Analysis of the Accuracy of CAM-type Deformity Resection on a Low-cost Arthroscopic Simulator in a Training Scenario.

Objective To evaluate surgeons' performance in resecting CAM-type deformities using a realistic arthroscopic surgery simulator. Methods An arthroscopic simulator was created using low-cost materials with the help of a GTMax Core A1 3D printer and the programs Invesalius and Meshmixer 2017, which were used to develop femoral head parts in ABS material, with the presence of a CAM-type deformity, to mimic a femoroacetabular impact situation. After the operations were performed by 16 surgeons, the femurs were compared to a previous model with deformity and another without, using Cloudcompare, and parameters such as the volumetric difference between the operated femurs, with and without deformity, the minimum and maximum distance between them, the percentage of the deformity resected, the estimated time for total resection of the deformity, as well as a qualitative analysis based on the images and graphs provided by the program representing the areas of the parts resected, were evaluated at the end. Results The average resection speed was 34.66 mm 3 /min (SD = 46 mm 3 /min, max = 147.33; min = -2.66). The average resection rate was 26.2% (SD = 34.7%, max = 111; min = -2). Qualitative analysis showed hyporesection of deformities and sometimes hyperresection of nondeformed areas. The simulator was highly rated by the surgeons, with a tactile sensation very similar to real surgery, according to them. Conclusion Arthroscopic simulators have proved very useful in training less experienced surgeons.

Modulation of FGF pathway signaling and vascular differentiation using designed oligomeric assemblies.

Many growth factors and cytokines signal by binding to the extracellular domains of their receptors and driving association and transphosphorylation of the receptor intracellular tyrosine kinase domains, initiating downstream signaling cascades. To enable systematic exploration of how receptor valency and geometry affect signaling outcomes, we designed cyclic homo-oligomers with up to 8 subunits using repeat protein building blocks that can be modularly extended. By incorporating a de novo-designed fibroblast growth factor receptor (FGFR)-binding module into these scaffolds, we generated a series of synthetic signaling ligands that exhibit potent valency- and geometry-dependent Ca2+ release and mitogen-activated protein kinase (MAPK) pathway activation. The high specificity of the designed agonists reveals distinct roles for two FGFR splice variants in driving arterial endothelium and perivascular cell fates during early vascular development. Our designed modular assemblies should be broadly useful for unraveling the complexities of signaling in key developmental transitions and for developing future therapeutic applications.

Computational design of non-porous pH-responsive antibody nanoparticles.

Programming protein nanomaterials to respond to changes in environmental conditions is a current challenge for protein design and is important for targeted delivery of biologics. Here we describe the design of octahedral non-porous nanoparticles with a targeting antibody on the two-fold symmetry axis, a designed trimer programmed to disassemble below a tunable pH transition point on the three-fold axis, and a designed tetramer on the four-fold symmetry axis. Designed non-covalent interfaces guide cooperative nanoparticle assembly from independently purified components, and a cryo-EM density map closely matches the computational design model. The designed nanoparticles can package protein and nucleic acid payloads, are endocytosed following antibody-mediated targeting of cell surface receptors, and undergo tunable pH-dependent disassembly at pH values ranging between 5.9 and 6.7. The ability to incorporate almost any antibody into a non-porous pH-dependent nanoparticle opens up new routes to antibody-directed targeted delivery.

Protein nanoparticle vaccines induce potent neutralizing antibody responses against MERS-CoV.

Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic betacoronavirus that causes severe and often lethal respiratory illness in humans. The MERS-CoV spike (S) protein is the viral fusogen and the target of neutralizing antibodies, and has therefore been the focus of vaccine design efforts. Currently there are no licensed vaccines against MERS-CoV and only a few candidates have advanced to Phase I clinical trials. Here we developed MERS-CoV vaccines utilizing a computationally designed protein nanoparticle platform that has generated safe and immunogenic vaccines against various enveloped viruses, including a licensed vaccine for SARS-CoV-2. Two-component protein nanoparticles displaying MERS-CoV S-derived antigens induced robust neutralizing antibody responses and protected mice against challenge with mouse-adapted MERS-CoV. Electron microscopy polyclonal epitope mapping and serum competition assays revealed the specificities of the dominant antibody responses elicited by immunogens displaying the prefusion-stabilized S-2P trimer, receptor binding domain (RBD), or N-terminal domain (NTD). An RBD nanoparticle vaccine elicited antibodies targeting multiple non-overlapping epitopes in the RBD, whereas anti-NTD antibodies elicited by the S-2P- and NTD-based immunogens converged on a single antigenic site. Our findings demonstrate the potential of two-component nanoparticle vaccine candidates for MERS-CoV and suggest that this platform technology could be broadly applicable to betacoronavirus vaccine development.

