Junction Stability Research Articles

First-Principles Study on the Effect of H, C, and N at the Interface on Austenite/Ferrite Homojunction

The homojunction provides an effective way to extend the properties of stainless steel, but also leaves more weak points for small atoms to penetrate. In this study, the effects of hydrogen, carbon, and nitrogen atoms at the interface on the austenite/ferrite homojunction were investigated using first principles. This study found that low concentrations of carbon/nitrogen are favorable for the pairing of FCC with BCC compared to hydrogen, which can effectively improve the bonding energy and stability of homogeneous junctions. However, at high concentrations, the interfacial hydrogen can partially act as a mediator for interfacial bonding, which results in a slower decrease in bonding energy. On the contrary, nitrogen causes a sharp decrease in interfacial matching due to excessive strengthening of austenite, which reduces both the binding energy and the stability of the overall system. This study provides valid data and a unique perspective on the development of the austenite/ferrite homojunction.

The Arp2/3 complex contributes to the regeneration of branched actin filament networks.

Actin filaments are one of the indispensable components of almost all cell types. These semi-flexible filaments constitute an important part of the cytoskeleton and give the cell its mechanical properties. Among various structures that actin filaments form, branched actin network plays a crucial role in many cellular processes such as cell motility and endocytosis. Branched actin filaments are formed when an activated Arp2/3 complex binds to the side of an existing filament (mother) and generates a new filament branching off the mother. These filaments are subjected to force when they are pushing against the membrane or as a result of myosin motors pulling on them. Furthermore, biochemical conditions, namely actin-binding proteins, can regulate the dynamics of branched actin networks, notably the stability of branch junctions. Here we use a microfluidics approach to reconstitute individual branched actin filaments from purified proteins in-vitro. Thanks to microfluidics, we can grow filaments branching off from the side of the mother filaments and pull on them with different orientations, varying continuously between 0 and 180 degrees. Such an approach allows us to characterize the effect of force orientation, as well as force magnitude, on debranching in the presence of different regulatory proteins. Furthermore, we are able to follow the fate of Arp2/3 after debranching. Contrary to previous observations, we find specific physiological conditions in which Arp2/3 can remain attached to the mother filament after branch departure. Unexpectedly, in this type of debranching, the remaining Arp2/3 complex is able to nucleate a new branch and contribute to the turnover of the network.

PEDF Protects Endothelial Barrier Integrity during Acute Myocardial Infarction via 67LR.

Maintaining the integrity and protecting the stability of tight junctions in endothelial cells is a potential therapeutic strategy against myocardial ischaemia. Laminin receptors (67LR) are highly expressed on endothelial cell membranes and are associated with endothelial barrier function. Herein, we sought to demonstrate the direct effects of pigment epithelial-derived factor (PEDF) on tight junctions between endothelial cells via 67LR during acute myocardial infarction (AMI) and elucidate its underlying mechanisms. We detected that PEDF directly increased the level of the tight junction protein zonula occludens protein 1 (ZO-1) after overexpression in vitro and in vivo using Western blotting. Evans Blue/TTC staining showed that PEDF significantly reduced the size of the infarcted myocardium. Immunofluorescence and the transwell cellular experiments suggested that PEDF significantly upregulated PI3K-AKT permeability and the distribution of ZO-1 between endothelial cells under OGD conditions. Interestingly, PEDF significantly upregulated the phosphorylation levels of PI3K-AKT-mTOR under oxygen and glucose deprivation conditions but had no significant effects on the total protein expression. The protective effect of PEDF on ZO-1 was significantly inhibited following the inhibition of PI3K-AKT-mTOR. The activation of phosphorylation of PI3K-AKT-mTOR by PEDF was blocked after silencing 67LR, as were the protective effects of PEDF on ZO-1. Therefore, we have reason to believe that PEDF increased ZO-1 expression through the 67LR-dependent PI3K-AKT-mTOR signaling pathway, thus maintaining tight junction stability and protecting cardiac function.

Surface Analysis of Ti-Alloy Micro-Grooved 12/14 Tapers Assembled to Non-Sleeved and Sleeved Ceramic Heads: A Comparative Study of Retrieved Hip Prostheses.

