Distal Domain Research Articles

Spatial distribution and possible origin of the high velocity lower crust in the northern margin of the South China Sea

AbstractHigh-velocity lower crust (HVLC) near the continent–ocean transition zone is significant for recording magmatic activity and early rifting history and is likely to be sourced from various types of magma supplies, including passive decompression melting, hot mantle plume, active small-scale mantle convection, and fertile source mantle. In the northern margin of the South China Sea (composed of the Dongsha Passive margin to the west and the SW Taiwan active margin to the east), a large amount of geophysical data, including refraction seismic data, enable us to prepare a map of the thickness distribution of the HVLC and estimate its volume. After revisiting published data, the volume of the HVLC in the northern margin of the South China Sea is found to be comparable but slightly less than that in continental large igneous provinces such as the Deccan Trap of west-central India or oceanic plateaus such as the Shatsky Rise in the northwest Pacific Ocean, probably indicating that they may have similar active source origins. Based on a more feature-based analysis (h–vp diagram; a scatter plot showing mean velocity of lower crust versus corresponding crustal thickness) to identify the source of the underplating material in a magma-involved margin, we tentatively suggest that the HVLC along the Dongsha passive margin showing no h–vp correlation is most likely dominated by small-scale mantle convection of the Peikang–Dongsha mantle convection cell, and the HVLC along the SW Taiwan active margin showing a negative h–vp correlation is more likely to be dominated by the pre-existing distal domain of the continental margin. We propose that rather than being dominated by a hot mantle plume, small-scale mantle convection may be more likely to be responsible for the opening of the South China Sea, and suggest that a more inclusive geodynamic model to reconcile geophysical and geochemical observations should be further investigated.

Palaeogeography and tectono‐stratigraphic evolution of the Aptian Ezanga‐Loémé evaporites along the proximal domain of the south Gabon‐Congo‐Cabinda margin

AbstractDuring the Early Cretaceous, massive evaporite accumulations formed in the opening South Atlantic. However, the depositional model of these salts is still poorly constrained at the scale of the West African margin. The present study focuses along the proximal domain of the south Gabon‐Congo‐Cabinda margin and is based on (i) log interpretations of 246 wells crossing undeformed to weakly deformed evaporite intervals and (ii) a structural characterization of the basement. The evaporites show 11 regional evaporite depositional cycles (CI–CXI) bounded by meter‐thick shale beds. The cycles display alternating meter‐scale carnallite‐halite beds that can be correlated over several hundred kilometres, and CVI, CVII, CVIIIa and CX culminate in localized tachyhydrite accumulations. Cross section correlations and isopach maps help to understand the palaeogeographical evolution of each cycle and depositional environments that evolved from relatively deep at the base of cycles, to very shallow at their top. CI formed a mosaic of halite‐prone depocenters deposited in pre‐salt topographic relief. CII and CIII were deposited uniformly over a flattened basin in a highly extended brine pond. From CIV to CVIIIa, the stratigraphic architecture of the salts was shaped by freshwater inflow sourced from the north and possible basement movements. This setting, together with an increased confinement of the proximal domain from the distal one with basin drawdown, favoured the development of depocenters with perennial subaqueous conditions and extreme salinities, in which more than 70 m of tachyhydrite accumulation could locally be preserved. From CVIIIb to CXI, the basin returned to a flat depositional setting without well‐developed depocenters and with increasing subsidence westwards. Marine influx increased starting from CX, allowing the deposition of sulphate beds. The salt section is capped by anhydrite deposits interbedded with clastic and dolomite, before the final marine invasion of the basin. For the first time, this study provides a large‐scale depositional tectonostratigraphic setting of the Aptian salts in the proximal domain of the West African margin. The results are of interest for K‐Mg salts exploration resources in the Aptian and pave the way for further investigation of the salt depositional environment in the distal domain of the margin.

Genetic analyses of embryo homology and ontogeny in the model grass Zea mays subsp. mays.

The homology of the single cotyledon of grasses and the ontogeny of the scutellum and coleoptile as the initial, highly modified structures of the grass embryo are investigated using leaf developmental genetics and targeted transcript analyses in the model grass Zea mays subsp. mays. Transcripts of leaf developmental genes are identified in both the initiating scutellum and the coleoptile, while mutations disrupting mediolateral leaf development also disrupt scutellum and coleoptile morphology, suggesting that these grass-specific organs are modified leaves. Higher-order mutations in WUSCHEL-LIKE HOMEOBOX3 (WOX3) genes, involved in mediolateral patterning of plant lateral organs, inform a model for the fusion of coleoptilar margins during maize embryo development. Genetic, RNA-targeting, and morphological evidence supports models for cotyledon evolution where the scutellum and coleoptile, respectively, comprise the distal and proximal domains of the highly modified, single grass cotyledon.

