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Nature Communications Jul 2024Mutations in the microtubule-associated motor protein KIF1A lead to severe neurological conditions known as KIF1A-associated neurological disorders (KAND). Despite...
Mutations in the microtubule-associated motor protein KIF1A lead to severe neurological conditions known as KIF1A-associated neurological disorders (KAND). Despite insights into its molecular mechanism, high-resolution structures of KIF1A-microtubule complexes remain undefined. Here, we present 2.7-3.5 Å resolution structures of dimeric microtubule-bound KIF1A, including the pathogenic P305L mutant, across various nucleotide states. Our structures reveal that KIF1A binds microtubules in one- and two-heads-bound configurations, with both heads exhibiting distinct conformations with tight inter-head connection. Notably, KIF1A's class-specific loop 12 (K-loop) forms electrostatic interactions with the C-terminal tails of both α- and β-tubulin. The P305L mutation does not disrupt these interactions but alters loop-12's conformation, impairing strong microtubule-binding. Structure-function analysis reveals the K-loop and head-head coordination as major determinants of KIF1A's superprocessive motility. Our findings advance the understanding of KIF1A's molecular mechanism and provide a basis for developing structure-guided therapeutics against KAND.
Topics: Kinesins; Microtubules; Humans; Cryoelectron Microscopy; Tubulin; Protein Binding; Mutation; Models, Molecular; Protein Conformation
PubMed: 38956021
DOI: 10.1038/s41467-024-48720-4 -
Environmental Science and Pollution... Jul 2024The removal of dyes from the aquatic ecosystem is necessary being a major threat to life. For enhanced remediation of methylene blue (MB) dye, a new ternary...
The removal of dyes from the aquatic ecosystem is necessary being a major threat to life. For enhanced remediation of methylene blue (MB) dye, a new ternary biopolymer-geopolymer-surfactant composite adsorbent is synthesized by combining phosphoric acid geopolymer (PAGP), calcium alginate (Alg), and sodium lauryl sulfate (SLS). During the synthesis of the composites, PAGP and SLS were mixed with the alginate matrix, producing porous hybrid beads. The PAGP-SLS-alginate (PSA) beads prepared were characterized using different analytical tools, i.e., scanning electron microscopy (SEM), Fourier transform infrared spectrophotometry (FTIR), X-ray diffractometry (XRD), surface area and porosimetery (SAP), and thermogravimetric analysis (TGA). To ascertain the ideal conditions for the adsorption process, a batch reactor procedure was used to investigate the effects of several parameters on MB adsorption, including pH (2, 4, 6, 8, 10), PSA adsorbent dosage (0.06-0.12 g), MB concentration (50-500 mg/L), contact time (15 to 300 min), and temperature (25, 35, and 45 °C). The SEM investigation indicated that ~ 1860 μm-sized PSA beads with 6-8 μm voids are generated. Based on XRD, FTIR, and SAP examinations, the material is amorphous, having numerous functional groups and an average pore size of 6.42 nm. Variation of pH has a little effect on the adsorption process, and the pH of 7.44 was found to be the pH of the PSA beads. According to the findings of the batch study, equilibrium adsorption was obtained in 270-300 min, showing that the adsorption process was moderately slow-moving and effective. The dye adsorption linearly increased with initial dye concentration over concentration range of 50-500 mg/L and reciprocally decreased with rise in temperature. 0.06 g adsorbent dose, 25 °C, pH10, and 270 min were found to be the better conditions for adsorption experiments. Langmuir isotherm fitted well compared to Freundlich, Temkin, and Dubinin-Radushkevich (DR) isotherm models on the experimental data, and the maximum adsorption capacity(q) calculated was 1666.6 mg. g. Pseudo-second-order (PSO) kinetics model and multi steps (two) intra particle diffusion (IPD) model fitted well on the adsorption kinetics data. The system's entropy, Gibbs free energy, and change in enthalpy were measured and found to be -109.171 J. mol. K, - 8.198 to - 6.014 kJ. mol, and - 40.747 kJ. mol. Thermodynamics study revealed that adsorption process is exothermic, energetically favorable and resulting in the decrease in randomness. Chemisorption is found to be the dominant mechanism as confirmed by pH effect, Langmuir isotherm, PSO kinetics, IPD model, and thermodynamics parameters. PSA beads were successfully regenerated using ethanol in a course of 120 min and re-used for five times. To sum up, the PSA adsorbent's impressive adsorption capability of 1666.66 mg/g highlights its potential as a successful solution for methylene blue removal. The results of this study add to the expanding corpus of information on sophisticated adsorption materials and demonstrate PSA's potential for real-world uses in wastewater treatment and environmental clean-up.
