A micromechanical investigation of diagenetically-induced changes to the anisotropic elastic properties of calcareous mudstones

Microscale diagenetic changes that occur during burial exert a profound influence on the elastic and mechanical properties of sediments—but are poorly quantified. The focus here is on how diagenesis influences the elastic properties of carbonate-rich mudstones, which are subject to a wide range of physical and chemical changes. Nanoindentation data for gas-window (180 C) Eagle Ford formation samples give intrinsic indentation moduli of the clay-sized calcite matrix of 40–50 GPa, which contrasts with 60–77 GPa for diagenetic calcite fills of foraminifera tests, closer to values for highly crystalline calcite. The matrix calcite is weakly anisotropic. Inverse analysis of immature (< 70 C) organic-rich chalks gives much lower intrinsic indentation moduli for biogenic calcite, between 17 and 30 GPa; the calcite is also more anisotropic, with values of 1.3. Diagenesis, which includes recrystallisation and pore-filling cementation, results in calcite becoming elastically stiffer and behaving in an increasingly isotropic manner, in agreement with grain scale studies using atomic force microscopy. The results demonstrate that nanoindentation can resolve diagenetic contributions to the mechanical response of mudstones, and suggest intrinsic structural changes to calcite, in addition to diagenetic cementation, need to be accounted for in rock-physics models of mud-rich sediments

Late diagenesis of illite-smectite in the Podhale Basin, southern Poland: Chemistry, morphology, and preferred orientation

Well-characterized samples from the Podhale Basin, southern Poland, formed the basis for exploring and illuminating subtle diagenetic changes to a mudstone toward the upper end of the diagenetic window, prior to metamorphism.Transmission electron microscopy (TEM) performed on dispersed grains and ion-beam thinned preparations, selected area diffraction patterns,and chemistry by TEM-EDS (energy dispersive spectra) augmented mineralogy and fabric data. The deepest samples show no change in their percent illite in illite-smectite (I-S), yet I-S–phase octahedral Fe3+ and Al3+ are statistically different between samples. A decrease in the Fe3+ concentration in the octahedral sheet correlates with an increase in I-S fabric intensity and apparent crystallinity. The D-statistic from the Kolmogorov-Smirnov test on TEM- EDS data describes statistical differences in the I-S chemistry. Previous work on these samples showed a significant increase in the preferred orientation of the I-S phase across the smectite to illite transition and a significant slowdown in the rate of development of preferred orientation beyond the termination of smectite illitization. Lattice fringe images describe an I-S morphology that coalesces into larger and tighter packets with increasing burial temperature and a decrease in I-S packet contact angle, yet some evidence for smectite collapse structures is retained. The deepest sample shows the thickest, most coherent I-S packets. We propose that the deepest samples in the Podhale Basin describe the precursor stage in phyllosilicate fabric preferred orientation increase from diagenesis into metamorphism, where continued evolution of crystallite packets and associated crystallinity create higher I-S fabric intensities as the structural formulae of I-S approach an end-member composition

Vertical effective stress as a control on quartz cementation in sandstones

Temperature-controlled precipitation kinetics has become the overwhelmingly dominant hypothesis for the control of quartz cementation in sandstones. Here, we integrate quantitative petrographic data, high spatial resolution oxygen isotope analyses of quartz cement, basin modelling and a kinetic model for quartz precipitation to suggest that the supply of silica from stress-sensitive intergranular pressure dissolution at grain contacts is in fact a key control on quartz cementation in sandstones. We present data from highly overpressured sandstones in which, despite the current burial temperature of 190 °C, quartz cement occurs in low amounts (4.6 ± 1.2% of bulk volume). In situ oxygen isotope data across quartz overgrowths suggest that cementation occurred over 100 Ma and a temperature range of 80–150 °C, during which time high fluid overpressures resulted in consistently low vertical effective stress. We argue that the very low amounts of quartz cement can only be explained by the low vertical effective stress which occurred throughout the burial history and which restricted silica supply as a result of a low rate of intergranular pressure dissolution at grain contacts

FOXD3 Regulates VISTA Expression in Melanoma.

