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The innate lymphoid cell (ILC) family is composed of heterogeneous innate effector and helper immune cells that preferentially reside in tissues where they promote tissue homeostasis. In cancer, they have been implicated in driving both pro- and anti-tumor responses. This apparent dichotomy highlights the need to better understand differences in the ILC composition and phenotype within different tumor types that could drive seemingly opposite anti-tumor responses. Here, we characterized the frequency and phenotype of various ILC subsets in melanoma metastases and primary epithelial ovarian tumors. We observed high PD-1 expression on ILC subsets isolated from epithelial ovarian tumor samples, while ILC populations in melanoma samples express higher levels of LAG-3. In addition, we found that the frequency of cytotoxic ILCs and NKp46+ILC3 in tumors positively correlates with monocytic cells and conventional type 2 dendritic cells, revealing potentially new interconnected immune cell subsets in the tumor microenvironment. Consequently, these observations may have direct relevance to tumor microenvironment composition and how ILC subset may influence anti-tumor immunity. [ABSTRACT FROM AUTHOR]

Participatory Research (PR), which places participants at the foreground, is an evolving field spanning several disciplines. This article critiques conceptual tensions surrounding the notion of participation itself and how PR tenets can be engaged with children (under 18). We propose an innovative heuristic framework to be used flexibly by the adult facilitator to aid decision-making when seeking balance between intervention and giving young participants room for expression. It serves as an aide memoire to facilitate cognisance of issues of control and directiveness that could potentially diminish children's voices, containing more explicit emphasis on the necessary movement between modes of adult facilitator involvement than existing PR models. Our proposed modes, 'organic and unbridled', 'organic yet enabling', 'enabled and shaping', are fleshed out and illustrated with reference to our own PR projects. We relate these modes to easily-called-to-mind notions, inspired by the metaphor of the choreography of dance: 'stepping-in'; 'stepping-out'; 'stepping-on-toes'. [ABSTRACT FROM AUTHOR]

Immunotherapeutic strategies aimed at enhancing tumor cell killing by tumor-specific T cells hold great potential for reducing tumor burden and prolonging survival of cancer patients. Although many potential tumor antigens have been described, identifying relevant targets when designing anti-cancer vaccines or targeted cell therapies remains a challenge. To identify novel, potentially immunogenic candidate tumor antigens, we performed integrated tumor transcriptomic, seromic, and proteomic analyses of high grade serous ovarian cancer (HGSC) patient tumor samples. We identified tumor neo-antigens and over-expressed antigens using whole exome and RNA sequencing and examined these in relation to patient-matched auto-antibody repertoires. Focusing on MHC class I epitopes recognized by CD8+ T cells, HLA-binding epitopes were identified or predicted from the highly expressed, mutated, or auto-antibody target antigen, or MHC-associated peptides (MAPs). Recognition of candidate antigenic peptides was assessed within the tumor-infiltrating T lymphocyte (TIL) population expanded from each patient. Known tumor-associated antigens (TAA) and cancer/testis antigens (CTA) were commonly found in the auto-antibody and MAP repertoires and CD8+ TILs recognizing epitopes from these antigens were detected, although neither expression level nor the presence of auto-antibodies correlated with TIL recognition. Auto-antibodies against tumor-mutated antigens were found in most patients, however, no TIL recognition of the highest predicted affinity neo-epitopes was detected. Using high expression level, auto-antibody recognition, and epitope prediction algorithms, we identified epitopes in 5 novel antigens (MOB1A, SOCS3, TUBB, PRKAR1A, CCDC6) recognized by HGSC patient TILs. Furthermore, selection of epitopes from the MAP repertoire identified 5 additional targets commonly recognized by multiple patient TILs. We find that the repertoire of TIL specificities includes recognition of highly expressed and immunogenic self-antigens that are processed and presented by tumors. These results indicate an ongoing autoimmune response against a range of self-antigens targeted by HGSC TILs. [ABSTRACT FROM AUTHOR]

The closure of school buildings due to COVID-19 created a challenge for parents and teachers supporting children's remote learning. This paper presents findings of a study that explored whether parents of children with special educational needs and disabilities (SEND) experienced an unusually challenging period and what obstacles they faced. An online survey was sent to parents during the first (March–June 2020) and second (January–March 2021) lockdowns in England: a total of 141 voluntary participants responded. Thematic data analysis identified three significant themes: Infrastructure (Quality and efficacy of resources; Access to school's virtual learning environment); Impact on parent (Perceived lack of ability or understanding; Relationships; Time; Mental health); Impact on child (Reduced stress and anxiety; Need for routine). Recommendations for schools include collaborating with parents to ensure children with SEND achieve greater equality and inclusivity in educational provision, by developing blended models for in-school and remote learning. [ABSTRACT FROM AUTHOR]