Ti6Al4V titanium alloy (Ti-alloy) sleeved ceramic heads have become widely used in revision surgery when the hip stem is left in situ. This solution guarantees a new junction between the bore of the ceramic head and the Ti-alloy sleeve, regardless of any possible, slight surface damage to the Ti-alloy taper of the stem. However, this solution introduces an additional Ti-alloy/Ti-alloy interface pairing, which is potentially susceptible to mechanically assisted crevice corrosion. This study evaluated both qualitatively and quantitatively the damage that occurred in vivo on Ti-alloy micro-grooved 12/14 tapers of (i) primary implants with non-sleeved ceramic heads (Group 1), (ii) secondary implants with non-sleeved ceramic heads (Group 2), and (iii) secondary implants with sleeved ceramic heads (Group 3). A total of 45 explants-15 for each group, including short-, medium- and long-neck heads-underwent optical evaluation for surface damage (Goldberg scoring), surface roughness analysis, and SEM/EDX analysis. The Goldberg scores did not reveal different patterns in the tapers' surface damage; surface damage was classified as absent or mild (surface damage score ≤2) in 94%, another 94%, and 92% of the analysed regions for Group 1, Group 2, and Group 3, respectively. Small but significant differences in morphological changes occurred in the tapers of the three groups: reductions no greater than a few percentage points in median values of roughness parameters were found in Group 1 and Group 2, while negligible changes were found in Group 3. SEM/EDX analysis revealed little (i.e., a slight increase in the oxygen content) to undetectable changes in the chemical composition on the Ti-alloy surface independently of the group. These results suggest that the Ti-alloy/Ti-alloy sleeve/taper junction is only mildly susceptible to mechanically assisted crevice corrosion. Assembling a sleeved ceramic head, with variable neck lengths up to a "long-neck", to a Ti-alloy micro-grooved 12/14 taper of a stem left in situ does not seem to increase the risk of revision due to trunnionosis, as long as junction stability (i.e., the proper seating of the sleeved ceramic head on the 12/14 taper) is achieved intraoperatively.

ArpC5 isoforms regulate Arp2/3 complex-dependent protrusion through differential Ena/VASP positioning.

Regulation of the Arp2/3 complex is required for productive nucleation of branched actin networks. An emerging aspect of regulation is the incorporation of subunit isoforms into the Arp2/3 complex. Specifically, both ArpC5 subunit isoforms, ArpC5 and ArpC5L, have been reported to fine-tune nucleation activity and branch junction stability. We have combined reverse genetics and cellular structural biology to describe how ArpC5 and ArpC5L differentially affect cell migration. Both define the structural stability of ArpC1 in branch junctions and, in turn, by determining protrusion characteristics, affect protein dynamics and actin network ultrastructure. ArpC5 isoforms also affect the positioning of members of the Ena/Vasodilator-stimulated phosphoprotein (VASP) family of actin filament elongators, which mediate ArpC5 isoform-specific effects on the actin assembly level. Our results suggest that ArpC5 and Ena/VASP proteins are part of a signaling pathway enhancing cell migration.

MBNL-dependent impaired development within the neuromuscular system in myotonic dystrophy type 1.

Myotonic dystrophy type I (DM1) is one of the most frequent muscular dystrophies in adults. Although DM1 has long been considered mainly a muscle disorder, growing evidence suggests the involvement of peripheral nerves in the pathogenicity of DM1 raising the question of whether motoneurons (MNs) actively contribute to neuromuscular defects in DM1. By using micropatterned 96-well plates as a coculture platform, we generated a functional neuromuscular model combining DM1 and muscleblind protein (MBNL) knock-out human-induced pluripotent stem cells-derived MNs and human healthy skeletal muscle cells. This approach led to the identification of presynaptic defects which affect the formation or stability of the neuromuscular junction at an early developmental stage. These neuropathological defects could be reproduced by the loss of RNA-binding MBNL proteins, whose loss of function in vivo is associated with muscular defects associated with DM1. These experiments indicate that the functional defects associated with MNs can be directly attributed to MBNL family proteins. Comparative transcriptomic analyses also revealed specific neuronal-related processes regulated by these proteins that are commonly misregulated in DM1. Beyond the application to DM1, our approach to generating a robust and reliable human neuromuscular system should facilitate disease modelling studies and drug screening assays.

