Damage-Based Assessment of the Fatigue Crack Initiation Site in High-Strength Steel Welded Joints Treated by HFMI

This study aimed to identify the fatigue crack initiation site of high-frequency mechanical impact (HFMI)-treated high-strength steel welded joints subjected to high peak stresses; the impact of HFMI treatment residual stress relaxation being of particular interest. First, the compressive residual stresses induced by HFMI treatment and their changes due to applied high peak stresses were quantified using advanced measurement techniques. Then, several features of crack initiation sites according to levels of applied peak stresses were identified through fracture surface observation of failed specimens. The relaxation behavior was simulated with finite element (FE) analyses incorporating the experimentally characterized residual stress field, load cycles including high peak load, improved weld geometry and non-linear material behavior. With local strain and local mean stress after relaxation, fatigue damage assessments along the surface of the HFMI groove were performed using the Smith–Watson–Topper (SWT) parameter to identify the critical location and compared with actual crack initiation sites. The obtained results demonstrate the shift of the crack initiation most prone position along the surface of the HFMI groove, resulting from a combination of stress concentration and residual stress relaxation effect.

ERBB3 Methylation and Immune Infiltration in Tumor Microenvironment of Cervical Cancer

ERBB3, a member of the ERBB family of receptor tyrosine kinases, plays an important role in cancer, despite its lack of intrinsic Carcinogenic mechanism of CESC. Research on bioinformatics methods through multi-omics, this work proves that ERBB3 gene mutation, methylation modification have extensive regulatory mechanisms on the CESC microenvironment. We found that ERBB3 is involved in carcinogenesis of cervical cancer and is not associated with its prognosis. The carcinogenic mechanism is mainly related to the suppression of the immune system between TILS and the methylation of the RNA level. Our study indicated ERBB3 is more likely to be a carcinogenic factor than a key prognostic factor for cervical cancer. Methylation of ERBB3 may work as a chekpoint immunotherapy target in CESC, DNA methylation modification of the 4480 base pair downstream of ERBB3 transcription initiation site was the highest.

VHCF Behavior of Welded Joints with HFMI Treatment under Moisture Conditions

Fatigue tests of cruciform welded joints made of Q355B steel at very-high-cycle fatigue (VHCF) regimes were carried out on as-welded specimens using highfrequency mechanical impact (HFMI) treatment in dry air and water-spray environments, respectively. The influence of the environment on fatigue life was more obvious in the VHCF regime. It was found that S-N curves became flat over the range of 106–108 cycles for as-welded specimens, while a continuously decreasing S-N curve existed for HFMI-treated specimens. Fatigue cracks initiated from the weld toe of the as-welded specimens in dry air and water-spray environments. Due to residual stress, the crack initiation site transition of HFMI-treated specimens from the weld toe to the weld root and base metal was observed at lower stress levels. Moreover, hydrogen-assisted quasi-cleavage and intergranular fracture were captured using a scanning electron microscope and a hydrogen permeation test.

Molecular Dissection of the Primase and Polymerase Activities of Deep-Sea Phage NrS-1 Primase-Polymerase

PrimPols are a class of primases that belong to the archaeo-eukaryotic primase (AEP) superfamily but have both primase and DNA polymerase activities. Replicative polymerase from NrS-1 phage (NrSPol) is a representative of the PrimPols. In this study, we identified key residues for the catalytic activity of NrSPol and found that a loop in NrSPol functionally replaces the zinc finger motif that is commonly found in other AEP family proteins. A helix bundle domain (HBD), conserved in the AEP superfamily, was recently reported to bind to the primase recognition site and to be crucial for initiation of primer synthesis. We found that NrSPol can recognize different primase recognition sites, and that the initiation site for primer synthesis is not stringent, suggesting that the HBD conformation is flexible. More importantly, we found that although the HBD-inactivating mutation impairs the primase activity of NrSPol, it significantly enhances the DNA polymerase activity, indicating that the HBD hinders the DNA polymerase activity. The conflict between the primase activity and the DNA polymerase activity in a single protein with the same catalytic domain may be one reason for why DNA polymerases are generally unable to synthesize DNA de novo.

De novo apical domain formation inside the Drosophila adult midgut epithelium

In the adult Drosophila midgut, basal intestinal stem cells give rise to enteroblasts that integrate into the epithelium as they differentiate into enterocytes. Integrating enteroblasts must generate a new apical domain and break through the septate junctions between neighboring enterocytes, while maintaining barrier function. We observe that enteroblasts form an apical membrane initiation site when they reach the septate junction between the enterocytes. Cadherin clears from the apical surface and an apical space appears above the enteroblast. New septate junctions then form laterally with the enterocytes and the AMIS develops into pre-apical compartment before it has a free apical surface in contact with the gut lumen. Finally, the enterocyte septate junction dissolves and the pre-enterocyte reaches the gut lumen with a fully-formed brush border. The process of enteroblast integration resembles lumen formation in mammalian epithelial cysts, highlighting the similarities between the fly midgut and mammalian epithelia.

