Large Beads Research Articles

Assessment of inhaled cationic antibiotics in an in vitro model of Pseudomonas aeruginosa lung biofilm

ObjectivesThis study aimed to evaluate the efficacy of inhaled cationic antibiotics, including tobramycin (TOB) and colistin (CST), using an in vitro alginate bead model that simulates Pseudomonas aeruginosa lung biofilms. MethodsBioluminescent P. aeruginosa were encapsulated within alginate beads and dispersed in either Mueller-Hinton broth (MHB) or artificial sputum medium (ASM). The impact of bead size and culture medium on TOB and CST efficacy was assessed by monitoring bioluminescence kinetics, followed by colony-forming unit (CFU/mL) measurements. Antibiotic efficacy was quantified using a Hill inhibitory model to analyze variations in CFU/mL in response to TOB and CST concentrations. ResultsThe TOB EC50 was found to be 8-fold higher when P. aeruginosa was encapsulated in larger beads (1200 μm) compared to smaller beads (60 μm). TOB efficacy further decreased twofold when larger beads were dispersed in ASM. In contrast, CST demonstrated superior efficacy, being four times more potent than TOB, with corresponding EC50 values of 20.5 ± 2.8 times MIC and 78.4 ± 10.2 times MIC, respectively. No change in MICs was observed for either antibiotic, even after exposing bacteria to 200 times the MIC. ConclusionsThis P. aeruginosa biofilm model highlights how alginate and mucus modulated the efficacy of TOB and CST, and suggested the superior efficacy of CST in eradicating pulmonary biofilms.

Plant-based marbled salami analogs: Emulsion-loaded microgels embedded within protein-polysaccharide hydrogel matrices

The inherent molecular differences between plant and animal proteins make it challenging to accurately simulate the optical and mechanical properties of real meat products using plant-derived ingredients. Soft matter physics approaches are therefore being employed to direct the assembly of plant proteins, polysaccharides, and lipids into meat-like structures. The main focus of the current study was to use soft matter physics approaches to create plant-based analogs of marbled meat products, such as salami, chorizo, or pepperoni. These products have whitish fatty regions embedded within reddish/brown lean regions. In this study, we mimicked the fatty regions in salami using emulsion-loaded alginate beads and the lean regions using composite hydrogels assembled from potato protein and gellan gum. Initially, the textural attributes of the plant-based lean regions were optimized by varying potato protein type (low or high isoelectric point) and concentration. As expected, the mechanical strength of the lean region analogs increased with increasing protein content and depended on protein type. Introducing alginate beads into the salami analogs had little impact on their textural attributes when measured under compression conditions but decreased their stiffness and fracture stress when measured under tensile conditions. In particular, introducing either small or large beads reduced the stiffness and breaking stress of the salami analogs, which was attributed to the introduction of weak points in the composite hydrogel. Finally, we showed that the appearance and textural attributes of real salami could be mimicked in plant-based salami by optimizing its composition and structure. This research provides valuable insights into methods of better simulating the properties of marbled meat products, like sausages and cold-cut meat products, thereby creating a more sustainable food supply.

Regulating the surface area increase of zero-valent iron (ZVI) during ball milling: A study of the mechanical mechanism and low-carbon milling

This study investigates the effect of milling parameters on the specific surface area (SSA) increase of zero-valent iron (ZVI) powder, which is crucial for its environmental applications. Combining experimental and orthogonal design methodologies, the research explores how the parameters modulate the SSA. The operational factors are analyzed from the perspectives of bead momentum, kinetic energy, and energy efficiency. A regression model is proposed for the precise fabrication of ZVI with predetermined SSA. Furthermore, the study identifies optimized parameters that minimize energy consumption and carbon emissions. The results suggest that the optimal milling time is 10 min, with larger beads (2.0 mm), higher quality beads (0.75 kg), and higher shaft rotational speed (2000 rpm) significantly enhancing SSA. Discrete Element Method (DEM) simulations reveal micro-mechanical interactions, supporting the parameter selection scheme for maximizing SSA. The findings contribute to optimizing the milling process of the ductile ZVI powder for environmental clean-up.

Characterization of integrins in cellular immunity of the oriental armyworm, Mythimna separata.

