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BioRxiv : the Preprint Server For... Jun 2024We present an efficient approach for synthesizing cationic poly(ethylene imine) derivatives using the multicomponent split-Ugi reaction to rapidly create a library of...
We present an efficient approach for synthesizing cationic poly(ethylene imine) derivatives using the multicomponent split-Ugi reaction to rapidly create a library of complex functional ionizable lipopolymers. We synthesized a diverse library of 155 polymers, formulated them into polyplexes to establish structure-activity relationships crucial for endosomal escape and efficient transfection. After discovering a lead structure, lipopolymer-lipid hybrid nanoparticles are introduced to preferentially deliver to and elicit effective mRNA transfection in lung endothelium and immune cells, including T cells with low toxicity. The lipopolymer-lipid hybrid nanoparticles showed 300-fold improvement in systemic mRNA delivery to the lung compared to -JetPEI . Lipopolymer-lipid hybrid nanoparticles demonstrated efficient delivery of mRNA-based therapeutics for treatment of two different disease models. Lewis Lung cancer progression was significantly delayed after treatment with loaded IL-12 mRNA in U155@lipids after repeated i.v. administration. Systemic delivery of human CFTR (hCFTR) mRNA resulted in production of functional form of CFTR protein in the lungs. The functionality of hCFTR protein was confirmed by restoration of CFTR- mediated chloride secretion in conductive airway epithelia in CFTR knockout mice after nasal instillation of hCFTR mRNA loaded U155@lipids. We further showed that, U155@lipids nanoparticles can deliver complex CRISPR-Cas9 based RNA cargo to the lung, achieving 5.6 ± 2.4 % gene editing in lung tissue. Moreover, we demonstrated successful PD-1 gene knockout of T cells . Our results highlight a versatile delivery platform for systemic delivering of mRNA of various sizes for gene therapy for a variety of therapeutics.
PubMed: 38915714
DOI: 10.1101/2024.06.11.598497 -
Scientific Reports Jun 2024As genomic databases expand and artificial intelligence tools advance, there is a growing demand for efficient characterization of large numbers of proteins. To this...
As genomic databases expand and artificial intelligence tools advance, there is a growing demand for efficient characterization of large numbers of proteins. To this end, here we describe a generalizable pipeline for high-throughput protein purification using small-scale expression in E. coli and an affordable liquid-handling robot. This low-cost platform enables the purification of 96 proteins in parallel with minimal waste and is scalable for processing hundreds of proteins weekly per user. We demonstrate the performance of this method with the expression and purification of the leading poly(ethylene terephthalate) hydrolases reported in the literature. Replicate experiments demonstrated reproducibility and enzyme purity and yields (up to 400 µg) sufficient for comprehensive analyses of both thermostability and activity, generating a standardized benchmark dataset for comparing these plastic-degrading enzymes. The cost-effectiveness and ease of implementation of this platform render it broadly applicable to diverse protein characterization challenges in the biological sciences.
Topics: Robotics; Escherichia coli; Protein Engineering; High-Throughput Screening Assays; Hydrolases; Polyethylene Terephthalates; Reproducibility of Results
PubMed: 38914665
DOI: 10.1038/s41598-024-64938-0 -
Scientific Reports Jun 2024The joint impact of tadalafil (Cilais) as a pharmaceutical residue and microplastics on fish is not well comprehended. The current study examined haematological,...
The joint impact of tadalafil (Cilais) as a pharmaceutical residue and microplastics on fish is not well comprehended. The current study examined haematological, biochemical, and antioxidant parameters, along with immunohistochemical and histological indications in tilapia (Oreochromis niloticus) after being exposed to tadalafil, polyethylene microplastics (PE-MPs), and their mixtures for 15 days. The fish were distributed into 1st group control group (The fish was maintained in untreated water without any supplements); 2nd group exposed to 10 mg/L PE-MPs;3rd group exposed to 20 mg/l tadalafil (Cilais); 4th group exposed to 20 mg/l tadalafil (Cilais) + 10 mg/LPE-MPs (in triplicate). The levels of creatinine, uric acid, glucose, AST, ALT, and albumin in fish treated with tadalafil alone or in combination with PE-MPs were significantly higher than those in the control group. Fish exposed to PE-MPs, tadalafil, and tadalafil plus PE-MPs showed significantly lower levels of RBCs, Hb, Ht, neutrophils, and lymphocytes compared to the control group. Serum levels of total antioxidant capacity and reduced glutathione (GSH) were notably lowered in fish groups subjected to PE-MPs, tadalafil, and tadalafil + PE-MPs combinations in comparison to the control group. Malondialdehyde (MDA) serum levels were notably elevated in fish groups subjected to PE-MPs, tadalafil, and tadalafil + PE-MPs combinations compared to the control group. The most severe impact was observed in the tadalafil + PE-MPs combination group. Interleukin-6 (IL-6) levels were significantly increased in liver tissues following exposure to both tadalafil and microplastics compared to tissues exposed to only one substance or the control group. Changes in the gills, liver, and renal tissues were seen following exposure to PE-MPs, tadalafil, and tadalafil + PE-MPs combination in comparison to the control group of fish. Ultimately, the mixture of tadalafil and PE-MPs resulted in the most detrimental outcomes. Tadalafil and PE-MPs exhibited showed greater adverse effects, likely due to tadalafil being absorbed onto PE-MPs.
