Selection Of Variants Research Articles

Case report: A case study of variant calling pipeline selection effect on the molecular diagnostics outcome

Next-generation sequencing technologies have not only defined a breakthrough in medical genetics, but also been able to enter routine clinical practice to determine individual genetic susceptibilities. Modern technological developments are routinely introduced to genetic analysis overtaking the established approaches, potentially raising a number of challenges. To what extent is the advantage of new methodologies in synthetic metrics, such as precision and recall, more important than stability and reproducibility? Could differences in the technical protocol for calling variants be crucial to the diagnosis and, by extension, the patient’s treatment strategy? A regulatory review process may delay the incorporation of potentially beneficial technologies, resulting in missed opportunities to make the right medical decisions. On the other hand, a blind adoption of new technologies based solely on synthetic metrics of precision and recall can lead to incorrect conclusions and adverse outcomes for the specific patient. Here, we use the example of a patient with a WHO-diagnosed desmoplastic/nodular SHH-medulloblastoma to explore how the choice of DNA variant search protocol affects the genetic diagnostics outcome.

Unravelling the impact of Al-Ti content on the microstructure-toughness relationship in the coarse-grained heated-affected zone of super-high-strength pipeline steels

This study aims to investigate the role of Al-Ti addition on particle modification in super-high-strength pipeline steels, and its following effect on microstructural evolution and impact toughness in the coarse-grained heated-affected zone (CGHAZ) is also explored. The steel plates with low and high Al-Ti content (0.009 wt-% Al + 0.006 wt-% Ti vs. 0.033 wt-% Al + 0.025 wt-% Ti), respectively, are subjected to gas metal arc welding with 6–7 kJ/cm heat input. The results reveal that the second-phase particles including micron-scale inclusion of Al-Mg oxide covered with CaS and nano-scale precipitate of titanium nitride are obtained in low Al-Ti steel, which are modified to the Al-Mg-Ti oxide core surrounded by CaS and TiN precipitate in high Al-Ti steel. The nano-scale TiN precipitate and micron-scale inclusion can restrict the austenite grain growth at high temperatures and induce acicular ferrite nucleation at medium temperatures, respectively, leading to the fine-grained mixed microstructure of bainite and acicular ferrite in CGHAZ of both steels. Because of the variant selection during the decomposition of austenite to bainite and acicular ferrite, those fine-grain structures with amounts of high-angle grain boundaries exhibit excellent CGHAZ toughness in both specimens. Compared with low Al-Ti steel, more precipitates induce smaller austenite grains, and the inclusions containing titanium increase the ability of inclusion to promote acicular ferrite nucleation in CGHAZ of high Al-Ti steel. Both of them resulted in smaller crystallographic grain and toughness improvement in CGHAZ of high Al-Ti steel than that in low Al-Ti steel.

Polymorphic insertions of DcSto miniature inverted-repeat transposable elements reveal genetic diversity structure within the cultivated carrot.

Miniature inverted-repeat transposable elements (MITEs) are a potent source of polymorphisms in plant genomes. A genotyping system, named DcS-ILP, based on polymorphic insertions of Stowaway MITEs (DcStos) localized in introns and identified in the reference genome DH1, has been developed for carrot. Here, we report an extension of the DcS-ILP genotyping system by incorporation of non-reference insertions identified in resequenced genomes representing the eastern gene pool. We genotyped 52 carrot accessions representing the eastern and western carrot gene pools with 92 markers developed previously (western DcS-ILP panel) together with 84 newly developed markers (eastern DcS-ILP panel). Overall, the DcS-ILP markers revealed a highly structured genetic diversity separating the eastern and the western carrot accessions at K = 2 and differentiating Indian breeding lines from the eastern accessions at K = 3. The eastern DcS-ILP panel proved to be more robust with respect to the eastern carrot gene pool, while it provided little information on the western accessions, as many of the DcSto insertions present in the eastern gene pool were absent in the western gene pool. As the western carrot accessions represent improved cultivars, DcSto insertional polymorphisms allowed detection of a selection-driven bottleneck at the improvement stage. Selection in the course of the improvement stage generally operated on standing variation, as the subset of DcSto insertions present in the western carrot likely originated from transposition events preceding the separation of both gene pools. However, occasional frequency shifts in the opposite direction were also revealed, possibly indicating selection for favorable variants associated with DcSto insertions.

