We analysed the reproducibility of Ki67 labelling index (LI) between two scorers using the International Ki67 Working Group (IKWG) global methods on an Android application (APP), correlated the APP ...and eyeball estimate (EBE) with digital image analysis (DIA) scores and determined the prognostic significance of Ki67LI. Global weighted (GW) and global unweighted (GUW) Ki67 app scores of hormone receptor‐positive and HER2 (human epidermal growth factor receptor 2)‐negative breast cancer patients were obtained. Reproducibility of Ki67LI between 2 scorers and correlation of APP and EBE scores with DIA scores were performed. The prognostic significance of APP scores and its correlation with other clinico‐pathologic variables were evaluated. The intra‐class correlation coefficient (ICC) between 2 scorers showed excellent reliability with both GW and GUW methods. ICC between DIA and APP scores was significantly greater than DIA versus EBE. The three categories of APP scores based on median value and cut points of 10%, 18% and 38% were significantly associated with poor DFS. On multivariate analysis, significant association between Ki67LI, tumour size, nodal involvement and DFS was noted. Our study shows that the visual Ki67 scoring app is effective in bringing consistency to KI67LI and APP scores showed significant correlation with DFS.
H2–D2 isotopic equilibration is often used to assess the involvement and reversibility of H2 dissociative chemisorption steps during hydrogenation surface catalysis under the premise that exchange ...turnovers require recombinative desorption of adatoms (H*, D*). Experimental and theoretical evidence reported here show instead that exchange occurs via non-competitive adsorption mechanisms that circumvent recombinative desorption steps at temperatures of catalytic relevance (<700 K) and that H2 and D2 adsorption enthalpies are similar, leading to equilibrium isotope effects near unity (0.7–1.0) on dispersed Pt nanoparticles. The kinetic effects of H2 and D2 pressures on exchange rates at 300–700 K are accurately described by elementary steps involving the non-competitive adsorption of H2 (or D2) and its reaction with D* (or H*), mediated via local displacements of adatoms to alternate binding sites in H*/D* adlayers but without requiring their intervening endothermic recombinative desorption. Recombination (H* + D*) routes become prevalent only above 700 K, leading to different kinetic trends with H2/D2 pressures and to higher activation barriers (78 vs 32 kJ mol–1) than for the single-site non-competitive adsorption routes that prevail at lower temperatures. Such conclusions are consistent with density functional theory (DFT) calculations on 4 × 4 Pt(111) models at saturation coverages (1 H*/Pts), which show that barriers for single-site metallocycle mechanisms (205 kJ mol–1) are much larger than those for routes involving H2 or D2 dissociation on “spaces” created by displacements of D*/H* adatoms (55 kJ mol–1) and the concomitant local supersaturation of the Pt surface. Such pathways weaken the binding of H* (or D*) at such locations, thus enabling the facile desorption and exchange of H2 and D2 under conditions that would otherwise preclude the measured rates of exchange events. Pt201 nanoparticle models allow greater adlayer relaxation and supramonolayer H* coverages and lead to even lower DFT-derived barriers for non-competitive adsorption routes (27 kJ mol–1 at 1.44 H*/Pts). Recombination routes exhibit higher barriers and become the predominant exchange pathways above 700 K; for reactions requiring such temperatures, exchange rates can provide a reliable assessment of the reversibility of dihydrogen dissociation events.
Ki67 has potential clinical importance in breast cancer but has yet to see broad acceptance due to inter-laboratory variability. Here we tested an open source and calibrated automated digital image ...analysis (DIA) platform to: (i) investigate the comparability of Ki67 measurement across corresponding core biopsy and resection specimen cases, and (ii) assess section to section differences in Ki67 scoring. Two sets of 60 previously stained slides containing 30 core-cut biopsy and 30 corresponding resection specimens from 30 estrogen receptor-positive breast cancer patients were sent to 17 participating labs for automated assessment of average Ki67 expression. The blocks were centrally cut and immunohistochemically (IHC) stained for Ki67 (MIB-1 antibody). The QuPath platform was used to evaluate tumoral Ki67 expression. Calibration of the DIA method was performed as in published studies. A guideline for building an automated Ki67 scoring algorithm was sent to participating labs. Very high correlation and no systematic error (p = 0.08) was found between consecutive Ki67 IHC sections. Ki67 scores were higher for core biopsy slides compared to paired whole sections from resections (p ≤ 0.001; median difference: 5.31%). The systematic discrepancy between core biopsy and corresponding whole sections was likely due to pre-analytical factors (tissue handling, fixation). Therefore, Ki67 IHC should be tested on core biopsy samples to best reflect the biological status of the tumor.