Serpentinization as a Tape Recorder of (Dis)Continuous Mantle Exhumation along the Alpine Tethys Ocean-Continent-Transition

Abstract Serpentinization has been widely documented and investigated at mid-ocean ridges (MOR) and subduction zones. In contrast, at magma-poor rifted margins serpentinization has received much less attention, despite its importance in controlling rheology and mass fluxes during breakup and establishing of a steady-state MOR. In this study, we present new petrological and geochemical data on subcontinental exhumed serpentinized peridotites from the spectacularly exposed Platta, Tasna and Totalp nappes in the Eastern Central Alps in SE Switzerland, belonging to the Alpine Tethys Ocean Continent Transition (OCT). The results testify of a complex history of fluid–rock interactions recorded by several serpentinization events starting with lizardite mesh and bastite textures (S1), subsequently followed by a succession of serpentine-filling veins with distinct textures and serpentine polysomes that include spherical polyhedral serpentine (S2); chrysotile ± polygonal ± lizardite banded veins (S3); lamellar antigorite veins and patches (S4) and chrysotile crack-seal (S5). The serpentinization sequence differs at proximal (i.e. continentwards) and distal (i.e. oceanwards) domains of the OCT. At proximal domains of the OCT (Upper Platta, Tasna) serpentinites record the complete serpentinization sequence (S1 to S5), whereas at distal domains (Lower Platta) serpentinization is restricted to pseudomorphic mesh and bastite (S1) and chrysotile crack-seal (S5). We attribute this discrepancy to contrasted mechanisms of mantle exhumation along the OCT. While at proximal domains mantle is unroofed along continuous and single large offset detachment faults allowing for the formation of all serpentine generations, mantle exhumation at distal domains is a more discontinuous process, controlled by sequential out-of-sequence detachment and flip-flop faults preventing the full development of all serpentine generations. In this frame, the nature and order of formation of the serpentine polysomes are directly controlled by the conditions of serpentinization (i.e. temperature, mantle composition and fluid/rock ratio). We propose that this new conceptual model can be extrapolated to serpentinization at slow to ultra-slow MORs, where close similarities in the serpentinization sequences have been recently reported.

Tracing Allostery in the Spliceosome Ski2-like RNA Helicase Brr2.

RNA ATPases/helicases remodel substrate RNA-protein complexes in distinct ways. The different RNA ATPases/helicases, taking part in the spliceosome complex, reshape the RNA/RNA-protein contacts to enable premature-mRNA splicing. Among them, the bad response to refrigeration 2 (Brr2) helicase promotes U4/U6 small nuclear (sn)RNA unwinding via ATP-driven translocation of the U4 snRNA strand, thus playing a pivotal role during the activation, catalytic, and disassembly phases of splicing. The plastic Brr2 architecture consists of an enzymatically active N-terminal cassette (N-cassette) and a structurally similar but inactive C-terminal cassette (C-cassette). The C-cassette, along with other allosteric effectors and regulators, tightly and timely controls Brr2's function via an elusive mechanism. Here, microsecond-long molecular dynamics simulations, dynamical network theory, and community network analysis are combined to elucidate how allosteric effectors/regulators modulate the Brr2 function. We unexpectedly reveal that U4 snRNA itself acts as an allosteric regulator, amplifying the cross-talk of distal Brr2 domains and triggering a conformational reorganization of the protein. Our findings offer fundamental understanding into Brr2's mechanism of action and broaden our knowledge on the sophisticated regulatory mechanisms by which spliceosome ATPases/helicases control gene expression. This includes their allosteric regulation exerted by client RNA strands, a mechanism that may be broadly applicable to other RNA-dependent ATPases/helicases.

Are hemodynamics responsible for inflammatory changes in venous vessel walls? A quantitative study of wall-enhancing intracranial arteriovenous malformation draining veins.

