Development Function Research Articles

Single cell RNAseq analysis reveals profound abnormalities in the distribution and diversity of thymic epithelial cells in Rag1 mutant mice

Abstract The thymus contains a heterogeneous population of stromal cells which orchestrate T cell development and selection. Recent works highlighted the complexity of thymic stromal cells, and medullary thymic epithelial cells (mTECs) in particular. However, the developmental origin, hierarchy, and function of these subpopulations remain ill-defined. Moreover, the characterization of cortical TECs (cTECs) has been largely restricted to the adult thymus. Here, we compared TEC distribution and gene expression in wild-type (WT) and in mice carrying Rag1 hypomorphic mutations observed in patients with immune deficiency and immune dysregulation. Single cell RNA-seq analysis of TECs isolated from adult Rag1 mutant mice revealed an excess of cTECs, which segregated in different clusters. The mTEC compartment, albeit reduced, showed a similar distribution of previously described subsets (mTEC I–IV), suggesting perturbation of mTEC development rather than mTEC differentiation into functional subsets. To address whether such abnormalities of cTEC and mTEC abundance and subset distribution in Rag1 mutant mice may reflect defects in TEC development, we extended scRNA-seq analysis to TECs from WT mice of neonatal age. A similar pattern was observed in TECs of adult Rag1 mutant mice and WT mice of neonatal age, indicating altered TEC development when Rag1 activity is perturbed. This defect correlates with a decreased number of double-positive thymocytes, suggesting that impaired lymphostromal cross-talk in the thymus of Rag1 mutant mice (and patients with similar defects) is associated with abnormalities of TEC composition which may contribute to altered immune tolerance that is often observed in these conditions.

A functionally informed evolutionary framework for the study of LRR-RLKs during stem cell maintenance.

Plants maintain populations of stem cells to generate new organs throughout the course of their lives. The pathways that regulate plant stem cell maintenance have garnered great interest over the past decades, as variation in these pathways contributes plant morphological diversity and can be harnessed for crop improvement. In order to facilitate cross-species comparisons of gene function and better understand how these stem cell regulatory pathways evolved, we undertook a functionally informed phylogenetic analysis of leucine-rich receptor-like kinases (LRR-RLK) and related proteins across diverse land plant model systems. Based on our phylogenetic analysis and on functional data, we propose a naming scheme for these stem cell signaling genes. We discovered evidence for frequent loss of protein domains in angiosperms but not in bryophytes. In addition, several clades of stem cell signaling genes are closely related to genes that function in immunity, although these distinct developmental and immune functions likely separated or after the divergence of lycophytes and angiosperms. Overall, the phylogenetic framework and evolutionary hypotheses we provide here will empower future research on cross-species comparisons of stem cell signaling pathways.

Polycomb group RING finger protein 5 influences several developmental signaling pathways during the in vitro differentiation of mouse embryonic stem cells.

Polycomb group (PcG) RING finger protein 5 (PCGF5) is a core component of the so-called Polycomb repressive complex 1.5 (PRC1.5), which is involved in epigenetic transcriptional repression. To explore the developmental function of Pcgf5, we generated Pcgf5 knockout (Pcgf5-/- ) mouse embryonic stem cell (mESC) lines with the help of CRISPR/Cas9 technology. We subjected the Pcgf5-/- and wild-type (WT) mESCs to a differentiation protocol toward mesodermal-cardiac cell types as aggregated embryoid bodies (EBs) and we found that knockout of Pcgf5 delayed the generation of the three germ layers, especially the ectoderm. Further, disruption of Pcgf5 impacted the epithelial-mesenchymal transition during EB morphogenesis and differentially affected the gene expression of essential developmental signaling pathways such as Nodal and Wnt. Finally, we also unveiled that loss of Pcgf5 induced the repression of genes involved in the Notch pathway, which may explain the enhancement of cardiomyocyte maturation and the dampening of ectodermal-neural differentiation observed in the Pcgf5-/- EBs.

A zebrafish hox gene acts before gastrulation to specify the hemangioblast.