Insect hemocytes eliminate foreign substances from the hemocoel through various immune reactions. Integrins, receptor proteins present on the cell membrane, are formed as a heterodimer from α and β subunits and are known to be involved in various immune reactions. To elucidate the role of integrins in the immunity of the lepidoptera Mythimna separata, genes encoding integrins were screened from the genome, resulting in the identification of eight α and four β integrin genes. The expression levels of the integrin genes did not change in response to the injection of small abiotic beads undergoing phagocytosis in M. separata larvae. However, significant inductions of some integrin gene expressions were observed in hemocytes that formed capsules around large abiotic beads during encapsulation, especially in MysIntα2. Under biotic stimulation, induction of the MysIntα2 was evident after exposures to Gram-negative bacteria (Escherichia coli) and entomopathogenic nematodes (Steinernema carpocapsae), but not to Gram-positive bacteria (Micrococcus luteus). Immunostaining analysis revealed that MysIntα2 was specifically localized to hemocytes surrounding the beads during the encapsulation reaction. Furthermore, the spreading and encapsulation abilities of hemocytes were significantly inhibited by incubation with MysIntα2 antibodies. Suppression of MysIntα2 expression in M. separata larvae by injecting double-stranded RNA also resulted in a decrease in encapsulation activity. Collectively, these results indicate that MysIntα2 plays pivotal roles in the cellular immune response of M. separata, particularly during encapsulation. This likely occurs through the regulation of hemocyte spreading activity, thereby facilitating the formation of multilayered capsules around large invaders.

Reproducing wrought grain structure in additive IN718 through nanosecond laser induced cavitation

Pulsed laser assisted additive manufacturing has been demonstrated as a promising technology for controlling grain structure in 3D-printing processes. The integration of a nanosecond laser onto a wire arc additive manufacturing tool has enabled the localized printing of Inconel 718 with grain sizes meeting ASTM 9 standards (average measured grain size of 13.7μm) for wrought material within a single bead under solidification conditions that would otherwise produce 340μm columnar grains. The observed grain refinement holds promise, provided scale up is possible, for overcoming the highly anisotropic mechanical properties and microcracking associated with large columnar grains of Inconel 718 that have long stood in the way of leveraging the advantages of direct energy deposition printing techniques of difficult to machine alloys. Experiments on large bead sizes allowed for decoupling surface versus bulk nanosecond laser/liquid metal interaction mechanisms to determine that the source of the observed grain refinement is the collapse of cavitation bubbles originating from acoustic waves generated by momentum transfer into the melt of an ablation plasma. Additionally, experiments that increased the cavitation bubble density within the mushy zone during solidification by tuning the nanosecond laser scan path went beyond the 25 times reduction in grain size to a 70 times factor of refinement with a minimum average grain diameter approaching 4μm.

Studies on Encapsulation of Gas Releasing Agent in Ca-Alginate Beads and Controlled Release Pattern of the Beads

Calcium alginate beads have been used to control release gases and volatile compounds for agricultural applications, drug delivery, waste treatment, and food product enhancement. The release rate of the gas and vapor depends on the diffusion of gas and vapor through the Ca-alginate gel matrix. However, the knowledge about the effect of process variables on the diffusion rate of gas and vapor still needs to be improved. Therefore, this study aims to investigate the effect of alginate concentration, gas release agent concentration, and bead size on the concentration of gas released from Ca-alginate beads. In this study, calcium carbonate was used as the model gas release agent, encapsulated in Ca-alginate beads to release carbon dioxide when the beads reacted with an acetic acid solution. The results showed that the concentration of carbon dioxide released by the beads depended on the bead diameter. The gas release rate was increased when the diameter of the beads was increased. The total amount of gas produced by the large beads was observed to be higher than that of small beads. Compared to small beads, large beads take longer to react completely to produce carbon dioxide gas. It was also found that the amount of carbon dioxide produced by the Ca-alginate beads was inversely proportional to alginate concentration but directly proportional to calcium carbonate concentration. The results indicated that the gas diffusion rate of the Ca-alginate beads can be tailored by adjusting the process variables of the Ca-alginate bead encapsulation process.