Topics: Animals; Tadalafil; Cichlids; Water Pollutants, Chemical; Microplastics; Antioxidants; Tilapia; Glutathione; Gills; Oxidative Stress
PubMed: 38914580
DOI: 10.1038/s41598-024-64282-3 -
Scientific Reports Jun 2024Nucleic acid amplification testing has great potential for point-of-need diagnostic testing with high detection sensitivity and specificity. Current sample preparation...
Nucleic acid amplification testing has great potential for point-of-need diagnostic testing with high detection sensitivity and specificity. Current sample preparation is limited by a tedious workflow requiring multiple steps, reagents and instrumentation, hampering nucleic acid testing at point of need. In this study, we present the use of mixed cellulose ester (MCE) paper for DNA binding by ionic interaction under molecular crowding conditions and fluid transport by wicking. The poly(ethylene) glycol-based (PEG) reagent simultaneously provides the high pH for alkaline lysis and crowding effects for ionic binding of the DNA under high salt conditions. In this study, we introduce Paper-based Abridged Solid-Phase Extraction with Alkaline Poly(ethylene) Glycol Lysis (PASAP). The anionic mixed cellulose ester (MCE) paper is used as solid phase and allows for fluid transport by wicking, eliminating the need for pipetting skills and the use of a magnet to retain beads. Following the release of DNA from the cells due to the lytic activity of the PASAP solution, the DNA binds to the anionic surface of the MCE paper, concentrating at the bottom while the sample matrix is transported towards the top by wicking. The paper was washed by dipping it in 40% isopropanol for 10 s. After air-drying for 30 s, the bottom section of the paper (3 mm × 4 mm) was snapped off using the cap of a PCR tube and immersed in the colourimetric loop-mediated isothermal amplification (cLAMP) solution for direct amplification and colourimetric detection. The total sample processing was completed in 15 min and ready for amplification. cLAMP enabled the detection of 10 CFU/mL of Escherichia coli (E. coli) from culture media and the detection of E. coli in milk < 10 CFU/mL (10 CFU) after incubation at 68 °C for 60 min, demonstrating applicability of the method to complex biological samples.
Topics: Nucleic Acid Amplification Techniques; Paper; Colorimetry; DNA; Solid Phase Extraction; Polyethylene Glycols; DNA, Bacterial; Molecular Diagnostic Techniques
PubMed: 38914553
DOI: 10.1038/s41598-024-59763-4 -
ACS Omega Jun 2024This study demonstrates a procedure for preparing gel-like cellulose nanofibers (CNFs) in polyethylene glycol (PEG) to toughen polylactic acid (PLA) nanocomposite films....
This study demonstrates a procedure for preparing gel-like cellulose nanofibers (CNFs) in polyethylene glycol (PEG) to toughen polylactic acid (PLA) nanocomposite films. A well-dispersed solution of CNFs in ethanol was produced from microcrystalline cellulose by using a high-pressure microfluidizer. The fiber diameter of CNFs was found to be in the range of 80-100 nm. Ethanol was replaced by PEG using a rotary evaporator to obtain gel-like CNFs/PEG. PLA/PEG/CNF films were prepared using the solvent casting method, with the CNF content varying from 0.15 to 5 phr. The effect of CNFs on the mechanical, morphological, and thermal properties of PLA nanocomposite films was investigated. The results demonstrate that the addition of CNFs improved Young's modulus and toughness of PLA/PEG films. In contrast, a slight decrease in mechanical properties was observed when the content of CNFs reached 0.83 phr. Considère's constructions are used to explain the neck phenomena and cold drawing of nanocomposite films. The crystallization and thermal stability of PLA nanocomposite films were enhanced, with a slight decrease in cold-crystalline temperature ( ) and an increase in decomposition temperature ( ).
