flexible 80 a shore resin vat polymerisation resin vat polymerisation sla model  (Formlabs Inc)

Journal: 3D Printing in Medicine

Article Title: Evaluation of 3D-printed vs. digital models in simulation-based training for iliac endovascular interventions

doi: 10.1186/s41205-026-00320-2

Figure Lengend Snippet: Overhead view of the 3D-printed aortic model set-up: ( a ) flexible 80 A (shore) resin vat polymerisation (SLA) model (Flexible 80 A Resin, Formlabs); ( b ) stiff clear resin vat polymerisation (SLA) model (Clear Resin V5, Formlabs); both connected to a fluid pump to replicate pulsatile circulation

Article Snippet: Fig. 1 Overhead view of the 3D-printed aortic model set-up: ( a ) flexible 80 A (shore) resin vat polymerisation (SLA) model (Flexible 80 A Resin, Formlabs); ( b ) stiff clear resin vat polymerisation (SLA) model (Clear Resin V5, Formlabs); both connected to a fluid pump to replicate pulsatile circulation In terms of model Fabrication, both 3D-printed models were derived from contrast-enhanced CT angiography datasets from institutional records.

Techniques:

Journal: bioRxiv

Article Title: Conformational dynamics of actin filaments crosslinked with alpha-actinin and their roles in suppressing cofilin-induced helical shortening and cluster formation

doi: 10.1101/2025.11.16.688667

Figure Lengend Snippet: ( A ) A longitudinal section profile illustrating for the analysis of two consecutive HHPs, the number of protomer pairs per HHP, and MADs in various actin filaments captured at different time points. The analytic methods used were developed in our previous study . ( B ) Still AFM images with time labels were shown for conditions with ATP present and after ATP depletion (primarily converted into ADP and inorganic phosphate in solution through ATPase activities of actin and S1) to observe the binding of S1 on actin filaments. As ATP levels gradually declined or nearly depleted due to ATP hydrolysis (ATP → ADP + Pi) during the actin-myosin reaction, S1 binding to actin filaments persisted for longer durations, enabling capture by HS-AFM at an imaging rate of 330 ms per frame. All actin filaments in the imaging field were aligned with the pointed end (PE) on the left and the barbed end (BE) on the right, reflecting the actual unipolar crosslinking configuration. As similarly done in our previous study by referring to the EM study , the polarity of actin filaments was confirmed by observing the tilted angles of S1, with its head pointing toward the PE, resembling an arrowhead along the filament. This characteristic orientation allowed differentiation between the PE and the BE. Representative yellow and cyan arrowheads indicate alpha-actinin and S1 molecules, respectively. Scale bars: 25 nm. Color scale: 0-12 nm. ( C ) Time-dependent changes in two consecutive HHPs, the number of protomer pairs per HHP, and MADs in actin filaments crosslinked by 50 nM alpha-actinin in the presence of 1 mM ATP, 0.1 mM CaCl₂, and 1 µM S1, but without MgCl 2 . Related to , Movie S1 .

Article Snippet: Movie S8 Molecular dynamics of unipolar-flexible crosslinking model obtained from all-atom MD simulations.

Techniques: Binding Assay, Imaging

Journal: bioRxiv

Article Title: Conformational dynamics of actin filaments crosslinked with alpha-actinin and their roles in suppressing cofilin-induced helical shortening and cluster formation

doi: 10.1101/2025.11.16.688667

Figure Lengend Snippet: ( A ) Experimental setup for HS-AFM observations of dynamic conformational changes in alpha-actinin-crosslinked actin filaments in the presence and absence of S1. ( B ) Representative AFM images of alpha-actinin-crosslinked actin filaments with and without S1. Yellow arrowheads representatively indicate alpha-actinin molecules; black arrowheads mark the longitudinal section points used to measure the HHP. Two parallelograms (ABCD and EFHG) represent the geometric framework used to measure the crosslinking angles of alpha-actinin and the crosslinking lengths between alpha-actinin-crosslinked actin filaments. ( C ) Histogram of HHP values for alpha-actinin-crosslinked actin filaments without S1. The mean HHP is 36.5 ± 2.8 nm (mean ± SD, N = 4270). F-buffer consists of 0.5 mM MgCl₂, 0.5 mM ATP, and 50 nM alpha-actinin at pH 6.8. ( D, E ) Ridgelines showing the distributions of alpha-actinin crosslinking angles and crosslinking lengths between filaments crosslinked with alpha-actinin, respectively. Noted that we focus solely on the four symmetric small crosslinking angles generated by two alpha-actinin molecules bridging two actin filaments, irrespective of their polarity and demonstrated in a parallelogram ABCD. Statistical significance was determined using a two-sample t-test , with differences considered significant at p ≤ 0.05. See Methods and for details.