The acquisition of sociolinguistic variation by multilingual merchants: the case of seventeenth-century Spanish

Abstract This article analyzes an unpublished corpus of 131 letters written in Spanish by four Dutch merchants during the seventeenth-century (1669–1677). The aim of the investigation is to determine whether the variation found in these letters was internal, external, or both, and to study the factors that favored each source. To do so, I will look at vowel variation in this corpus. Since standard vowel variants were being selected in seventeenth-century Spanish, the research questions addressed in this work are the following: Did multilingual merchants acquire the emerging standard of the moment? Did multilingual merchants contribute to the selection of standard variants? Did multilingual merchants acquire the sociolinguistic variation of that period? How did multilingual merchants differ from L1 speakers and why? How did linguistic and social factors influence this variation? The results revealed a great amount of intra- and inter-speaker vowel variation, and this variability was a matter of both internal and external factors. Therefore, this study argues that, while multilingual merchants acquired some of the standard variants that were being selected in the emerging standard Spanish of the seventeenth century, they reported linguistic variation that can be explained considering the context of use and the socio-economic conditions underlying their acquisition of linguistic repertoires.

Effect of Cooling Rate on α Variant Selection and Microstructure Evolution in TB17 Titanium Alloy

The α variant selection and microstructure evolution in a new metastable β titanium alloy TB17 were studied in depth by DTA, microhardness, XRD, SEM, and EBSD characterization methods. Under the rapid cooling rate conditions (150 °C/min–400 °C/min), only a very small amount of granular αWM (α Widmanstatten precipitates within the grains) precipitated within the grains. The secondary α phase precipitated in the alloy changed from granular to fine needle-like at moderate cooling rates (15 °C/min–20 °C/min). When continuing to slow down the cooling rates (10 °C/min and 1 °C/min), the αGB (α precipitates along the β grain boundaries), αWGB (α Widmanstatten precipitates that developed from β grain boundaries or αGB) and αWM grew rapidly. Moreover, the continuous cooling transformation (CCT) diagram illustrated the effect of cooling rate on the β/α phase transition. EBSD analysis revealed that the variants selection of α near the original β grain boundary is mainly divided into three categories. (i) The double-BOR (Burgers orientation relationship) αWGB colonies within neighboring β grains grow in different directions but have the same crystallographic orientation. (ii) The double-BOR αWGB colonies within neighboring β grains have different growth directions and different crystallographic orientations. (iii) The double-BOR αWGB colonies within the same grain have the same growth direction, but different crystallographic directions. And these double-BOR αWGB colonies correspond to two variants of the given {0001}α//{110}β.

Effect of austenitizing temperature on variant selection of martensite transformation in a 2 GPa grade steel

The correlation of prior austenite grain (PAG) size, mechanical properties, and the variant selection in martensite transformation of a 2 GPa grade automobile steel has been systematically investigated by using a combination of scaning electron microscopy (SEM), electron backscatter diffraction (EBSD), and uniaxial tesile tests. As the austenitizing temperature decreases from 970 °C to 870 °C, the as-quenched microstructure of the studied steel is fully lath martensite. Meanwhile, the average size of PAG is refined from 7.2 ± 3.9 μm to 2.1 ± 1.2 μm, and the volume fraction of high angle grain boundaries (HAGB) is increased from 59.8% to 72.3%, accompanied with an increased number fraction of grains belonging to closely spaced planes (CP) group. With the refinement of the PAGs, the martensitic variant selectivity becomes strong, and the Bain group variant selection discretely transites to the CP group, resulting in a dominate CP group. Consequently, a tensile strength of 2014 ± 33 MPa, a tensile elongtation of 12.9 ± 0.5% and a maximum strength × ductility production of 26.0 ± 1.4 GPa·%, are obtained when the austenitization temperature is at 920 °C.