Hox genes encode transcription factors that have been implicated in embryonic, adult and disease processes. The earliest developmental program known to be directed by Hox genes is the timing of ingression of presumptive axial mesoderm during gastrulation. We previously used morpholino (MO)-based knockdown to implicate the zebrafish hoxd4a gene in the specification of the hemangioblast, an event occurring at pre-gastrulation stages, well before the earliest known Hox gene function. The precise time at which hoxd4a function is required for this specification is not defined. We therefore fused the hoxd4a coding region to the human estrogen receptor (hERT2 ). Following co-injection of anti-hoxd4a MO with mRNA encoding the Hoxd4a-ERT2 fusion protein, hemangioblast specification was fully rescued when embryos were exposed to the estrogen analog 4-hydroxy-tamoxifen (4-OHT) at 4 hr post-fertilization (hpf), but only poorly at 6 hpf and not at all at 8 hpf, thereby defining a pre-gastrulation role for Hoxd4a, the earliest developmental function of a vertebrate Hox gene so far described. Both DNA binding and interaction with cofactor Pbx were further shown to be required for rescue of the morphant phenotype. Confirmation of the morphant phenotype was sought via the generation of hoxd4a null mutants using CRISPR/Cas9 technology. Null mutants of hoxd4a up to the third generation (F3 ) failed to recapitulate the morphant phenotype, and were largely refractory to the effects of injected anti-hoxd4a MO suggesting the action of genetic compensation.

A pan-cancer study of class-3 semaphorins as therapeutic targets in cancer

BackgroundInitially characterized as axon guidance factors, semaphorins also have been implicated to have critical roles in multiple physiological and developmental functions, including the regulation of immune responses, angiogenesis, organ formation, and the etiology of multiple forms of cancer. Moreover, their contribution in immunity and the regulation of tumour microenvironment is becoming increasingly recognized. Here, we provide a comprehensive analysis of class-3 semaphorins, the only secreted family of genes among veterbrate semaphorins, in terms of their expression profiles and their association with patient survival. We also relate their role with immune subtypes, tumour microenvironment, and drug sensitivity using a pan-cancer study.ResultsExpression profiles of class-3 semaphorins (SEMA3s) and their association with patient survival and tumour microenvironment were studied in 31 cancer types using the TCGA pan-cancer data. The expression of SEMA3 family varies in different cancer types with striking inter- and intra- cancer heterogeneity. In general, our results show that SEMA3A, SEMA3C, and SEMA3F are primarily upregulated in cancer cells, while the rest of SEMA3s are mainly down-regulated in the tested tumours. The expression of SEMA3 family members was frequently associated with patient overall survival. However, the direction of the association varied with regards to the particular SEMA3 isoform queried and the specific cancer type tested. More specifically, SEMA3A and SEMA3E primarily associate with a poor prognosis of survival, while SEMA3G typically associates with survival advantage. The rest of SEMA3s show either survival advantage or disadvantage dependent on cancer type. In addition, all SEMA3 genes show significant association with immune infiltrate subtypes, and they also correlate with level of stromal cell infiltration and tumour cell stemness with various degrees. Finally, our study revealed that SEMA3 genes, especially SEMA3C and SEMA3F may contribute to drug induced cancer cell resistance.ConclusionsOur systematic analysis of class-3 semaphorin gene expression and their association with immune infiltrates, tumour microenvironment and cancer patient outcomes highlights the need to study each SEMA3 member as a separate entity within each specific cancer type. Also our study validated the identification of class-3 semaphorin signals as promising therapeutic targets in cancer although further laboratory validation still needed.

Spatiotemporal Patterning of ABC Multidrug‐Resistance Transporters During Germ Layer Specification

Xenobiotic transporters are important for the protection of differentiated somatic cells, yet less is known about the role of these proteins in rapidly changing embryonic cells. A surprisingly large number of ABC transporters (80%) are expressed from fertilized egg to gastrulation, but their spatial and temporal patterns of expression during early embryogenesis remain incompletely understood. Here I report a pattern of ABC transporter expression that emerges during the early differentiation of mesoderm, endoderm, and ectoderm of the sea urchin. These included orthologs of six major xenobiotic transporters, including ABCB1, ABCB4, ABCC1, ABCC4, ABCC5, and ABCG2. In‐situ hybridization (ISH) revealed that transporters followed one of three patterns of early developmental expression: The first, seen with protective and homeostatic genes such as ABCB1 and ABCC1, was the early and ubiquitous expression across all three germ layers, and later enrichment in ectoderm‐derived sensory and feeding structures of the larva. The second, seen with the transporters ABCC4 and ‐C5 that are known to transport signal molecules, is a spatially restricted expression domain within a ring of mesodermal cells of the early blastula; ABCC4 then becomes restricted to primordial germ cells (PGCs) and their embryonic niche during gastrulation. Third, ABCB4 and ABCG2 show expression only in endoderm fated cells, particularly in the hindgut (presumptive intestines) at the onset of gut development; both genes eventually expand to the midgut region (presumptive stomach). To further refine the ISH map, I used fluorescent substrates to assay the functional outputs of transporter mRNA localization. ABCC1 and ‐C4 specific substrates are strongly effluxed out of the ring of mesodermal cells. This overlaps with robust ABCC5 transport activity in blastulae, suggesting a potential ABCC‐mediated signaling center in early mesoderm, where the PGCs are born. In the endoderm, ABCB4 and ABCG2‐specific efflux activity begins several days before the larva feeds. Collectively, this work is the first systems‐level description of the emergence of xenobiotic transporter expression patterns across the germ layers during embryogenesis. The resulting map of transport reveals unanticipated developmental functions for these genes and provides a framework for understanding the genetic regulatory controls of transporter expression. This will have additional impact on predicting and mitigating the effects of xenobiotic exposures during vulnerable windows of early development.Support or Funding InformationSupported by NIH R01ES027921 and R01ES030318 to AH, and NIH F32ES029843 to CSS.

