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Variance in the properties of optical mesoscopic probes is often a limiting factor in applications. In the thermodynamic limit, the smaller the probe, the larger the relative variance. However, specific viral protein cages can assemble efficiently outside the bounds of statistical fluctuations at equilibrium through a process that is characterized by intrinsic quality-control and self-limiting capabilities. In this paper, an approach is described that leverages stoichiometric and structural accuracy in the murine polyoma virus capsid assembly to demonstrate bright, narrowly distributed fluorescence intensity from multichromophore particles that surpass state-of-the-art fluorescent nanosphere probes. Charge-detection mass spectrometry analysis demonstrated that proteins resulting from the fusion of superfolding green fluorescent protein (sfGFP) murine polyoma virus coat proteins self-assemble in vitrointo virus-like particles that have similar stoichiometry as virus-like particles formed from wild-type virus coat proteins. Single-particle analysis by total internal reflection fluorescence microscopy provided evidence for a narrow fluorescence intensity that reflects stoichiometric accuracy of the construct.

The deformation mode during ECAP is often idealized as simple shear, whereas experimental data and numerical simulations indicate the presence of a more complex deformation zone associated with strain gradients. As plastic flow is largely influenced by a material's processing history, especially in terms of grain size, texture and pre-deformation, the evolution of local material flow during multiple passes of ECAP is of significant interest. In this study, we use a method for visio-plastic analysis of the local deformation during ECAP of the aluminum alloy AA6060 via different processing routes in a friction-optimized tool with a tool angle of 90°: Prior to the final pass, the billets are cut lengthwise and prepared with a grid of indents on the inside. After interrupting the deformation during the final ECAP pass, both billet halves are extracted from the modular tool. The positions of the indents along several paths, i.e., flow lines, are analyzed from the partially deformed billets using optical images and a graphics software. For the analysis of the material flow we use a phenomenological model that describes the material path along the flow line based on a super-ellipse function, with only one parameter defining the evolution of curvature along the flow line. The resulting data allow for a detailed analysis of strain distributions and gradients inside billets after different numbers of passes. Through comprehensive microstructural and mechanical characterization of the processed material, we discuss changes in macroscopic properties and their relationship to microstructural evolution and local strain accumulation during ECAP.

AbstractObjectiveThis study explored the structure validity and sex and country measurement invariance for Filipino and U.S. adults using the Five Factor Wellness Inventory (FFWEL-A2).MethodConfirmatory factor analysis and measurement invariance were conducted to determine consistency of responses across sex and country categories. Factorial ANOVAs were used to determine differences by sex, race, and country.ResultsThe hypothesized 5-factor and second-order factor structure of the FFWEL provided equivalent but poor to adequate fit to the data in a large sample (N = 2,071) of U.S. and a smaller sample (N = 269) of Filipino counselors and counselors-in-training. Coefficients αwere >0.80 for almost all domain scores, but only five of the 17 facet scores met the 0.80 criterion. Weak measurement invariance was evident for men (n = 502) and women (N = 1838) across both the U.S. White and Filipino group comparisons.ConclusionsCounselors should take care to explore the way they and clients understand wellness and consider that not everyone will have the same understanding of the individual factors as outlined in the FFWEL-A2, especially among male, female, and diverse people from different countries.Significance statementWellness is an ethical and professional imperative for counselors to avoid professional impairment and burn out. In this study, scores on the FFWEL-A2, a prominent measure of wellness, displayed partially biased measures of male and female scores across U.S. and Filipino samples and across diverse racial groups, probably due to complexity of the wellness model.

Nucleic acid nanoparticles (NANPs) are increasingly used in preclinical investigations as delivery vectors. Tools that can characterize assembly and assess quality will accelerate their development and clinical translation. Standard techniques used to characterize NANPs, like gel electrophoresis, lack the resolution for precise characterization. Here, we introduce the use of charge detection mass spectrometry (CD-MS) to characterize these materials. Using this technique, we determined the mass of NANPs varying in size, shape, and molecular mass, NANPs varying in production quality due to formulations lacking component oligonucleotides, and NANPs functionalized with protein and nucleic acid-based secondary molecules. Based on these demonstrations, CD-MS is a promising tool to precisely characterize NANPs, enabling more precise assessments of the manufacturing and processing of these materials.