PubMed: 38955975
DOI: 10.1007/s11356-024-33573-7 -
Environmental Science and Pollution... Jul 2024Highly active FeO/GO/Au composite nanomaterial was fabricated as a substrate of surface-enhanced Raman spectroscopy (SERS) and applied for pesticide residue detection....
Highly active FeO/GO/Au composite nanomaterial was fabricated as a substrate of surface-enhanced Raman spectroscopy (SERS) and applied for pesticide residue detection. The three-layer multifunctional FeO/GO/Au nanoparticles (NPs) were designed by facile method, with high hotspots, and were characterized by various techniques, including ultraviolet spectrophotometry (UV), X-ray diffraction (XRD), infrared absorption spectrometer (IR), and transmission electron microscopy (TEM). The performance of FeO/GO/Au was evaluated by Raman spectroscopy with R6G as a probe molecule to verify its enhancement effect. It exhibited a strong Raman signal with 10 M of R6G. Furthermore, the presence of FeO/GO/Au nanohybrid enabled the SERS-based method to detect mancozeb and showed an excellent linear relationship in the range of 0.25-25 ppm, with a low limit of detection (0.077 ppm), satisfactory EF, stability, and repeatability. In addition, the mechanism of SERS enhancement with electromagnetic mechanism (EM) and chemical mechanism (CM) was discussed in detail. Therefore, the proposed SERS approach holds promise as an auxiliary technique for screening contaminated agricultural products, environmental sample, and food in the future.
PubMed: 38955969
DOI: 10.1007/s11356-024-34191-z -
Tissue Engineering and Regenerative... Jul 2024This study aimed to identify glycine analogs conducive to the formation of cell-absorbable nanocomplexes, enhancing collagen synthesis and subsequent osteogenesis in...
Glycinamide Facilitates Nanocomplex Formation and Functions Synergistically with Bone Morphogenetic Protein 2 to Promote Osteoblast Differentiation In Vitro and Bone Regeneration in a Mouse Calvarial Defect Model.
BACKGROUND
This study aimed to identify glycine analogs conducive to the formation of cell-absorbable nanocomplexes, enhancing collagen synthesis and subsequent osteogenesis in combination with BMP2 for improved bone regeneration.
METHODS
Glycine and its derivatives were assessed for their effects on osteogenic differentiation in MC3T3-E1 cells and human bone marrow mesenchymal stem cells (BMSCs) under osteogenic conditions or with BMP2. Osteogenic differentiation was assessed through alkaline phosphatase staining and real-time quantitative polymerase chain reaction (RT-qPCR). Nanocomplex formation was examined via scanning electron microscopy, circular dichroism, and ultraviolet-visible spectroscopy. In vivo osteogenic effects were validated using a mouse calvarial defect model, and bone regeneration was evaluated through micro-computed tomography and histomorphometric analysis.
RESULTS
Glycine, glycine methyl ester, and glycinamide significantly enhanced collagen synthesis and ALP activity in conjunction with an osteogenic medium (OSM). GA emerged as the most effective inducer of osteoblast differentiation marker genes. Combining GA with BMP2 synergistically stimulated ALP activity and the expression of osteoblast markers in both cell lines. GA readily formed nanocomplexes, facilitating cellular uptake through strong electrostatic interactions. In an in vivo calvarial defect mouse model, the GA and BMP2 combination demonstrated enhanced bone volume, bone volume/tissue volume ratio, trabecular numbers, and mature bone formation compared to other combinations.