Article Snippet: Movie S8 Molecular dynamics of unipolar-flexible crosslinking model obtained from all-atom MD simulations.

Techniques: Generated

Journal: bioRxiv

Article Title: Conformational dynamics of actin filaments crosslinked with alpha-actinin and their roles in suppressing cofilin-induced helical shortening and cluster formation

doi: 10.1101/2025.11.16.688667

Figure Lengend Snippet:

Article Snippet: Movie S8 Molecular dynamics of unipolar-flexible crosslinking model obtained from all-atom MD simulations.

Techniques: T-Test

Journal: bioRxiv

Article Title: Conformational dynamics of actin filaments crosslinked with alpha-actinin and their roles in suppressing cofilin-induced helical shortening and cluster formation

doi: 10.1101/2025.11.16.688667

Figure Lengend Snippet: ( A ) Experimental setup for HS-AFM observations cofilin-induced helical shortening, cofilin binding and clustering in bare actin filaments versus alpha-actinin-crosslinked-actin filaments. ( B ) Time-dependent changes in the peak heights and HHPs of actin filaments incubated with only 600 nM hs cofilin 1 and 1mM ATP. A ∼25% decrease in HHP and the formation of mature cofilin clusters on actin filaments in the absence of alpha-actinin served as positive controls, providing a basis for comparison with the cofilin clusters and HHP shown in C . Red and green arrowheads indicate the peaks in mature cofilin clusters and bare actin segments (served as negative controls), respectively. Time label indicates the time after adding cofilin. Scale bars: 25 nm. Color scale: 0-12 nm. ( C ) Time-dependent changes in the peak heights and HHPs of actin filaments incubated with 400 nM alpha-actinin and 1 mM ATP, and subsequently added with 600 nM cofilin. The cofilin binding and formation of mature cofilin clusters, and the shortening of HHP were significantly suppressed by binding and crosslinking of alpha-actinin. Red arrowheads denote the peaks in mature cofilin clusters within the supertwisted half helices of actin segments (cofilactin) without alpha-actinin, and blue arrowheads indicate those in immature cofilin clusters within the long half helices of actin segments crosslinked with alpha-actinin (cofilactin-actinin), respectively. A representative yellow arrowhead denotes alpha-actinin. Scale bars: 25 nm. Color scale: 0-13 nm. ( D ) Histograms of the peak height and HHP measured from various actin filaments with and without alpha-actinin binding and crosslinking, showing the presence of both mature and immature cofilin clusters. These clusters corresponded to short and normal HHPs, respectively. Notably, their categorization as mature or immature clusters was based on their differences in peak heights and length of corresponding HHPs, and their comparisons with bare actin (i.e., peak height of 8.6 ± 0.8 nm, HHP of 36.8 ± 4.3 nm), as reported in our previous studies . The mean ± SD heights and lengths are 10.7 ± 1.0 nm (N = 425) and 27.9 ± 3.0 nm (N= 300) for mature clusters of cofilactin segment without alpha-actinin, and 9.9 ± 1.1 nm (N = 2514) and 33.0 ± 4.6 nm (N = 2003) for immature clusters of cofilactin segment with alpha-actinin crosslinking (cofilactin-actinin). A two-population t-test ( p < 0.05) indicates that the differences in height and HHP distribution between the clusters of cofilactin segments without and with alpha-actinin crosslinking are significant ( p = 2.9×10 -38 for height, p = 8.5×10 -68 for HHP).

