High-affinity antibodies specific to the core region of the tau protein exhibit diagnostic and therapeutic potential for Alzheimer's disease.

BACKGROUND: Recent advances in blood-based biomarker discovery are paving the way for simpler, more accessible diagnostic tools that can detect early signs of Alzheimer's disease (AD). Recent successes in the development of amyloid-targeting immunotherapy approaches mark an important advancement in providing new options for the treatment of AD. We have developed a set of high-affinity monoclonal antibodies (mAbs) to tau protein that have the potential as tools for diagnosis and treatment of AD. METHODS: Sheep were immunised with either full-length tau (1-441) or truncated paired helical filament (PHF)-core tau (297-391). A stringent bio-panning and epitope selection strategy, with a particular focus directed to epitopes within the disease-relevant PHF-core tau, was used to identify single-chain antibodies (scAbs). These scAbs were ranked by affinity for each epitope class, with leads converted to high-affinity mAbs. These antibodies and their potential utility were assessed by their performance in tau immunoassays, as well as their ability to prevent tau aggregation and propagation. Further characterisation of these antibodies was performed by immunohistochemical staining of brain sections and immuno-gold electronmicroscopy of isolated PHFs. RESULTS: Our work resulted in a set of high-affinity antibodies reacting with multiple epitopes spanning the entire tau protein molecule. The tau antibodies directed against the core tau unit of the PHF inhibited pathological aggregation and seeding using several biochemical and cell assay systems. Through staining of brain sections and PHFs, the panel of antibodies revealed which tau epitopes were available, truncated, or occluded. In addition, highly sensitive immunoassays were developed with the ability to distinguish between and quantify various tau fragments. CONCLUSION: This article introduces an alternative immunodiagnostic approach based on the concept of a "tauosome" - the diverse set of tau fragments present within biological fluids. The development of an antibody panel that can distinguish a range of different tau fragments provides the basis for a novel approach to potential diagnosis and monitoring of disease progression. Our results further support the notion that tau immunotherapy targeting the PHF-core needs to combine appropriate selection of both the target epitope and antibody affinity to optimise therapeutic potential.

Characterizing the Effect of Filament Moisture on Tensile Properties and Morphology of Fused Deposition Modeled Polylactic Acid/Polybutylene Succinate Parts.

Moisture absorption into hygroscopic/hydrophilic materials used in fused deposition modeling (FDM) can diminish desired mechanical properties. Sensitivity to moisture is dependent on material properties and environmental factors and needs characterization. In this article, moisture sensitivity of four grades of polylactic acid (PLA) filaments and four different ratios of PLA/polybutylene succinate (PBS) blended filaments were characterized through FDM printed American society for testing and materials (ASTM-D638) test samples after conditioning the filaments at different relative humidity levels. The tensile testing and scanning electron microscopy (SEM) of the samples' fracture surfaces revealed that PLA 4043D was the most moisture-sensitive among the chosen grades of PLA filaments. Through filament tension test and melt flow index (MFI) testing it was observed that moisture had a significant detrimental effect (20% reduction in tensile strength and 50% increase in MFI) on PLA 4043D filaments. Samples from moisture-conditioned PLA/PBS 75/25 blended filaments displayed a significant reduction (10%) in tensile strength. Moreover, the MFI of 75/25 filaments was increased with subsequent increases in moisture level. Investigation of tensile properties of ASTM samples made from four grades of PLA filaments exposed to room temperature and humidity conditions for 3 months showed an even more significant decrease in strength (ranging from 24% to 36%).

Small-molecule mediated MuRF1 Inhibition protects from Doxorubicin-induced Cardiac Atrophy and Contractile Dysfunction.

