38948236
DOI: 10.7717/peerj.17596 -
PeerJ 2024Burmese amber preserves a diverse assemblage of Cretaceous arachnids, and among pseudoscorpions (Arachnida: Pseudoscorpiones), ten species in five families have already...
Burmese amber preserves a diverse assemblage of Cretaceous arachnids, and among pseudoscorpions (Arachnida: Pseudoscorpiones), ten species in five families have already been named. Here, we describe a new fossil species from Burmese amber in the pseudoscorpion family Hyidae, providing detailed measurements, photographs and 3D-models from synchrotron scanning. Based on morphology, the new fossil, sp. nov. is placed in the genus , and is nearly identical to extant species in the genus, except for the position of trichobothrium on the pedipalpal chela, thereby indicating extreme morphological stasis in this invertebrate lineage over the last 99 million years. represents the first described fossil species in Hyidae, and the third described Burmese fossil in the superfamily Neobisioidea. It also joins the garypinid, , in representing the oldest fossil records for extant pseudoscorpion genera. Considering proposed divergence dates, the newly described fossil species bolsters a Gondwanan origin for Hyidae, and provides evidence for the "" hypothesis for the Burma Terrane, in which this landmass rifted from Gondwana in the Late Jurassic and collided with Eurasia by the Cretaceous/Eocene. Like species today, likely inhabited humicolous microhabitats in tropical forests on the Burma Terrane, supporting ecological niche stasis for this family since the Mesozoic.
Topics: Animals; Fossils; Arachnida; Amber; Biological Evolution; Myanmar; Phylogeny
PubMed: 38948233
DOI: 10.7717/peerj.17515 -
PeerJ 2024Plasmodesmata are transmembrane channels embedded within the cell wall that can facilitate the intercellular communication in plants. Plasmodesmata callose-binding...
Plasmodesmata are transmembrane channels embedded within the cell wall that can facilitate the intercellular communication in plants. Plasmodesmata callose-binding (PDCB) protein that associates with the plasmodesmata contributes to cell wall extension. Given that the elongation of cotton fiber cells correlates with the dynamics of the cell wall, this protein can be related to the cotton fiber elongation. This study sought to identify PDCB family members within the genome and to elucidate their expression profiles. A total of 45 distinct family members were observed through the identification and screening processes. The analysis of their physicochemical properties revealed the similarity in the amino acid composition and molecular weight across most members. The phylogenetic analysis facilitated the construction of an evolutionary tree, categorizing these members into five groups mainly distributed on 20 chromosomes. The fine mapping results facilitated a tissue-specific examination of group V, revealing that the expression level of peaked five days after flowering. The VIGS experiments resulted in a marked decrease in the gene expression level and a significant reduction in the mature fiber length, averaging a shortening of 1.43-4.77 mm. The results indicated that played a pivotal role in the cotton fiber development and served as a candidate for enhancing cotton yield.
Topics: Gossypium; Plasmodesmata; Cotton Fiber; Plant Proteins; Phylogeny; Gene Expression Regulation, Plant; Glucans; Multigene Family; Cell Wall; Carrier Proteins
PubMed: 38948221
DOI: 10.7717/peerj.17625 -
Macromolecules Jun 2024The arrangement of crystalline domains in semicrystalline polymers is key to understanding how to optimize the nanostructured morphology for enabling better properties....
The arrangement of crystalline domains in semicrystalline polymers is key to understanding how to optimize the nanostructured morphology for enabling better properties. For example, in polystyrene--poly(ethylene oxide) (PS--PEO), the degree of crystallinity and arrangement of the crystallites within the PEO phase plays a crucial role in determining the physical properties of the electrolyte. Here, we used four-dimensional scanning transmission electron microscopy to directly visualize the crystal domains within the PEO-rich region of the PS--PEO block copolymer and show the relative angle of the domain with respect to the PEO-PS interface. As demonstrated here, our analysis method is applicable to other electron-beam sensitive materials, especially semicrystalline polymers, to unveil their local phase condition and distribution.
PubMed: 38948181
DOI: 10.1021/acs.macromol.3c02231 -
Frontiers in Oral Health 2024To determine which components in a new restorative material (Renewal MI) improve its ability to form resin tags within demineralized dentine.
OBJECTIVES
To determine which components in a new restorative material (Renewal MI) improve its ability to form resin tags within demineralized dentine.