A novel variational approach for multiphoton microscopy image restoration: from PSF estimation to 3D deconvolution

In multi-photon microscopy (MPM), a recent in-vivo fluorescence microscopy system, the task of image restoration can be decomposed into two interlinked inverse problems: firstly, the characterization of the point spread function (PSF) and subsequently, the deconvolution (i.e. deblurring) to remove the PSF effect, and reduce noise. The acquired MPM image quality is critically affected by PSF blurring and intense noise. The PSF in MPM is highly spread in 3D and is not well characterized, presenting high variability with respect to the observed objects. This makes the restoration of MPM images challenging. Common PSF estimation methods in fluorescence microscopy, including MPM, involve capturing images of sub-resolution beads, followed by quantifying the resulting ellipsoidal 3D spot. In this work, we revisit this approach, coping with its inherent limitations in terms of accuracy and practicality. We estimate the PSF from the observation of relatively large beads (approximately 1 μm in diameter). This goes through the formulation and resolution of an original non-convex minimization problem, for which we propose a proximal alternating method along with convergence guarantees. Following the PSF estimation step, we then introduce an innovative strategy to deal with the high level multiplicative noise degrading the acquisitions. We rely on a heteroscedastic noise model for which we estimate the parameters. We then solve a constrained optimization problem to restore the image, accounting for the estimated PSF and noise, while allowing a minimal hyper-parameter tuning. Theoretical guarantees are given for the restoration algorithm. These algorithmic contributions lead to an end-to-end pipeline for 3D image restoration in MPM, that we share as a publicly available Python software. We demonstrate its effectiveness through several experiments on both simulated and real data.

Optical polarization studies of latex beads in aqueous solution: An analog for radar scattering in icy regolith

This study presents low phase angle 0°–5° measurements of polarimetric properties of icy planetary regolith analog materials acquired using the custom-built Multi-Axis- Goniometer-Instrument (MAGI). We present same sense (SC), and opposite sense (OC) backscatter circular polarization coefficients, circular polarization ratio (CPR), and degree linear polarization (DLP) of spherical latex (non-dye) beads of varying sizes and volume concentrations (v/v) in aqueous solutions (λ=0.8μm) in water. We also present measurements of alumina powder in air at λ=1.064μm. Measurements showed that at a low incidence angle (i=0°), backscatter is dominated by surface specular single-bounce scattering, that hides other scattering processes. At high (i=15°) incidence angle, surface single-bounce surface scattering becomes negligible, allowing for the detection of diffuse, dihedral (multiple bounces) scattering. We find that classical Mie alumina particles (2.1μm,4.0μm) enhance subsurface scattering due to a larger void space relative to larger Mie particles (30μm), that cause the radar signal to scatter forward off small imperfections, maintaining the polarization properties of the signal and generating high >1CPR. Latex beads, representing impurities, demonstrate the impact of isotropic and anisotropic scattering on radar signatures. This study also found that the scattering medium’s anisotropy correlates to the size of the beads, while the void space of the medium inversely correlates with the bead size and the volume concentration (v/v) of the beads. Rayleigh beads, due to isotropic scattering from the reduced scattering cross-section and higher transparency relative to larger impurities, generate subsurface single bounce scattering from the sample platform, producing OC≫SC and a low (<0.5)CPR across all v/v. Rayleigh impurities in transparent water-ice simulate single bounce scattering from underlying layers. Conversely, classical and large Mie beads generate anisotropic scattering that intensifies scattering in the forward direction with high CPR, inversely proportional to the volume concentration. This study aids in interpreting radar observations of icy bodies in the solar system, providing insights into the interplay between radar waves and icy regolith compositions.

AIEgens-enhanced rapid sensitive immunofluorescent assay for SARS-CoV-2 with digital microfluidics

BackgroundSensitive and rapid antigen detection is critical for the diagnosis and treatment of infectious diseases, but conventional ELISAs including chemiluminescence-based assays are limited in sensitivity and require many operation steps. Fluorescence immunoassays are fast and convenient but often show limited sensitivity and dynamic range. ResultsTo address the need, an aggregation-induced emission fluorgens (AIEgens) enhanced immunofluorescent assay with beads-based quantification on the digital microfluidic (DMF) platform was developed. Portable DMF devices and chips with small electrodes were fabricated, capable of manipulating droplets within 100 nL and boosting the reaction efficiency. AIEgen nanoparticles (NPs) with high fluorescence and photostability were synthesized to enhance the test sensitivity and detection range. The integration of AIEgen probes, transparent DMF chip design, and the large magnetic beads (10 μm) as capture agents enabled rapid and direct image-taking and signal calculation of the test result. The performance of this platform was demonstrated by point-of-care quantification of SARS-CoV-2 nucleocapsid (N) protein. Within 25 min, a limit of detection of 5.08 pg mL−1 and a limit of quantification of 8.91 pg mL−1 can be achieved using <1 μL sample. The system showed high reproducibility across the wide dynamic range (10–105 pg mL−1), with the coefficient of variance ranging from 2.6% to 9.8%. SignificanceThis rapid, sensitive AIEgens-enhanced immunofluorescent assay on the DMF platform showed simplified reaction steps and improved performance, providing insight into the small-volume point-of-care testing of different biomarkers in research and clinical applications.