Immune checkpoint inhibitors have improved patient survival in melanoma, but the innate resistance of many patients necessitates the investigation of alternative immune targets. Many immune checkpoint proteins lack proper characterization, including V-domain Ig suppressor of T cell activation (VISTA). VISTA expression on immune cells can suppress T cell activity; however, few studies have investigated its expression and regulation in cancer cells. In this study, we observe that VISTA is expressed in melanoma patient samples and cell lines. Tumor cell-specific expression of VISTA promotes tumor onset in vivo, associated with increased intratumoral T regulatory cells, and enhanced PDL-1 expression on tumor-infiltrating macrophages. VISTA transcript levels are regulated by the stemness factor Forkhead box D3 (FOXD3). BRAF inhibition upregulates FOXD3 and reduces VISTA expression. Overall, this study demonstrates melanoma cell expression of VISTA and its regulation by FOXD3, contributing to the rationale for therapeutic strategies that combine targeted inhibitors with immune checkpoint blockade

High-throughput chemogenetic drug screening reveals PKC-RhoA/PKN as a targetable signaling vulnerability in GNAQ-driven uveal melanoma

Uveal melanoma (UM) is the most prevalent cancer of the eye in adults, driven by activating mutation of GNAQ/GNA11; however, there are limited therapies against UM and metastatic UM (mUM). Here, we perform a high-throughput chemogenetic drug screen in GNAQ-mutant UM contrasted with BRAF-mutant cutaneous melanoma, defining the druggable landscape of these distinct melanoma subtypes. Across all compounds, darovasertib demonstrates the highest preferential activity against UM. Our investigation reveals that darovasertib potently inhibits PKC as well as PKN/PRK, an AGC kinase family that is part of the "dark kinome." We find that downstream of the Gαq-RhoA signaling axis, PKN converges with ROCK to control FAK, a mediator of non-canonical Gαq-driven signaling. Strikingly, darovasertib synergizes with FAK inhibitors to halt UM growth and promote cytotoxic cell death in vitro and in preclinical metastatic mouse models, thus exposing a signaling vulnerability that can be exploited as a multimodal precision therapy against mUM.</p

Evidence of prokineticin dysregulation in fallopian tube from women with ectopic pregnancy

OBJECTIVE: To demonstrate expression and regulation of prokineticins (PROKs) and their receptors (PROKRs) in Fallopian tube (FT) from non-pregnant women and women with ectopic pregnancy (EP). DESIGN: Tissue analysis. SETTING: Large UK teaching hospital PATIENTS: Women undergoing hysterectomy for benign gynecological conditions (n=15) and surgery for EP (n=16). INTERVENTIONS: Quantitative RT-PCR and immunohistochemistry were used to determine FT PROK/PROKR mRNA expression and protein localization, respectively. PROK/PROKR levels were measured in tubal explant cultures stimulated with estrogen and progestogen. MAIN OUTCOME MEASURES: Differential expression of PROK and PROKR. RESULTS: FT PROK2 and PROKR1 mRNA levels were upregulated during the progesterone-dominant mid-luteal phase of the menstrual cycle. Increased PROKR1 expression was observed in tubal explant cultures treated with medroxyprogesterone acetate. PROK and PROKR proteins were localized to the epithelium and smooth muscle layers of the FT. PROKR1 and PROKR2 mRNA levels were lower in FT from women with EP compared to non-pregnant FT from the mid-luteal phase. CONCLUSION: These data suggest a potential role for PROKs in FT function. PROKs are known to affect smooth muscle contraction in the gut. Dysregulated PROK expression in FT could affect FT smooth muscle contractility and embryo-tubal transport, providing a potential cause for EP

Assembling Neurospheres: Dynamics of Neural Progenitor/Stem Cell Aggregation Probed Using an Optical Trap

Optical trapping (tweezing) has been used in conjunction with fluid flow technology to dissect the mechanics and spatio-temporal dynamics of how neural progenitor/stem cells (NSCs) adhere and aggregate. Hitherto unavailable information has been obtained on the most probable minimum time (∼5 s) and most probable minimum distance of approach (4–6 µm) required for irreversible adhesion of proximate cells to occur. Our experiments also allow us to study and quantify the spatial characteristics of filopodial- and membrane-mediated adhesion, and to probe the functional dynamics of NSCs to quantify a lower limit of the adhesive force by which NSCs aggregate (∼18 pN). Our findings, which we also validate by computational modeling, have important implications for the neurosphere assay: once aggregated, neurospheres cannot disassemble merely by being subjected to shaking or by thermal effects. Our findings provide quantitative affirmation to the notion that the neurosphere assay may not be a valid measure of clonality and “stemness”. Post-adhesion dynamics were also studied and oscillatory motion in filopodia-mediated adhesion was observed. Furthermore, we have also explored the effect of the removal of calcium ions: both filopodia-mediated as well as membrane-membrane adhesion were inhibited. On the other hand, F-actin disrupted the dynamics of such adhesion events such that filopodia-mediated adhesion was inhibited but not membrane-membrane adhesion