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The recent uptick in the approval of ex vivo cell therapies highlights the relevance of lentivirus (LV) as an enabling viral vector of modern medicine. As labile biologics, however, LVs pose critical challenges to industrial biomanufacturing. In particular, LV purification—currently reliant on filtration and anion-exchange or size-exclusion chromatography—suffers from long process times and low yield of transducing particles, which translate into high waiting time and cost to patients. Seeking to improve LV downstream processing, this study introduces peptides targeting the enveloped protein Vesicular stomatitis virus G (VSV-G) to serve as affinity ligands for the chromatographic purification of LV particles. An ensemble of candidate ligands was initially discovered by implementing a dual-fluorescence screening technology and a targeted in silico approach designed to identify sequences with high selectivity and tunable affinity. The selected peptides were conjugated on Poros resin and their LV binding-and-release performance was optimized by adjusting the flow rate, composition, and pH of the chromatographic buffers. Ligands GKEAAFAA and SRAFVGDADRD were selected for their high product yield (50%–60% of viral genomes; 40%–50% of HT1080 cell-transducing particles) upon elution in PIPES buffer with 0.65 M NaCl at pH 7.4. The peptide-based adsorbents also presented remarkable values of binding capacity (up to 3·109 TU per mL of resin, or 5·1011 vp per mL of resin, at the residence time of 1 min) and clearance of host cell proteins (up to a 220-fold reduction of HEK293 HCPs). Additionally, GKEAAFAA demonstrated high resistance to caustic cleaning-in-place (0.5 M NaOH, 30 min) with no observable loss in product yield and quality. 

Most researchers assume minimal impact of pretreatment on strontium isotope ratios ( 87 Sr/ 86 Sr) for bones and teeth, and methods vary tremendously. We compared 14 pretreatment methods, including no prep other than powdering enamel, ashing, soaking in water, an oxidizing agent (bleach or hydrogen peroxide) or acetic acid (0.1 M, 1.0 M, and 1.0 M buffered with calcium acetate), and a combination of these steps. We prepared and analyzed aliquots of powdered molar enamel from three proboscideans (one modern captive Indian elephant, Elephas maximus indicus ; one Pleistocene mastodon, Mammut americanum ; and one Miocene gomphothere, Afrochoerodon kisumuensis ). Each pretreatment was performed in triplicate and we measured 87 Sr/ 86 Sr, Sr concentration, and uranium (U) concentration, using the same lab space and instrumentation for all samples. Variability in 87 Sr/ 86 Sr and Sr and U concentrations was considerable across pretreatments. Mean 87 Sr/ 86 Sr across methods ranged from 0.70999 to 0.71029 for the modern tooth, 0.71458 to 0.71502 for the Pleistocene tooth, and 0.70804 to 0.70817 for the Miocene tooth. The modern tooth contained the least Sr and negligible U. The Pleistocene tooth contained slightly more Sr and measurable amounts of U, and the Miocene tooth had approximately 5x more Sr and U than the Pleistocene tooth. For all three teeth, variance in 87 Sr/ 86 Sr, Sr concentrations, and U concentrations among replicates was statistically indistinguishable across pretreatments, but there were apparent differences among pretreatments for the modern and Pleistocene teeth. Both contained relatively little Sr, and it is possible that small amounts of exogenous Sr from reagents, building materials or dust affected some replicates for some pretreatments. For the modern tooth, median 87 Sr/ 86 Sr varied considerably (but statistically insignificantly) across pretreatments. For the Pleistocene tooth, variability in median 87 Sr/ 86 Sr was also considerable; some pretreatments were statistically distinct but there were no obvious patterns among methods. For the Miocene tooth, variability in median 87 Sr/ 86 Sr was much smaller, but there were significant differences among pretreatments. Most pretreatments yielded 87 Sr/ 86 Sr and Sr concentrations comparable to, or lower than, untreated powder, suggesting selective removal of exogenous material with high 87 Sr/ 86 Sr. Further evaluation of the mechanisms driving isotopic variability both within and among pretreatment methods is warranted. Researchers should clearly report their methods and avoid combining data obtained using different methods. Small differences in 87 Sr/ 86 Sr could impact data interpretations, especially in areas where isotopic variability is low. 

The method of regularised stokeslets is widely used to model microscale biological propulsion. The method is usually implemented with only the single-layer potential, the double-layer potential being neglected, despite this formulation often not being justified a priori due to nonrigid surface deformation. We describe a meshless approach enabling the inclusion of the double layer which is applied to several Stokes flow problems in which neglect of the double layer is not strictly valid: the drag on a spherical droplet with partial-slip boundary condition, swimming velocity and rate of working of a force-free spherical squirmer, and trajectory, swimmer-generated flow and rate of working of undulatory swimmers of varying slenderness. The resistance problem is solved accurately with modest discretisation on a notebook computer with the inclusion of the double layer ranging from no-slip to free-slip limits; the neglect of the double-layer potential results in up to 24% error, confirming the importance of the double layer in applications such as nanofluidics, in which partial slip may occur. The squirming swimmer problem is also solved for both velocity and rate of working to within a small percent error when the double-layer potential is included, but the error in the rate of working is above 250% when the double layer is neglected. The undulating swimmer problem by contrast produces a very similar value of the velocity and rate of working for both slender and nonslender swimmers, whether or not the double layer is included, which may be due to the deformation’s ‘locally rigid body’ nature, providing empirical evidence that its neglect may be reasonable in many problems of interest. The inclusion of the double layer enables us to confirm robustly that slenderness provides major advantages in efficient motility despite minimal qualitative changes to the flow field and force distribution. 