Signal enhancement of vascular walls on vessel wall MRI might be a biomarker for inflammation. It has been theorized that contrast enhancement on vessel wall imaging (VWI) in draining veins of intracranial arteriovenous malformations (AVMs) may be associated with disease progression and development of venous stenosis. The aim of this study was to investigate the relationship between vessel wall enhancement and hemodynamic stressors along AVM draining veins. Eight AVM patients with 15 draining veins visualized on VWI were included. Based on MR venography data, patient-specific 3D surface models of the venous anatomy distal to the nidus were segmented. The enhanced vascular wall regions were manually extracted and mapped onto the venous surface models after registration of image data. Using image-based blood flow simulations applying patient-specific boundary conditions based on phase-contrast quantitative MR angiography, hemodynamics were investigated in the enhanced vasculature. For the shear-related parameters, time-averaged wall shear stress (TAWSS), oscillatory shear index (OSI), and relative residence time (RRT) were calculated. Velocity, oscillatory velocity index (OVI), and vorticity were extracted for the intraluminal flow-related hemodynamics. Visual observations demonstrated overlap of enhancement with local lower shear stresses resulting from decreased velocities. Thus, higher RRT values were measured in the enhanced areas. Furthermore, nonenhancing draining veins showed on average slightly higher flow velocities and TAWSS. Significant decreases of 55% (p = 0.03) for TAWSS and of 24% (p = 0.03) for vorticity were identified in enhanced areas compared with near distal and proximal domains. Velocity magnitude in the enhanced region showed a nonsignificant decrease of 14% (p = 0.06). Furthermore, increases were present in the OSI (32%, p = 0.3), RRT (25%, p = 0.15), and OVI (26%, p = 0.3) in enhanced vessel sections, although the differences were not significant. This novel multimodal investigation of hemodynamics in AVM draining veins allows for precise prediction of occurring shear- and flow-related phenomena in enhanced vessel walls. These findings may suggest low shear to be a local predisposing factor for venous stenosis in AVMs.

Gain and loss of upper limb abilities in Duchenne muscular dystrophy patients: A 24-month study

Duchenne muscular dystrophy (DMD) is a neuromuscular condition characterized by muscle weakness. The Performance of upper limb (PUL) test is designed to evaluate upper limb function in DMD patients across three domains. The aim of this study is to identify frequently lost or gained PUL 2.0 abilities at distinct functional stages in DMD patients. This retrospective study analyzed prospectively collected data on 24-month PUL 2.0 changes related to ambulatory function. Ambulant patients were categorized based on initial 6MWT distance, non-ambulant patients by time since ambulation loss. Each PUL 2.0 item was classified as shift up, no change, or shift down. The study's cohort incuded 274 patients, with 626 paired evaluations at the 24-month mark. Among these, 55.1 % had activity loss, while 29.1 % had gains. Ambulant patients showed the lowest loss rates, mainly in the shoulder domain. The highest loss rate was in the shoulder domain in the transitioning subgroup and in elbow and distal domains in the non-ambulant patients. Younger ambulant patients demonstrated multiple gains, whereas in the other functional subgroups there were fewer gains, mostly tied to singular activities. Our findings highlight divergent upper limb domain progression, partly linked to functional status and baseline function.

A c- MPL Splice Variant up-Regulated in Myeloproliferative Neoplasms Modulates Thrombopoietin Signaling and Induces Cycling of Hematopoietic Stem Cells and Inflammatory Progenitors