Life Cycle Greenhouse Gas Emissions of CO2-Enabled Sedimentary Basin Geothermal.

The expansion of renewable energy and the large-scale deployment of carbon dioxide (CO2) capture and storage (CCS) can decarbonize the power sector. The use of CO2 to extract geothermal heat from naturally porous and permeable sedimentary basins to generate electricity (CO2-plume geothermal (CPG) system) presents an opportunity to simultaneously generate renewable energy and geologically store CO2. In this study, we estimate the life cycle greenhouse gas (GHG) impacts of CPG systems through 12 scenarios in which CPG systems are combined with one of six CO2 sources (e.g., bioenergy with carbon capture and storage (BECCS) and iron and steel facilities) and operate in two geological settings. We find the life cycle GHG emissions of CPG systems ranging from -0.25 to -6.18 kg CO2eq/kWh. CPG systems can achieve the highest emissions reductions when utilizing the CO2 captured from BECCS. We evaluate uncertainty through a Monte Carlo simulation, demonstrating consistent net reductions in life cycle emissions and a local, one-parameter-at-a-time sensitivity analysis that identifies the CO2 capture capacity as the high-impact parameter of the results. Through the production of electricity, CPG systems can provide additional environmental benefits to the deployment of large-scale CCS.

Whole-brain gray matter maturation trajectories associated with autistic traits from adolescence to early adulthood.

A growing number of evidence supports a continued distribution of autistic traits in the general population. However, brain maturation trajectories of autistic traits as well as the influence of sex on these trajectories remain largely unknown. We investigated the association of autistic traits in the general population, with longitudinal gray matter (GM) maturation trajectories during the critical period of adolescence. We assessed 709 community-based adolescents (54.7% women) at age 14 and 22. After testing the effect of sex, we used whole-brain voxel-based morphometry to measure longitudinal GM volumes changes associated with autistic traits measured by the Social Responsiveness Scale (SRS) total and sub-scores. In women, we observed that the SRS was associated with slower GM volume decrease globally and in the left parahippocampus and middle temporal gyrus. The social communication sub-score correlated with slower GM volume decrease in the left parahippocampal, superior temporal gyrus, and pallidum; and the social cognition sub-score correlated with slower GM volume decrease in the left middle temporal gyrus, the right ventromedial prefrontal and orbitofrontal cortex. No longitudinal association was found in men. Autistic traits in young women were found to be associated with specific brain trajectories in regions of the social brain and the reward circuit known to be involved in Autism Spectrum Disorder. These findings support both the hypothesis of an earlier GM maturation associated with autistic traits in adolescence and of protective mechanisms in women. They advocate for further studies on brain trajectories associated with autistic traits in women.

Mental Well-being InSciEd Out: Health Partnerships with the Boys & Girls Clubs of Puerto Rico.

BACKGROUND: Mental health care is a top clinical concern for modern Puerto Rico, especially given a dramatically changing economic landscape paired with recurrent natural disasters. Youth are particularly at-risk due to long-term impacts of toxic stress and adverse childhood experiences on health and development. OBJECTIVES: Here we present a novel clinician-community-educator-scientist partnership to address Puerto Rican youth mental well-being and wellness. We deployed pilot health workshops within the Boys & Girls Clubs of Puerto Rico to build youth mental health conceptual understanding and competencies in stress recognition and management. The work in progress herein evaluates acceptability and feasibility of our curricular model. METHODS: Dialogue with community stakeholders guided curricular design of workshops for youth ages 6 to 13 and older. Prior to implementation, educators and volunteers attended a 1-day training on educational strategies. Workshop success was evaluated using qualitative approaches (i.e., narrative feedback, educator and volunteer reflections, youth Talking Drawings) to assess youth engagement, youth conceptual health understanding, and educator/volunteer impressions of feasibility and impact. RESULTS: Initial findings indicate high acceptability and feasibility of our curricular model. Youth engagement and enthusiasm were noted in educator feedback and continue to be sustained post-workshop. Preliminary analysis shows accompanying increases in youth conceptual mental health understanding, particularly for 6- to 12-year-olds in recognition of stress and healthy coping mechanisms. Reciprocal gains were observed for volunteers. CONCLUSIONS: Activities have evolved into a formal partnership called Semilla, which features expanded analysis of mental well-being and wellness outcomes. Our collaborative model continues to engage Puerto Rican youth in the science of their well-being.