Simulation of fiber-induced melt pressure fluctuations within large scale polymer composite deposition beads

The process-structure-property relationship in Large Area Additive Manufacturing (LAAM) technology is an ongoing area of research as the inherent microstructural properties (chiefly fibers and voids) affect the performance of printed parts. Unfortunately, we currently lack adequate understanding of micro void nucleation and evolution during the LAAM and fused deposition modelling (FDM) processes. Modeling of the polymer melt flow during the extrusion process is important in understanding the underlying microstructural formation and associated properties of the print, that determines the part performance in service conditions. In this paper we compute fiber-induced local pressure fluctuations which may promote void formation in the bead’s microstructure. On a macro-scale, we determine flow fields of a purely viscous, Newtonian planar polymer deposition flow through a LAAM nozzle which are utilized on a micro-scale model where we simulate the evolution of a single ellipsoidal fiber along streamlines obtained from the macro-model. On the micro-scale, we determine instantaneous values of the translational and rotational velocities of the rigid ellipsoidal fiber that satisfies a balance of hydrodynamic forces and couples on the fiber’s surface based on a Newton Raphson algorithm and we track the fiber’s motion along the flow path via an explicit numerical iterative algorithm. Model verification is achieved by benchmarking results with solutions from well-known Jeffery’s equation of motion of a particle in homogeneous simple shear flow. We account for rotary diffusivity due to short-range fiber-fiber interaction in the FEA simulation by determining an effective fluid domain size representative of the interaction coefficient of the melt flow through a correlation analysis that yields an equivalent steady state orientation based on the Advani-Tucker equation. We also consider different possible motions of the fiber along individual LAAM flow paths from a given set of random initial fiber conditions to determine pressure bounds on the fiber surface along each streamline. For improved computational efficiency, calculations are carried out with respect to the fiber’s local coordinate axes to overcome the rigor of adaptive remeshing during the quasi-transient analysis. Results show low pressure extremes near the fiber’s surface which varies across the printed bead as well as through its thickness. Discussion is provided to gain insight into the effect of low-pressure extremes on micro void formation, particularly at the nozzle exit and during die swell/expansion.

Effect of microplastic particles on the population growth rate and clearance rate of selected ciliates (Protista, Ciliophora).

Microplastics (MPs), due to their micro size, which overlaps with the typical food size of various aquatic organisms, can be ingested and move up the food chain, accumulating in the bodies of organisms at higher trophic levels. Few studies have focused on the uptake of MPs by ciliates, which are an important element of the microbial cycle. Three different ciliate species were used in this study: Blepharisma japonicum, Euplotes sp., and Spirostomum teres, as well as polystyrene beads with diameters of 1 and 2µm at two concentrations (106 and 107 beads × mL-1). The results of the experiments showed that MPs have a variable, species-specific effect on the population growth rate of ciliates, which is directly dependent on their concentration in the environment (P < 0.01). It was also observed that the number of MPs ingested changed over time depending on their concentration and size. On average, the highest number of ingested MPs (883.11 ± 521.47) was recorded at 60min of exposure to a low concentration of small beads in B. japonicum. The lowest number of beads was ingested after 5min of exposure to a low concentration of large beads in the same species. The rate of MP uptake by the ciliate species was significantly dependent on their concentration, exposure time, and size (P < 0.001). The highest clearance rate was observed in the fifth minute of the experiment in the environment with the lowest MP concentration.