The main capsid protein (CP) of norovirus, the leading cause of gastroenteritis, is expected to self-assemble into virus-like particles with the same structure as the wild-type virus, a capsid with 180 CPs in a T= 3 icosahedron. Using charge detection mass spectrometry (CD-MS), we find that the norovirus GI.1 variant is structurally promiscuous, forming a wide variety of well-defined structures, some that are icosahedral capsids and others that are not. The structures that are present evolve with time and vary with solution conditions. The presence of icosahedral T= 3 and T= 4 capsids (240 CPs) under some conditions was confirmed by cryo-electron microscopy (cryo-EM). The cryo-EM studies also confirmed the presence of an unexpected prolate geometry based on an elongated T= 4 capsid with 300 CPs. In addition, CD-MS measurements indicate the presence of well-defined peaks with masses corresponding to 420, 480, 600, and 700 CPs. The peak corresponding to 420 CPs is probably due to an icosahedral T= 7 capsid, but this could not be confirmed by cryo-EM. It is possible that the T= 7 particles are too fragile to survive vitrification. There are no mass peaks associated with the T= 9 and T= 12 icosahedra with 540 and 720 CPs. The larger structures with 480, 600, and 700 CPs are not icosahedral; however, their measured charges suggest that they are hollow shells. The use of CD-MS to monitor virus-like particles assembly may have important applications in vaccine development and quality control.

AbstractObjectiveCounseling Center Assessment of Psychological Symptoms (CCAPS-34) scores were studied for longitudinal bias-free construct evidence.MethodA sample of 4,696 university students referred to a university counseling center were assessed twice for evidence of longitudinal measurement invariance.ResultsAdequate or marginal longitudinal measurement invariance (LMI) of all subscales except for Eating Concerns was confirmed. Coefficients alpha and omega suggested that the internal consistencies for both initial and subsequent administrations were largely adequate for screening level test scores (≥ .80), except for the Hostility and Alcohol Use subscales (≥ .73). Second session effect sizes (Cohen’s d) were small but detectable and statistically significant for most subscales (-0.02–.17).ConclusionsCounselors can use most CCAPS-34 subscale scores for tracking psychological symptoms and constructs over time with confidence. The Generalized Anxiety, Social Anxiety, Academic Concerns, and Alcohol Use subscales can be used with great confidence. Some confidence is warranted from the LMI evidence when using the Depression and Hostility subscales, but caution in warranted when using the Eating Concerns subscale.

During extrusion, the material is subjected to a heterogeneous deformation that results in the formation of gradients of microstructure and thus mechanical properties. In this study, using continuously cast commercially pure aluminum as a reference material, we show how a cast structure with coarse columnar grains affects the formation of these gradients during extrusion at room temperature. Characterization of the initial material as well as the extruded round bars by optical and electron microscopy, X-ray diffraction as well as by means of mechanical testing documents the formation of four characteristic annular sections. For extrusion along the casting direction there is a <100>/<111> double-fiber textured center with lowest hardness, followed by a <111> single-fiber textured ring with a hardness plateau, a double-fiber textured region with grains arranged alternatingly in an iris-like shape, and an (ultra-)fine grained surface layer with highest hardness. For extrusion opposite to the casting direction we find similar characteristics of the center section. With increasing distance from the center follow another double-fiber textured section with increasing hardness, a <111> single-fiber textured ring with nearly homogeneous hardness and a surface layer with a slightly rotated <111> fiber and highest hardness. The key finding is that the macroscopic anisotropy of the cast material, resulting from the grain size, crystal orientation and growth direction of the columnar grains, determines the local material flow during extrusion and thus leads to the formation of these different complex multi-gradient macrostructures that may provide unique properties for complex applications such as light-weight safety-relevant components.

Caudal malocclusions in cats may result in a variety of traumatic lesions affecting the soft tissues of the ipsilateral mandible such as fovea, gingival cleft, and proliferative lesions. Fifty-one cats diagnosed with a traumatic caudal malocclusion were compared with a control hospital population and evaluated for prevalence with respect to breed and sex. Twenty-two cats that were treated had radiographic, clinical findings, and the outcome of treatment (extraction or odontoplasty) recorded. Maine Coon, Persian, and male neutered cats were overrepresented while Domestic Shorthairs were underrepresented within the study population. Radiographically, 50% of the fovea lesions had an area of decreased bone density in the region of the lesion and none of these had evidence of periodontal disease. All gingival cleft lesions had radiographic changes consistent with periodontal disease. 15.4% of proliferative lesions presented with radiographic changes, with only half of those presenting with both radiographic and clinical evidence of periodontal disease. Eleven cats were treated by odontoplasty and eleven by extraction. One cat treated by odontoplasty developed new lesions caudally, and another had persistence of the initial lesions. Two cats in the extraction group developed new lesions rostral to the extracted teeth. In most instances, odontoplasty or extraction resulted in successful soft tissue lesion resolution. In rare cases, additional treatment was necessary due to either persistence or development of new lesions.