Prevalence of Drug Resistance Associated Substitutions in Persons With Chronic Hepatitis C Infection and Virological Failure Following Initial or Re-treatment With Pan-genotypic Direct-Acting Antivirals: A Systematic Review and Meta-analysis.

The advent of short-course, curative treatment with direct-acting antivirals (DAA) has given promise for the global elimination of hepatitis C virus (HCV) infections by 2030. Virological failure occurs in 2%-12% of persons receiving curative DAA treatment and may be presaged by pre-existing polymorphisms or result from selection of drug resistant variants during therapy. We conducted a systematic review to assess the prevalence of HCV resistance associated substitutions (RAS) among individuals with chronic hepatitis C infection who had virological failure following initial or re-treatment with pan-genotypic DAA regimens. We included 34 and 22 studies assessing RAS in people with virological failure published between January 2014 and July 2023. Pooled RAS prevalence was estimated using random-effects meta-analysis. The pooled prevalence of RAS in people with virological failure following initial DAA treatment was 78.0% (95% confidence interval [CI]: 62.0-92.0) for sofosbuvir/velpatasvir, 81.0% (95% CI: 67.0-93.0) for sofosbuvir/daclatasvir, and 79.0% (95% CI: 70.0-87.0) for glecaprevir/pibrentasvir, with a high prevalence of resistance to the NS5A inhibitors. Among those with virological failure following re-treatment regimens, RAS were present in 93.0% (95% CI: 83.0-99.0) for sofosbuvir/velpatasvir/voxilepravir and in 100% (95% CI: 92.0-100) for glecaprevir/pibrentasvir, with resistance driven by RAS to NS5A inhibitors. At least 1 RAS is present in a high proportion of the few individuals with virological failure following initial or re-treatment with pan-genotypic DAA regimens. There is a need for ongoing surveillance for DAA-associated resistance, to assess risk factors for their development and clinical impact to inform best practice strategies for re-treatment.

Analysis of Genetic Characteristics Associated with Reduced Bedaquiline Susceptibility in Multidrug-Resistant Mycobacterium tuberculosis

Bedaquiline (BDQ) has shown efficacy in shortening treatment duration and enhancing treatment success rates for multidrug-resistant tuberculosis (MDR-TB), thereby prompting widespread adoption. However, resistance to BDQ has emerged. This study aimed to identify genetic characteristics associated with decreased susceptibility to BDQ, using a public database to aid in the detection of resistant strains. Seventy-one BDQ-resistant and 929 BDQ-susceptible isolates from the open-source CRyPTIC database were selected for analysis. Variant calling was conducted via the clockwork pipeline. Univariate logistic regression was performed for each gene mutation, followed by LASSO regression for further variant selection. Ultimately, a multiple linear regression model was developed using log2-transformed Minimum Inhibitory Concentration values as the dependent variable, with variant selection refined through stepwise regression based on the Akaike Information Criterion. Ten gene mutations were significantly associated with reduced BDQ susceptibility, including two key gene mutations: Rv0678_141_ins_1 and Rv1979c_D249E, with effect estimates of 1.76 (95% CI: 0.67 - 2.84) and 1.69 (95% CI: 0.22 - 3.17), respectively. Other implicated genes included Rv2699c_-84_del_1, hsaB_I179T, mmpL9_T241A, pncA_C14R, Rv0373c_G621S, Rv0893c_L27F, Rv1770_A4D, and Rv3428c_S327C. This study identified ten gene mutations linked to decreased susceptibility to BDQ, providing a reference for developing a comprehensive catalog of BDQ-resistant genes.

