BRCA1 Interactors - Rad50 and BRIP1: As Prognostic and Predictive Molecules for the Severity of Triple-Negative Breast Cancer

BRIP1 is one of the major interacting partner of BRCA1 which plays an important role in repair by homologous recombination (HR). This gene is mutated in around 4% cases of breast cancer, however, its mechanism of action is unclear. In this study, we presented the fundamental role of BRCA1 interactors BRIP1 and RAD50 in the development of differential severity in Triple-Negative Breast Cancer(TNBC) among various affected individuals. We showed that in some TNBC lines like MDA-MB-231 the functioning of both BRCA1/TP53 is compromised. Furthermore, the sensing of DNA damage is affected, depicted through the low expression of damage sensing molecule Rad50 and reduced formation of H2AX foci. Due to less damage sensing capability and low availability of BRCA1 at the damage sites, the repair by HR becomes inefficient leading to more damage. Accumulation of damage sends a signal for over activation of NHEJ repair pathways. Over expressed NHEJ molecules with compromised HR and checkpoint conditions lead to higher proliferation and error-prone repair, which increases the mutation rate and corresponding tumor severity. The severity phenotypes were more in cells having compromised BRCA1-BRIP1 functioning. The in silico analysis of the TCGA-UCSC xena datasets with genes expression in deceased population shows a significant correlation of BRCA1 expression with OS in TNBCs (0.0272). The association of BRCA1 with OS becomes stronger with the addition of BRIP1expression (0.000876**). Since the overall survival(OS) is directly proportional to the extent of severity, the data analysis hints at the role of BRIP1 in controlling the severity of TNBC.

Inflammasome-independent functions of NAIPs and NLRs in the intestinal epithelium

The gut relies on the complex interaction between epithelial, stromal and immune cells to maintain gut health in the face of food particles and pathogens. Innate sensing by the intestinal epithelium is critical for maintaining epithelial barrier function and also orchestrating mucosal immune responses. Numerous innate pattern recognition receptors (PRRs) are involved in such sensing. In recent years, several Nucleotide-binding-domain and Leucine-rich repeat-containing receptors (NLRs) have been found to partake in pathogen or damage sensing while also being implicated in gut pathologies, such as colitis and colorectal cancer (CRC). Here, we discuss the current literature focusing on NLR family apoptosis inhibitory proteins (NAIPs) and other NLRs that have non-inflammasome roles in the gut. The mechanisms behind NLR-mediated protection often converges on similar signalling pathways, such as STAT3, MAPK and NFκB. Further understanding of how these NLRs contribute to the maintenance of gut homeostasis will be important for understanding gut pathologies and developing new therapies.

In-situ damage sensing in intra-ply glass/carbon laminate composites under interlaminar shear loading

An experimental study is preformed to investigate the in-situ damage sensing capabilities of intra-ply hybrid carbon/glass laminate and epoxy composites under quasi-static interlaminar shear loading. A three-dimensional electrical sensory network is generated inside the composites through embedded carbon nanotubes (CNTs) in an epoxy matrix along with the carbon fibers in the intra-ply hybrid laminates. CNTs are dispersed in the epoxy matrix using a combination of ultrasonication and shear mixing techniques. Four circumferential ring probes are used to examine the electrical response under interlaminar shear load. The effect of four different intra-ply orientations (((0–90)C, where carbon fibers are oriented along the loading direction), ((0–90)G, where glass fibers are oriented along the loading direction), ((45/−45, where glass and carbon fibers are oriented at 45o/−45o and the laminates are repeated), and ((45/−45)A, where glass and carbon fibers are oriented at 45o/−45o and the laminates are alternated)) on the shear constitutive behavior and the damage detection are discussed. Intra-ply orientations of (45/−45) and (45/−45)A showed higher interlaminar shear strength and shear strain at break compared to (0/90)C and (0/90)G orientations. Out of all four orientations, (45/−45)A provided a better resolution of electrical response for damage sensing applications.

Making Connections: Integrative Signaling Mechanisms Coordinate DNA Break Repair in Chromatin

DNA double-strand breaks (DSBs) are hazardous to genome integrity and can promote mutations and disease if not handled correctly. Cells respond to these dangers by engaging DNA damage response (DDR) pathways that are able to identify DNA breaks within chromatin leading ultimately to their repair. The recognition and repair of DSBs by the DDR is largely dependent on the ability of DNA damage sensing factors to bind to and interact with nucleic acids, nucleosomes and their modified forms to target these activities to the break site. These contacts orientate and localize factors to lesions within chromatin, allowing signaling and faithful repair of the break to occur. Coordinating these events requires the integration of several signaling and binding events. Studies are revealing an enormously complex array of interactions that contribute to DNA lesion recognition and repair including binding events on DNA, as well as RNA, RNA:DNA hybrids, nucleosomes, histone and non-histone protein post-translational modifications and protein-protein interactions. Here we examine several DDR pathways that highlight and provide prime examples of these emerging concepts. A combination of approaches including genetic, cellular, and structural biology have begun to reveal new insights into the molecular interactions that govern the DDR within chromatin. While many questions remain, a clearer picture has started to emerge for how DNA-templated processes including transcription, replication and DSB repair are coordinated. Multivalent interactions with several biomolecules serve as key signals to recruit and orientate proteins at DNA lesions, which is essential to integrate signaling events and coordinate the DDR within the milieu of the nucleus where competing genome functions take place. Genome architecture, chromatin structure and phase separation have emerged as additional vital regulatory mechanisms that also influence genome integrity pathways including DSB repair. Collectively, recent advancements in the field have not only provided a deeper understanding of these fundamental processes that maintain genome integrity and cellular homeostasis but have also started to identify new strategies to target deficiencies in these pathways that are prevalent in human diseases including cancer.