Single-Cell Quantification of the Mechanical Stability of Cell-Cell Adherens Junction Using Glass Micropipettes.

Micropipette-based methods have been widely used for the manipulation of cells and characterization of the mechanical properties at the cell or tissue level. Here, we introduce the glass micropipette-based mechanical assays for the stability of cell-cell adhesion. A probing microbead coated with specific adhesion ligands, captured by a glass micropipette, is manipulated to form the adhesion complexes with the corresponding receptors on a single cell. Once the cell is moving away from the micropipette, forces are generated from 20 pN to 100 nN to the adhesion complexes, which are quantified in real-time based on the bending of the glass micropipette. We specifically emphasize the principle and method to probe the rupturing forces of the adhesion complexes at controlled force loading rates, the ligand coating on the probe microbeads, the force calibration of the glass micropipette, and the applications of the method to probe the E-cadherin-based cell-cell adhesions. The principles can be broadly applied to other cell adhesions such as cell-matrix adhesions, neuronal synapses, and bacterial-cell adhesions.

Formation and stability of mucogingival junction on teeth without clinically detectable mucogingival junction secondary to vestibular extension procedure done for multiple adjacent teeth: A clinical study.

There are very limited data on the postsurgical formation of a mucogingival junction (MGJ) on teeth without its clinical detectability. The purpose of this study was to assess the formation and stability of MGJ on teeth without clinically detectable MGJ secondary to vestibular extension procedures for multiple adjacent teeth evaluated 6 months postoperatively. This prospective interventional single-arm clinical study was conducted in the department of Periodontology, which was approved by the institutional ethical committee, MUHS, Nashik, and registered with the Clinical Trial Registry of India. This trial included 22 participants aged between 18 and 50 years of either gender, including teeth without clinically detectable MGJ along with adjacent teeth having detectable MGJs. The following clinical parameters were taken at baseline, presurgical, immediate postsurgical, 1-month and 6-month follow-ups: plaque index, gingival index, and position of MGJ. gingival margin level, probing depth, width of keratinized gingiva, width of attached gingiva, clinical attachment level, and vestibular depth. Descriptive statistics included mean, median, mode, etc., and the inferential statistics done were analysis of variance along with post hoc Tukey and independent sample tests. Apical shift of MGJ was observed from baseline to 6 months secondary to split-full-split repositioning MGJ with vestibular extension procedure, which was statistically significant (P < 0.05). The formation of MGJ was delineated by clinical and biochemical methods at sites with nondetectable MGJ. The coronal migration of MGJ at 6 months as compared to 1 month was not statistically significant (P > 0.05). The MGJ remained stable at 6 months postoperatively at detectable and nondetectable sites. Within the limitations of this study, we can conclude that there is a definite formation of MGJ in participants without clinically detectable MGJ treated with "split-full-split MGJ-repositioning vestibular extension procedure." The MGJ, which formed apically at a 1-month postsurgical visit compared to the presurgical position, remained stable for 6 months to 1-year follow-up period at both detectable and nondetectable sites.

How an electrical current can stabilize a molecular nanojunction.

The stability of molecular junctions under transport is of the utmost importance for the field of molecular electronics. This question is often addressed within the paradigm of current-induced heating of nuclear degrees of freedom or current-induced forces acting upon the nuclei. At the same time, an essential characteristic of the failure of a molecular electronic device is its changing conductance - typically from a finite value for the intact device to zero for a device that lost its functionality. In this publication, we focus on the current-induced changes in the molecular conductance, which are inherent to molecular junctions at the limit of mechanical stability. We employ a numerically exact framework based on the hierarchical equations of motion approach, which treats both electronic and nuclear degrees of freedom on an equal footing and does not impose additional assumptions. Studying generic model systems for molecular junctions with dissociative potentials for a wide range of parameters spanning the adiabatic and the nonadiabatic regime, we find that molecular junctions that exhibit a decrease in conductance upon dissociation are more stable than junctions that are more conducting in their dissociated state. This represents a new mechanism that stabilizes molecular junctions under current. Moreover, we identify characteristic signatures in the current of breaking junctions related to the interplay between changes in the conductance and the nuclear configuration and show how these are related to properties of the leads rather than characteristics of the molecule itself.