Introduction: Myeloproliferative neoplasms (MPN) are clonal disorders caused by mutations that activate JAK/STAT signaling through the thrombopoietin receptor, MPL, leading to overproduction of myeloid lineages. MPL is the only hematopoietic growth factor receptor expressed on the surface of hematopoietic stem cells (HSC) where it maintains their quiescence by tethering HSCs to osteoblasts in the endosteal niche. MPL is also expressed on megakaryocytes and platelets. Individuals born with loss of function mutations in the distal MPL cytokine receptor homology domain (CRHD) develop thrombocytopenia and progressive bone marrow failure. How MPL functions in MPN remains unclear. Here, we describe a novel MPL splice variant ( MPL-SV) that is present in CD34 + cells and platelets from healthy controls, but overexpressed in the MPN. We therefore sought to: 1) define MPL-SV function in hematopoiesis and MPN, and, 2) elucidate mechanisms underlying MPL-SV. Methods: After identifying a novel MPL-SV transcript from JAK2 V617F MPNplatelets, we compared its expression to MPL transcripts lacking the deletion in MPN patients and healthy controls. To assess its function, we generated transgenic (Tg) mice expressing varied levels of MPL-SV in HSC. Next, we examined responses of the Tg mice to thrombopoietin (THPO), hematopoietic stem progenitor cell (HSPC) frequency by flow cytometry, and HSPC function by colony-forming assays (CFA). To elucidate mechanisms, we performed single cell RNA sequencing (scRNAseq) of bone marrow (BM)-derived Lin -, Sca +, c-kit + (LSK) cells followed by pathway analysis. Results: Using primers to amplify the MPL CRHD in platelets from patients with JAK2 V617F MPN, we discovered an MPL- SV lacking 7 amino acids in the MPL amino-terminal domain, a common hotspot for activating MPN mutations. MPL-SV is present in CD34 + cells and platelets of healthy controls, but overexpressed in MPN relative to MPL transcripts lacking the deletion. To examine its function, we generated transgenic (Tg) mice expressing MPL-SV in HSC. Tg miceexpressing a single copy of MPL-SV have unperturbed hematopoiesis, whereas Tg mice expressing 6 copies of the transgene ( MPL-SV-6) develop marked thrombocytopenia, mild splenomegaly, and a modest increase in red cells by 9-12 weeks of age. Baseline serum THPO levels are elevated in Tg MPL-SV-6 mice and they fail to increase platelet counts following exogenous THPO administration, suggesting defective function of MPL-SV in binding and/or internalizing THPO. To further assess MPL-SV function, we compared CFA from HSPC of Tg MPL-SV-6 and wildtype mice, which show a decrease in total colony forming units (CFU) from BM or spleen-derived MPL-SV-6 HSPC. Accordingly, there was a decrease in LSK cells, multi-potent progenitors (MPPs), and megakaryocytes (Mk) in BM by flow cytometry. By contrast, spleens show increases in LK cells and short-term HSC. Further analysis by scRNAseq of BM-derived LSK cells reveal a marked decrease in both quiescent HSC and HSC with a Mk-bias associated with a relative expansion in more proliferative HSC, including those with a bias towards erythroid and mature myeloid differentiation. Pathway analysis reveal that MPL-SV activates transcriptional networks involved in TNF-α signaling via NF-κB and oxidative phosphorylation. Analysis of cell-cell interactions by CellChat show decreased signaling through intercellular adhesion molecule 1 (ICAM-1) in MPL-SV-6 HSC compared to controls. Importantly, ICAM1 maintains quiescent HSC within their BM niche. Similarly, cell cycle analysis from scRNAseq indicate that quiescent HSC are stimulated to cycle and generate HSC primed to differentiate into erythroid and myeloid progeny. Conclusions: We discovered a novel MPL-SV that is up-regulated in MPN. In Tg mice, MPL-SV impairs responses to THPO and disrupts hematopoiesis by depleting quiescent HSC and BM megakaryocytes. Single cell transcriptomics suggest that MPL-SV drives expansion of cycling HSC and HSC primed to differentiate into progenitors involved in inflammation, including myeloid cells. Mechanistically, HSPC expressing MPL-SV-6 activate transcriptional networks involved in TNFα signaling via NF-κB pathways. Our results highlight the central role for MPL in hematopoiesis and suggest that aberrant MPL splicing contributes to MPN pathogenesis.

Clinical features, functional consequences, and rescue pharmacology of missense GRID1 and GRID2 human variants.

GRID1 and GRID2 encode the enigmatic GluD1 and GluD2 proteins, which form tetrameric receptors that play important roles in synapse organization and development of the central nervous system. Variation in these genes has been implicated in neurodevelopmental phenotypes. We evaluated GRID1 and GRID2 human variants from the literature, ClinVar, and clinical laboratories and found that many of these variants reside in intolerant domains, including the amino terminal domain of both GRID1 and GRID2. Other conserved regions, such as the M3 transmembrane domain, show different intolerance between GRID1 and GRID2. We introduced these variants into GluD1 and GluD2 cDNA and performed electrophysiological and biochemical assays to investigate the mechanisms of dysfunction of GRID1/2 variants. One variant in the GRID1 distal amino terminal domain resides at a position predicted to interact with Cbln2/Cbln4, and the variant disrupts complex formation between GluD1 and Cbln2, which could perturb its role in synapse organization. We also discovered that, like the lurcher mutation (GluD2-A654T), other rare variants in the GRID2 M3 domain create constitutively active receptors that share similar pathogenic phenotypes. We also found that the SCHEMA schizophrenia M3 variant GluD1-A650T produced constitutively active receptors. We tested a variety of compounds for their ability to inhibit constitutive currents of GluD receptor variants and found that pentamidine potently inhibited GluD2-T649A constitutive channels (IC50 50nM). These results identify regions of intolerance to variation in the GRID genes, illustrate the functional consequences of GRID1 and GRID2 variants, and suggest how these receptors function normally and in disease.