Local structural flexibility drives oligomorphism in computationally designed protein assemblies.

Many naturally occurring protein assemblies have dynamic structures that allow them to perform specialized functions. For example, clathrin coats adopt a wide variety of architectures to adapt to vesicular cargos of various sizes. Although computational methods for designing novel self-assembling proteins have advanced substantially over the past decade, most existing methods focus on designing static structures with high accuracy. Here we characterize the structures of three distinct computationally designed protein assemblies that each form multiple unanticipated architectures, and identify flexibility in specific regions of the subunits of each assembly as the source of structural diversity. Cryo-EM single-particle reconstructions and native mass spectrometry showed that only two distinct architectures were observed in two of the three cases, while we obtained six cryo-EM reconstructions that likely represent a subset of the architectures present in solution in the third case. Structural modeling and molecular dynamics simulations indicated that the surprising observation of a defined range of architectures, instead of non-specific aggregation, can be explained by constrained flexibility within the building blocks. Our results suggest that deliberate use of structural flexibility as a design principle will allow exploration of previously inaccessible structural and functional space in designed protein assemblies.

Accurate computational design of three-dimensional protein crystals.

Protein crystallization plays a central role in structural biology. Despite this, the process of crystallization remains poorly understood and highly empirical, with crystal contacts, lattice packing arrangements and space group preferences being largely unpredictable. Programming protein crystallization through precisely engineered side-chain-side-chain interactions across protein-protein interfaces is an outstanding challenge. Here we develop a general computational approach for designing three-dimensional protein crystals with prespecified lattice architectures at atomic accuracy that hierarchically constrains the overall number of degrees of freedom of the system. We design three pairs of oligomers that can be individually purified, and upon mixing, spontaneously self-assemble into >100 µm three-dimensional crystals. The structures of these crystals are nearly identical to the computational design models, closely corresponding in both overall architecture and the specific protein-protein interactions. The dimensions of the crystal unit cell can be systematically redesigned while retaining the space group symmetry and overall architecture, and the crystals are extremely porous and highly stable. Our approach enables the computational design of protein crystals with high accuracy, and the designed protein crystals, which have both structural and assembly information encoded in their primary sequences, provide a powerful platform for biological materials engineering.

Precisely patterned nanofibres made from extendable protein multiplexes.

Molecular systems with coincident cyclic and superhelical symmetry axes have considerable advantages for materials design as they can be readily lengthened or shortened by changing the length of the constituent monomers. Among proteins, alpha-helical coiled coils have such symmetric, extendable architectures, but are limited by the relatively fixed geometry and flexibility of the helical protomers. Here we describe a systematic approach to generating modular and rigid repeat protein oligomers with coincident C2 to C8 and superhelical symmetry axes that can be readily extended by repeat propagation. From these building blocks, we demonstrate that a wide range of unbounded fibres can be systematically designed by introducing hydrophilic surface patches that force staggering of the monomers; the geometry of such fibres can be precisely tuned by varying the number of repeat units in the monomer and the placement of the hydrophilic patches.

Multivalent antigen display on nanoparticle immunogens increases B cell clonotype diversity and neutralization breadth to pneumoviruses.

Nanoparticles for multivalent display and delivery of vaccine antigens have emerged as a promising avenue for enhancing B cell responses to protein subunit vaccines. Here, we evaluated B cell responses in rhesus macaques immunized with prefusion-stabilized respiratory syncytial virus (RSV) F glycoprotein trimer compared with nanoparticles displaying 10 or 20 copies of the same antigen. We show that multivalent display skews antibody specificities and drives epitope-focusing of responding B cells. Antibody cloning and repertoire sequencing revealed that focusing was driven by the expansion of clonally distinct B cells through recruitment of diverse precursors. We identified two antibody lineages that developed either ultrapotent neutralization or pneumovirus cross-neutralization from precursor B cells with low initial affinity for the RSV-F immunogen. This suggests that increased avidity by multivalent display facilitates the activation and recruitment of these cells. Diversification of the B cell response by multivalent nanoparticle immunogens has broad implications for vaccine design.