Process development activities required to develop a robust and scalable synthetic route to AZD7594 (1) are disclosed. Included in this paper is the rationale for the selection of a palladium-mediated Buchwald–Hartwig coupling to form a key C–N bond. Reaction optimization and understanding were performed to minimize impurity formation and to develop a robust control strategy. The optimized synthetic sequence was used to manufacture 220 kg of AZD7594 (1) for clinical trials.

Large surface areas are important for enhancing mass and energy transfer in biological and technological processes. Bicontinuous interfacially jammed emulsion gels (bijels) increase the surface area between two fluids by intertwining them into particle stabilized networks. To facilitate efficient mass and energy exchange viathe bijels’ high surface area, the fluid networks need to be connected to their respective bulk phases. Here, we generate bijels between two bulk fluids and investigate the connections the bijel makes. We analyze these connections by investigating the colloidal stability, interfacial rheology and mass transfer dynamics during bijel formation. To this end, we employ confocal and electron microscopy, as well as dynamic light scattering, pendant drop analysis, electrophoretic mobility measurements and diffusion simulations. We find that the connections the bijel makes to the bulk fluid can be disrupted by severe colloidal aggregation and interruptions of the bicontinuous fluid network. However, the addition of alcohol to the bulk fluid moderates aggregation and allows undisturbed fluid network formation, facilitating open connections between bijel and bulk fluid. The unprecedented control of bijel pore connections from this research will be crucial for the application of bijels as separation membranes, electrochemical energy storage materials and chemical reactors.

A novel thermodynamically consistent macroscopic magnetic hysteresis model is presented. Magnetization is calculated from the reversible part of the magnetic field, while the evolution law of the irreversible part involves physically meaningful material constants: the coercive field and the initial susceptibilities of the hysteretic and anhysteretic curves. It is possible to invert the model through an iterative procedure, allowing either the magnetic field or the flux density as an input to the model. The model is tested on both soft and hard magnetic materials for major and minor loops.

Hysteresis model of soft magnetic material enables an accurate estimation of the losses in electrical application. In rotating machine, the flux distribution can magnetize many directions of the iron core. The combination of the rotational and the alternating flux density appears between the slot and the yoke region. Even if the alternating flux density appears in the middle of each tooth, the magnetic properties varies with the magnetized direction with respect to the rolling direction. Whereas the losses strictly increases with the amplitude of an alternating flux density, the hysteresis loss presents a maximum under rotational flux density condition. Although this phenomenon is properly described by the coherent rotation of the domains at the microscopic scale, the macroscopic representation of the material should represent this specific feature. In this article, a general vectorization technique is applied to the Jiles–Atherton model of hysteresis. The anisotropic anhysteresis behavior is derived from the analysis of a cubic crystal. This anhysteresis model drives the coherent rotation of the hysteresis model. Even if the model parameters are identified with alternating flux condition, the rotational flux density can be predicted properly.

Die Rhizarthrose führt zu einer wesentlichen Einschränkung der Handfunktion, da das Daumensattelgelenk die für die Greiffunktion entscheidende Oppositionsfähigkeit des Daumens ermöglicht. Im Anfangsstadium der Erkrankung stehen zunächst wirksame konservative Therapieoptionen und später gelenkerhaltende Eingriffe zur Verfügung. Im fortgeschrittenen Erkrankungsstadium, nach Ausschöpfen der konservativen Therapie und bei entsprechendem Leidensdruck hat sich die Resektionsarthroplastik des Daumensattelgelenks mit ihren Varianten als maßgeblicher Standardeingriff bewährt, da sie zuverlässig und langfristig mit einer niedrigen Komplikationsrate zu einer deutlichen Schmerzlinderung, einer sehr hohen Patientenzufriedenheit und einer guten Wiederherstellung der Handfunktion führt. Daneben stehen für besondere Indikationen weitere gelenkresezierende Eingriffe wie etwa der endoprothetische Ersatz oder die Arthrodese des Daumensattelgelenks zur Verfügung.