CONCLUSION
GA and BMP2 synergistically promoted in vitro osteoblast differentiation and in vivo bone regeneration through nanocomplex formation. This combination holds therapeutic promise for individuals with bone defects, showcasing its potential for clinical intervention.
PubMed: 38955905
DOI: 10.1007/s13770-024-00657-x -
The AAPS Journal Jul 2024The selection of skin is crucial for the in vitro permeation test (IVPT). The purpose of this study was to investigate the influence of different freezing-thawing...
The selection of skin is crucial for the in vitro permeation test (IVPT). The purpose of this study was to investigate the influence of different freezing-thawing processes on the barrier function of skin and the transdermal permeability of granisetron and lidocaine. Rat and hairless mouse skins were thawed at three different conditions after being frozen at -20℃ for 9 days: thawed at 4℃, room temperature (RT), and 32℃. There were no significant differences in the steady-state fluxes of drugs between fresh and thawed samples, but compared with fresh skin there were significant differences in lag time for the permeation of granisetron in rat skins thawed at RT and 32℃. Histological research and scanning electron microscopy images showed no obvious structural damage on frozen/thawed skin, while immunohistochemical staining and enzyme-linked immunosorbent assay for the tight junction (TJ) protein Cldn-1 showed significantly impaired epidermal barrier. It was concluded that the freezing-thawing process increases the diffusion rate of hydrophilic drugs partly due to the functional degradation of TJs. It's recommended that hairless, inbred strains and identical animal donors should be used, and the selected thawing method of skin should be validated prior to IVPT, especially for hydrophilic drugs.
Topics: Animals; Freezing; Skin; Mice; Mice, Hairless; Skin Absorption; Rats; Permeability; Male; Administration, Cutaneous; Lidocaine; Rats, Sprague-Dawley
PubMed: 38955873
DOI: 10.1208/s12248-024-00941-7 -
Mikrochimica Acta Jul 2024CoFe@C was first prepared by calcining the precursor of CoFe-metal-organic framework-74 (CoFe-MOF-74), then an electrochemical sensor for the determination of...
CoFe@C was first prepared by calcining the precursor of CoFe-metal-organic framework-74 (CoFe-MOF-74), then an electrochemical sensor for the determination of neohesperidin dihydrochalcone (NHDC) was constructed, which was stemmed from the novel CoFe@C/Nafion composite film modified glassy carbon electrode (GCE). The CoFe@C/Nafion composite was verified by field-emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM). Electrochemical impedance spectroscopy (EIS) was used to evaluate its electrical properties as a modified material for an electrochemical sensor. Compared with CoFe-MOF-74 precursor modified electrode, CoFe@C/Nafion electrode exhibited a great synergic catalytic effect and extremely increased the oxidation peak signal of NHDC. The effects of various experimental conditions on the oxidation of NHDC were investigated and the calibration plot was tested. The results bespoken that CoFe@C/Nafion GCE has good reproducibility and anti-interference under the optimal experimental conditions. In addition, the differential pulse current response of NHDC was linear with its concentration within the range 0.08 ~ 20 µmol/L, and the linear regression coefficient was 0.9957. The detection limit was as low as 14.2 nmol/L (S/N = 3). In order to further verify the feasibility of the method, it was successfully used to determine the content of NHDC in Chinese medicine, with a satisfactory result, good in accordance with that of high performance liquid chromatography (HPLC).
Topics: Electrodes; Cobalt; Metal-Organic Frameworks; Limit of Detection; Chalcones; Electrochemical Techniques; Drugs, Chinese Herbal; Hesperidin; Fluorocarbon Polymers; Oxidation-Reduction; Carbon; Reproducibility of Results; Iron
PubMed: 38955844
DOI: 10.1007/s00604-024-06525-8 -
Current Microbiology Jul 2024Antimicrobial resistance is a global health issue, in which microorganisms develop resistance to antimicrobial drugs, making infections more difficult to treat. This...