Genotypic and phenotypic characterisation of respiratory syncytial virus after nirsevimab breakthrough infections: a large, multicentre, observational, real-world study

Nirsevimab, a long-acting monoclonal antibody, has been approved for the prevention of respiratory syncytial virus (RSV) infection in infants. In France, more than 210 000 single doses were administered in infants younger than 1 year during the 2023-24 season. In this context, the selection and spread of escape variants might be a concern. Here, we aimed to characterise RSV associated with breakthrough infection. We did a multicentre, national, observational study in France during the 2023-24 RSV season in RSV-infected infants (aged <1 year) who either received or did not receive a dose of nirsevimab before their first RSV season. We excluded infants with insufficient information about nirsevimab treatment or without parental consent. We used respiratory samples collected in each laboratory for full-length RSV RNA sequencing to analyse changes in the nirsevimab binding site Ø. We tested clinical RSV isolates for neutralisation by nirsevimab. We analysed F candidate substitutions by fusion-inhibition assay. Of the 695 RSV infected infants, we analysed 545 (78%) full-length RSV genome sequences: 260 (48%) from nirsevimab-treated breakthrough infections (236 [91%] RSV-A and 24 [9%] RSV-B) and 285 (52%) from untreated RSV-infected infants (236 [83%] RSV-A and 49 [17%] RSV-B). Analysis of RSV-A did not reveal any substitution in site Ø known to be associated with resistance to nirsevimab. Two (8%) of 24 RSV-B breakthrough infections had resistance-associated substitutions: F:N208D (dominant resistance-associated substitution) and a newly described F:I64M plus F:K65R combination (minority resistance-associated substitution), both of which induced high levels of resistance in the fusion-inhibition assay. This study is, to the best of our knowledge, the largest genotypic and phenotypic surveillance study of nirsevimab breakthrough infections to date. Nirsevimab breakthrough variants remain very rare despite the drug's widespread use. The detection of resistance-associated substitutions in the RSV-B F protein highlights the importance of active molecular surveillance. ANRS Maladies Infectieuses Emergentes and the French Ministry of Health and Prevention.

Human IL-22 receptor-targeted small protein antagonist suppress murine DSS-induced colitis

BackgroundHuman interleukin-22 (IL-22) is known as a “dual function” cytokine that acts as a master regulator to maintain homeostasis, structural integrity of the intestinal epithelial barrier, and shielding against bacterial pathogens. On the other hand, the overexpression of IL-22 is associated with hyper-proliferation and recruitment of pathologic effector cells, leading to tissue damage and chronic inflammation in specific diseases including inflammatory bowel disease (IBD). To study a role of IL-22-mediated signaling axis during intestinal inflammation, we generated a set of small protein blockers of IL-22R1 and verified their inhibitory potential on murine model of colitis.MethodsWe used directed evolution of proteins to identify binders of human IL-22 receptor alpha (IL-22R1), designated as ABR ligands. This approach combines the assembly of a highly complex combinatorial protein library derived from small albumin-binding domain scaffold and selection of promising protein variants using ribosome display followed by large-scale ELISA screening. The binding affinity and specificity of ABR variants were analyzed on transfected HEK293T cells by flow cytometry and LigandTracer. Inhibitory function was further verified by competition ELISA, HEK-Blue IL-22 reporter cells, and murine dextran sulfate sodium (DSS)-induced colitis.ResultsWe demonstrate that ABR specifically recognizes transgenic IL-22R1 expressed on HEK293T cells and IL-22R1 on TNFα/IFNγ-activated HaCaT cells. Moreover, some ABR binders compete with the IL-22 cytokine and function as IL-22R1 antagonists in HEK-Blue IL22 reporter cells. In a murine model of DSS-induced acute intestinal inflammation, daily intraperitoneal administration of the best IL-22R1 antagonist, ABR167, suppressed the development of clinical and histological markers of colitis including prevention of mucosal inflammation and architecture deterioration. In addition, ABR167 reduces the DSS-induced increase in mRNA transcript levels of inflammatory cytokines such as IL-1β, IL-6, IL-10, and IL-17A.ConclusionsWe developed small anti-human IL-22R1 blockers with antagonistic properties that ascertain a substantial role of IL-22-mediated signaling in the development of intestinal inflammation. The developed ABR blockers can be useful as a molecular clue for further IBD drug development.