Management of the Vertebral Artery in Craniovertebral Junction Stabilization Surgery.

The vertebral artery (VA) has an intimate relationship with the bones of the craniovertebral junction. An exact understanding of the VA anatomy in general and in the specific surgical case in particular is absolutely necessary in order to avoid intraoperative vascular injury. The course of the VA on the inferior aspect of the superior facet of the C2 vertebra makes it susceptible to damage during transarticular and interarticular fixation with the screw insertion in the adjacent lateral mass. The consequences of the intraoperative VA injury will depend on the patency of other arteries supplying the brain. In case of this complication, quick decision-making is essential to avoid excessive blood loss and to preserve adequate cerebral blood flow.

Annexin A2 modulates phospholipid membrane composition upstream of Arp2 to control angiogenic sprout initiation.

The intersection of protein and lipid biology is of growing importance for understanding how cells address structural challenges during adhesion and migration. While protein complexes engaged with the cytoskeleton play a vital role, support from the phospholipid membrane is crucial for directing localization and assembly of key protein complexes. During angiogenesis, dramatic cellular remodeling is necessary for endothelial cells to shift from a stable monolayer to invasive structures. However, the molecular dynamics between lipids and proteins during endothelial invasion are not defined. Here, we utilized cell culture, immunofluorescence, and lipidomic analyses to identify a novel role for the membrane binding protein Annexin A2 (ANXA2) in modulating the composition of specific membrane lipids necessary for cortical F-actin organization and adherens junction stabilization. In the absence of ANXA2, there is disorganized cortical F-actin, reduced junctional Arp2, excess sprout initiation, and ultimately failed sprout maturation. Furthermore, we observed reduced filipin III labeling of membrane cholesterol in cells with reduced ANXA2, suggesting there is an alteration in phospholipid membrane dynamics. Lipidomic analyses revealed that 42 lipid species were altered with loss of ANXA2, including an accumulation of phosphatidylcholine (16:0_16:0). We found that supplementation of phosphatidylcholine (16:0_16:0) in wild-type endothelial cells mimicked the ANXA2 knock-down phenotype, indicating that ANXA2 regulated the phospholipid membrane upstream of Arp2 recruitment and organization of cortical F-actin. Altogether, these data indicate a novel role for ANXA2 in coordinating events at endothelial junctions needed to initiate sprouting and show that proper lipid modulation is a critical component of these events.

Calcium-dependent cAMP mediates the mechanoresponsive behaviour of endothelial cells to high-frequency nanomechanostimulation

The endothelial junction plays a central role in regulating intravascular and interstitial tissue permeability. The ability to manipulate its integrity therefore not only facilitates an improved understanding of its underlying molecular mechanisms but also provides insight into potential therapeutic solutions. Herein, we explore the effects of short-duration nanometer-amplitude MHz-order mechanostimulation on interendothelial junction stability and hence the barrier capacity of endothelial monolayers. Following an initial transient in which the endothelial barrier is permeabilised due to Rho–ROCK-activated actin stress fibre formation and junction disruption typical of a cell's response to insults, we observe, quite uniquely, the integrity of the endothelial barrier to not only spontaneously recover but also to be enhanced considerably—without the need for additional stimuli or intervention. Central to this peculiar biphasic response, which has not been observed with other stimuli to date, is the role of second messenger calcium and cyclic adenosine monophosphate (cAMP) signalling. We show that intracellular Ca2+, modulated by the high frequency excitation, is responsible for activating reorganisation of the actin cytoskeleton in the barrier recovery phase, in which circumferential actin bundles are formed to stabilise the adherens junctions via a cAMP-mediated Epac1–Rap1 pathway. Despite the short-duration stimulation (8 min), the approximate 4-fold enhancement in the transendothelial electrical resistance (TEER) of endothelial cells from different tissue sources, and the corresponding reduction in paracellular permeability, was found to persist over hours. The effect can further be extended through multiple treatments without resulting in hyperpermeabilisation of the barrier, as found with prolonged use of chemical stimuli, through which only 1.1- to 1.2-fold improvement in TEER has been reported. Such an ability to regulate and enhance endothelial barrier capacity is particularly useful in the development of in vitro barrier models that more closely resemble their in vivo counterparts.