Similarly slow diffusion of BAM and SecYEG complexes in live E. coli cells observed with 3D spt-PALM

The β-barrel assembly machinery (BAM) complex is responsible for inserting outer membrane proteins (OMPs) into the Escherichia coli outer membrane. The SecYEG translocon inserts inner membrane proteins into the inner membrane and translocates both soluble proteins and nascent OMPs into the periplasm. Recent reports describe Sec possibly playing a direct role in OMP biogenesis through interactions with the soluble polypeptide transport-associated (POTRA) domains of BamA (the central OMP component of BAM). Here we probe the diffusion behavior of these protein complexes using photoactivatable super-resolution localization microscopy and single-particle tracking in live E. coli cells of BAM and SecYEG components BamA and SecE and compare them to other outer and inner membrane proteins. To accurately measure trajectories on the highly curved cell surface, three-dimensional tracking was performed using double-helix point-spread function microscopy. All proteins tested exhibit two diffusive modes characterized by “slow” and “fast” diffusion coefficients. We implement a diffusion coefficient analysis as a function of the measurement lag time to separate positional uncertainty from true mobility. The resulting true diffusion coefficients of the slow and fast modes showed a complete immobility of full-length BamA constructs in the time frame of the experiment, whereas the OMP OmpLA displayed a slow diffusion consistent with the high viscosity of the outer membrane. The periplasmic POTRA domains of BamA were found to anchor BAM to other cellular structures and render it immobile. However, deletion of individual distal POTRA domains resulted in increased mobility, suggesting that these domains are required for the full set of cellular interactions. SecE diffusion was much slower than that of the inner membrane protein PgpB and was more like OMPs and BamA. Strikingly, SecE diffused faster upon POTRA domain deletion. These results are consistent with the existence of a BAM-SecYEG trans-periplasmic assembly in live E. coli cells.

Nucleolar structure connects with global nuclear organization.

The nucleolus is a multifunctional nuclear body. To tease out the roles of nucleolar structure without resorting to the use of multi-action drugs, we knocked down the RNA polymerase I subunit RPA194 in HeLa cells by siRNA. Loss of RPA194 resulted in nucleolar-structural segregation and effects on both nucleolus-proximal and distal-nuclear components. The perinucleolar compartment was disrupted, centromere clustering around nucleoli was significantly reduced, and the intranuclear locations of specific genomic loci were altered. Moreover, Cajal bodies, distal from nucleoli, underwent morphological and some compositional changes. In comparison, when the preribosomal RNA-processing factor, UTP4, was knocked down, neither nucleolar segregation nor the intranuclear effects were observed, demonstrating that the changes of nucleolar proximal and distal nuclear domains in RPA194 knockdown cells unlikely arise from a cessation of ribosome synthesis, rather from the consequence of nucleolar-structure alteration. These findings point to a commutative system that links nucleolar structure to the maintenance and spatial organization of certain nuclear domains and genomic loci.

The effects of subsalt relief on gravity-driven salt tectonics: Results from analogue modelling

Gravity-driven salt-related deformation on passive margins is commonly interpreted as kinematically-linked domains of updip extension and downdip contraction with an intermediate, undeformed zone of translation. Our study uses analogue models to show how salt flow is affected by the subsalt geometry resulting in complex deformation and the alternation of extensional and contractional domains in space and time in translational salt provinces. In our work, the analogue models incorporate subsalt relief associated with half-graben and graben that are realistic geometries along passive margins and combine the effects of gravity spreading in which deformation is driven by differential sedimentary loading, and an early-stage of gravity gliding induced by basin tilt. This approach allows for analysis of how salt flow and overburden deformation are affected by: (1) smoothly dipping subsalt relief; (2) subsalt relief characterized by half-graben with a gently landward dipping extensional fault; (3) subsalt half-graben connected laterally with a distal symmetric graben. For each of these models, we evaluate: 1) the interplay between slipping deformation corresponding to gravity gliding, gravity spreading mechanisms, and subsalt geometry; 2) the spatial and temporal distribution of salt-related structural domains; and 3) the contrasting styles of salt tectonics for different subsalt geometries. In addition, some of the experiments have been repeated considering thicker salt and pre-kinematic overburden and different sedimentation rates. We show that the structures of the subsalt relief variably resist salt flow producing flow mismatch. This variable resistance controls the orientation of the extension and contraction supra-salt structures. Salt surplus and contraction occur over landward dipping base-salt ramps, and a zone of salt depletion and subsidence limited by extensional and contractional hinges occurs over the basinward dipping base-salt ramp. Variations in width and the dip of these base-salt ramps result in stronger flow mismatches and deformation complexity. We also discuss variations in salt flow profile in response to the gravitational forces that drive deformation, allowing a good first-order approximation of viscous salt flow at a regional scale that can be directly compared to examples from various salt-bearing continental margins, particularly the relatively less understood but extremely economically important distal domains of Brazil's Campos and Santos basins.