Cancer chemotherapy induces cell stress in rapidly dividing cancer cells to trigger their growth arrest and apoptosis. However, adverse effects related to cardiotoxicity underpinned by a limited regenerative potential of the heart limits clinical application: In particular, chemotherapy with doxorubicin (DOXO) causes acute heart injury that can transition to persisting cardiomyopathy (DOXO-CM). Here, we tested if MuRF1 inhibition ("MuRFi") was able to attenuate DOXO-CM. To mimic DOXO chemotherapy, we treated mice over four weeks with five DOXO injection and a cumulative dosage of 25 mg/kg. At day 28, mice had lower body and heart weights, reduced cardiac cross-sectional myofibrillar areas (CSAs), and functional ejection fractions (EFs) and fractional shortenings (FS) as indicated by echocardiography (ECHO). In contrast, mice with a 1g/kg Myomed#205 spiked diet, a previously described experimental MuRFi therapy, showed lower DOXO-CM at day 28, and also reduced acute DOXO cardiac injury at day 7 (single DOXO dose; 15 mg/kg). Underlying molecular signatures using Western blot (WB) studies showed at day 28 reduced phospho-AKT (AKTp) and phospo-4EBP1 (4EBP1p) levels following DOXO that were normalized following MuRFi treatment. Taken together, our data suggest that MuRFi treatment is suitable to attenuate DOXO-CM by preserving AKTp and 4EBP1p levels in DOXO stressed cardiomyocytes, thereby supporting de novo protein translation and cardiomyocyte survival under translational arrest stress.

An alternative filament fabrication method as the basis for 3D-printing personalized implants from elastic ethylene vinyl acetate copolymer.

In this work, a novel tool for small-scale filament production is presented. Unlike traditional methods such as hot melt extrusion (HME), the device (i) allows filament manufacturing from small material amounts as low as three grams, (ii) ensures high diameter stability almost independent of the viscoelastic behavior of the polymer melt, and (iii) enables processing of materials with rheological profiles specifically tailored toward fused filament fabrication (FFF). Hence, novel materials, previously difficult to process due to HME limitations, become easily accessible for FFF for the first time. Here, we showcase the production of highly flexible drug-free, and drug-loaded filaments based on ethylene-vinyl acetate polymers with a vinyl acetate content of 28 w% (EVA28) and unprecedented high melt flow rates of up to 400 g/10 min. Owing to their low viscosity, FFF with low print nozzle sizes of 250 µm was achieved for the first time for EVA28. These small nozzle diameters facilitate 3D-printing of high-resolution structures in small-dimensional dosage forms such as subcutaneous implantable drug delivery systems, which can later be used for personalization. Consequently, the material portfolio for FFF is tremendously broadened, allowing material and formulation optimization toward FFF, independent of a preliminary extrusion process.

Finite Difference Modeling and Experimental Investigation of Cyclic Thermal Heating in the Fused Filament Fabrication Process.

Fused filament fabrication (FFF) is one of the most popular additive manufacturing (AM) processes due to its simplicity and low initial and maintenance costs. However, good printing results such as high dimensionality, avoidance of cooling cracks, and warping are directly related to heat control in the process and require precise settings of printing parameters. Therefore, accurate prediction and understanding of temperature peaks and cooling behavior in a local area and in a larger part are important in FFF, as in other AM processes. To analyze the temperature peaks and cooling behavior, we simulated the heat distribution, including convective heat transfer, in a cuboid sample. The model uses the finite difference method (FDM), which is advantageous for parallel computing on graphics processing units and makes temperature simulations also of larger parts feasible. After the verification process, we validate the simulation with an in situ measurement during FFF printing. We conclude the process simulation with a parameter study in which we vary the function of the heat transfer coefficient and part size. For smaller parts, we found that the print bed temperature is crucial for the temperature gradient. The approximations of the heat transfer process play only a secondary role. For larger components, the opposite effect can be observed. The description of heat transfer plays a decisive role for the heat distribution in the component, whereas the bed temperature determines the temperature distribution only in the immediate vicinity of the bed. The developed FFF process model thus provides a good basis for further investigations and can be easily extended by additional effects or transferred to other AM processes.

Characterization of Chemically Treated Flexible Body-Centered Cubic Lattice Structures Fabricated by Fused Filament Fabrication Process.

Fused filament fabrication (FFF) has opened new opportunities for the effortless fabrication of complex structures at low cost. The additively manufactured lattice structures have been widely used in different sectors. However, the parts fabricated through FFF suffered from poor surface and dimensional characteristics. These disadvantages have been overcome by using different post-processing techniques. The present investigation has been focused on the post-processing of flexible lattice structures through chemical treatment methods. The flexible lattice structures have been fabricated by using thermoplastic polyurethane material. Body-centered cubic lattice structures have been chosen for the present study. The fabricated lattice structures have been post-processed using dimethyl sulfoxide solvent through the chemical immersion method. The response characteristics chosen for the present study were surface roughness, compressive strength, and dimensional accuracy. The measurement has been taken before and after the chemical treatment method for comparison purpose. The results of experimental studies depicted that the proposed methodology significantly enhanced the surface quality and dimensional accuracy, whereas compressive strength has been observed to be slightly reduced after the post-processing method.