PubMed: 38911786
DOI: 10.1021/acsomega.4c01594 -
Macromolecules May 2024Polymer nanocomposites exhibit a heterogeneous mechanical behavior that is strongly dependent on the interaction between the polymer matrix and the nanofiller. Here, we...
Polymer nanocomposites exhibit a heterogeneous mechanical behavior that is strongly dependent on the interaction between the polymer matrix and the nanofiller. Here, we provide a detailed investigation of the mechanical response of model polymer nanocomposites under deformation, across a range of temperatures, from the glassy regime to the liquid one, via atomistic molecular dynamics simulations. We study the poly(ethylene oxide) matrix with silica nanoparticles (PEO/SiO) as a model polymer nanocomposite system with attractive polymer/nanofiller interactions. Probing the properties of polymer chains at the molecular level reveals that the effective mass density of the matrix and interphase regions changes during deformation. This decrease in density is much more pronounced in the glassy state. We focus on factors that govern the mechanical response of PEO/SiO systems by investigating the distribution of the (local) mechanical properties, focusing on the polymer/nanofiller interphase and matrix regions. As expected when heating the system, a decrease in Young's modulus is observed, accompanied by an increase in Poisson's ratio. The observed differences regarding the rigidity between the interphase and the matrix region decrease as the temperature rises; at temperatures well above the glass-transition temperature, the rigidity of the interphase approaches the matrix one. To describe the nonlinear viscoelastic behavior of polymer chains, the elastic modulus of the PEO/SiO systems is further calculated as a function of the strain for the entire nanocomposite, as well as the interphase and matrix regions. The elastic modulus drops dramatically with increasing strain for both the matrix and the interphase, especially in the small-deformation regime. We also shed light on characteristic structural and dynamic attributes during deformation. Specifically, we examine the rearrangement behavior as well as the segmental and center-of-mass dynamics of polymer chains during deformation by probing the mobility of polymer chains in both axial and radial motions under deformation. The behavior of the polymer motion in the axial direction is dominated by the deformation, particularly at the interphase, whereas a more pronounced effect of the temperature is observed in the radial directions for both the interphase and matrix regions.
PubMed: 38911610
DOI: 10.1021/acs.macromol.4c00537 -
Industrial & Engineering Chemistry... Jun 2024A novel investigation of the effects of the hydrophilic and hydrophobic segments of hydrophobically modified ethoxylated urethanes (HEURs) on the rheological properties...
A novel investigation of the effects of the hydrophilic and hydrophobic segments of hydrophobically modified ethoxylated urethanes (HEURs) on the rheological properties of their aqueous solutions, latex-based emulsions, and waterborne paints is demonstrated. Different HEUR thickeners were produced by varying the poly(ethylene glycol) (PEG) molecular weight and terminal hydrophobic size. Results reveal that the strength of hydrophobic associations and, consequently, the rheological properties of HEUR formulations can be effectively controlled by modifying the structure of the hydrophobic segment, specifically, the combination of diisocyanate and monoalcohol. This allows for the on-demand attainment of diverse rheological behaviors ranging from predominantly Newtonian profiles exhibiting lower viscosities to markedly pseudoplastic behaviors with significantly higher viscosities. The length of the hydrophilic group appears to affect viscosity only marginally up to a molecular weight of 23,000 g/mol, with more notable effects at 33,000 g/mol. Additionally, it was indicated that the rheological responses observed in water solutions provide a reliable forecast of their behavior in latex-based emulsions and waterborne paints. Coarse-grained molecular dynamics (CG-MD) simulations were also applied to gain insight into HEUR micelle dynamics in aqueous solutions. Guided by the DBSCAN algorithm, the simulations successfully captured the concentration-dependent behavior and the impact of hydrophilic chain length, aligning with the experimental viscosity trends. Various metrics were employed to provide a comprehensive analysis of the micellization process, including the hydrophobic cluster volume, the total micellar volume, the aggregation number, and the number of chains interconnecting with other micelles.
PubMed: 38911482
DOI: 10.1021/acs.iecr.4c00253 -
Cell Reports. Physical Science May 2024Recreating tissue environments with precise control over mechanical, biochemical, and cellular organization is essential for next-generation tissue models for drug...
Recreating tissue environments with precise control over mechanical, biochemical, and cellular organization is essential for next-generation tissue models for drug discovery, development studies, and the replication of disease environments. However, controlling these properties at cell-scale lengths remains challenging. Here, we report the development of printing approaches that leverage polyethylene glycol diacrylate (PEGDA) hydrogels containing photocaged oligonucleotides to spatially program material characteristics with non-destructive, non-ultraviolet light. We further integrate this system with a perfusion chamber to allow us to alter the composition of PEGDA hydrogels while retaining common light-activatable photocaged DNAs. We demonstrate that the hydrogels can capture DNA functionalized materials, including cells coated with complementary oligonucleotides with spatial control using biocompatible wavelengths. Overall, these materials open pathways to orthogonal capture of any DNA functionalized materials while not changing the sequences of the DNA.