Crystallographic Study of Transformation Products of Heat-Affected Zone and Correlation with Properties of FH690 Heavy-Gauge Marine Steel by Multi-Pass Submerged Arc Welding

In this work, the microstructure–property relationship of the heat-affected zone (HAZ) of a FH690 ultra-heavy marine steel plate was investigated based on insight of microstructure and crystallographic features. After multi-pass welding with a heat input of ~30 kJ/cm, an ~8 mm wide HAZ was obtained with a coarse grain HAZ (CGHAZ) of ~3.8 mm, fine grain HAZ (FGHAZ) of ~3.4 mm, and intercritical HAZ (ICHAZ) of ~1 mm. High impact toughness values of ~120 and 140 J at -60 °C were obtained for coarse grain HAZ and fine grain HAZ, respectively. The microstructure of the CGHAZ and FGHAZ was fine lath bainite. Although the average prior austenite grain size for the CGHAZ was ~75 μm, which was five times that of the FGHAZ (15 μm), a high density of high-angle grain boundaries (HAGBs) with misorientation higher than 45° was obtained in the CGHAZ. This is the underlying reason for the excellent low-temperature toughness of the HAZ. Thermo-dynamic calculations indicated that the high density of HAGBs in the CGHAZ was attributed to the decreased bainitic transformation temperature due to the reduced phase transformation driving force via the high nickel addition, leading to weak variant selection. In addition, the high nickel addition offered high hardenability for high hardness in the FGHAZ. The outcome of this study could provide an experimental and fundamental basis for designing high-strength ultra-heavy steel plates with excellent weldability.

Biosensor-Guided Engineering of a Baeyer-Villiger Monooxygenase for Aliphatic Ester Production.

Esters are valuable aroma compounds and can be produced enzymatically by Baeyer-Villiger monooxygenases (BVMOs) from (aliphatic) ketone precursors. However, a genetically encoded biosensor system for the assessment of BVMO activity and the detection of reaction products is missing. In this work, we assembled a synthetic enzyme cascade - featuring an esterase, an alcohol dehydrogenase, and LuxAB - in the heterologous host Escherichia coli. Target esters are produced by a BVMO, subsequently cleaved, and the corresponding alcohol oxidized through the artificial pathway. Ultimately, aldehyde products are detected in vivo by LuxAB, a luciferase from Photorhabdus luminescens that emits bioluminescence upon the oxidation of aldehydes to the corresponding carboxylates. This biosensor system greatly accelerated the screening and selection of active BVMO variants from a focused library, omitting commonly used low-throughput chromatographic analysis. Engineered enzymes accepted linear aliphatic ketones such as 2-undecanone and 2-dodecanone and exhibited improved ester formation.

Quantitative evaluation of local strain distribution induced by deformation twins and triggering effect of deformation twinning on neighboring grains in polycrystalline AZ31 magnesium alloys

{101‾2}<1‾011> deformation twinning commonly occurs in Mg/Mg alloys. In their polycrystals, however, twinning is not always activated at locations with high Schmid factor for {101‾2}<1‾011> twinning. It is believed that twinning behavior in polycrystalline Mg/Mg alloys can be understood with a consideration of interaction between adjacent twins as well as local deformation behavior on a micro- and meso‑structural scale. In this study, we theoretically estimate local plastic strain within twins by transforming coordinates and then compare it with that experimentally obtained from digital image correlation (DIC) analysis for understanding the reason for non-Schmid twinning in polycrystalline-Mg alloys. The theoretically estimated strain of twins was found to be in good agreement with the DIC-strain value. Furthermore, it was observed that the variant selection of twin can be explained by the compatibility parameter m’, rather than Schmid factor, suggesting that twin activation in polycrystalline-Mg is strongly influenced by adjacently appeared twins.