Objective: Necrotizing enterocolitis (NEC) is the most common and lethal gastrointestinal disease affecting preterm infants. NEC develops suddenly and is characterized by gut barrier destruction, an inflammatory response, intestinal necrosis and multi-system organ failure. There is currently no method for early NEC detection, and the pathogenesis of NEC remains unclear. Design: To further understand the molecular mechanisms that support NEC, we used solution phase hybridization and next-generation DNA sequencing of bisulfite converted DNA to perform targeted genome-wide analysis of DNA methylation at high read depth. Results: We found that ileal samples from surgical NEC infants (n = 5) exist in a broadly hypermethylated state relative to their non-NEC counterparts (n = 9). These trends were not uniform, with hypermethylation being most consistently observed outside CpG islands and promoters. We further identified several biologically interesting gene promoters that displayed differential methylation in NEC and a number of biological pathways that appear dysregulated in NEC. We also found that DNA methylation patterns identified in ileal NEC tissue were correlated with those found and published previously in stool samples from NEC-affected infants. Conclusion: We confirmed that surgical NEC is associated with broad DNA hypermethylation in the ileum, and this may be detectable in stool samples of affected individuals. Thus, an epigenomic liquid biopsy of stool may have significant potential as a biomarker with respect to the diagnostic/predictive detection of NEC. Our findings, along with recent similar observations in colon, suggest that epigenomic dysregulation is a significant feature of surgical NEC. These findings motivate future studies which will involve the longitudinal screening of samples obtained prior to the onset of NEC. Our long-term goal is the development of novel screening, diagnostic and phenotyping methods for NEC. [ABSTRACT FROM AUTHOR]

The intellectual disability found in people with Down syndrome is associated with numerous changes in early brain development, including the proliferation and differentiation of neural progenitor cells (NPCs) and the formation and maintenance of myelin in the brain. To study how early neural precursors are affected by trisomy 21, we differentiated two isogenic lines of induced pluripotent stem cells derived from people with Down syndrome into brain-like and spinal cord-like NPCs and promoted a transition towards oligodendroglial fate by activating the Sonic hedgehog (SHH) pathway. In the spinal cord-like trisomic cells, we found no difference in expression of OLIG2 or NKX2.2, two transcription factors essential for commitment to the oligodendrocyte lineage. However, in the brain-like trisomic NPCs, OLIG2 is significantly upregulated and is associated with reduced expression of NKX2.2. We found that this gene dysregulation and block in NPC transition can be normalized by increasing the concentration of a SHH pathway agonist (SAG) during differentiation. These results underscore the importance of regional and cell type differences in gene expression in Down syndrome and demonstrate that modulation of SHH signaling in trisomic cells can rescue an early perturbed step in neural lineage specification. [ABSTRACT FROM AUTHOR]

This paper presents a unique perspective into challenges faced by kindergarten teachers in engaging children in Science, Technology, Engineering, Arts and Mathematics (STEAM) education in Vietnam. It reports on teachers' perspectives of an adaptation of immersive learning experiences (ILE), to develop understanding of pedagogical approaches to STEAM education in real-world contexts. The study adopted a qualitative and interpretivist methodology, engaging 105 teachers in informal and collaborative activities. Their perceptions were gathered via participatory tools; photographic images recorded how they engaged with ILE. Data analysis occurred using de Freitas's four-dimensional framework. The study responds to evidence that STEAM education is not prevalent in Vietnamese schools, and reflects on the top-down siloed approach ubiquitous within its early childhood education (ECE) sector. Findings indicate the necessity for a prolonged period of training using ILE that enables teachers' development of pedagogical approaches to STEAM education. Significantly, it provides new insight for a variety of stakeholders (including teacher trainers and policy makers) into the under-researched field of Vietnamese teachers' pedagogical approaches in ECE, and extends its findings to countries adopting more formalised approaches to education. [ABSTRACT FROM AUTHOR]

Whilst the importance of engaging with children's voices is now more widely recognized, there is still a dearth of representation of younger children's voices specifically. Visual methodological tools, such as photo-elicitation and photo-voice are used in research; however, there remains a lack of clarity about what they mean, how they are used, and whether they are effective in providing valuable insights into younger children's everyday worlds. This paper draws on the findings of two research projects utilizing the tools to engage with the voices of 56 children aged four to five years, in the classes of three schools in the UK. It identifies the differences between photo-elicitation and photo-voice and appraises their advantages and limitations when engaging with child voice. It explores whether through careful, critical and conscientious application, these tools might advance understandings of younger children's perceptions of matters affecting them. The definition of photo-elicitation is the insertion of a photograph by the researcher into a research interview to evoke information, feelings, and memories due to the photograph's particular form of representation; photo-voice is defined as a technique that enables people to record their own photographs. [ABSTRACT FROM AUTHOR]

In the digital era, we witness the increasing use of artificial intelligence (AI) to solve problems, while improving productivity and efficiency. Yet, inevitably costs are involved with delegating power to algorithmically based systems, some of whose workings are opaque and unobservable and thus termed the "black box". Central to understanding the "black box" is to acknowledge that the algorithm is not mendaciously undertaking this action; it is simply using the recombination afforded to scaled computable machine learning algorithms. But an algorithm with arbitrary precision can easily reconstruct those characteristics and make life-changing decisions, particularly in financial services (credit scoring, risk assessment, etc.), and it could be difficult to reconstruct, if this was done in a fair manner reflecting the values of society. If we permit AI to make life-changing decisions, what are the opportunity costs, data trade-offs, and implications for social, economic, technical, legal, and environmental systems? We find that over 160 ethical AI principles exist, advocating organisations to act responsibly to avoid causing digital societal harms. This maelstrom of guidance, none of which is compulsory, serves to confuse, as opposed to guide. We need to think carefully about how we implement these algorithms, the delegation of decisions and data usage, in the absence of human oversight and AI governance. The paper seeks to harmonise and align approaches, illustrating the opportunities and threats of AI, while raising awareness of Corporate Digital Responsibility (CDR) as a potential collaborative mechanism to demystify governance complexity and to establish an equitable digital society. [ABSTRACT FROM AUTHOR]