De novo design of highly selective miniprotein inhibitors of integrins αvß6 and αvß8.

The RGD (Arg-Gly-Asp)-binding integrins αvß6 and αvß8 are clinically validated cancer and fibrosis targets of considerable therapeutic importance. Compounds that can discriminate between homologous αvß6 and αvß8 and other RGD integrins, stabilize specific conformational states, and have high thermal stability could have considerable therapeutic utility. Existing small molecule and antibody inhibitors do not have all these properties, and hence new approaches are needed. Here we describe a generalized method for computationally designing RGD-containing miniproteins selective for a single RGD integrin heterodimer and conformational state. We design hyperstable, selective αvß6 and αvß8 inhibitors that bind with picomolar affinity. CryoEM structures of the designed inhibitor-integrin complexes are very close to the computational design models, and show that the inhibitors stabilize specific conformational states of the αvß6 and the αvß8 integrins. In a lung fibrosis mouse model, the αvß6 inhibitor potently reduced fibrotic burden and improved overall lung mechanics, demonstrating the therapeutic potential of de novo designed integrin binding proteins with high selectivity.

De novo design of highly selective miniprotein inhibitors of integrins αvß6 and αvß8.

The RGD (Arg-Gly-Asp)-binding integrins αvß6 and αvß8 are clinically validated cancer and fibrosis targets of considerable therapeutic importance. Compounds that can discriminate between the two closely related integrin proteins and other RGD integrins, stabilize specific conformational states, and have sufficient stability enabling tissue restricted administration could have considerable therapeutic utility. Existing small molecules and antibody inhibitors do not have all of these properties, and hence there is a need for new approaches. Here we describe a method for computationally designing hyperstable RGD-containing miniproteins that are highly selective for a single RGD integrin heterodimer and conformational state, and use this strategy to design inhibitors of αvß6 and αvß8 with high selectivity. The αvß6 and αvß8 inhibitors have picomolar affinities for their targets, and >1000-fold selectivity over other RGD integrins. CryoEM structures are within 0.6-0.7Å root-mean-square deviation (RMSD) to the computational design models; the designed αvß6 inhibitor and native ligand stabilize the open conformation in contrast to the therapeutic anti-αvß6 antibody BG00011 that stabilizes the bent-closed conformation and caused on-target toxicity in patients with lung fibrosis, and the αvß8 inhibitor maintains the constitutively fixed extended-closed αvß8 conformation. In a mouse model of bleomycin-induced lung fibrosis, the αvß6 inhibitor potently reduced fibrotic burden and improved overall lung mechanics when delivered via oropharyngeal administration mimicking inhalation, demonstrating the therapeutic potential of de novo designed integrin binding proteins with high selectivity.

Eurytrematosis in cattle in southern Espírito Santo State, Brazil - case report.

This work reports an outbreak of eurytrematosis in cattle in the municipality of Ibitirama, southern Espírito Santo State, Brazil. Six cattle were necropsied from August to December 2019, with finding of Eurytrema coelomaticum in the pancreas. A survey of epidemiological data was carried out on the farms along with coproparasitological examination of cattle from the same herd. Parasites were found in all necropsied animals, with different degrees of parasitism, ranging from mild to massive infection (6 - 2000 specimens). Macroscopic analyses of the pancreas revealed changes in 83.33% (5/6) of the cases, and by microscopy, pancreatic fibrosis ranging from Grade I to Grade III was observed. Inspection of the grazing areas confirmed the presence of two intermediate hosts, a terrestrial snail of the Bradybaena genus, with larval forms of the trematode in histological findings, and a grasshopper of the Conocephalus genus. Although none of the cattle showed clinical signs in the coproparasitological examination, 73.80% (31/42) tested positive for E. coelomaticum eggs. This is the first record of an outbreak of eurytrematosis in cattle in Espírito Santo State, indicating the importance of carrying out diagnosis based on epidemiology and necroscopic and parasitological examinations in animals in the region so that appropriate control measures can be adopted.