Dual localization of receptor-type adenylate cyclases and cAMP response protein 3 unveils the presence of two putative signaling microdomains in Trypanosoma cruzi.

Trypanosoma cruzi is the etiologic agent of Chagas disease, a leading cause of disability and premature death in the Americas. This parasite spends its life between a triatomine insect and a mammalian host, transitioning between developmental stages in response to microenvironmental changes. Among the second messengers driving differentiation in T. cruzi, cAMP has been shown to mediate metacyclogenesis and response to osmotic stress, but this signaling pathway remains largely unexplored in this parasite. Adenylate cyclases (ACs) catalyze the conversion of ATP to cAMP. They comprise a multigene family encoding putative receptor-type ACs in T. cruzi. Using protein sequence alignment, we classified them into five groups and chose a representative member from each group to study their localization (TcAC1-TcAC5). We expressed an HA-tagged version of each protein in T. cruzi and performed immunofluorescence analysis. A peculiar dual localization of TcAC1 and TcAC2 was observed in the flagellar distal domain and in the contractile vacuole complex (CVC), and their enzymatic activity was confirmed by gene complementation in yeast. Furthermore, TcAC1 overexpressing parasites showed an increased metacyclogenesis, a defect in host cell invasion, and a reduced intracellular replication, highlighting the importance of this protein throughout T. cruzi life cycle. These mutants were more tolerant to hypoosmotic stress and showed a higher adhesion capacity during in vitro metacyclogenesis, whereas the wild-type phenotype was restored after disrupting TcAC1 localization. Finally, TcAC1 was found to interact with cAMP response protein 3 (TcCARP3), co-localizing with this protein in the flagellar tip and CVC. IMPORTANCE We identified three components of the cAMP signaling pathway (TcAC1, TcAC2, and TcCARP3) with dual localization in Trypanosoma cruzi: the flagellar distal domain and the CVC, structures involved in cell adhesion and osmoregulation, respectively. We found evidence on the role of TcAC1 in both cellular processes, as well as in metacyclogenesis. Our data suggest that TcACs act as signal sensors and transducers through cAMP synthesis in membrane microdomains. We propose a model in which TcACs sense the harsh conditions in the triatomine hindgut (nutrient deprivation, acidic pH, osmotic stress, ionic composition, hydrophobic interactions) and become active. Synthesis of cAMP then triggers cell adhesion prior completion of metacyclogenesis, while mediating a response to osmotic stress in the parasite. These results shed light into the mechanisms driving cAMP-mediated cell differentiation in T. cruzi, while raising new questions on the activation of TcACs and the role of downstream components of this pathway.

Qualitative exploration of the visual function impairments and impacts on vision-dependent activities of daily living in Retinitis Pigmentosa and Leber Congenital Amaurosis: content validation of the ViSIO-PRO and ViSIO-ObsRO measures

BackgroundRetinitis Pigmentosa (RP) and Leber Congenital Amaurosis (LCA) are rare inherited retinal degenerative disorders. The associated visual impairments have significant impacts on patients’ vision-dependent activities of daily living (ADL), mobility, and distal health-related quality of life (HRQoL). To adequately capture patient and caregiver perspectives in clinical trials, patient and observer-reported outcome instruments must demonstrate sufficient evidence of content validity in the target population. This study aimed to explore the patient experience of RP/LCA and assess the content validity of the Visual Symptom and Impact Outcomes PRO (ViSIO-PRO) and ObsRO (ViSIO-ObsRO) instruments in RP/LCA.MethodsA total of 66 qualitative, combined concept elicitation (CE) and cognitive debriefing (CD) interviews were conducted (33 adults, 10 adolescents, 8 children and 15 caregivers of children) in the US, France, Germany, and Canada. Patients had a clinical and genetic diagnosis of RP/LCA from a range of genotypes. CE results were used to further inform the development of a conceptual model and CD interviews assessed the relevance and understanding of the 44-item ViSIO-PRO and 26-item ViSIO-ObsRO instruments. Interviews were conducted across two iterative rounds to allow item modifications.ResultsFindings were consistent across RP/LCA genotypes. Night blindness, reduced peripheral vision, vision in very bright lighting and light/dark adaptation were the most frequently reported visual function symptoms impacting vision-dependent ADL and mobility. Impacts on distal HRQoL domains were also reported. The ViSIO-PRO and ObsRO items were well understood by participants and relevant across genotypes. The instructions, 7-day recall period and response scales were well understood and endorsed. Participant and expert clinician feedback supported modifications to item wording, the addition of six new ViSIO-PRO items and one new ViSIO-ObsRO item, and the removal of one ViSIO-PRO item due to lack of relevance.ConclusionsFindings support the content validity of the ViSIO-PRO and ViSIO-ObsRO instruments for use across RP/LCA genotypes. Ongoing research to evaluate the psychometric validity of the instruments will support future use of the instruments as efficacy endpoints in clinical trials and in general clinical practice to track disease severity and impact of disease on functioning.