Elimination of pores and microstructural characterization in Ti-6Al-4V alloy welds using fast-frequency double pulse TIG welding

This study utilizes a fast-frequency double pulse tungsten inert gas (FFDP TIG) process to weld 2 mm thick Ti-6Al-4V alloy, with the objective of investigating the influence of a large fast-frequency current amplitude on porosity, microstructure, and tensile properties of the weld joint. Unlike conventional TIG welding, the FFDP TIG welding process led to a reduction and elimination of porosity, and refined the prior β grain and α phase by 34.3 % and 33.3 %, respectively, while also refining the martensitic α′ structure. Stirring of the molten pool resulted in improved uniformity of grain orientation distribution, accompanied by decreased variant selection of the α phase and reduced texture strength of the α phases. Moreover, three-variant clusters of α/α′ phases with triangular morphology and micro- and nanoscale recrystallized α grains appear in the FFDP TIG weld. Concurrently, the FFDP TIG process significantly enhanced the tensile strength and ductility by 14.5 % and 114 %, respectively, compared to the conventional TIG process.

From the "One-Molecule, One-Target, One-Disease" Concept towards Looking for Multi-Target Therapeutics for Treating Non-Polio Enterovirus (NPEV) Infections.

Non-polio enteroviruses (NPEVs), namely coxsackieviruses (CV), echoviruses (E), enteroviruses (EV), and rhinoviruses (RV), are responsible for a wide variety of illnesses. Some infections can progress to life-threatening conditions in children or immunocompromised patients. To date, no treatments have been approved. Several molecules have been evaluated through clinical trials without success. To overcome these failures, the multi-target directed ligand (MTDL) strategy could be applied to tackle enterovirus infections. This work analyzes registered clinical trials involving antiviral drugs to highlight the best candidates and develops filters to apply to a selection for MTDL synthesis. We explicitly stated the methods used to answer the question: which solution can fight NPEVs effectively? We note the originality and relevance of this proposal in relation to the state of the art in the enterovirus-inhibitors field. Several combinations are possible to broaden the antiviral spectrum and potency. We discuss data related to the virus and data related to each LEAD compound identified so far. Overall, this study proposes a perspective on different strategies to overcome issues identified in clinical trials and evaluate the "MTDL" potential to improve the efficacy of drugs, broaden the antiviral targets, possibly reduce the adverse effects, drug design costs and limit the selection of drug-resistant virus variants.

Influence of prior austenite grain size on martensite/austenite constituents in low-carbon bainitic steel

We elucidated here the influence of prior austenite grain (PAG) size on bainitic transformation products from the perspective of kinetics. The refinement of PAGs provided more sites for grain boundary nucleation and accelerated bainitic transformation kinetics. M/A constituents were also refined in refined PAGs with a higher carbon concentration, contributing to higher stability and even formation of retained austenite. The smaller M/A constituents exhibit stronger variant selection and localized strain.

Direct quenching and tempering to achieve high strength and toughness of GPa-Grade nano-precipitated steel: The effect of precipitation behavior and variant selectivity

In order to satisfy the application requirements for strength and toughness of hydropower steels for large pumped storage power plants. In this study, based on the synergy of deformation and phase transition, the trade-off between strength and toughness was broken, and the GPa-grade nano-precipitated steel was prepared by direct quenching and tempering. The effects of Nb–V–Ti composite nano-precipitation behavior and martensite variant selectivity on the strengthening and toughening mechanisms were highlighted. The results show that directly quenched steel has the optimal comprehensive mechanical properties after tempering at 550 °C. Combining the contributions of various strengthening mechanisms, the increase in the yield strength relative to off-line quenching and tempering is mainly attributed to additional dislocation strengthening (∼33 MPa) and precipitation strengthening (∼174 MPa). Direct quenching enhances the homogeneous nucleation of finer mono- and binary precipitates and their stable growth during tempering, while the ternary precipitates do not suffer from elemental inhomogeneities due to lack of re-dissolution. Furthermore, the high-frequency variants of the dominant close-packed plane group tend to form high-angle grain boundaries with different orientations and planar alignments, derived from the refinement of Bain groups induced by the self-plasticity modulation of martensite transition due to the accumulation of deformed geometrically necessary dislocations and finite dislocation slip.