Development of a model to study visual categorization learning in chickens (&lt;i&gt;Gallus gallus domesticus&lt;/i&gt;)

Categorization is a cognitive process that enables individuals to recognize similar yet distinct stimuli as equivalent [1–3]. To categorize an object, agents must identify key features of the new object by applying what they have learned from previous interactions with objects in that category [4]. Thus, categorization eliminates the need for the agent to repeatedly investigate each new object, thereby significantly expanding its adaptive capabilities. However, the nervous mechanisms that regulate this process are still not well understood. The primary aim of the present study is to establish an experimental behavioral model that will facilitate the investigation of the neurobiological mechanisms underlying visual categorization learning. Chickens (Gallus gallus domesticus) were selected as a visual learning model. The study of categorization used the chickens’ innate tendency to peck at new small objects and remember their characteristics. For this purpose, a chick was placed in a cage that resembled a house cage with beads affixed to the floor. The “bead floor” consisted of more than 100 beads of different colors and food scattered throughout [5]. In the developed model, pecking solely on the beads was deemed inaccurate as opposed to pecking on the food. For training purposes, the chick was given 80 peckings and evaluated based on how many nibbles it used to create categories for “beads” vs. “food”. If the chick did not make at least 5 cues or was unable to make 80 cues within 10 minutes during the training session, it was excluded from the study. First, the study investigated the effect of simultaneously presenting two new categories of “food” and “beads” on chicks’ behavior. The group that formed both “beads” and “new food” categories made more errors during the learning period, but performed as well as the group that only formed the “bead” category during the subsequent test. The study analyzed whether chicks could categorize objects (beads) of different sizes into a unified group. The chicks were trained with a floor of small beads and later tested with a different floor of larger beads. Results revealed that the chicks did not transfer their categorization ability from small to large beads. In the opposite scenario, where the chicks were presented with large beads during their training, and then offered small beads during the test, they refrained from pecking the smaller ones. In the next stage, we examined how the object’s color affected the development of a novel category. The results showed that when the chicks were presented with the floor containing an additional color of beads, specifically yellow, they made the majority of errors while pecking the “new type of beads”. Then, the study evaluated whether the chicks could transition the classification of “beads” between color set № 1 (blue, pink, green, yellow, and black silver) and color set № 2 (light green, beige, blue, red, gold, and white), and vice versa. The findings indicate that the chickens do form the categories of “beads” in both versions of the task, and after training, do not peck at beads of the new color. Chickens are capable of categorizing “beads” and “food”, and can generalize within these categories. Thus, this experimental study established the formation of categories in chickens during fast learning. Future application of the model will enable the investigation of the underlying neurobiological mechanisms in categorization learning.

Prehistoric Stone Ornaments from Phromtin Tai, Central Thailand: New Perspectives on Workshop Traditions through the Study of Drilling Methods

The multi-component site of Phromtin Tai, Thailand is notable for its long occupation from the Late Bronze Age (ca. 700 - 500 B.C.E.) through early historic periods (ca. C.E. 500 - 900). Multiple field seasons of excavation in burial and habitation areas at the site have recovered a large number of glass and stone beads. Here we present the study of 57 stone beads from the site using qualitative and quantitative methods that demonstrate the presence of multiple stone ornament workshop traditions. Examination of perforations from beads and pendants made from regionally available raw materials of marble, nephrite, serpentine, and other undetermined stone have identified the use of metal drills with abrasives and diamond drills. These drilling techniques along with the distinctive shapes of some finely made beads, as well as the reworking of older broken beads suggests the presence of different scales and organization of local production in Southeast Asia. The different shapes and production processes of carnelian and banded agate beads also may represent various workshop traditions. Some of these latter beads may have originated in South Asia, while others may have been made in different regions of Southeast Asia. This study demonstrates that careful examination of ornament production techniques, and especially bead perforation technology, can be used to identify the presence of different workshop traditions allowing for more fine-grained consideration of inter- and intra-regional bead exchange networks in Southeast Asia. The presence of many semi-precious stone beads of different materials and having morphologically and technologically distinct features at Phromtin Tai demonstrate the active consumption of the beads by social elites at the site. These diversified and exotic status markers represent an intensification and acceleration of the economy and social complexity at Phromtin Tai. KEYWORDS: stone beads, bead drilling, bead production, Central Thailand, Iron Age.