Article Snippet: Movie S8 Molecular dynamics of unipolar-flexible crosslinking model obtained from all-atom MD simulations.

Techniques: Binding Assay, Incubation, Comparison

Journal: bioRxiv

Article Title: Conformational dynamics of actin filaments crosslinked with alpha-actinin and their roles in suppressing cofilin-induced helical shortening and cluster formation

doi: 10.1101/2025.11.16.688667

Figure Lengend Snippet: ( A ) The unipolar and bipolar crosslinking models of the actin filaments with alpha-actinin. The unipolar crosslinking configuration occurs when the actin filaments share the same polarity, whereas the bipolar crosslinking configuration appears when the actin filaments align with opposite polarities. The quantitative analyses were performed using conformations obtained from all-atom MD simulations for 300 ns. ( B ) Quantitative analysis of the crosslinking lengths and alpha-actin crosslinking angles relative to the filament axis in the unipolar-flexible, bipolar-flexible, and bipolar-fixed models obtained from the MD simulations. ( C ) Analysis of the HHP, rise, and AD in the bare actin filaments and in the unipolar-flexible, bipolar-flexible, and bipolar-fixed models using the conformations derived from the MD simulations. ( D, E ) Characterization of the residue-level interactions between the alpha-actinin ABDs and actin protomers during crosslinking in the unipolar-flexible and bipolar-flexible models. In our model, if an atom of one residue (Cα) was in contact within 2 Å of an atom of another residue (Cα), these two residues were considered to be interacting. Our MD simulations identified key alpha-actinin–actin residue pairs with contact using probability density function (nm -1 ), which can be greater than 1 since there are two alpha-actinin ends interacting actin filaments. Thus, the maximum probability value is 2.

Article Snippet: Movie S8 Molecular dynamics of unipolar-flexible crosslinking model obtained from all-atom MD simulations.

Techniques: Derivative Assay, Residue

Journal: bioRxiv

Article Title: Conformational dynamics of actin filaments crosslinked with alpha-actinin and their roles in suppressing cofilin-induced helical shortening and cluster formation

doi: 10.1101/2025.11.16.688667

Figure Lengend Snippet: ( A, B ) Twisted and flattened conformational transitions between G-actin, F-actin, and C-actin structures have been previously linked to actin polymerization and depolymerization and cofilin clustering. Specifically, twisted G-actin polymerizes to form flattened actin filament protomers (F-actin form) and vice versa , while cofilin-induced clustering leads to twisted protomers (C-actin), and subsequently shortening the HHP in actin filament accompanied by a reduction in the number of protomers per HHP , , . Our current study deciphers that alpha-actinin binding and crosslinking help preserve the normal canonical helical structure of actin filaments, with actin protomers most likely adopting a flattened conformation (F-actin). This conformation facilitates S1 binding but prevents cofilin binding or clustering. In contrast, the twisted conformation of actin protomers induced by cofilin binding and clustering induces the shortening of helical twists, which supports cofilin’s cooperative binding and clustering , . We suggest that the process of cooperativity of cofilin binding and clustering likely follows the sequence: initial cofilin binding and clustering onto shortened ADP-bound HHP → protomer twisting → HHP shortening accompanied by a reduction in the number of protomers per HHP → cooperative binding and clustering. However, there is currently no atomic structural evidence to confirm whether the twisted actin protomers within the shortened bare helices could inhibit S1 binding, as suggested from previous fluorescence-based observation , despite the known mechanism of direct competition for binding sites between cofilin and S1 on actin filaments . This remains an area for further investigation in future studies. PE: pointed end; BE: barbed end. ( C ) Illustration depicting alpha-actinin-crosslinked actin filaments under the unipolar and bipolar conformations with normal and rigid HHPs resistant to cofilin binding and/or clustering, highlighting their distinct localizations and functions within cells. This Figure was created using BioRender.com. Related to .

Article Snippet: Movie S8 Molecular dynamics of unipolar-flexible crosslinking model obtained from all-atom MD simulations.

Techniques: Binding Assay, Sequencing, Fluorescence