This paper presents findings from a research study, which sought to illuminate the perceived notions of inclusion of four to five year old children in pedagogical activities, in the early years classes of two schools in the North of England. It employed a qualitative methodology to gather extensive data with forty children over a six-week period in each school. This included collecting fieldnotes; undertaking observations of children in pedagogical activities; and conducting group and individual interviews. Central to the research aim was the use of participative tools to engage with children's voices; these included photographs and drawings. Children's perceived notions of inclusion resonated with two dimensions: belonging and relationships (with practitioner and/or child) and democratic pedagogies. The findings advance the conceptualisation of the notion of inclusion and bring to the fore the voices of a young group of children that has not been studied before. Engaging with children in meaningful ways can enable practitioners to better understand young children's perceived, multi-faceted notions of inclusion as they experience it within pedagogical activities. [ABSTRACT FROM AUTHOR]

Biological vision incorporates intelligent cooperation between the sensory and the motor systems, which is facilitated by the development of motor skills that help to shape visual information that is relevant to a specific vision task. In this article, we seek to explore an approach to active vision inspired by biological systems, which uses limited constraints for motor strategies through progressive adaptation via an evolutionary method. This type of approach gives freedom to artificial systems in the discovery of eye-movement strategies that may be useful to solve a given vision task but are not known to us. In the experiment sections of this article, we use this type of evolutionary active vision system for more complex natural images in both two-dimensional (2D) and three-dimensional (3D) environments. To further improve the results, we experiment with the use of pre-processing the visual input with both the uniform local binary patterns (ULBP) and the histogram of oriented gradients (HOG) for classification tasks in the 2D and 3D environments. The 3D experiments include application of the active vision system to object categorisation and indoor versus outdoor environment classification. Our experiments are conducted on the iCub humanoid robot simulator platform. [ABSTRACT FROM AUTHOR]

Objective: We explored the potential of genome‐wide epigenomic liquid biopsy for the comprehensive analysis of cell‐free DNA (cfDNA) methylation signatures in maternal plasma in early gestation. Method: We used solution phase hybridization for targeted region capture of bisulfite‐converted DNA obtained from plasma of pregnant women in early gestation and nonpregnant female controls. Results: Targeted sequencing of ~80.5 Mb of the plasma methylome generated an average read depth across all 17 plasma samples of ~42x. We used these data to explore the pregnancy‐specific characteristics of cfDNA methylation in plasma and found that pregnancy resulted in clearly detectable global alterations in DNA methylation patterns that were influenced by genomic location. We analyzed similar, previously published, data from first‐trimester maternal leukocyte populations and gestational age–matched chorionic villus (CV) and confirmed that tissue‐specific DNA methylation signatures in these samples had a significant influence on global and gene‐specific methylation in the plasma of pregnant women. Conclusion: We describe an approach for targeted epigenomic liquid biopsy in pregnancy and discuss our findings in the context of noninvasive prenatal testing with respect to phenotypic pregnancy monitoring and the early detection of complex gestational phenotypes such as preeclampsia and preterm birth. [ABSTRACT FROM AUTHOR]

Background: Necrotizing enterocolitis (NEC) remains one of the overall leading causes of death in premature infants, and the pathogenesis is unpredictable and not well characterized. The aim of our study was to determine the molecular phenotype of NEC via transcriptomic and epithelial cell-specific epigenomic analysis, with a specific focus on DNA methylation. Methods: Using laser capture microdissection, epithelial cell-specific methylation signatures were characterized by whole-genome bisulfite sequencing of ileal and colonic samples at the time of surgery for NEC and after NEC had healed at reanastomosis (n = 40). RNA sequencing was also performed to determine the transcriptomic profile of these samples, and a comparison was made to the methylome data. Results: We found that surgical NEC has a considerable impact on the epigenome by broadly increasing DNA methylation levels, although these effects are less pronounced in genomic regions associated with the regulation of gene expression. Furthermore, NEC-related DNA methylation signatures were influenced by tissue of origin, with significant differences being noted between colon and ileum. We also identified numerous transcriptional changes in NEC and clear associations between gene expression and DNA methylation. Conclusions: We have defined the intestinal epigenomic and transcriptomic signatures during surgical NEC, which will advance our understanding of disease pathogenesis and may enable the development of novel precision medicine approaches for NEC prediction, diagnosis and phenotyping. [ABSTRACT FROM AUTHOR]