Antimicrobial resistance is a global health issue, in which microorganisms develop resistance to antimicrobial drugs, making infections more difficult to treat. This threatens the effectiveness of standard medical treatments and necessitates the urgent development of new strategies to combat resistant microbes. Studies have increasingly explored natural sources of new antimicrobial agents that harness the rich diversity of compounds found in plant species. This pursuit holds promise for the discovery of novel treatments for combating antimicrobial resistance. In this context, the chemical composition, antibacterial, and antibiofilm activities of the essential oil from Croton urticifolius Lam. leaves (CuEO) were evaluated. CuEO was extracted via hydrodistillation, and its chemical constituents were identified via gas chromatography-mass spectrometry (GC/MS). The antibacterial activity of CuEO was evaluated in a 96-well plate via the microdilution method, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were determined. The effect of CuEO on biofilm formation was assessed by quantifying the biomass using crystal violet staining and viable cell counting. In addition, alterations in the cellular morphology of biofilms treated with CuEO were examined using scanning electron microscopy (SEM) and laser confocal microscopy. GC/MS analysis identified 26 compounds, with elemicine (39.72%); eucalyptol (19.03%), E-caryophyllene (5.36%), and methyleugenol (4.12%) as the major compounds. In terms of antibacterial activity, CuEO showed bacteriostatic effects against Staphylococcus aureus ATCC 700698, S. aureus ATCC 25923, Staphylococcus epidermidis ATCC 12228, and Escherichia coli ATCC 11303, and bactericidal activity against S. aureus ATCC 700698. In addition, CuEO significantly inhibited bacterial biofilm formation. Microscopic analysis showed that CuEO damaged the bacterial membrane by leaching out the cytoplasmic content. Therefore, the results of this study show that the essential oil of C. urticifolius may be a promising natural alternative for preventing infections caused by bacterial biofilms. This study is the first to report the antibiofilm activity of C. urticifolius essential oil.
Topics: Biofilms; Oils, Volatile; Croton; Anti-Bacterial Agents; Microbial Sensitivity Tests; Plant Leaves; Staphylococcus aureus; Gas Chromatography-Mass Spectrometry; Plant Extracts; Cell Membrane
PubMed: 38955831
DOI: 10.1007/s00284-024-03779-6 -
Microscopy (Oxford, England) Jul 2024Scanning Transmission Electron Microscopy (STEM) enables direct determination of atomic arrangements in materials and devices. However, materials such as battery...
Scanning Transmission Electron Microscopy (STEM) enables direct determination of atomic arrangements in materials and devices. However, materials such as battery components are weak for electron beam irradiation and low electron doses are required to prevent beam-induced damages. Noise removal is thus essential for precise structural analysis of electron beam sensitive materials at atomic resolution. Total square variation (TSV) regularization is an algorithm that exhibits high noise removal performance. However, the use of the TSV regularization term leads to significant image blurring and intensity reduction. To address these problems, we here propose a new approach adopting L2 norm regularization based on higher-order total variation. An atomic-resolution STEM image can be approximated as a set of smooth curves represented by quadratic functions. Since the third-degree derivative of any quadratic function is 0, total third-degree variation (TTDV) is suitable for a regularization term. The application of TTDV for denoising the atomic-resolution STEM image of CaF2 observed along the [001] zone axis is shown, where we can clearly see the Ca and F atomic columns without compromising image quality.
PubMed: 38955673
DOI: 10.1093/jmicro/dfae031 -
The Journal of Prosthetic Dentistry Jul 2024Three Co-Cr alloy types (Co-Cr-Mo, Co-Cr-W, and Co-Cr-Mo-W) have been commonly used in the fabrication of dental prostheses. These alloys can be manufactured using...
STATEMENT OF PROBLEM
Three Co-Cr alloy types (Co-Cr-Mo, Co-Cr-W, and Co-Cr-Mo-W) have been commonly used in the fabrication of dental prostheses. These alloys can be manufactured using either conventional casting or selective laser melting (SLM) techniques. Nevertheless, research that directly compares these materials and/or manufacturing processes in terms of their microstructural and mechanical characteristics is sparse.