Generation of Customized Bone Implants from CT Scans Using FEA and AM.

Additive manufacturing (AM) allows the creation of customized designs for various medical devices, such as implants, casts, and splints. Amongst other AM technologies, fused filament fabrication (FFF) facilitates the production of intricate geometries that are often unattainable through conventional methods like subtractive manufacturing. This study aimed to develop a methodology for substituting a pathological talus bone with a personalized one created using additive manufacturing. The process involved generating a numerical parametric solid model of the specific anatomical region using computed tomography (CT) scans of the corresponding healthy organ from the patient. The healthy talus served as a mirrored template to replace the defective one. Structural simulation of the model through finite element analysis (FEA) helped compare and select different materials to identify the most suitable one for the replacement bone. The implant was then produced using FFF technology. The developed procedure yielded commendable results. The models maintained high geometric accuracy, while significantly reducing the computational time. PEEK emerged as the optimal material for bone replacement among the considered options and several specimens of talus were successfully printed.

An Investigation into Mechanical Properties of 3D Printed Thermoplastic-Thermoset Mixed-Matrix Composites: Synergistic Effects of Thermoplastic Skeletal Lattice Geometries and Thermoset Properties.

This study aims to develop thermoplastic (TP) and thermoset (TS) based mixed matrix composite using design dependent physical compatibility. Using thermoplastic-based (PLA) skeletal lattices with diverse patterns (gyroid and grid) and different infill densities (10% and 20%) followed by infiltration of two different thermoset resin systems (epoxy and polyurethane-based) using a customized FDM 3D printer equipped with a resin dispensing unit, the optimised design and TP-TS material combination was established for best mechanical performance. Under uniaxial tensile stress, the failure modes of TP gyroid structures with polyurethane-based composites included 'fiber pull-out', interfacial debonding and fiber breakage, while epoxy based mixed matrix composites with all design variants demonstrated brittle failure. Higher elongation (higher area under curve) was observed in 20% infilled gyroid patterned composite with polyurethane matrix indicating the capability of operation in mechanical shock absorption application. Electron microscopy-based fractography analysis revealed that thermoset matrix properties governed the fracture modes for the thermoplastic phase. This work focused on the strategic optimisation of both toughness and stiffness of mixed matrix composite components for rapid fabrication of construction materials.

Advancements in carbon nanotube-polymer composites: Enhancing properties and applications through advanced manufacturing techniques.

Carbon nanotube (CNT)-polymer composites exhibit significant advancements in mechanical, electrical, and thermal properties, enabling numerous promising applications. This review delves into recent research on manufacturing methods, filament extrusion, additive manufacturing (AM), properties, and applications of CNT polymer composites. Factors like processing conditions, polymer types, and CNT concentrations determine the ultimate properties of the composite material. The dispersion of CNT within various manufacturing techniques, such as melt mixing, solution mixing, and in-situ polymerization, significantly impacts the properties of the composite material. These composite materials are extensively used in AM, particularly in 3D printing, where filament blends are extruded and printed to custom-shaped objects. The finding underscores the effect of CNT content on the properties of CNT-polymer composite material in different applications. However, gaps remain in optimizing manufacturing processes and AM techniques, essential for tailoring these composites to specific application needs. Future research should focus on developing cost-effective and scalable manufacturing methods to unlock the full potential of CNT-polymer composites in various industries.

Cardiac Myosin and Thin Filament as Targets for Lead and Cadmium Divalent Cations.