Our goal was to undertake a genome‐wide epigenomic liquid biopsy of cerebrospinal fluid (CSF) for the comprehensive analysis of cell‐free DNA (cfDNA) methylation signatures in the human central nervous system (CNS). Solution‐phase hybridization and massively parallel sequencing of bisulfite converted human DNA was employed to compare methylation signatures of cfDNA obtained from CSF with plasma. Recovery of cfDNA from CSF was relatively low (68–840 pg/mL) compared to plasma (2720–8390 pg/mL) and cfDNA fragments from CSF were approximately 20 bp shorter than their plasma‐derived counterparts. Distributions of CpG methylation signatures were significantly altered between CSF and plasma, both globally and at the level of functional elements including exons, introns, CpG islands, and shores. Sliding window analysis was used to identify differentially methylated regions. We found numerous gene/locus‐specific differences in CpG methylation between cfDNA from CSF and plasma. These loci were more frequently hypomethylated in CSF compared to plasma. Differentially methylated CpGs in CSF were identified in genes related to branching of neurites and neuronal development. Using the GTEx RNA expression database, we found clear association between tissue‐specific gene expression in the CNS and cfDNA methylation patterns in CSF. Ingenuity pathway analysis of differentially methylated regions identified an enrichment of functional pathways related to neurobiology. In conclusion, we present a genome‐wide analysis of DNA methylation in human CSF. Our methods and the resulting data demonstrate the potential of epigenomic liquid biopsy of the human CNS for molecular phenotyping of brain‐derived DNA methylation signatures. [ABSTRACT FROM AUTHOR]

This paper develops understandings of how being publicly identified and consequently labelled as 'looked after' can have damaging consequences for young people, particularly in how they are perceived by their peers in the context of schooling. Based on qualitative research in northern England utilising participatory approaches with young people and interviews with support staff, we explore barriers that inhibit young people's sense of belonging. We highlight how the very processes and practices set up to support the young people can often have unintended consequences by routinely positioning them as Other, before considering the implications for education and schooling in particular. [ABSTRACT FROM AUTHOR]

Major depressive disorder (MDD) affects approximately 15 million Americans. Approximately 2 million of these are classified as being refractory to treatment (TR‐MDD). Because of the lack of available therapies for TR‐MDD, and the high risk of suicide, there is interest in identifying new treatment modalities and diagnostic methods. Understanding of the impact of genomic copy number variation in the etiology of a variety of neuropsychiatric phenotypes is increasing. Low copy repeat elements at 15q13.3 facilitate non‐allelic homologous recombination, resulting in recurrent copy number variants (CNVs). Numerous reports have described association between microdeletions in this region and a variety of neuropsychiatric phenotypes, with CHRNA7 implicated as a candidate gene. However, the pathogenicity of 15q13.3 duplications is less clear. As part of an ongoing study, in which we have identified a number of metabolomic anomalies in spinal fluid from TR‐MDD patients, we also evaluated genomic copy number variation in patients (n = 125) and controls (n = 26) via array‐based copy number genomic hybridization (CGH); the case frequency was compared with frequencies reported in a prior study as well as a larger population‐sized cohort. We identified five TR‐MDD patients with microduplications involving CHRNA7. CHRNA7 duplications are the most common CNVs identified by clinical CGH in this cohort. Therefore, this study provides insight into the potential involvement of CHRNA7 duplications in the etiology of TR‐MDD and informs those involved with care of affected individuals. [ABSTRACT FROM AUTHOR]

Background/objective: We communicate high-read-depth bisulfite sequencing analysis of the chorionic villus (CV) DNA methylome from samples obtained between 11 and 13 weeks gestation and samples of gestationally age-matched maternal blood cells (MBC).Methods: This was achieved through solution-phase targeted region capture (84 Mb) of bisulfite converted human DNA.Results: We identified biphasic distribution of methylation in CV and MBC genomes. We found greater numbers of intermediate methylated sites (20%-80% methylated) in CV and greater number of high methylation sites in MBC and investigated distributions of these in promoters, introns, exons, CpG islands, CpG islands shores, and enhancers. We identified differentially methylated sites distinguishing CV and MBC. These are less likely to occur in CpG islands (CGIs), particularly those that exist outside promoters, exons, and introns. We found that gene promoter and gene body methylation patterns are associated with mRNA transcriptional profiles in CV. Despite the relative hypomethylation of CV genomes, we found that these contain DM regions that are more likely to be hypermethylated in CV relative to MBC.Conclusions: Our data provide novel insight into the structure and organization of the CV epigenome, which may inform future studies of placental biology and noninvasive prenatal phenotyping. [ABSTRACT FROM AUTHOR]