Although an important biomarker in breast cancer, Ki67 lacks scoring standardization, which has limited its clinical use. Our previous study found variability when laboratories used their own scoring ...methods on centrally stained tissue microarray slides. In this current study, 16 laboratories from eight countries calibrated to a specific Ki67 scoring method and then scored 50 centrally MIB-1 stained tissue microarray cases. Simple instructions prescribed scoring pattern and staining thresholds for determination of the percentage of stained tumor cells. To calibrate, laboratories scored 18 'training' and 'test' web-based images. Software tracked object selection and scoring. Success for the calibration was prespecified as Root Mean Square Error of scores compared with reference <0.6 and Maximum Absolute Deviation from reference <1.0 (log2-transformed data). Prespecified success criteria for tissue microarray scoring required intraclass correlation significantly >0.70 but aiming for observed intraclass correlation ≥0.90. Laboratory performance showed non-significant but promising trends of improvement through the calibration exercise (mean Root Mean Square Error decreased from 0.6 to 0.4, Maximum Absolute Deviation from 1.6 to 0.9; paired t-test: P=0.07 for Root Mean Square Error, 0.06 for Maximum Absolute Deviation). For tissue microarray scoring, the intraclass correlation estimate was 0.94 (95% credible interval: 0.90-0.97), markedly and significantly >0.70, the prespecified minimum target for success. Some discrepancies persisted, including around clinically relevant cutoffs. After calibrating to a common scoring method via a web-based tool, laboratories can achieve high inter-laboratory reproducibility in Ki67 scoring on centrally stained tissue microarray slides. Although these data are potentially encouraging, suggesting that it may be possible to standardize scoring of Ki67 among pathology laboratories, clinically important discrepancies persist. Before this biomarker could be recommended for clinical use, future research will need to extend this approach to biopsies and whole sections, account for staining variability, and link to outcomes.
Carbonic anhydrase IX (CAIX) is a hypoxia and HIF-1-inducible protein that regulates intra- and extracellular pH under hypoxic conditions and promotes tumor cell survival and invasion in hypoxic ...microenvironments. Interrogation of 3,630 human breast cancers provided definitive evidence of CAIX as an independent poor prognostic biomarker for distant metastases and survival. shRNA-mediated depletion of CAIX expression in 4T1 mouse metastatic breast cancer cells capable of inducing CAIX in hypoxia resulted in regression of orthotopic mammary tumors and inhibition of spontaneous lung metastasis formation. Stable depletion of CAIX in MDA-MB-231 human breast cancer xenografts also resulted in attenuation of primary tumor growth. CAIX depletion in the 4T1 cells led to caspase-independent cell death and reversal of extracellular acidosis under hypoxic conditions in vitro. Treatment of mice harboring CAIX-positive 4T1 mammary tumors with novel CAIX-specific small molecule inhibitors that mimicked the effects of CAIX depletion in vitro resulted in significant inhibition of tumor growth and metastasis formation in both spontaneous and experimental models of metastasis, without inhibitory effects on CAIX-negative tumors. Similar inhibitory effects on primary tumor growth were observed in mice harboring orthotopic tumors comprised of lung metatstatic MDA-MB-231 LM2-4(Luc+) cells. Our findings show that CAIX is vital for growth and metastasis of hypoxic breast tumors and is a specific, targetable biomarker for breast cancer metastasis.
Colony-stimulating factor-1 receptor (CSF-1R) signaling promotes an immune suppressive microenvironment enriched in M2 macrophages. Given that CSF-1R inhibitors are under investigation in clinical ...trials, including in breast cancer, CSF-1R expression and association with immune biomarkers could identify patients who derive greater benefit from combination with immunotherapies. TIMER2.0 and bc-GenExMiner v4.7 were used to assess the correlation of CSF1R mRNA with immune infiltrates and prognosis. Following a prespecified training–validation approach, an optimized immunohistochemistry assay was applied to assess CSF-1R on carcinoma cells and macrophages on breast cancer tissue microarray series representing 2384 patients, coupled to comprehensive clinicopathological, biomarker, and outcome data. Significant positive correlations were observed between CSF1R mRNA and immune infiltrates. High carcinoma CSF-1R correlated with grade 3 tumors >2 cm, hormone receptor negativity, high Ki67, immune checkpoint biomarkers, and macrophages expressing CSF-1R and CD163. High carcinoma CSF-1R was significantly associated with poor survival in univariate and multivariate analyses. Adverse prognostic associations were retained in ER+ cases regardless of the presence of CD8+ T cells. CSF-1R+ macrophages were not prognostic. High carcinoma CSF-1R is associated with aggressive breast cancer biology and poor prognosis, particularly in ER+ cases, and identifies patients in whom biomarker-directed CSF-1R therapies may yield superior therapeutic responses.