Enhanced comprehensive mechanical properties of laser additive manufactured TC17 by design of new heat treatment based on continuous cooling transition

Heat treatment is critical for enhancing the mechanical properties of high strength titanium alloys, especially for exploiting the potential of laser additive manufactured titanium alloys. In this work, the influence of the cooling rate of continuous cooling transition on microstructure evolution and mechanical behavior was investigated in TC17 titanium alloy fabricated by laser directed energy deposition (LDED) technology. It was found that the number density and orientation characteristics of grain boundary α phases (αGB) are jointly influenced by the cooling rate and the structure of β/β grain boundaries (GBs). The average number density (λavg) of αGB has a consistent trend with the degree of variant selection (DVS) for precipitated α clusters subsequently, which is attributed to the autocatalytic effect of the pre-existing α on the post-precipitated α phases. The largest λavg of αGB and the highest DVS of α clusters could be simultaneously obtained at a suitable cooling rate (4 °C/min). In that case, plenty of α/β phase interfaces and dominant variant type ensure high strength, meanwhile, the combinations of activated multi-slip systems and varied crack propagation paths extend work-hardening to maintain greater plastic deformation. This paper provides a novel thought for designing customized heat treatments of LDEDed high strength titanium alloys, and more importantly, promotes the engineering applications of large and complex components prepared by additive manufacturing technology.

Rational selection of TbpB variants elucidates a bivalent vaccine formulation with broad spectrum coverage against Neisseria gonorrhoeae.

Neisseria gonorrhoeae is the causative agent of gonorrhea, an on-going public health problem due in part to the lack of success with efforts to develop an efficacious vaccine to prevent this sexually transmitted infection. An attractive candidate vaccine antigen because of its essential function and surface exposure, the gonococcal transferrin binding protein B (TbpB) exhibits high levels of antigenic variability which poses a significant obstacle in evoking a broadly protective vaccine composition. Here, we utilize phylogenetic information to rationally select TbpB variants for inclusion into a potential gonococcal vaccine and identify two TbpB variants that when formulated together elicit a highly cross-reactive antibody response in both rabbits and mice against a diverse panel of TbpB variants and clinically relevant gonococcal strains. Further, this formulation performed well in experimental proxies of real-world usage, including eliciting bactericidal activity against 8 diverse gonococcal strains and decreasing the median duration of colonization after vaginal infection in female mice by two heterologous strains of N. gonorrhoeae . Together, these data support the use of a combination of TbpB variants for a broadly protective gonococcal vaccine.

Disrupting variant selection memory effect in laser powder bed fusion to improve strength-ductility synergy of Ti-6Al-4V alloys

Texture formation is frequently observed in parts produced by Laser Powder Bed Fusion (L-PBF), which can induce anisotropy and may potentially degrade plasticity. In this study, we introduce a laser remelting strategy to mitigate these adverse effects. By employing experimental observations and numerical simulations, we established the relationship between melt pool thermal history, variant selection, and mechanical properties. Our results indicate that the strengthening of texture can be prevented by disrupting the variant selection memory effect when there is a difference in scanning speeds between the printing and remelting lasers. The achieved random variant orientation is attributed to the altered cooling rates and temperature gradient directions during solidification across different layers. The optimized Ti-6Al-4V alloy demonstrates high strength (1211.5 ± 13 MPa) and significant elongation (12.3% ± 0.8%), exhibiting a superior strength-ductility synergy compared to samples produced by direct printing or laser remelting with consistent parameters, as well as most reported L-PBF processed Ti-6Al-4V alloys. Our findings provide new insights into phase transformation kinetics in L-PBF of Ti-6Al-4V alloys and facilitate the optimization of this process for manufacturing high-performance components.