Background: B7-H3 and B7-H4 are highly expressed by many human malignancies making them attractive immunotherapeutic targets. However, their expression patterns and immune contexts in epithelial ovarian cancer have not been well characterized. Methods: We used flow cytometry, immunohistochemistry, and genomic analyses to determine the patterns of B7-H3, B7-H4, and PD-L1 expression by tumor, stromal, and immune cells in the ovarian tumor microenvironment (TME). We analyzed immune cell frequency and expression of PD-1, TIM3, LAG3, ICOS, TIA-1, granzyme B, 2B4, CD107a, and GITR on T cells; CD20, CD22, IgD, BTLA, and CD27 on B cells; CD16 on monocytes; and B7-H3, B7-H4, PD-L1, PD-L2, ICOSL, CD40, CD86, and CLEC9a on antigen-presenting cells by flow cytometry. We determined intratumoral cellular location of immune cells using immunohistochemistry. We compared differences in immune infiltration in tumors with low or high tumor-to-stroma ratio and in tumors from the same or unrelated patients. Results: On non-immune cells, B7-H4 expression was restricted to tumor cells whereas B7-H3 was expressed by both tumor and stromal cells. Stromal cells of the ovarian TME expressed high levels of B7-H3 compared to tumor cells. We used this differential expression to assess the tumor-to-stroma ratio of ovarian tumors and found that high tumor-to-stroma ratio was associated with increased expression of CD16 by monocytes, increased frequencies of PD-1high CD8+ T cells, increased PD-L1 expression by APCs, and decreased CLEC9a expression by APCs. We found that expression of PD-L1 or CD86 on APCs and the proportion of PD-1high CD4+ T cells were strongly correlated on immune cells from tumors within the same patient, whereas expression of CD40 and ICOSL on APCs and the proportion of PD-1high CD8+ T cells were not. Conclusions: This study provides insight into the expression patterns of B7-H3 and B7-H4 in the ovarian TME. Further, we demonstrate an association between the tumor-to-stroma ratio and the phenotype of tumor-infiltrating immune cells. We also find that some but not all immune parameters show consistency between peritoneal metastatic sites. These data have implications for the design of immunotherapies targeting these B7 molecules in epithelial ovarian cancer. [ABSTRACT FROM AUTHOR]

B7-H4, an immune suppressive member of the B7 family, is highly expressed in a wide variety of human malignancies making it an attractive immunotherapeutic target. However, the association between B7-H4 expression in the tumor microenvironment and the immune infiltrate has not been comprehensively examined. To evaluate the immune tumor microenvironment, we analyzed epithelial ovarian tumors from 28 patients using flow cytometry, immunohistochemistry, functional, and genomic analyses. We determined B7-H4 expression patterns and compared the immune infiltrates of tumors with high and low surface expression of B7-H4. Frequencies and phenotypes of tumor and immune cells were determined using multiple flow cytometry panels. Immunohistochemistry was used to analyze cellular infiltration and location. Publicly available datasets were interrogated to determine intratumoral cytokine and chemokine expression. We found that B7-H4 was predominantly expressed by tumor cells in the epithelial ovarian tumor microenvironment. Surface expression of B7-H4 on tumor cells was correlated with higher levels of infiltrating mature antigen-presenting cells. Further, expression of CXCL17, a monocyte and dendritic cell chemoattractant, correlated strongly with B7-H4 expression. T cells expressed activation markers, but T cells expressing a combination of markers associated with T cell activation/exhaustion phenotype were not prevalent. Overall, our data suggest that B7-H4 is associated with a pro-inflammatory tumor microenvironment. [ABSTRACT FROM AUTHOR]

The paper seeks to extend the knowledge and understanding of how engaging with young children's voices in a meaningful way, can alter practitioners' pedagogical practice and thus create environments for learning that are more inclusive. It draws on the findings of a research study that explored practitioners' (teachers, nursery nurses and teaching assistants) views about engaging with young children's voices of perceived notions of inclusion in pedagogical activities in the early years classes of two schools in the North of England. It adopted a qualitative methodological approach that operates within constructivist and interpretivist paradigms. The study revealed that practitioners retain some resistance to responding to the voices of young children and that internal and external pressures influence their decision-making. Moreover, it signifies the necessity for greater emphasis on the importance of engaging with children's voices in the training of newly qualified teachers, and the ongoing professional development for all practitioners in early childhood education. [ABSTRACT FROM AUTHOR]

In light of the updated Eatwell Guide and the corresponding change in the consumption of fruit smoothies, the aim of this study was to measure the glycaemic index and load of two commercial fruit smoothies and to investigate the retention of dietary fibre following production. In vitro analysis was performed to identify fibre material (cellulose and pectins) using calcofluor staining and immunocytochemical labelling. A repeated measures cross-over study was conducted (n 10) to determine the glycaemic index (GI) and glycaemic load (GL) of the smoothies. Results showed that dietary fibre was still present in the smoothies after processing (16.9-17.5% cellular material by dry weight). The GI was low for both smoothies (39 and 36), whereas the GL was medium and borderline-low, respectively (11.4 and 9.7). The retention of fibre in these smoothies may have a potential positive effect on glycaemic response and may contribute to daily fibre requirements. [ABSTRACT FROM AUTHOR]

Higher aspirin doses may be inferior in ticagrelor-treated acute coronary syndrome (ACS) patients and reducing bleeding risk whilst maintaining antithrombotic benefits could improve outcomes. We characterized the pharmacodynamics of a novel dual-antiplatelet-therapy regimen consisting of very-low-dose twice-daily (BD) aspirin with standard-dose ticagrelor. A total of 20 ticagrelor-treated ACS patients entered a randomized crossover to take aspirin 20 mg BD (12-hourly) during one 14-day period and 75 mg once-daily (OD) in the other. After 14 days of treatment, serum thromboxane (TX)B2 and light-transmittance aggregometry were assessed pre- and 2 h post-morning-dose, bleeding time was measured post-dose, and TXA2 and prostacyclin stable metabolites were measured in urine collected 2 h post-morning-dose. Data are expressed as mean ± SD. After 14 days treatment, serum TXB2 levels were significantly greater 2 h post-dosing with aspirin 20 mg BD vs. 75 mg OD (3.0 ± 3.6 ng/mL vs. 0.8 ± 1.9 ng/mL; p = 0.018) whereas pre-dosing levels were not significantly different (3.5 ± 4.1 ng/mL vs. 2.5 ± 3.1 ng/mL, p = 0.23). 1-mmol/L arachidonic acid-induced platelet aggregation was similarly inhibited by both regimens pre-dose (8.5 ± 14.3% vs. 5.1 ± 3.6%, p = 0.24) and post-dose (8.7 ± 14.2% vs. 6.6 ± 5.3%; p = 0.41). Post-dose bleeding time was shorter with 20 mg BD (680 ± 306 s vs. 834 ± 386 s, p = 0.02). Urinary prostacyclin and TX metabolite excretion were not significantly different. In conclusion, compared to aspirin 75 mg OD, aspirin 20 mg BD provided consistent inhibition of platelet TXA2 release and aggregation, and improved post-dose hemostasis, in ticagrelor-treated ACS patients. Further studies are warranted to assess whether this regimen improves the balance of clinical efficacy and safety. [ABSTRACT FROM AUTHOR]