Impact of size and shape in the transport of microplastics by a springtail species

ABSTRACT We investigated the impact of shape (beads, fibers and films) and size (0.5 and 1.0 mm; diameter or length) of polypropylene microplastics (MPs) on MP transport by a springtail species, Folsomia candida in dishes. The percentages of transported beads, fibers and films were 9.10%, 3.18% and 4.10%, respectively. For 1.0 mm MPs, the number of MPs transported was significantly higher for beads than for fibers and films, whereas, for 0.5 mm MPs, the number was significantly higher for fibers than for the other MP types. Additionally, the number of MPs transported was higher for small fibers than for large fibers, whereas it was higher for large beads than for small beads. These results indicate that the effects of F. candida on MP migration depend on MP shape and size. Our results highlight the importance of considering the physical characteristics of MPs while elucidating the interaction between soil fauna and MPs.

Bead-on-fibre morphology in shear-thinning flow

Thin-film flow down a fibre exhibits rich dynamics and is relevant to applications such as desalination, fibre coating and fog harvesting. These flows are subject to instabilities that result in dynamic bead-on-fibre patterns. We perform an experimental study of shear-thinning flow down fibres using 20 different xanthan gum solutions as our working liquid. The bead-on-fibre morphology can be oriented either symmetrically or asymmetrically on the fibre, and this depends upon the surface tension, fibre diameter and liquid rheology, as defined by the Ostwald power-law index. For highly shear-thinning liquids, it is possible for the pattern to be complex and exhibit simultaneously both asymmetric large beads and symmetric small beads in the isolated and convective flow regimes. We quantify the transition between flow regimes and bead dynamics for the asymmetric morphology, and compare with Newtonian flow, as it depends upon the experimental parameters. Finally, the dimensionless bead frequency is shown to scale with the Bond number for all of our experimental data (symmetric and asymmetric).

Enhanced removal of cesium from hydrobiotite using polyacrylonitrile (PAN)-based nickel ferrocyanide beads

The desorption of cesium (Cs) from contaminated clay minerals remains challenging because of the restricted efficiency of the removal process. Therefore, in the present study, a bead-type adsorbent was added during the conventional acid washing process to improve the removal of Cs+ from a clay mineral. As the Cs+ adsorbent, polyacrylonitrile-based nickel potassium hexacyanoferrate (NiFC-PAN) was used to selectively adsorb Cs+ in a strongly acidic solution containing competing ions. To prevent erosion of the adsorbent under harsh environmental conditions and to facilitate the separation of clay particles, PAN was deliberately constructed as large beads. The synthesized adsorbent (NiFC/PAN in a 2:1 ratio) showed high selectivity for Cs+, with a maximum capacity for Cs+ adsorption of 162.78 mg/g in 0.5 M HNO3 solution. Because the NiFC-PAN demonstrated greater Cs+ selectivity than the clay mineral (hydrobiotite, HBT), the addition of NiFC-PAN during the acid washing significantly increased Cs+ desorption (73.3%) by inhibiting the re-adsorption of Cs+ on the HBT. The radioactivity of 137Cs-HBT was substantially decreased from 209 to 27 Bq/g by the acid treatment in the presence of NiFC-PAN, corresponding to a desorption efficiency of 87.1%. Therefore, these results suggest that the proposed technique is a potentially useful and effective method for decontamination of radioactive clay.

Extremely Long Chains of Magnetic Particles via Large Plastic Beads Observed in Bimodal Magnetic Elastomers

The relationship between the magnetorheology of bimodal magnetic elastomers with high concentrations (60 vol %) of plastic beads with diameters of 8 or 200 μm and the meso-structure of the particles was investigated. Dynamic viscoelasticity measurements revealed that the change in storage modulus of the bimodal elastomer with 200 μm beads was 2.8 × 105 Pa at a magnetic field of 370 mT. The change in the storage modulus for monomodal elastomer without beads was 4.9 × 104 Pa. The bimodal elastomer with 8 μm beads hardly responded to the magnetic field. In-situ observation for the particle morphology was performed using synchrotron X-ray CT. For the bimodal elastomer with 200 μm beads, a highly aligned structure of magnetic particles was observed in the gaps between the beads when the magnetic field was applied. On the other hand, for the bimodal elastomer with 8 μm beads, no chain structure of magnetic particles was observed. The orientation angle between the long axis of the aggregation of magnetic particles and the magnetic field direction was determined by an image analysis in three dimensions. The orientation angle varied from 56° to 11° for the bimodal elastomer with 200 μm beads and from 64° to 49° for that with 8 μm beads by applying the magnetic field. The orientation angle of the monomodal elastomer without beads changed from 63° to 21°. It was found that the addition of beads with a diameter of 200 μm linked the chains of magnetic particles, while beads with a diameter of 8 μm prevented the chain formation of the magnetic particles.