Lead and cadmium are heavy metals widely distributed in the environment and contribute significantly to cardiovascular morbidity and mortality. Using Leadmium Green dye, we have shown that lead and cadmium enter cardiomyocytes, distributing throughout the cell. Using an in vitro motility assay, we have shown that sliding velocity of actin and native thin filaments over myosin decreases with increasing concentrations of Pb2+ and Cd2+. Significantly lower concentrations of Pb2+ and Cd2+ (0.6 mM) were required to stop sliding of thin filaments over myosin compared to the stopping actin sliding over the same myosin (1.1-1.6 mM). Lower concentration of Cd2+ (1.1 mM) needed to stop actin sliding over myosin compared to the Pb2++Cd2+ combination (1.3 mM) and lead alone (1.6 mM). There were no differences found in the effects of lead and cadmium cations on relative force developed by myosin heads or number of actin filaments bound to myosin. Sliding velocity of actin over myosin in the left atrium, right and left ventricles changed equally when exposed to the same dose of the same metal. Thus, we have demonstrated for the first time that Pb2+ and Cd2+ can directly affect myosin and thin filament function, with Cd2+ exerting a more toxic influence on myosin function compared to Pb2+.

Small organic osmolytes accelerate actin filament assembly and stiffen filaments.

Actin filament assembly and mechanics are crucial for maintenance of cell structure, motility, and division. Actin filament assembly occurs in a crowded intracellular environment consisting of various types of molecules, including small organic molecules known as osmolytes. Ample evidence highlights the protective functions of osmolytes such as trimethylamine-N-oxide (TMAO), including their effects on protein stability and their ability to counteract cellular osmotic stress. Yet, how TMAO affects individual actin filament assembly dynamics and mechanics is not well understood. We hypothesize that, owing to its protective nature, TMAO will enhance filament dynamics and stiffen actin filaments due to increased stability. In this study, we investigate osmolyte-dependent actin filament assembly and bending mechanics by measuring filament elongation rates, steady-state filament lengths, and bending persistence lengths in the presence of TMAO using total internal reflection fluorescence microscopy and pyrene assays. Our results demonstrate that TMAO increases filament elongation rates as well as steady-state average filament lengths, and enhances filament bending stiffness. Together, these results will help us understand how small organic osmolytes modulate cytoskeletal protein assembly and mechanics in living cells.

Three-dimensional cellular architecture of the sigmoid filament in Trichomonas vaginalis.

Trichomonas vaginalis is a parasite protozoan that causes human trichomoniasis, a sexually transmitted infection (STI) that affects more than 156 million people worldwide. T. vaginalis contains an uncommon and complex cytoskeleton constituting the mastigont system, formed by several fibers and proteinaceous structures associated with basal bodies. Among these structures is the pelta-axostylar complex made of microtubules and striated filaments such as the costa and the parabasal filaments. In addition, some structures are poorly known and studied, such as the sigmoid filament and the X-filament. Here, we have isolated the Trichomonas vaginalis cytoskeleton and used UHR-SEM (ultra-high resolution scanning electron microscopy), tomography, immunofluorescence, immunolabeling, and backscattered electrons on SEM, negative staining to model the three-dimensional architecture and possible function of the sigmoid.

Biomedical Composites of Polycaprolactone/Hydroxyapatite for Bioplotting: Comprehensive Interpretation of the Reinforcement Course.

Robust materials in medical applications are sought after and researched, especially for 3D printing in bone tissue engineering. Poly[ε-caprolactone] (PCL) is a commonly used polymer for scaffolding and other medical uses. Its strength is a drawback compared to other polymers. Herein, PCL was mixed with hydroxyapatite (HAp). Composites were developed at various concentrations (0.0-8.0 wt. %, 2.0 step), aiming to enhance the strength of PCL with a biocompatible additive in bioplotting. Initially, pellets were derived from the shredding of filaments extruded after mixing PCL and HAp at predetermined quantities for each composite. Specimens were then manufactured by bioplotting 3D printing. The samples were tested for their thermal and rheological properties and were also mechanically, morphologically, and chemically examined. The mechanical properties included tensile and flexural investigations, while morphological and chemical examinations were carried out employing scanning electron microscopy and energy dispersive spectroscopy, respectively. The structure of the manufactured specimens was analyzed using micro-computed tomography with regard to both their dimensional deviations and voids. PCL/HAp 6.0 wt. % was the composite that showed the most enhanced mechanical (14.6% strength improvement) and structural properties, proving the efficiency of HAp as a reinforcement filler in medical applications.

Assessing the contribution of sewing threads to microfiber release during domestic laundering.