The landscape of genetic testing in ovarian cancer patients has changed dramatically in recent years. The therapeutic benefits of poly ADP-ribose polymerase (PARP) inhibitors in treatment of BRCA1/2-related ovarian cancers has resulted in an increased demand and urgency for genetic testing results, while technological developments have led to widespread use of multigene cancer panels and development of tumour testing protocols. Traditional genetic counselling models are no longer sustainable and must evolve to match the rapid evolution of genetic testing technologies and developments in personalized medicine. Recently, representatives from oncology, clinical genetics, molecular genetics, pathology, and patient advocacy came together to create a national multi-disciplinary Canadian consortium. By aligning stakeholder interests, the BRCA Testing to Treatment (BRCA TtoT) Community of Practice aims to develop a national strategy for tumour and germline BRCA1/2 testing and genetic counselling in women with ovarian cancer. This article serves to provide an overview of the recent evolution of genetic assessment for BRCA1/2-associated gynecologic malignancies and outline a Canadian roadmap to facilitate change, improve genetic testing rates, and ultimately improve outcomes for hereditary ovarian cancer patients and their families. [ABSTRACT FROM AUTHOR]

Examining the different stages of learning through play in humans during early life has been a topic of interest for various scholars. Play evolves from practice to symbolic and then later to play with rules. During practice play, infants go through a process of developing knowledge while they interact with the surrounding objects, facilitating the creation of new knowledge about objects and object related behaviors. Such knowledge is used to form schemas in which the manifestation of sensorimotor experiences is captured. Through subsequent play, certain schemas are further combined to generate chains able to achieve behaviors that require multiple steps. The chains of schemas demonstrate the formation of higher level actions in a hierarchical structure. In this work we present a schema-based play generator for artificial agents, termed Dev-PSchema. With the help of experiments in a simulated environment and with the iCub robot, we demonstrate the ability of our system to create schemas of sensorimotor experiences from playful interaction with the environment. We show the creation of schema chains consisting of a sequence of actions that allow an agent to autonomously perform complex tasks. In addition to demonstrating the ability to learn through playful behavior, we demonstrate the capability of Dev-PSchema to simulate different infants with different preferences toward novel vs. familiar objects. [ABSTRACT FROM AUTHOR]

Abstract: Uterine carcinosarcoma, also known as Malignant Mixed Müllerian Tumour, is a high‐grade biphasic neoplasm composed of sarcomatous elements thought to originate via transdifferentiation from high‐grade endometrial carcinoma. To identify molecular factors contributing to the histogenesis of this tumour, we analyzed DNA extracted from matched carcinoma and sarcoma components from 12 cases of carcinosarcoma by a molecular inversion probe microarray to assess genomic copy number alterations (CNAs) and allelic imbalances. Widespread CNAs were identified in tumours with serous histology in the carcinoma component (9/12), while the remaining three cases with endometrioid carcinoma were near‐diploid. Quantification of the extent of genomic aberrations revealed a significant increase in sarcoma relative to carcinoma in tumours with well‐delineated histologic components. Focal amplification of 13q31.3 was identified in 6/12 profiled tumours, of which four harboured the aberration exclusively in the sarcoma component. This result was verified by fluorescence in situ hybridization against GPC5, the only gene situated within the minimal region of amplification. In a validation cohort composed of 97 carcinosarcomas and other uterine sarcomas, amplification of GPC5 (GPC5/CEP13 ratio ≥ 2.2) was identified in 11/97 (11.3%) cases (9/64 carcinosarcoma, 1/3 rhabdomyosarcoma, 1/21 leiomyosarcoma, 0/8 adenosarcoma, 0/1 undifferentiated endometrial sarcoma) and an additional 4 (2.8%) cases had low level gains (GPC5/CEP13 ratio ≥1.5 but <2.2). The functional relevance of Glypican‐5, the gene product of GPC5, in regulating differentiation and lineage commitment was demonstrated in an endometrial carcinoma cell line in vitro. In conclusion, we identified GPC5 amplification as a molecular event mediating epithelial‐mesenchymal transdifferentiation in a subset of uterine carcinosarcomas. [ABSTRACT FROM AUTHOR]