METHODS
Varied components included polylysine (PLS), monocalcium phosphate (MCP), powder to liquid ratio (PLR), 4-methacryloyloxyethyl trimellitate anhydride (4META), and polypropylene glycol dimethacrylate (PPGDMA). Urethane dimethacrylate (UDMA), containing PPGDMA (24 wt%) and 4META (3 wt%), was mixed with glass filler with MCP (8 wt%) and PLS (5 wt%). PLR was 3:1 or 5:1. Reducing MCP and/or PLS to 4 and 2 wt% respectively or fully removing MCP, PLS, 4META or PPGDMA gave 16 formulations in total. Renewal MI, Z250 (with or without Scotchbond Universal adhesive) and Activa were used as commercial comparators. Collagen discs were obtained by totally demineralizing 2 mm thick, human, premolar, coronal dentine discs by immersion in formic acid (4M) for 48 h. The restorative materials were then applied on top ( = 3), before dissolving the collagen in sodium hypochlorite (15%). SEM/EDX was employed to determine resin tags length, composition, and surface coverage.
RESULTS
Tags were >400, 20 and 200 µm and covered 62, 55 and 39% of the adhesion interface for Renewal MI, Scotchbond and Activa, respectively. With experimental formulations, they were 200 and >400 µm long with high vs. low PLR and composed primarily of polymerized monomers. Percentages of the adhesion interface covered varied between 35 and 84%. Reducing PLS or MCP caused a decline in coverage that was linear with their concentrations. Reducing MCP had lesser effect when PLS or PLR were low. Removal of 4META caused a greater reduction in coverage than PPGDMA removal.
CONCLUSION
PLS, MCP, 4META, PPGDMA and low PLR together enhance Renewal MI tags formation in, and thereby sealing of, demineralized dentine.
PubMed: 38948090
DOI: 10.3389/froh.2024.1420541 -
Theranostics 2024The repair of osteoporotic bone defects remains challenging due to excessive reactive oxygen species (ROS), persistent inflammation, and an imbalance between...
The repair of osteoporotic bone defects remains challenging due to excessive reactive oxygen species (ROS), persistent inflammation, and an imbalance between osteogenesis and osteoclastogenesis. Here, an injectable H-releasing hydrogel (magnesium@polyethylene glycol-poly(lactic-co-glycolic acid), Mg@PEG-PLGA) was developed to remodel the challenging bone environment and accelerate the repair of osteoporotic bone defects. This Mg@PEG-PLGA gel shows excellent injectability, shape adaptability, and phase-transition ability, can fill irregular bone defect areas via minimally invasive injection, and can transform into a porous scaffold to provide mechanical support. With the appropriate release of H and magnesium ions, the 2Mg@PEG-PLGA gel (loaded with 2 mg of Mg) displayed significant immunomodulatory effects through reducing intracellular ROS, guiding macrophage polarization toward the M2 phenotype, and inhibiting the IκB/NF-κB signaling pathway. Moreover, experiments showed that the 2Mg@PEG-PLGA gel inhibited osteoclastogenesis while promoting osteogenesis. Most notably, in animal experiments, the 2Mg@PEG-PLGA gel significantly promoted the repair of osteoporotic bone defects by scavenging ROS and inhibiting inflammation and osteoclastogenesis. Overall, our study provides critical insight into the design and development of H-releasing magnesium-based hydrogels as potential implants for repairing osteoporotic bone defects.
Topics: Animals; Magnesium; Reactive Oxygen Species; Mice; Polyethylene Glycols; Hydrogels; Osteoporosis; Osteogenesis; Hydrogen; RAW 264.7 Cells; Bone Regeneration; Immunomodulation; Tissue Scaffolds; Macrophages; Polyesters
PubMed: 38948054
DOI: 10.7150/thno.97412 -
ACS Omega Jun 2024Carbon-based nanopowders have been used as ionization materials for laser desorption ionization-mass spectrometry (LDI-MS) and are very efficient at detection in low /...
Carbon-based nanopowders have been used as ionization materials for laser desorption ionization-mass spectrometry (LDI-MS) and are very efficient at detection in low / regions. In this study, we aimed to develop a new sheet-type graphite material that possessed a randomly grooved nanostructured surface consisting of developed sp-conjugated atomic carbon to facilitate the desorption/ionization of small compounds in LDI-MS. The graphite sheet exhibited higher UV absorption and provided higher ionization efficiency and survival yield in the LDI-MS detection of a thermometer ion, 4-chloro-benzopyridinium, than those of highly oriented graphite plates. These properties demonstrate that the present graphite sheet is suited for use as an LDI-MS material. Graphite sheet-assisted LDI-MS successfully detected various substances, including amino acids, peptides, and polyethylene glycol polymers, with higher ion intensities and less noise than those associated with conventional organic matrix-assisted LDI-MS (MALDI-MS). Furthermore, graphite sheet-assisted LDI-MS analysis provided more peaks (252 peaks) derived from soy sauce than those obtained by MALDI-MS (36 peaks) and required fewer preparation processes (dilution and air-dried) compared with previously established graphite carbon black-assisted LDI-MS (171 peaks) in the positive mode. This study demonstrates that graphite sheet-assisted LDI-MS has the potential for small organic compound analyses in the biomedical and food science fields.
PubMed: 38947851
DOI: 10.1021/acsomega.4c04524