Transcriptome analysis of the response provided by Lasiopodomys mandarinus to severe hypoxia includes enhancing DNA repair and damage prevention

BackgroundSevere hypoxia induces a series of stress responses in mammals; however, subterranean rodents have evolved several adaptation mechanisms of energy metabolisms and O2 utilization for hypoxia. Mammalian brains show extreme aerobic metabolism. Following hypoxia exposure, mammals usually experience irreversible brain damage and can even develop serious diseases, such as hypoxic ischemic encephalopathy and brain edema. To investigate mechanisms underlying the responses of subterranean rodents to severe hypoxia, we performed a cross-species brain transcriptomic analysis using RNA sequencing and identified differentially expressed genes (DEGs) between the subterranean rodent Lasiopodomys mandarinus and its closely related aboveground species L. brandtii under severe hypoxia (5.0% O2, 6 h) and normoxia (20.9% O2, 6 h).ResultsWe obtained 361 million clean reads, including 69,611 unigenes in L. mandarinus and 69,360 in L. brandtii. We identified 359 and 515 DEGs by comparing the hypoxic and normoxia groups of L. mandarinus and L. brandtii, respectively. Gene Ontology (GO) analysis showed that upregulated DEGs in both species displayed similar terms in response to severe hypoxia; the main difference is that GO terms of L. brandtii were enriched in the immune system. However, in the downregulated DEGs, GO terms of L. mandarinus were enriched in cell proliferation and protein transport and those of L. brandtii were enriched in nuclease and hydrolase activities, particularly in terms of developmental functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that upregulated DEGs in L. mandarinus were associated with DNA repair and damage prevention as well as angiogenesis and metastasis inhibition, whereas downregulated DEGs were associated with neuronal synaptic transmission and tumor-associated metabolic pathways. In L. brandtii, upregulated KEGG pathways were enriched in the immune, endocrine, and cardiovascular systems and particularly in cancer-related pathways, whereas downregulated DEGs were associated with environmental information processing and misregulation in cancers.ConclusionsL. mandarinus has evolved hypoxia adaptation by enhancing DNA repair, damage prevention, and augmenting sensing, whereas L. brandtii showed a higher risk of tumorigenesis and promoted innate immunity toward severe hypoxia. These results reveal the hypoxic mechanisms of L. mandarinus to severe hypoxia, which may provide research clues for hypoxic diseases.

Experimental Demyelination of the Lateral Olfactory Tract and Anterior Commissure

Demyelination significantly affects brain function. Several experimental methods, each inducing varying levels of myelin and neuronal damage, have been developed to understand the process of myelin loss and to find new therapies to promote remyelination. The present work investigates the effect of one such method, lysolecithin administration, on the white matter tracts in the olfactory system. The olfactory forebrain contains two distinct tracts with differing developmental histories, axonal composition, and function: the lateral olfactory tract (LOT), which carries ipsilateral olfactory information from the olfactory bulb to olfactory cortex, and the anterior commissure (AC), which interconnects olfactory regions across hemispheres. The effects of lysolecithin injections were assessed in two ways: (1) the expression of myelin basic protein, a component of compacted myelin sheaths, was quantified using immunohistochemistry and (2) electron microscopy was used to obtain measurements of myelin thickness of individual axons as well as qualitative descriptions of the extent of damage to myelin and surrounding tissue. Data were collected at 7, 14, 21, and 30 days post-injection (dpi). While both the LOT and AC exhibited significant demyelination at 7 dpi and had returned to control levels by 30 dpi, the process differed between the two tracts. Remyelination occurred more rapidly in the LOT: substantial recovery was observed in the LOT by 14 dpi, but not in the AC until 21 dpi. The findings indicate that (a) the LOT and AC are indeed suitable tracts for studying lysolecithin-induced de- and remyelination and (b) experimental demyelination proceeds differently between the two tracts.