Many high-grade serous carcinomas (HGSCs) of the pelvis are thought to originate in the distal portion of the fallopian tube. Serous tubal intra-epithelial carcinoma (STIC) lesions are the putative precursor to HGSC and identifiable in ~ 50% of advanced stage cases. To better understand the molecular etiology of HGSCs, we report a multi-center integrated genomic analysis of advanced stage tumors with and without STIC lesions and normal tissues. The most significant focal DNA SCNAs were shared between cases with and without STIC lesions. The RNA sequence and the miRNA data did not identify any clear separation between cases with and without STIC lesions. HGSCs had molecular profiles more similar to normal fallopian tube epithelium than ovarian surface epithelium or peritoneum. The data suggest that the molecular features of HGSCs with and without associated STIC lesions are mostly shared, indicating a common biologic origin, likely to be the distal fallopian tube among all cases. [ABSTRACT FROM AUTHOR]

A high proportion of ovarian cancers from women who carry germline mutations in mismatch repair (MMR) genes demonstrate microsatellite instability (MSI). The utility of pre-screening ovarian cancer specimens for MSI to identify potential patients for germline screening for MMR mutations is uncertain. 656 women with malignant ovarian cancer underwent both MSI testing and germline mutation testing for large rearrangements in three MMR genes, MLH1, MSH2 and MSH6. Germline DNA sequencing data for the same genes was available. Among the 656 women, only four (0.6%) carried a clearly pathogenic MMR mutation. All four cancers from patients with mutations had loss of two or more microsatellite markers (MSI-high). Eighty-four of 652 (13.0%) women without a mutation had MSI-high ovarian cancers. Using MSI-high as a prescreening criterion, the sensitivity of MSI testing to identify germline MMR gene mutations was 100% and the positive predictive value was 4.5%. Germline mutations in MLH1, MSH2 and MSH6 are rare among unselected cases of ovarian cancer. Patients with germline mutations often will have MSI-positive cancers and pre-screening of ovarian cancer specimens may be an efficient way of identifying patients with Lynch syndrome. [ABSTRACT FROM AUTHOR]

Minimally Invasive Karyotyping (MINK) was communicated in 2009 as a novel method for the non-invasive detection of fetal copy number anomalies in maternal plasma DNA. The original manuscript illustrated the potential of MINK using a model system in which fragmented genomic DNA obtained from a trisomy 21 male individual was mixed with that of his karyotypically normal mother at dilutions representing fetal fractions found in maternal plasma. Although it has been previously shown that MINK is able to non-invasively detect fetal microdeletions, its utility for aneuploidy detection in maternal plasma has not previously been demonstrated. The current study illustrates the ability of MINK to detect common aneuploidy in early gestation, compares its performance to other published third party methods (and related software packages) for prenatal aneuploidy detection and evaluates the performance of these methods across a range of sequencing read inputs. Plasma samples were obtained from 416 pregnant women between gestational weeks 8.1 and 34.4. Shotgun DNA sequencing was performed and data analyzed using MINK RAPIDR and WISECONDOR. MINK performed with greater accuracy than RAPIDR and WISECONDOR, correctly identifying 60 out of 61 true trisomy cases, and reporting only one false positive in 355 normal pregnancies. Significantly, MINK achieved accurate detection of trisomy 21 using just 2 million aligned input reads, whereas WISECONDOR required 6 million reads and RAPIDR did not achieve complete accuracy at any read input tested. In conclusion, we demonstrate that MINK provides an analysis pipeline for the detection of fetal aneuploidy in samples of maternal plasma DNA. [ABSTRACT FROM AUTHOR]

Aims Molecular investigation of small-cell carcinoma of the ovary, hypercalcaemic type ( SCCOHT) has revealed that it is a monogenetic tumour characterized by alteration of SMARCA4 ( BRG1), encoding a member of the switch/sucrose non-fermentable ( SWI/ SNF) chromatin remodelling complex. A large majority of cases show loss of expression of the corresponding SMARCA4/ BRG1 protein. Furthermore, three cases of SCCOHT with retained SMARCA4 protein expression showed loss of SMARCB1/ INI1 expression. The aim of this study was to assess the sensitivity and specificity of loss of SMARCA4 expression as a diagnostic test for SCCOHT. Methods and results We performed SMARCA4 and SMARCB1 staining in 245 tumours, many of which were potentially in the differential diagnosis of SCCOHT. We also stained 56 cases of SCCOHT for SMARCA4 and 37 of these for SMARCB1. Fifty-four of the SCCOHT cases showed complete absence of SMARCA4 expression. The two cases with retained expression showed molecular alteration of SMARCA4. Of the 217 other neoplasms with interpretable staining, all retained SMARCA4 expression. Although the majority showed diffuse, strong nuclear expression, a heterogeneous, typically weak staining pattern was present in 13% of cases. All 37 cases of SCCOHT tested and all other neoplasms, apart from three malignant rhabdoid tumours, showed retained nuclear SMARCB1 expression. Loss of SMARCA4 expression had a sensitivity of 96.55% and specificity of 100%. Conclusions Loss of SMARCA4 expression is sensitive and specific for SCCOHT. Although some mimics show heterogeneous expression, there is retention of nuclear staining in at least a part of the tumour; therefore, only complete loss of staining should be regarded as being supportive of SCCOHT. [ABSTRACT FROM AUTHOR]