Environmental science studies from the past decade have emphasized that microplastics in aquatic environments are mostly caused by domestic laundering of synthetic textiles. Although many studies have explored the microfiber release behavior of fabrics washed in laundry, attempts to witness microfiber release from sewing threads, which are an inevitable part of any finished garment, are meager. With this research gap, this study attempted to analyze the potential of sewing threads to release microfibers during washing and the extent to which they can contribute to the overall microfiber release during domestic laundering. The study's findings revealed an average release of 2.65 ± 0.70 (n = 33) microfibers/m from the sewing thread sewn on the fabric during laundering. The sewing process was noted to cause damage to the sewing thread, which led to a comparatively higher microfiber release (∼114%) compared with the sewing threads that were washed before sewing. Among the selected sewing threads, higher microfiber emissions were reported with spun threads, followed by twistless filaments, and twisted filament threads. The results showed that coarser sewing threads with higher Tex values released more microfibers than finer Tex threads. Compared to the 20 Tex spun thread, the 80 Tex spun thread showed a 22-150% increase in microfiber release. In the case of filament sewing thread, a similar impact was noted, whereas the role of twist was found to be efficient in reducing microfiber emission. Compared to the untwisted filaments, the ply twisted filaments exhibited approximately 76% lower microfiber emissions. The findings of this study showed that sewing thread contributed approximately 1.09% of the total microfiber emissions from apparel during laundry.

3D-printed immediate release solid dosage forms: a patent evaluation of US11622940B2.

Three-dimensional (3D) printing is one of the most flexible technologies for preparing tablets, offering controlled drug release profiles. The current patent describes the preparation of immediate-release 3D-printed tablets of hydrochlorothiazide to improve disintegration and dissolution profile. The patent involves the preparation of drug-loaded filament via hot-melt extrusion and utilizing the same filaments for printing 3D-printed tablets using fused deposition modeling. The tablets were printed with different shapes and sizes by incorporating channels within the tablet spaces, termed as gaplets. The introduction of channels within the tablet design improves the disintegration and dissolution profile of the drug significantly. The morphological characteristic of 3D-printed tablets was studied by using scanning electron microscopy and revealed the presence of gaplets in the tablets.

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Advancements in reliability of mechanical performance of 3D PRINTED Ag-doped bioceramic antibacterial scaffolds for bone tissue engineering.

The current gold-standard approach for addressing bone defects in load-bearing applications sees the use of either autographs or allographs. These solutions, however, have limitations as autographs and allographs carry the risk of additional trauma, the threat of disease transmission, and potential donor rejection. An attractive candidate for overcoming the challenges associated with the use of autographs and allographs is a 3D porous scaffold displaying the needed mechanical competency for use in load-bearing applications that can stimulate bone tissue regeneration and provide antibacterial capabilities. To date, no reports document a 3D porous scaffold that fully meets the criteria specified above. In this work, we show how the use of fused filament fabrication (FFF) 3D printing technology in combination with a bimodal distribution of Ag-doped bioactive glass-ceramic (Ag-BG) micro-sized particles can successfully deliver porous 3D scaffolds with attractive and reliable mechanical performance characteristics capable of stimulating bone tissue regeneration and the ability to provide inherent antibacterial properties. To characterize the reliability of the mechanical performance of the FFF-printed Ag-BG scaffolds, Weibull statistics were evaluated for both the compressive (N = 25; m = 13.6 ± 0.9) and flexural (N = 25; m = 7.3 ± 0.7) strengths. Methicillin-resistant Staphylococcus aureus (MRSA) was used both in planktonic and biofilm forms to highlight the advanced antibacterial characteristics of the FFF-printed Ag-BG scaffolds. Biological performance was evaluated in vitro through indirect exposure to human marrow stromal cells (hMSCs), where the FFF-printed Ag-BG scaffolds were found to provide an attractive environment for cell infiltration and mineralization. Our work demonstrates how fused filament fabrication technology can be used with bioactive and antibacterial materials such as Ag-BG to deliver mechanically competent porous 3D scaffolds capable of stimulating bone tissue regeneration while simultaneously providing antibacterial performance capabilities.

Architecture of CTPS filament networks revealed by cryo-electron tomography.