NANOENGINEERED GLASS FIBER REINFORCED COMPOSITE LAMINATES WITH INTEGRATED MULTIFUNCTIONALITY

Combining one or more functional capabilities of subsystems within a structure can provide system-level savings, particularly for weight-critical applications such as air and space vehicles. Nanoengineering presents a significant opportunity for additional functionalities on the nanoscale without the necessity to modify shape, design, or load carrying capacity of the structure. Here, an integrated-multifunctional nano-engineered system was preliminarily studied in composite laminate structures. The study would support the exploration of a system designed to serve independent yet synergistic functionalities in life-cycle enhancements, energy savings during manufacturing, in-situ cure (manufacturing) monitoring, and in-service damage sensing. For the preliminary study, an integrated multifunctional composite (IMC) laminate was created via aligned nanofiber introduction into the composite interlaminar region and the laminate surfaces of Hexcel E-glass/913 unidirectional glass fiber prepreg. Various heights ranging from 10 - 40 μm-tall vertically aligned carbon nanotube (VA-CNT) arrays, as well as patterned and buckled VA-CNT architectures, were used to reinforce the weak interlaminar regions within the laminates showing a ~ 4 - 5% increase in short beam strength of VA-CNT reinforced specimens hence demonstrating interlaminar enhancement for life-cycle advancements. The same layers, being electrically conductive, can provide several additional multifunctionalities.

A Modern Genotoxicity Testing Paradigm: Integration of the High-Throughput CometChip® and the TGx-DDI Transcriptomic Biomarker in Human HepaRG™ Cell Cultures

Higher-throughput, mode-of-action-based assays provide a valuable approach to expedite chemical evaluation for human health risk assessment. In this study, we combined the high-throughput alkaline DNA damage-sensing CometChip® assay with the TGx-DDI transcriptomic biomarker (DDI = DNA damage-inducing) using high-throughput TempO-Seq®, as an integrated genotoxicity testing approach. We used metabolically competent differentiated human HepaRG™ cell cultures to enable the identification of chemicals that require bioactivation to cause genotoxicity. We studied 12 chemicals (nine DDI, three non-DDI) in increasing concentrations to measure and classify chemicals based on their ability to damage DNA. The CometChip® classified 10/12 test chemicals correctly, missing a positive DDI call for aflatoxin B1 and propyl gallate. The poor detection of aflatoxin B1 adducts is consistent with the insensitivity of the standard alkaline comet assay to bulky lesions (a shortcoming that can be overcome by trapping repair intermediates). The TGx-DDI biomarker accurately classified 10/12 agents. TGx-DDI correctly identified aflatoxin B1 as DDI, demonstrating efficacy for combined used of these complementary methodologies. Zidovudine, a known DDI chemical, was misclassified as it inhibits transcription, which prevents measurable changes in gene expression. Eugenol, a non-DDI chemical known to render misleading positive results at high concentrations, was classified as DDI at the highest concentration tested. When combined, the CometChip® assay and the TGx-DDI biomarker were 100% accurate in identifying chemicals that induce DNA damage. Quantitative benchmark concentration (BMC) modeling was applied to evaluate chemical potencies for both assays. The BMCs for the CometChip® assay and the TGx-DDI biomarker were highly concordant (within 4-fold) and resulted in identical potency rankings. These results demonstrate that these two assays can be integrated for efficient identification and potency ranking of DNA damaging agents in HepaRG™ cell cultures.

Functional Coatings for Damage Detection in Aerospace Structures

The future of aerospace structures is highly dependent on the advancement of reliable and high-performance materials, such as composite materials and metals. Innovation in high resolution non-invasive evaluation of these materials is needed for their qualification and monitoring for structural integrity. Aluminum oxide (or α-alumina) nanoparticles present photoluminescent properties that allow stress and damage sensing via photoluminescence piezospectroscopy. This work describes how these nanoparticles are added into a polymer matrix to create functional coatings that monitor the damage of the underlying composite or metallic substrates. Different volume fractions of α-alumina nanoparticles in the piezospectroscopic coatings were studied for determining the sensitivity of the coatings and successful damage detection was demonstrated for an open-hole tension composite substrate as well as 2024 aluminum tensile substrates with a subsurface notch.