How post‐salt sediment flux and progradation rate influence salt tectonics on rifted margins: Insights from geodynamic modelling

AbstractContinental rifted margins can be associated with widespread and thick salt deposits, which are often formed during the final stages of rifting, prior to breakup. These salt‐bearing margins are typically characterized by pronounced post‐rift salt tectonics with variable and complex structural styles and evolution. We use a lithosphere‐scale geodynamic numerical model to investigate the role of varying post‐rift sediment fluxes and progradation rates on rifted margin salt tectonics. We focus on a single, intermediate, rifted margin type and salt basin geometry to explore scenarios with different: (i) constant and (ii) time‐varying post‐salt sediment fluxes. We demonstrate that these promote significant contrasts in the style and magnitude of salt tectonics in the proximal, transitional and distal margin domains. The differences are primarily controlled by the relationship between the rates of sediment progradation (Vprog) and salt flow (Vs). When Vprog > Vs, the salt is rapidly buried and both vertical and lateral salt flow are suppressed across the entire margin. When Vprog < Vs, the salt flows vertically and seaward faster than sediments prograde producing major diapirism in the proximal domain and major distal nappe advance, but only moderate overburden extension and distal diapirism. When Vprog ~ Vs, there is moderate proximal diapirism and distal nappe advance, but major updip extension and downdip shortening, which produces major distal diapirism. Modelling results are comparable to various natural systems and help improve our understanding of the controls and dynamics of salt tectonics along salt‐bearing rifted margins.

Binding domain on CD22 molecules contributing to the biological activity of T cell-engaging bispecific antibodies

CD22, as the B-cell malignancies antigen, has been targeted for immunotherapies through CAR-T cells, antibody-drug conjugates (ADCs) and immunotoxins via interaction of antibodies with binding domains on the receptor. We hypothesized that avidity and binding domain of antibody to target cells may have significant impact on the biological function in tumor immunotherapy, and T cell-engaging bispecific antibody (TCB) targeting CD22 could be used in the therapy of hematologic malignancies. So, to address the question, we utilized the information of six previously reported CD22 mAbs to generate CD22-TCBs with different avidity to different domains on CD22 protein. We found that the avidity of CD22-TCBs to protein was not consistent with the avidity to target cells, indicating that TCBs had different binding mode to the protein and cells. In vitro results indicated that CD22-TCBs mediated cytotoxicity depended on the avidity of antibodies to target cells rather than to protein. Moreover, distal binding domain of the antigen contributed to the avidity and biological activity of IgG-[L]-scfv-like CD22-TCBs. The T cells' proliferation, activation, cytotoxicity as well as cytokine release were compared, and G5/44 BsAb was selected for further in vivo assessment in anti-tumor activity. In vivo results demonstrated that CD22-TCB (G5/44 BsAb) significantly inhibited the tumors growth in mice. All these data suggested that CD22-TCBs could be developed as a promising candidate for B-cell malignancies therapy through optimizing the design with avidity and binding domain to CD22 target in consideration.

Rapid nanobody-based imaging of mesothelin expressing malignancies compatible with blocking therapeutic antibodies.

Mesothelin (MSLN) is overexpressed in a wide variety of cancers with few therapeutic options and has recently emerged as an attractive target for cancer therapy, with a large number of approaches currently under preclinical and clinical investigation. In this respect, developing mesothelin specific tracers as molecular companion tools for predicting patient eligibility, monitoring then response to mesothelin-targeting therapies, and tracking the evolution of the disease or for real-time visualisation of tumours during surgery is of growing importance. We generated by phage display a nanobody (Nb S1) and used enzymatic approaches were used to site-directed conjugate Nb S1 with either ATTO 647N fluorochrome or NODAGA chelator for fluorescence and positron emission tomography imaging (PET) respectively. We demonstrated that Nb S1 displays a high apparent affinity and specificity for human mesothelin and demonstrated that the binding, although located in the membrane distal domain of mesothelin, is not impeded by the presence of MUC16, the only known ligand of mesothelin, nor by the therapeutic antibody amatuximab. In vivo experiments showed that both ATTO 647N and [68Ga]Ga-NODAGA-S1 rapidly and specifically accumulated in mesothelin positive tumours compared to mesothelin negative tumours or irrelevant Nb with a high tumour/background ratio. The ex vivo biodistribution profile analysis also confirmed a significantly higher uptake of Nb S1 in MSLN-positive tumours than in MSLNlow tumours. We demonstrated for the first time the use of an anti-MSLN nanobody as PET radiotracer for same day imaging of MSLN+ tumours, targeting an epitope compatible with the monitoring of amatuximab-based therapies and current SS1-derived-drug conjugates.