Background: The clinical utility of molecular profiling of tumor tissue to guide treatment of patients with advanced solid tumors is unknown. Our objectives were to evaluate the frequency of genomic alterations, clinical "actionability" of somatic variants, enrollment in mutation-targeted or other clinical trials, and outcome of molecular profiling for advanced solid tumor patients at the Princess Margaret Cancer Centre (PM). Methods: Patients with advanced solid tumors aged ≥18 years, good performance status, and archival tumor tissue available were prospectively consented. DNA from archival formalin-fixed paraffin-embedded tumor tissue was tested using a MALDI-TOF MS hotspot panel or a targeted next generation sequencing (NGS) panel. Somatic variants were classified according to clinical actionability and an annotated report included in the electronic medical record. Oncologists were provided with summary tables of their patients' molecular profiling results and available mutationspecific clinical trials. Enrolment in genotype-matched versus genotype-unmatched clinical trials following release of profiling results and response by RECIST v1.1 criteria were evaluated. Results: From March 2012 to July 2014, 1893 patients were enrolled and 1640 tested. After a median follow-up of 18 months, 245 patients (15 %) who were tested were subsequently treated on 277 therapeutic clinical trials, including 84 patients (5 %) on 89 genotype-matched trials. The overall response rate was higher in patients treated on genotypematched trials (19 %) compared with genotype-unmatched trials (9 %; p < 0.026). In a multi-variable model, trial matching by genotype (p = 0.021) and female gender (p = 0.034) were the only factors associated with increased likelihood of treatment response. Conclusions: Few advanced solid tumor patients enrolled in a prospective institutional molecular profiling trial were treated subsequently on genotype-matched therapeutic trials. In this non-randomized comparison, genotypeenrichment of early phase clinical trials was associated with an increased objective tumor response rate. Trial registration: NCT01505400 (date of registration 4 January 2012). [ABSTRACT FROM AUTHOR]

Our goal was to test the hypothesis that inter-individual genomic copy number variation in control samples is a confounding factor in the non-invasive prenatal detection of fetal microdeletions via the sequence-based analysis of maternal plasma DNA. The database of genomic variants (DGV) was used to determine the “Genomic Variants Frequency” (GVF) for each 50kb region in the human genome. Whole genome sequencing of fifteen karyotypically normal maternal plasma and six CVS DNA controls samples was performed. The coefficient of variation of relative read counts (cv.RTC) for these samples was determined for each 50kb region. Maternal plasma from two pregnancies affected with a chromosome 5p microdeletion was also sequenced, and analyzed using the GCREM algorithm. We found strong correlation between high variance in read counts and GVF amongst controls. Consequently we were unable to confirm the presence of the microdeletion via sequencing of maternal plasma samples obtained from two sequential affected pregnancies. Caution should be exercised when performing NIPT for microdeletions. It is vital to develop our understanding of the factors that impact the sensitivity and specificity of these approaches. In particular, benign copy number variation amongst controls is a major confounder, and their effects should be corrected bioinformatically. [ABSTRACT FROM AUTHOR]

This paper reports a small-scale research project which took place in one primary school in the north-east of England. The study aimed to listen to children’s views about how the practices of teachers helped and/or hindered their sense of inclusion in classrooms. Inclusion was understood here in a broad sense rather than specifically relating to children with special educational needs. Participatory research tools were used as part of group interviews with children from three different year groups. Even though the children were mostly happy with their school experience, it was noticeable that there were some areas for concern for some children that related to four interconnecting themes: unfairness, shouting, loneliness and seating plans. All of these themes seemed to be connected with children’s interpersonal relationships – with teachers and with each other – and can be seen as crucial in terms of understanding inclusion in schools and further developing existing practices. [ABSTRACT FROM PUBLISHER]

Objective Previously, it has been shown that KIF14 mRNA is overexpressed in ovarian cancer (OvCa), regardless of histological subtype. KIF14 levels are independently predictive of poor outcome and increased rates of recurrence in serous OvCa patients. Furthermore, it has been shown that KIF14 also controls the in vivo tumorigenicity of OvCa cell lines. In this study, we evaluate the potential of KIF14 as a therapeutic target through selective inhibition of KIF14 in primary high-grade serous patient-derived OvCa cells. Methods To assess the dependence of primary serous OvCa cultures on KIF14, protein levels in 11 prospective high grade serous ovarian cancer samples were increased (KIF14 overexpression by transfection) or decreased (anti-KIF14 shRNA) in vitro, and proliferative capacity, anchorage independence and xenograft growth were assessed. Results Seven of eleven samples demonstrated increased/decreased in vitro proliferation in response to KIF14 overexpression/knockdown, respectively. When examining in vitro tumorigenicity (colony formation) and in vivo growth (subcutaneous xenografts) in response to KIF14 manipulation, none of the samples demonstrated growth in soft agar (11 samples), or xenograft growth (4 samples). Conclusions Although primary high-grade serous OvCa cells may depend on KIF14 for in vitro proliferation we were unable to demonstrate a role for KIF14 on tumorigenicity or develop an in vivo model for assessment. We have, however developed an effective in vitro method to evaluate the effect of target gene manipulation on the proliferative capacity of primary OvCa cultures. [ABSTRACT FROM AUTHOR]