ObjectiveMucosal T cells play a major role in inflammatory bowel disease (IBD). However, their immunometabolism during intestinal inflammation is poorly understood. Due to its impact on cellular metabolism and proinflammatory immune cell function, we here focus on the enzyme ATP citrate lyase (ACLY) in mucosal T cell immunometabolism and its relevance for IBD.DesignACLY expression and its immunometabolic impact on colitogenic T cell function were analysed in mucosal T cells from patients with IBD and in two experimental colitis models.ResultsACLY was markedly expressed in colon tissue under steady-state conditions but was significantly downregulated in lamina propria mononuclear cells in experimental dextran sodium sulfate-induced colitis and in CD4+and to a lesser extent in CD8+T cells infiltrating the inflamed gut in patients with IBD. ACLY-deficient CD4+T cells showed an impaired capacity to induce intestinal inflammation in a transfer colitis model as compared with wild-type T cells. Assessment of T cell immunometabolism revealed that ACLY deficiency dampened the production of IBD-relevant cytokines and impaired glycolytic ATP production but enriched metabolites involved in the biosynthesis of phospholipids and phosphatidylcholine. Interestingly, the short-chain fatty acid butyrate was identified as a potent suppressor of ACLY expression in T cells, while IL-36α and resolvin E1 induced ACLY levels. In a translational approach, in vivo administration of the butyrate prodrug tributyrin downregulated mucosal infiltration of ACLYhighCD4+T cells and ameliorated chronic colitis.ConclusionACLY controls mucosal T cell immunometabolism and experimental colitis. Therapeutic modulation of ACLY expression in T cells emerges as a novel strategy to promote the resolution of intestinal inflammation.

AbstractPsychometric characteristics of CESD-R scores were explored on a sample of 966 undergraduate students. Internal consistency (α = .92), external convergent and discriminant validity, and response bias were adequate to excellent. Strong measurement invariance was evident for gender and race comparisons, and the unidimensional model fit the data best.

Despite the significant role of the gut microbiota in infant health and development, little is known about the ecological processes determining gut microbial community assembly. According to ecology theory, the timing and order of arrival of microbial species into an ecosystem affect microbial community assembly, a phenomenon termed priority effects. Bifidobacteriumspecies are recognized as highly abundant early colonizers of the infant’s gut, partly due to their ability to selectively utilize human milk oligosaccharides (HMOs) from breast milk. However, the role of priority effects in Bifidobacteriumcommunity assembly remains unclear. Here, we investigated the Bifidobacteriumcommunity assembly in the gut of 25 breastfed Danish infants longitudinally sampled throughout the first 6 months of life. Our results showed that the breastfed infants were often initially, but temporarily, dominated by suboptimal HMO-utilizing Bifidobacteriumtaxa, such as B. longumsubsp. longum, before more efficient HMO-utilizers such as B. longumsubsp. infantis, replaced the first colonizer as the dominant Bifidobacteriumtaxon. Subsequently, we validated this observation using gnotobiotic mice sequentially colonized with B. longumsubsp. longumand B. longumsubsp. infantisor vice versa, with or without supplementation of HMOs in the drinking water. The results showed that in the absence of HMOs, order of arrival determined dominance. Yet, when mice were supplemented with HMOs the strength of priority effects diminished, and B. longumsubsp. infantisdominated regardless of colonization order. Our data demonstrate that the arrival order of Bifidobacteriumtaxa and the deterministic force of breast milk-derived HMOs, dictate Bifidobacteriumcommunity assembly in the infant’s gut.

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AbstractThe Social Justice Scale (SJS) and various subscales were studied on a sample of 414 human services university undergraduates. Internal consistency (α = .93), discriminant validity, and response bias were adequate. The 4-factor model demonstrated an excellent fit. Implications were discussed for use of the SJS in practice and research.

Ultrafast imaging modes, such as coherent plane-wave compounding (CPWC), increase image uniformity and reduce grating lobe artifacts by dynamic receive apertures. The focal length and the desired aperture width maintain a given ratio, which is called the ${F}$ -number. Fixed ${F}$ -numbers, however, exclude useful low-frequency components from the focusing and reduce the lateral resolution. Herein, this reduction is avoided by a frequency-dependent ${F}$ -number. This ${F}$ -number derives from the far-field directivity pattern of a focused aperture and can be expressed in closed form. The ${F}$ -number, at low frequencies, widens the aperture to improve the lateral resolution. The ${F}$ -number, at high frequencies, narrows the aperture to avoid lobe overlaps and suppress grating lobes. Phantom and in vivo experiments with a Fourier-domain beamforming algorithm validated the proposed ${F}$ -number in CPWC. The lateral resolution, which was measured by the median lateral full-widths at half-maximum of wires, improved by up to 46.8% and 14.9% in a wire and a tissue phantom, respectively, in comparison to fixed ${F}$ -numbers. Grating lobe artifacts, which were measured by the median peak signal-to-noise ratios of wires, reduced by up to 9.9 dB in comparison to the full aperture. The proposed ${F}$ -number thus outperformed ${F}$ -numbers that were recently derived from the directivity of the array elements.