Este trabalho objetivou relatar um caso de euritrematose em bovinos no município de Ibitirama, Sul do Estado do Espírito Santo, Brasil. Foram necropsiados seis bovinos de agosto a dezembro de 2019, que apresentaram Eurytrema coelomaticum no pâncreas. Foi realizado levantamento de dados epidemiológicos nas propriedades e exames coproparasitológico em bovinos do mesmo plantel. Em todos os animais necropsiados foram encontrados parasitos, com diferentes graus de parasitismo, variando de infeção branda a maciça (6 - 2000 exemplares). Análises macroscópicas dos pâncreas revelaram alterações em 83,33% (5/6) dos casos e, na microscopia, observou-se fibrose pancreática variando de Grau I a Grau III. A inspeção das áreas de pastejo constatou a presença dos dois hospedeiros intermediários, moluscos terrestres do gênero Bradybaena com formas larvares do trematoda em achados histológicos e gafanhotos do gênero Conocephalus. Nenhum dos bovinos apresentou sinais clínicos, no entanto, no exame coproparasitológico, 73,80% (31/42) testaram positivo para ovos de E. coelomaticum. Este é o primeiro registro de surto de euritrematose em bovinos no estado do Espírito Santo, mostrando a importância da realização do diagnostico a partir da epidemiologia e de exames necroscópicos e parasitológicos em animais da região para que sejam adotadas medidas adequadas de controle.

Twelve tips for implementing and teaching anti-racism curriculum in medical education.

Racism has implicit and explicit manifestations that perpetuate disparities and negatively influence patient-centered health outcomes. Subsequently, a list of action items was provided to assist medical schools in becoming anti-racist institutions. A deep subject matter knowledge, beliefs, and reflections were a driving force for the management of medical schools or faculty members involved in undergraduate and postgraduate medical education to move forward toward inclusion of anti-racism in traditional medical curriculum or adapting existing training modules on diversity, equity, and inclusion. This paper proposes twelve practical and specific tips for implementing and teaching anti-racism in medical education. These twelve tips elaborate on the proposed actions for leaders in undergraduate and postgraduate medical education, valuable for designing future curricula and educational activities.

Computational design of non-porous, pH-responsive antibody nanoparticles.

Programming protein nanomaterials to respond to changes in environmental conditions is a current challenge for protein design and important for targeted delivery of biologics. We describe the design of octahedral non-porous nanoparticles with the three symmetry axes (four-fold, three-fold, and two-fold) occupied by three distinct protein homooligomers: a de novo designed tetramer, an antibody of interest, and a designed trimer programmed to disassemble below a tunable pH transition point. The nanoparticles assemble cooperatively from independently purified components, and a cryo-EM density map reveals that the structure is very close to the computational design model. The designed nanoparticles can package a variety of molecular payloads, are endocytosed following antibody-mediated targeting of cell surface receptors, and undergo tunable pH-dependent disassembly at pH values ranging between to 5.9-6.7. To our knowledge, these are the first designed nanoparticles with more than two structural components and with finely tunable environmental sensitivity, and they provide new routes to antibody-directed targeted delivery.

Improving the secretion of designed protein assemblies through negative design of cryptic transmembrane domains.

Computationally designed protein nanoparticles have recently emerged as a promising platform for the development of new vaccines and biologics. For many applications, secretion of designed nanoparticles from eukaryotic cells would be advantageous, but in practice, they often secrete poorly. Here we show that designed hydrophobic interfaces that drive nanoparticle assembly are often predicted to form cryptic transmembrane domains, suggesting that interaction with the membrane insertion machinery could limit efficient secretion. We develop a general computational protocol, the Degreaser, to design away cryptic transmembrane domains without sacrificing protein stability. The retroactive application of the Degreaser to previously designed nanoparticle components and nanoparticles considerably improves secretion, and modular integration of the Degreaser into design pipelines results in new nanoparticles that secrete as robustly as naturally occurring protein assemblies. Both the Degreaser protocol and the nanoparticles we describe may be broadly useful in biotechnological applications.

Modulation of FGF pathway signaling and vascular differentiation using designed oligomeric assemblies.