PubMed: 38911357
DOI: 10.1016/j.xcrp.2024.101922 -
Cureus May 2024Soft tissue inflammatory responses to metal debris from prostheses, categorised as adverse reactions to metal debris (ARMD), are frequent complications of total hip...
Soft tissue inflammatory responses to metal debris from prostheses, categorised as adverse reactions to metal debris (ARMD), are frequent complications of total hip arthroplasty (THA) and often result in implant failure. Introducing modular implant designs in modern orthopaedics has brought benefits to total hip replacements but has also increased patients' susceptibility to corrosion-related risks. ARMD can develop from various metal articulating surfaces, including ceramic-on-polyethylene (CoP), ceramic-on-ceramic (CoC), metal-on-metal (MoM), and metal-on-polyethylene (MoP) configurations. In this case study, a 68-year-old male who underwent a MoP implant for osteoarthritis of the right hip 16 years ago presented with pain and difficulty walking, exacerbated over the past three months. Clinical examination revealed tenderness around the implant and a limited range of motion. Imaging studies, including X-rays and ultrasound-guided aspiration, coupled with normal serum and urinary cobalt (Co) and chromium (Cr) levels, confirmed the diagnosis of ARMD. Given the severity of symptoms and radiographic findings, surgical intervention was warranted, leading to a two-stage revision with implant augmentation using a Burch-Schneider cage. Three months post operation, the patient experienced significant improvements in pain levels, range of motion (ROM), and hip function. This case underscores the importance of vigilant surveillance for ARMD in patients undergoing non-MoM THA, even years post surgery. Prompt recognition and management of ARMD are crucial to mitigate the risk of long-term complications and optimise patient outcomes. Further research is needed to understand the risk factors and mechanisms underlying ARMD in MoP THA, aiding in developing preventive strategies and refined treatment protocols.
PubMed: 38910664
DOI: 10.7759/cureus.60810 -
Bone & Joint Open Jun 2024In metal-on-metal (MoM) hip arthroplasties and resurfacings, mechanically induced corrosion can lead to elevated serum metal ions, a local inflammatory response, and...
AIMS
In metal-on-metal (MoM) hip arthroplasties and resurfacings, mechanically induced corrosion can lead to elevated serum metal ions, a local inflammatory response, and formation of pseudotumours, ultimately requiring revision. The size and diametral clearance of anatomical (ADM) and modular (MDM) dual-mobility polyethylene bearings match those of Birmingham hip MoM components. If the acetabular component is satisfactorily positioned, well integrated into the bone, and has no surface damage, this presents the opportunity for revision with exchange of the metal head for ADM/MDM polyethylene bearings without removal of the acetabular component.
METHODS
Between 2012 and 2020, across two centres, 94 patients underwent revision of Birmingham MoM hip arthroplasties or resurfacings. Mean age was 65.5 years (33 to 87). In 53 patients (56.4%), the acetabular component was retained and dual-mobility bearings were used (DM); in 41 (43.6%) the acetabulum was revised (AR). Patients underwent follow-up of minimum two-years (mean 4.6 (2.1 to 8.5) years).
RESULTS
In the DM group, two (3.8%) patients underwent further surgery: one (1.9%) for dislocation and one (1.9%) for infection. In the AR group, four (9.8%) underwent further procedures: two (4.9%) for loosening of the acetabular component and two (4.9%) following dislocations. There were no other dislocations in either group. In the DM group, operating time (68.4 vs 101.5 mins, p < 0.001), postoperative drop in haemoglobin (16.6 vs 27.8 g/L, p < 0.001), and length of stay (1.8 vs 2.4 days, p < 0.001) were significantly lower. There was a significant reduction in serum metal ions postoperatively in both groups (p < 0.001), although there was no difference between groups for this reduction (p = 0.674 (cobalt); p = 0.186 (chromium)).
CONCLUSION
In selected patients with Birmingham MoM hips, where the acetabular component is well-fixed and in a satisfactory position with no surface damage, the metal head can be exchanged for polyethylene ADM/MDM bearings with retention of the acetabular prosthesis. This presents significant benefits, with a shorter procedure and a lower risk of complications.
PubMed: 38910515
DOI: 10.1302/2633-1462.56.BJO-2023-0165.R1