The management of sacral tumours.

Tumours of the sacrum are difficult to manage. The sacrum provides the structural connection between the torso and lower half of the body and is subject to both axial and rotational forces. Thus, tumours or their treatment can compromise the stability of the spinopelvic junction. Additionally, nerves responsible for lower limb motor groups as well as bowel, bladder, and sexual function traverse or abut the sacrum. Preservation or sacrifice of these nerves in the treatment of sacral tumours has profound implications on the function and quality of life of the patient. This annotation will discuss current treatment protocols for sacral tumours.Cite this article: Bone Joint J2022;104-B(12):1284-1291.

Stabilization of DNA fork junctions by Smc5/6 complexes revealed by single-molecule imaging.

SMC complexes play key roles in genome maintenance, where they ensure efficient genome replication and segregation. The SMC complex Smc5/6 is a crucial player in DNA replication and repair, yet many molecular features that determine its roles are unclear. Here, we use single-molecule microscopy to investigate Smc5/6's interaction with DNA. We find that Smc5/6 forms oligomers that dynamically redistribute on dsDNA by 1D diffusion and statically bind to ssDNA. Using combined force manipulation and single-molecule microscopy, we generate ssDNA-dsDNA junctions that mimic structures present in DNA repair intermediates or replication forks. We show that Smc5/6 accumulates at these junction sites, stabilizes the fork, and promotes the retention of RPA. Our observations provide a model for the complex's enrichment at sites of replication stress and DNA lesions from where it coordinates the recruitment and activation of downstream repair proteins.

Feedback between mechanosensitive signaling and active forces governs endothelial junction integrity

The formation and recovery of gaps in the vascular endothelium governs a wide range of physiological and pathological phenomena, from angiogenesis to tumor cell extravasation. However, the interplay between the mechanical and signaling processes that drive dynamic behavior in vascular endothelial cells is not well understood. In this study, we propose a chemo-mechanical model to investigate the regulation of endothelial junctions as dependent on the feedback between actomyosin contractility, VE-cadherin bond turnover, and actin polymerization, which mediate the forces exerted on the cell-cell interface. Simulations reveal that active cell tension can stabilize cadherin bonds, but excessive RhoA signaling can drive bond dissociation and junction failure. While actin polymerization aids gap closure, high levels of Rac1 can induce junction weakening. Combining the modeling framework with experiments, our model predicts the influence of pharmacological treatments on the junction state and identifies that a critical balance between RhoA and Rac1 expression is required to maintain junction stability. Our proposed framework can help guide the development of therapeutics that target the Rho family of GTPases and downstream active mechanical processes.

Order of magnitude reduction in Joule heating of single molecular junctions between graphene electrodes.

Maintaining stability of single-molecular junctions (SMJs) in the presence of current flow is a prerequisite for their potential device applications. However, theoretical understanding of nonequilibrium heat transport in current-carrying SMJs is a challenging problem due to the different kinds of nonlinear interactions involved, including electron-vibration and anharmonic vibrational coupling. Here, we overcome this challenge by accelerating Langevin-type current-induced molecular dynamics using machine-learning potential derived from density functional theory. We show that SMJs with graphene electrodes generate an order of magnitude less heating than those with gold electrodes. This is rooted in the better phonon spectral overlap of graphene with molecular vibrations, rendering harmonic phonon heat transport being dominant. In contrast, in a spectrally mismatched junction with gold electrodes, anharmonic coupling becomes important to transport heat away from the molecule to surrounding electrodes. Our work paves the way for studying current-induced heat transport and energy redistribution in realistic SMJs.