Growth factors and cytokines signal by binding to the extracellular domains of their receptors and drive association and transphosphorylation of the receptor intracellular tyrosine kinase domains, initiating downstream signaling cascades. To enable systematic exploration of how receptor valency and geometry affects signaling outcomes, we designed cyclic homo-oligomers with up to 8 subunits using repeat protein building blocks that can be modularly extended. By incorporating a de novo designed fibroblast growth-factor receptor (FGFR) binding module into these scaffolds, we generated a series of synthetic signaling ligands that exhibit potent valency- and geometry-dependent Ca2+ release and MAPK pathway activation. The high specificity of the designed agonists reveal distinct roles for two FGFR splice variants in driving endothelial and mesenchymal cell fates during early vascular development. The ability to incorporate receptor binding domains and repeat extensions in a modular fashion makes our designed scaffolds broadly useful for probing and manipulating cellular signaling pathways.

The Current State of Antiracism Curricula in Undergraduate and Graduate Medical Education: A Qualitative Study of US Academic Health Centers.

PURPOSE: We undertook a study to evaluate the current state of pedagogy on antiracism, including barriers to implementation and strengths of existing curricula, in undergraduate medical education (UME) and graduate medical education (GME) programs in US academic health centers. METHODS: We conducted a cross-sectional study with an exploratory qualitative approach using semistructured interviews. Participants were leaders of UME and GME programs at 5 institutions participating in the Academic Units for Primary Care Training and Enhancement program and 6 affiliated sites from November 2021 to April 2022. RESULTS: A total of 29 program leaders from the 11 academic health centers participated in this study. Three participants from 2 institutions reported the implementation of robust, intentional, and longitudinal antiracism curricula. Nine participants from 7 institutions described race and antiracism-related topics integrated into health equity curricula. Only 9 participants reported having "adequately trained" faculty. Participants mentioned individual, systemic, and structural barriers to implementing antiracism-related training in medical education such as institutional inertia and insufficient resources. Fear related to introducing an antiracism curriculum and undervaluing of this curriculum relative to other content were identified. Through learners and faculty feedback, antiracism content was evaluated and included in UME and GME curricula. Most participants identified learners as a stronger voice for transformation than faculty; antiracism content was mainly included in health equity curricula. CONCLUSIONS: Inclusion of antiracism in medical education requires intentional training, focused institutional policies, enhanced foundational awareness of the impact of racism on patients and communities, and changes at the level of institutions and accreditation bodies.

Ataque de nervios: The impact of sociodemographic, health history, and psychological dimensions on Puerto Rican adults.

Introduction: Ataque de nervios (ADN) is a cultural syndrome prevalent in Puerto Ricans characterized as an episode of intense emotional upset due to overwhelming stress. Methods: The Ataque de Nervios Questionnaire, developed at the Center for the Study and Treatment for Fear and Anxiety (CETMA), served as the diagnostic tool for this retrospective secondary data analysis. We evaluated three models regarding ADN's function as a marker of (1) sociodemographic vulnerability, (2) health history risk, and (3) psychological vulnerability. This last model was subdivided to assess the scores of screening tests regarding anxiety (Anxiety Sensitivity Inventory, Beck Anxiety Inventory, and State-Trait Anxiety Inventory), affect (Beck Depression Inventory, Emotional Dysregulation Scale, Positive and Negative Affective Schedule), personality (NEO Five-Factor Inventory), and trauma (considering the responses to the Childhood Trauma Questionnaire and the Life Event Checklist). Results: Our study sample had a total of 121 Puerto Rican adult patients from CETMA out of which 75% had ADN. We differentiated subjects according to their ADN status with t-tests and Mann-Whitney U tests and evaluated our models using logistic regressions. People with ADN showed more anxiety, depressive symptoms, emotional dysregulation, and negative affect than those without ADN. They also revealed lower positive affect and agreeableness. Highly extraverted but minimally agreeable personalities related to ADN. Living with a partner and being employed were risk factors for ADN. Having higher educational levels showed the strongest effect size: it greatly reduced the odds of an ataque. Discussion: These characteristics suggest a distinct profile of ADN seen in employed, educated, adult Puerto Ricans living on the Island experiencing anxiety. Our study provides clinical tools to comprehend our patients' ADN experience, enriching our practice as culturally competent health providers.