Repurposing the inhibitors of MMP-9 and SGLT-2 against ubiquitin specific protease 30 in Parkinson’s disease: computational modelling studies

Ubiquitin specific protease 30 (USP30) has been attributed to mitochondrial dysfunction and impediment of mitophagy in Parkinson’s disease (PD). This happens once ubiquitin that supposed to bind with deformed mitochondria at the insistence of Parkin, it’s been recruited by USP30 via the distal ubiquitin binding domain. This is a challenge when PINK1 and Parkin loss their functions due to mutation. Although, there are reports on USP30s’ inhibitors but no study on the repurposing of inhibitors approved against MMP-9 and SGLT-2 as potential inhibitors of USP30 in PD. Thus, the highlight therein, is to repurpose approved inhibitors of MMP-9 and SGLT-2 against USP30 in PD using extensive computational modelling framework. 3D structures of Ligands and USP30 were obtained from PubChem and protein database (PDB) servers respectively, and were subjected to molecular docking, ADMET evaluation, DFT calculation, molecular dynamics simulation (MDS) and free energy calculations. Out of the 18 drugs, 2 drugs showed good binding affinity to the distal ubiquitin binding domain, moderate pharmacokinetic properties and good stability. The findings showed canagliflozin and empagliflozin as potential inhibitors of USP30. Thus, we present these drugs as repurposing candidates for the treatment of PD. However, the findings in this current study needs to be validated experimentally. Communicated by Ramaswamy H. Sarma

Red Sea rifting in central Egypt: constraints from the offshore Quseir province

The formation of oceanic crust in the southern and central Red Sea is generally accepted to have started at c. 5 Ma. However, the nature of the crust in the northern Red Sea is still debated. This paper describes the rift architecture, dynamics and evolution of the northern Red Sea and identifies domains that relate to first-order geodynamic processes. The proximal margin domain is located onshore and is characterized by latest Oligocene–Miocene half-graben basins. New seismic interpretations show that the offshore region is a necking domain dominated by low-angle, high-offset extensional faults, which led to the exhumation of lower crustal gabbros at Brothers Islands. Two-dimensional forward models suggest that the necking domain passes into a distal margin domain, where the continental crust thins to <10 km at 120 km from the coast. Sensitivity testing of interpretations for the distal domain indicates a probable scenario where exhumed lower continental crust or serpentinized mantle is present. A comprehensive rift model for the northern Red Sea in the Quseir province accounts for magmatic underplating accompanied by half-graben development at c. 25 Ma, followed by Early Miocene crustal thinning accommodated by an east-dipping detachment fault. A Late Miocene phase with a flip of the detachment geometry led to the present day configuration.

Cytochrome P450 Surface Domains Prevent the β-Carotene Monohydroxylase CYP97H1 of Euglena gracilis from Acting as a Dihydroxylase.

Molecular biodiversity results from branched metabolic pathways driven by enzymatic regioselectivities. An additional complexity occurs in metabolites with an internal structural symmetry, offering identical extremities to the enzymes. For example, in the terpene family, β-carotene presents two identical terminal closed-ring structures. Theses cycles can be hydroxylated by cytochrome P450s from the CYP97 family. Two sequential hydroxylations lead first to the formation of monohydroxylated β-cryptoxanthin and subsequently to that of dihydroxylated zeaxanthin. Among the CYP97 dihydroxylases, CYP97H1 from Euglena gracilis has been described as the only monohydroxylase. This study aims to determine which enzymatic domains are involved in this regioselectivity, conferring unique monohydroxylase activity on a substrate offering two identical sites for hydroxylation. We explored the effect of truncations, substitutions and domain swapping with other CYP97 members and found that CYP97H1 harbours a unique N-terminal globular domain. This CYP97H1 N-terminal domain harbours a hydrophobic patch at the entrance of the substrate channel, which is involved in the monohydroxylase activity of CYP97H1. This domain, at the surface of the enzyme, highlights the role of distal and non-catalytic domains in regulating enzyme specificity.