PURPOSE
The purpose of this in vitro study was to conduct microstructural and mechanical analysis via X-ray interpretation, optical microscopy, scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS), image analysis, X-ray diffraction (XRD), instrumented indentation testing (IIT), and 3-point bending testing to characterize Co-Cr-Mo, Co-Cr-W, and Co-Cr-Mo-W alloys produced through conventional casting and SLM.
MATERIAL AND METHODS
Six Co-Cr-based alloys were analyzed and divided into 3 types based on their elemental composition (Co-Cr-Mo, Co-Cr-W, and Co-Cr-Mo-W). Additionally, each group was categorized based on the manufacturing process used (casting or SLM). X-ray scans were used to assess porosity. The microstructures of the specimens were assessed through SEM/EDS examination and XRD analysis. IIT was used to determine the Martens hardness (HM) and elastic index (η), while the elastic modulus (E) was estimated through the 3-point bending test. The mechanical properties were statistically analyzed using 2-way analysis of variance (ANOVA) and the Tukey multiple comparison post hoc test, with alloy type and manufacturing process as discriminating variables (α=.05).
RESULTS
All cast groups exhibited gross porosity, while no pores or other flaws were found in the SLM groups. Based on the XRD results, the crystalline structure of all Co-Cr specimens consisted of the face-centered cubic γ phase (γ-fcc), along with the hexagonal close-packed ε phase (ε-hcp) and CrC carbide. Different microstructures were identified between the cast and SLM alloys. Significant differences were identified for the mean standard deviation HM (ranging from 2601 ±94 N/mm to 3633 ±61 N/mm) and mean ±standard deviation η (ranging from 16.8 ±0.3% to 20.9 ±0.3%) among alloys prepared by the same manufacturing process, while all SLM alloys had statistically higher HM and η results than their cast counterparts (P<.05). No statistically significant differences were identified for the mean ±standard deviation E (ranging from 170 ±25 GPa to 244 ±36 GPa) among the groups prepared with the same manufacturing process (P>.05), but the SLM alloys had significantly higher results (P<.05) than the cast alloys.
CONCLUSIONS
In general, the manufacturing procedure significantly affected the porosity, microstructure, and mechanical properties of the tested Co-Cr alloys. SLM decreased the internal porosity, provided a uniform microstructure, and improved the mechanical properties for all the tested alloy types.
PubMed: 38955601
DOI: 10.1016/j.prosdent.2024.06.015 -
Pharmacological Reviews Jul 2024The class F of G protein-coupled receptors (GPCRs) consists of ten Frizzleds (FZD) and Smoothened (SMO). FZDs bind and are activated by secreted lipoglycoproteins of the...
The class F of G protein-coupled receptors (GPCRs) consists of ten Frizzleds (FZD) and Smoothened (SMO). FZDs bind and are activated by secreted lipoglycoproteins of the Wingless/Int-1 (WNT) family and SMO is indirectly activated by the Hedgehog (Hh) family of morphogens acting on the transmembrane protein Patched (PTCH). The advance of our understanding of FZDs and SMO as dynamic transmembrane receptors and molecular machines, which emerged during the past 14 years since the first class F GPCR IUPHAR nomenclature report, justifies an update. This article focuses on the advances in molecular pharmacology and structural biology providing new mechanistic insight into ligand recognition, receptor activation mechanisms, signal initiation and signal specification. Furthermore, class F GPCRs continue to develop as drug targets, and novel technologies and tools such as genetically encoded biosensors and CRISP/Cas9 edited cell systems have contributed to refined functional analysis of these receptors. Also, advances in crystal structure analysis and cryogenic electron microscopy contribute to a rapid development of our knowledge about structure-function relationships providing a great starting point for drug development. Despite the progress questions and challenges remain to fully understand the complexity of the WNT/FZD and Hh/SMO signaling systems. The recent years of research have brought about substantial functional and structural insight into mechanisms of activation of Frizzleds and Smoothened. While the advance furthers our mechanistic understanding of ligand recognition, receptor activation, signal specification and initiation, broader opportunities emerge that allow targeting class F GPCRs for therapy and regenerative medicine employing both biologics and small molecule compounds.
PubMed: 38955509
DOI: 10.1124/pharmrev.124.001062