Origin of variable propensity for anomalous slip in body-centered cubic metals

Many transition metals crystalizing in the body-centered cubic (bcc) structure exhibit anomalous slip on low-stressed planes at low homologous temperatures, which cannot be reconciled with the Schmid law. Specifically, for uniaxial loading in the center of the stereographic triangle, this is manifested by and screw dislocations moving on low-stressed planes. While the anomalous slip is often attributed to non-planar cores of screw dislocations or to the tendency for their networks to glide easily, it remains unclear why it dominates the plastic deformation in some bcc metals, whereas it is weak or even absent in others. Using molecular statics simulations at 0 K, we demonstrate that the anomalous slip in bcc metals is intimately linked with the stability of screw junctions between two intersecting screw dislocations under stress (for example, and screws giving rise to the [100] junction). Our atomic-level studies show that in nearly all bcc metals of the 5th and 6th groups these junctions cannot be broken by the applied stress and the three dislocations can only move on the common plane (in the above example on the plane). On the other hand, these junctions are found to be unstable in alkali metals, tantalum, and iron, where the application of stress results in unzipping of the two dislocations and their further glide on the planes predicted for isolated dislocations. These results also suggest that the experimentally observed increased propensity for the anomalous slip in further stages of plastic deformation may be explained by reduced curvatures of screw dislocations in dense networks.

KRIT1: A Traffic Warden at the Busy Crossroads Between Redox Signaling and the Pathogenesis of Cerebral Cavernous Malformation Disease.

Significance: KRIT1 (Krev interaction trapped 1) is a scaffolding protein that plays a critical role in vascular morphogenesis and homeostasis. Its loss-of-function has been unequivocally associated with the pathogenesis of Cerebral Cavernous Malformation (CCM), a major cerebrovascular disease of genetic origin characterized by defective endothelial cell-cell adhesion and ensuing structural alterations and hyperpermeability in brain capillaries. KRIT1 contributes to the maintenance of endothelial barrier function by stabilizing the integrity of adherens junctions and inhibiting the formation of actin stress fibers. Recent Advances: Among the multiple regulatory mechanisms proposed so far, significant evidence accumulated over the past decade has clearly shown that the role of KRIT1 in the stability of endothelial barriers, including the blood-brain barrier, is largely based on its involvement in the complex machinery governing cellular redox homeostasis and responses to oxidative stress and inflammation. KRIT1 loss-of-function has, indeed, been demonstrated to cause an impairment of major redox-sensitive mechanisms involved in spatiotemporal regulation of cell adhesion and signaling, which ultimately leads to decreased cell-cell junction stability and enhanced sensitivity to oxidative stress and inflammation. Critical Issues: This review explores the redox mechanisms that influence endothelial cell adhesion and barrier function, focusing on the role of KRIT1 in such mechanisms. We propose that this supports a novel model wherein redox signaling forms the common link between the various pathogenetic mechanisms and therapeutic approaches hitherto associated with CCM disease. Future Directions: A comprehensive characterization of the role of KRIT1 in redox control of endothelial barrier physiology and defense against oxy-inflammatory insults will provide valuable insights into the development of precision medicine strategies. Antioxid. Redox Signal. 38, 496-528.

Recent insights into gap junction biogenesis in the cochlea.

In the cochlea, connexin 26 (Cx26) and connexin 30 (Cx30) co-assemble into two types of homomeric and heteromeric gap junctions between adjacent non-sensory epithelial cells. These channels provide a mechanical coupling between connected cells, and their activity is critical to maintain cochlear homeostasis. Many of the mutations in GJB2 or GJB6, which encode Cx26 and Cx30 in humans, impair the formation of membrane channels and cause autosomal syndromic and non-syndromic hearing loss. Thus, deciphering the connexin trafficking pathways in situ should represent a major step forward in understanding the pathogenic significance of many of these mutations. A growing body of evidence now suggests that Cx26/Cx30 heteromeric and Cx30 homomeric channels display distinct assembly mechanisms. Here, we review the most recent advances that have been made toward unraveling the biogenesis and stability of these gap junctions in the cochlea.

Study of the Thermal Stability of Copper Contact Junctions in Si/SiO2 Substrates

Study of the Thermal Stability of Copper Contact Junctions in Si/SiO2 Substrates