Investigation of Zohr Hydraulic Flying Lead Reaction Drive Shafts

The HFLs for the Zohr Phase 1 project contains a cobra head at each end that incorporates the female couplers and the locking mechanism. Beginning in February 2020, and with the most recent incident recorded in September 2020, a total of 4 supplied HFL reaction drive shafts (RDS) failed subsea, resulting in partial separation of the HFL cobra heads from their respective fixed stabplates with a loss of hydraulic supply pressure and subsequent automatic well shut ins. HFL failures occurred on both the XT and HIPPS side of the HFLs on 3 different well sites. A further RDS tested at the laboratory from the UTA end of an HFL showed signs of microscopic cracking consistent with the failed specimens suggesting it may have had the potential to lead to a failure in the future. The failed HFLs were retrieved and returned onshore, the HFL locking mechanism was stripped down to gain access to the failed ends of the RDS and a visual inspection was performed. The initial inspection after partial disassembly to reveal the inside of the HFL locking mechanism identified that the RDS had completely failed at a location on the threaded portion of the RDS. Surface deposits were collected from each probe surfaces and analysed using scanning electron microscopy (SEM), together with energy- dispersive X-ray (EDX). A piece 10mm long was taken from each of the four probes for quantitative chemical analysis. Standard tensile and Charpy V-notch impact and Vickers hardness surveys have been conducted. Each of the failed probe exhibited an intergranular fracture surface morphology. This was confirmed through metallography/EBSD. No single initiation site was located on fracture surfaces, although some regions showed a mixed fractographic morphology, with some small areas of micro-void coalescence. Secondary intergranular cracking and corrosion was apparent at various locations, in each of the failed probes, including in thread roots, in samples 183 and 188, and just above the thread, in sample 052. These observations points towards an environmentally assisted cracking mechanism (i.e. stress corrosion cracking). Metallography revealed two layers within surface films, both in cracks and on the fracture surface: an inner layer, rich in nickel, sulphur and aluminium, and an outer, rich in copper and sulphur. Mechanical testing and chemical analysis revealed consistent results across the probes. The probe material was specified as Nibron Special (CuNi14Al3/DIN 2.1504) with a size of 2inch. Would be challenging to get the full root cause of using this material for subsea applications as it is resistant to seawater. Another factor contributed allows risk of material failure which should be eliminated for all subsea industry or taken into consideration to avoid further failures.

mCherry contains a fluorescent protein isoform that interferes with its reporter function

Fluorescent proteins are essential reporters in cell biology and molecular biology. Here, we reveal that red-fluorescent proteins possess an alternative translation initiation site that produces a short functional protein isoform. The short isoform creates significant background fluorescence that biases the outcome of expression studies. Our investigation identifies the short protein isoform, traces its origin, and determines the extent of the issue within the family of red fluorescent protein. Our analysis shows that the short isoform defect of the red fluorescent protein family may affect the interpretation of many published studies. Finally, we provide a re-engineered mCherry variant that lacks background expression as an improved tool for imaging and protein expression studies.

Selective packaging of HIV-1 RNA genome is guided by the stability of 5′ untranslated region polyA stem

To generate infectious virus, HIV-1 must package two copies of its full-length RNA into particles. HIV-1 transcription initiates from multiple, neighboring sites, generating RNA species that only differ by a few nucleotides at the 5′ end, including those with one (1G) or three (3G) 5′ guanosines. Strikingly, 1G RNA is preferentially packaged into virions over 3G RNA. We investigated how HIV-1 distinguishes between these nearly identical RNAs using in-gel chemical probing combined with recently developed computational tools for determining RNA conformational ensembles, as well as cell-based assays to quantify the efficiency of RNA packaging into viral particles. We found that 1G and 3G RNAs fold into distinct structural ensembles. The 1G RNA, but not the 3G RNA, primarily adopts conformations with an intact polyA stem, exposed dimerization initiation site, and multiple, unpaired guanosines known to mediate Gag binding. Furthermore, we identified mutants that exhibited altered genome selectivity and packaged 3G RNA efficiently. In these mutants, both 1G and 3G RNAs fold into similar conformational ensembles, such that they can no longer be distinguished. Our findings demonstrate that polyA stem stability guides RNA-packaging selectivity. These studies also uncover the mechanism by which HIV-1 selects its genome for packaging: 1G RNA is preferentially packaged because it exposes structural elements that promote RNA dimerization and Gag binding.

Development of Electrochemical Surface Treatment to Visualize Critical Corrosion-Inducing Inclusions of Zr in Chloride Environments

An electrochemical surface treatment was developed to visualize the corrosion-inducing inclusions of Zr in chloride environments. Pure Zr and Zr alloy (Zr-0.5O-0.5C) were evaluated in this study. The electrochemical surface treatment consisted of repeated galvanostatic anodic polarization and potentiostatic cathodic polarization. After the electrochemical surface treatment, only one brittle and non-conductive shell of Zr oxide was observed at the corrosion initiation site on the tested surface. The corrosion-inducing inclusions were found inside the corrosion pit under the optimum polarization conditions. At the corrosion initiation site on pure Zr, the area inside the corrosion pit contained larger amounts of O, C, and Si than the surrounding matrix. In the case of the Zr-0.5O-0.5C alloy, relatively larger inclusions were observed after the treatment. Extremely large amounts of Si, together with O and C, were present in the inclusions. The inclusions that induced localized corrosion on the pure Zr and Zr-0.5O-0.5C specimens were found to be precipitated, involving the specific aggregation of Si. The surface treatment developed in this study is expected to be utilized as a powerful tool to elucidate the localized corrosion mechanism of Zr in chloride environments.