The cytoophidium is a novel type of membraneless organelle, first observed in the ovaries of Drosophila using fluorescence microscopy. In vitro, purified Drosophila melanogaster CTPS (dmCTPS) can form metabolic filaments under the presence of either substrates or products, and their structures that have been analyzed using cryo-electron microscopy (cryo-EM). These dmCTPS filaments are considered the fundamental units of cytoophidia. However, due to the resolution gap between light and electron microscopy, the precise assembly pattern of cytoophidia remains unclear. In this study, we find that dmCTPS filaments can spontaneously assemble in vitro, forming network structures that reach micron-scale dimensions. Using cryo-electron tomography (cryo-ET), we reconstruct the network structures formed by dmCTPS filaments under substrate or product binding conditions and elucidate their assembly process. The dmCTPS filaments initially form structural bundles, which then further assemble into larger networks. By identifying, tracking, and statistically analyzing the filaments, we observed distinct characteristics of the structural bundles formed under different conditions. This study provides the first systematic analysis of dmCTPS filament networks, offering new insights into the relationship between cytoophidia and metabolic filaments.

Enhanced Feedstock Processability for the Indirect Additive Manufacturing of Metals by Material Extrusion through Ethylene-Propylene Copolymer Modification.

Filament-based material extrusion (MEX) represents one of the most commonly used additive manufacturing techniques for polymer materials. In a special variation of this process, highly filled polymer filaments are used to create metal parts via a multi-step process. The challenges associated with creating a dense final part are versatile due to the different and partly contrary requirements of the individual processing steps. Especially for processing in MEX, the compound must show sufficiently low viscosity, which is often achieved by the addition of wax. However, wax addition also leads to a significant reduction in ductility. This can cause filaments to break, which leads to failure of the MEX process. Therefore, the present study investigates the influence of different ethylene-propylene copolymers (EPCs) with varying ethylene contents as a ductility-enhancing component within the feedstock to improve filament processing behavior. The resulting feedstock materials are evaluated regarding their mechanical, thermal and debinding behavior. In addition, the processability in MEX is assessed. This study shows that a rising ethylene content within the EPC leads to a higher ductility and an enhanced filament flexibility while also influencing the crystallization behavior of the feedstock. For the MEX process, an ethylene fraction of 12% within the EPC was found to be the optimum regarding processability for the highly filled filaments in MEX and the additional processing steps to create sintered metal parts.

Mechanical Properties and Economic Analysis of Fused Filament Fabrication Continuous Carbon Fiber Reinforced Composites.

Additive manufacturing of composites offers advantages over metals since composites are lightweight, fatigue and corrosion-resistant, and show high strength and stiffness. This work investigates the tensile and flexural performance of continuous carbon-fiber reinforced (CCF) composites with different guide angles and number of layers. The cost and printing time analyses were also conducted. Tensile specimens with a contour-only specimen and one CCF layer with a 0° guide angle exhibited nearly comparable strength values. Increasing the number of CCF layers enhances the tensile properties. For the identical cost and reinforcement amount, 0°/0° provides a higher tensile strength and elastic modulus compared with 15°/-15°. The same phenomenon was observed for 15°/0°/-15° and 0°/0°/0°. The samples with one and two reinforcement layers had similar stiffness and maximum load values for flexural tests. For the samples with four layers, there was a considerable improvement in stiffness but a minor decrease in the maximum load.

Enhancing Clay-Based 3D-Printed Mortars with Polymeric Mesh Reinforcement Techniques.

Additive manufacturing (AM) technologies, including 3D mortar printing (3DMP), 3D concrete printing (3DCP), and Liquid Deposition Modeling (LDM), offer significant advantages in construction. They reduce project time, costs, and resource requirements while enabling free design possibilities and automating construction processes, thereby reducing workplace accidents. However, AM faces challenges in achieving superior mechanical performance compared to traditional methods due to poor interlayer bonding and material anisotropies. This study aims to enhance structural properties in AM constructions by embedding 3D-printed polymeric meshes in clay-based mortars. Clay-based materials are chosen for their environmental benefits. The study uses meshes with optimal geometry from the literature, printed with three widely used polymeric materials in 3D printing applications (PLA, ABS, and PETG). To reinforce the mechanical properties of the printed specimens, the meshes were strategically placed in the interlayer direction during the 3D printing process. The results show that the 3D-printed specimens with meshes have improved flexural strength, validating the successful integration of these reinforcements.