Bead size has a greater effect on in vitro elution from antimicrobial-impregnated calcium sulfate beads than drug concentration

To compare the elution characteristics of amikacin-impregnated calcium sulfate (CaSO4) beads based on different drug concentrations and bead size configurations. Six groups of amikacin-impregnated CaSO4 beads and one negative control group. Amikacin-impregnated CaSO4 beads were formed with either 500 mg (low-concentration) or 1 g (high-concentration) of amikacin per 15 g CaSO4 hemihydrate powder. The number of beads necessary to approximate 150 mg of amikacin for each of the 3 bead sizes (3 mm, 5 mm, and 7 mm) at both low and high concentrations were placed in 6 mL of phosphate-buffered saline. The saline was sampled at 14 time points over 28 days. Amikacin concentrations were determined using liquid chromatography-mass spectrometry. Smaller beads reached higher mean peak concentrations than larger beads (P < .0006). Peak concentrations for the low- and high-concentration groups were 20.5 mg/mL and 27.4 mg/mL, 13.1 mg/mL and 14.0 mg/mL, and 8.85 mg/mL and 6.75 mg/mL for the 3 mm, 5 mm, and 7 mm beads, respectively. Bead size also affected the length of therapeutic duration, lasting 6 days for the 3 mm and 5 mm beads and 9 days for the 7 mm beads. However, this was only statistically evident among the high-concentration beads (P < .044). Antimicrobial concentration within the same bead sizes did not affect elution. Amikacin-impregnated CaSO4 beads achieved extreme supratherapeutic eluent concentrations. While additional studies are needed, bead size significantly affected elution with smaller beads reaching higher peak concentrations and 7 mm, high-concentration beads demonstrating a longer therapeutic duration than smaller beads.

MINFLUX dissects the unimpeded walking of kinesin-1.

We introduce an interferometric MINFLUX microscope that records protein movements with up to 1.7 nanometer per millisecond spatiotemporal precision. Such precision has previously required attaching disproportionately large beads to the protein, but MINFLUX requires the detection of only about 20 photons from an approximately 1-nanometer-sized fluorophore. Therefore, we were able to study the stepping of the motor protein kinesin-1 on microtubules at up to physiological adenosine-5'-triphosphate (ATP) concentrations. We uncovered rotations of the stalk and the heads of load-free kinesin during stepping and showed that ATP is taken up with a single head bound to the microtubule and that ATP hydrolysis occurs when both heads are bound. Our results show that MINFLUX quantifies (sub)millisecond conformational changes of proteins with minimal disturbance.

Fabrication and characterization of a cellulose monolith-like particle for virus purification

Monolith-like particles (MLP) are a new base bead architecture optimized for industrial chromatographic downstream processing of very large targets such as whole viral particles (5–300 nm diameter). They have been designed with large pore size though-channels to facilitate access of large diameter targets to the intraparticle chromatographic surface. Herein we describe a novel MLP bead architecture that have been fabricated and characterized for virus purification. This is the first study to demonstrate cellulose-based MLP beads applied for bioseparation of viral particles. The MLP was produced from cellulose acetate by a combined concept of thermally induced phase separation and oil-in-water emulsification. Hydrolyzation, cross-linking, and surface modification with dextran sulfate were carried out for pseudo-affinity chromatographic applications. Scanning electric microscopy analysis showed the presence of large pores that penetrated from the outer surface to within the intraparticle structure. Mercury intrusion porosimetry analysis estimated the mode pore radius to be 1.5 µm. For influenza A virus purification, the MLP surface modified with dextran sulfate exhibited significant higher dynamic binding capacity compared with commercial resins, Cellufine Sulfate and Capto DeVirS. These results demonstrate that viral particles can access the intraparticle surface within these novel large through-pore MLP beads. The novel MLP base architecture described in this study will be a useful support matrix for giant biomolecule and whole viral particle purification.