Transcription factors associate with architectural proteins to regulate genome organization and three-dimensional gene regulation. 

Charge density waves (CDWs) have been observed in nearly all families of copper-oxide superconductors. But the behavior of these phases across different families has been perplexing. In La-based cuprates, the CDW wavevector is an increasing function of doping, exhibiting the so-called Yamada behavior, while in Y- and Bi-based materials the behavior is the opposite. Here, we report a combined resonant soft X-ray scattering (RSXS) and neutron scattering study of charge and spin density waves in isotopically enriched La 1.8 − x Eu 0.2 Sr x CuO 4 over a range of doping 0.07 ≤ x ≤ 0.20 . We find that the CDW amplitude is temperature independent and develops well above experimentally accessible temperatures. Further, the CDW wavevector shows a nonmonotonic temperature dependence, exhibiting Yamada behavior at low temperature with a sudden change occurring near the spin ordering temperature. We describe these observations using a Landau–Ginzburg theory for an incommensurate CDW in a metallic system with a finite charge compressibility and spin-CDW coupling. Extrapolating to high temperature, where the CDW amplitude is small and spin order is absent, our analysis predicts a decreasing wavevector with doping, similar to Y and Bi cuprates. Our study suggests that CDW order in all families of cuprates forms by a common mechanism. 

We have conducted a paleomagnetic study of Holocene sediments from Lake Victoria in order to develop a high-resolution record of paleomagnetic secular variation (PSV). This study has recovered PSV records from two cores (V95-1P and V95-7P) in northern Lake Victoria (0.5°S). The PSV is recorded in fine-grained detrital magnetite/titanomagnetite grains, but the rock magnetic data suggest that significant magnetic mineral dissolution has occurred, which limits our paleomagnetic studies to the uppermost ~5 m of both cores. Detailed alternating field (af) demagnetization of the natural remanence (NRM) shows that a distinctive characteristic remanence (ChRM) is demagnetized from ~10 to 40 mT, which decreases simply toward the origin. The resulting directional PSV records for 1P and 7P are correlatable with 22 distinct inclination features and 19 declination features. Radiocarbon dating of the cores is based on eight radiocarbon dates from core 1P, which can be correlated into core 7P using both the PSV and rock magnetic/environmental measurements. The final PSV time series cover the last 11,000 years with an average sediment accumulation rate of ~40 cm/kyr. The Lake Victoria PSV records can be correlated with new PSV records from Lake Malawi. Comparison of the correlatable PSV feature ages between the two lakes indicates that the PSV records are not significantly different in age, although Lake Victoria PSV ages might average ~100 years younger. We think that the Lake Victoria and Lake Malawi PSV records, together, provide the most accurate, well-dated, and consistent record of Holocene PSV for Africa yet developed.

 

Adeno‐associated viruses (AAVs) have acquired a central role in modern medicine as delivery agents for gene therapies targeting rare diseases. While new AAVs with improved tissue targeting, potency, and safety are being introduced, their biomanufacturing technology is lagging. In particular, the AAV purification pipeline hinges on protein ligands for the affinity‐based capture step. While featuring excellent AAV binding capacity and selectivity, these ligands require strong acid (pH <3) elution conditions, which can compromise the product's activity and stability. Additionally, their high cost and limited lifetime has a significant impact on the price tag of AAV‐based therapies. Seeking to introduce a more robust and affordable affinity technology, this study introduces a cohort of peptide ligands that (i) mimic the biorecognition activity of the AAV receptor (AAVR) and anti‐AAV antibody A20, (ii) enable product elution under near‐physiological conditions (pH 6.0), and (iii) grant extended reusability by withstanding multiple regenerations. A20‐mimetic CYIHFSGYTNYNPSLKSC and AAVR‐mimetic CVIDGSQSTDDDKIC demonstrated excellent capture of serotypes belonging to distinct clones/clades – namely, AAV1, AAV2, AAV5, AAV6, AAV8, and AAV9. This corroborates the in silico models documenting their ability to target regions of the viral capsid that are conserved across all serotypes. CVIDGSQSTDDDKIC‐Toyopearl resin features binding capacity (≈10¹⁴vp mL⁻¹) and product yields (≈60%–80%) on par with commercial adsorbents, and purifies AAV2 from HEK293 and Sf9 cell lysates with high recovery (up to 78%), reduction of host cell proteins (up to 700‐fold), and high transduction activity (up to 65%).

 

Through billions of years of evolution, microorganisms mastered unique swimming behaviors to thrive in complex fluid environments. Limitations in nanofabrication have thus far hindered the ability to design and program synthetic swimmers with the same abilities. Here we encode multi-behavioral responses in microscopic self-propelled tori using nanoscale 3D printing. We show experimentally and theoretically that the tori continuously transition between two primary swimming modes in response to a magnetic field. The tori also manipulated and transported other artificial swimmers, bimetallic nanorods, as well as passive colloidal particles. In the first behavioral mode, the tori accumulated and transported nanorods; in the second mode, nanorods aligned along the toriʼs self-generated streamlines. Our results indicate that such shape-programmed microswimmers have a potential to manipulate biological active matter, e.g. bacteria or cells.