Charge detection mass spectrometry (CDMS) was examined as a means of studying proteasomes. To this end, the following masses of the 20S, 19S, 26S, and 30S proteasomes from Saccharomyces cerevisiae(budding yeast) were measured: m(20S) = 738.8 ± 2.9 kDa, m(19S) = 926.2 ± 4.8 kDa, m(26S) = 1,637.0 ± 7.6 kDa, and m(30S) = 2,534.2 ± 10.8 kDa. Under some conditions, larger (20S)x(where x= 1 to ∼13) assemblies are observed; the 19S regulatory particle also oligomerizes, but to a lesser extent, forming (19S)xcomplexes (where x= 1 to 4, favoring the x= 3 trimer). The (20S)xoligomers are favored in vitro, as the pH of the solution is lowered (from 7.0 to 5.4, in a 20 mM ammonium acetate solution) and may be related to in vivo proteasome storage granules that are observed under carbon starvation. From measurements of m(20S)x(x= 1 to ∼13) species, it appears that each multimer retains all 28 proteins of the 20S complex subunit. Several types of structures that might explain the formation of (20S)xassemblies are considered. We stress that each structural type [hypothetical planar, raft-like geometries (where individual proteasomes associate through side-by-side interactions); elongated, rodlike geometries (where subunits are bound end-to-end); and geometries that are roughly spherical (arising from aggregation through nonspecific subunit interactions)] is highly speculative but still interesting to consider, and a short discussion is provided. The utility of CDMS for characterizing proteasomes and related oligomers is discussed.

Recombinant adeno-associated virus (rAAV) is a leading gene therapy vector. However, neutralizing antibodies reduce its efficacy. Traditional methods used to investigate antibody binding provide limited information. Here, charge detection mass spectrometry (CD-MS) was used to investigate the binding of monoclonal antibody ADK8 to AAV serotype 8 (AAV8). CD-MS provides a label-free approach to antibody binding. Individual binding events can be monitored as each event is indicated by a shift of the antibody–antigen complex to a higher mass. Unlike other methods, the CD-MS approach reveals the distribution of antibodies bound on capsids, allowing AAV8 subpopulations with different affinities to be identified. The charge state generated by the electrospray of large ions is normally correlated with the structure, and the charge is expected to increase when an antibody binds to the capsid exterior. Surprisingly, binding of the first ADK8 to AAV8 causes a substantial decrease in the charge, suggesting that the first antibody binding event causes a significant structural change. The charge increases for subsequent binding events. Finally, high ADK8 concentrations cause agglutination, where ADK8 links AAV capsids to form dimers and higher order multimers.

AbstractThis study explored the psychometric characteristics of the Alcohol Use Disorders Identification Test (AUDIT) on a sample of 414 university undergraduate students (Mage = 19.07, SD = 0.95). Internal consistency (α = .83), external divergent validity, and internal structural validity were adequate; the 2-factor model provided the best fit to the data.

Medical liability insurance covers physicians’ liability, and its price could affect physicians’ practice. In this article, we use a unique county-level dataset to study how medical liability insurance prices of three specialties, internal medicine, general surgery, and obstetrics–gynecology (OB-GYN), changed after the Affordable Care Act (ACA) elective Medicaid expansion provision. The Medicaid expansion has largely increased the demand for health care services and potentially exposed physicians to higher medical liability risks. With higher expected losses, insurers could react by increasing medical malpractice insurance prices. We first study all counties in states that elected to expand Medicaid and compare them to counties in nonexpansion states. Then we narrow our analysis to consider differential effects in bordering counties with different Medicaid expansion statuses over the period 2010–2018. In both samples, we find significantly higher medical liability insurance prices 2 years after the expansion (on average) in expansion states in comparison to nonexpansion states, and the difference is larger for physicians practicing internal medicine (6–8% at 2 years after expansion) and general surgery (12–16% at 2 years after expansion) but less so for OB-GYN. Our OB-GYN results are likely because significant numbers of births were already covered under Medicaid and were not affected by the expansion. Our finding suggests that the expansion of health insurance increases liability costs to medical practitioners.