Our aim was to characterize the pathological, molecular and clinical outcomes of clear cell carcinoma of the endometrium (CCC).
CCC underwent ProMisE (Proactive Molecular Risk Classifier for ...Endometrial Cancer) classification identifying four molecular subtypes: i) ‘POLEmut’ for ECs with pathogenic POLE mutations, ii) ‘MMRd’, if there is loss of mismatch repair proteins by immunohistochemistry (IHC), iii) ‘p53wt’ or iv) ‘p53abn’ based on p53 IHC staining. Clinicopathologic parameters, immune markers (CD3, CD8, CD79a, CD138, PD-1), ER, L1CAM, and outcomes were assessed.
52 CCCs were classified, including 1 (2%) POLEmut, 5 (10%) MMRd, 28 (54%) p53wt and 18 (35%) p53abn. Women with p53abn and p53wt CCCs were older than women with MMRd and POLEmut subtypes. p53wt CCC were distinct from typical p53wt endometrioid carcinomas; more likely to arise in older, thinner women, with advanced stage disease, LVSI and lymph node involvement, and a higher proportion ER negative, L1CAM overexpressing tumors with markedly worse outcomes. High levels of immune infiltrates (TILhigh) were observed in 75% of the CCC cohort. L1CAM overexpression was highest within p53abn and p53wt subtypes of CCC.
CCC is a heterogeneous disease encompassing all four molecular subtypes and a wide range of clinical outcomes. Outcomes of patients with POLEmut, MMRd and p53abn CCC are not distinguishable from those of other patients with these respective subtypes of EC; p53wt CCC, however, differ from endometrioid p53wt EC in clinical, pathological, molecular features and outcomes. Thus, p53wt CCC of endometrium appear to be a distinct clinicopathological entity within the larger group of p53wt ECs.
•There is molecular heterogeneity within clear cell carcinoma (CCC) of the endometrium.•P53wt CCC of endometrium show aggressive features and are distinct from other p53wt endometrial cancers.•Molecular classification of CCC of endometrium elicits prognostic parameters that are not apparent with histology alone.
•Children of a parent with schizophrenia experience multiple deficits.•Potential alleviating factors still need to be tested and confirmed.•Evidence-based and child-centered interventions still need ...to be developed.
As well as having a higher genetic vulnerability to psychiatric problems, children of a parent with schizophrenia suffer a significantly poorer quality of life than those with healthy parents. In mental healthcare settings, the well-being of these children is still overlooked. It is crucial to develop child-centered interventions for them. This scoping review focuses specifically on children of a parent with schizophrenia to identify the likely impacts on their life and development, the factors and strategies that may alleviate negative impacts, and available interventions.
We applied a systematic approach to search the following databases: PsycINFO, MEDLINE, Embase, Google Scholar, CNKI and CEPS to identify relevant English and Chinese publications focusing on children. Quality assessments of quantitative and qualitative studies were undertaken, using the Downs and Black instrument and the CASP Checklist respectively.
After screening, thirty-three studies were included for review. The existing evidence indicates that children of a parent with schizophrenia experience multiple deficits. Although various factors have been identified that can potentially alleviate their negative experiences, few are well supported with solid empirical evidence that confirm causal effects. The needs of these children are commonly neglected: little professional support has been provided, and the usefulness of the available support has yet to be determined.
Based on the review, we argue that effective means should be implemented so that children of a parent with schizophrenia needing help can be identified and experts can overcome barriers to providing help. The potential modifiable factors that can alleviate the negative impacts of having a parent with schizophrenia on youngsters need to be tested and confirmed. Interventions should be evidence-based, schizophrenia-specific, and child-centered.
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