PiRNAs Regulated by Mitochondria Variation Linked With Reproduction and Aging in Caenorhabditis elegans.

In Caenorhabditis elegans, the binding of Piwi protein to a non-coding RNA form, called piRNA, has been found to be important to both reproductive and aging processes. As the biosynthesis of piRNA is modulated by mitochondrial function, it is likely that the interaction between mitochondrial function and piRNA expression plays an unknown, yet important, role in reproductive and aging processes because both processes are known to be affected by declines in mitochondrial quality and activity. While the relationship between reproduction and longevity is not characterized in full, the optimality theory of aging and the disposable soma theory suggest that a trade-off between energy and resources is needed for reproductive and aging maintenance. In this study, the influence of mitochondrial variations, via a respiratory chain complex IV (COX1) polymorphism, on piRNA expression was examined in relation to the reproductive and aging outcomes of C. elegans. The COX1 polymorphism in mitochondria was found to affect the number of piRNAs expressed, the development of germ cells, and the length of the lifespan of the nematodes. Interestingly, more than two-thirds of the piRNA expression changes associated with the mitochondrial variation were found to also be affected by age. A gene ontology analysis of the altered piRNA species found that the piRNAs affected by mitochondrial variation and age were linked to genes known to have roles in reproductive and developmental function. Moreover, a piRNA-lncRNA-mRNA regulatory network based on the differential expression patterns of piRNA related to the mitochondrial variation was constructed to further identify potential gene targets with functional interactions. Similarly, this network identified genes involved in reproduction, development, and aging processes. These findings provide new insight into understanding how mitochondrial variations may regulate piRNA expression and may influence the underlying molecular mechanisms that affect reproduction and aging.

Comparative Transcriptomic Analyses of Developing Melanocortin Neurons Reveal New Regulators for the Anorexigenic Neuron Identity.

Despite their opposing actions on food intake, POMC and NPY/AgRP neurons in the arcuate nucleus of the hypothalamus (ARH) are derived from the same progenitors that give rise to ARH neurons. However, the mechanism whereby common neuronal precursors subsequently adopt either the anorexigenic (POMC) or the orexigenic (NPY/AgRP) identity remains elusive. We hypothesize that POMC and NPY/AgRP cell fates are specified and maintained by distinct intrinsic factors. In search of them, we profiled the transcriptomes of developing POMC and NPY/AgRP neurons in mice. Moreover, cell-type-specific transcriptomic analyses revealed transcription regulators that are selectively enriched in either population, but whose developmental functions are unknown in these neurons. Among them, we found the expression of the PR domain-containing factor 12 (Prdm12) was enriched in POMC neurons but absent in NPY/AgRP neurons. To study the role of Prdm12 in vivo, we developed and characterized a floxed Prdm12 allele. Selective ablation of Prdm12 in embryonic POMC neurons led to significantly reduced Pomc expression as well as early-onset obesity in mice of either sex that recapitulates symptoms of human POMC deficiency. Interestingly, however, specific deletion of Prdm12 in adult POMC neurons showed that it is no longer required for Pomc expression or energy balance. Collectively, these findings establish a critical role for Prdm12 in the anorexigenic neuron identity and suggest that it acts developmentally to program body weight homeostasis. Finally, the combination of cell-type-specific genomic and genetic analyses provides a means to dissect cellular and functional diversity in the hypothalamus whose neurodevelopment remains poorly studied.SIGNIFICANCE STATEMENT POMC and NPY/AgRP neurons are derived from the same hypothalamic progenitors but have opposing effects on food intake. We profiled the transcriptomes of genetically labeled POMC and NPY/AgRP neurons in the developing mouse hypothalamus to decipher the transcriptional codes behind the versus orexigenic neuron identity. Our analyses revealed 29 transcription regulators that are selectively enriched in one of the two populations. We generated new mouse genetic models to selective ablate one of POMC-neuron enriched transcription factors Prdm12 in developing and adult POMC neurons. Our studies establish a previously unrecognized role for Prdm12 in the anorexigenic neuron identity and suggest that it acts developmentally to program body weight homeostasis.

Notch3 in Development, Health and Disease.

Notch3 is one of four mammalian Notch proteins, which act as signalling receptors to control cell fate in many developmental and adult tissue contexts. Notch signalling continues to be important in the adult organism for tissue maintenance and renewal and mis-regulation of Notch is involved in many diseases. Genetic studies have shown that Notch3 gene knockouts are viable and have limited developmental defects, focussed mostly on defects in the arterial smooth muscle cell lineage. Additional studies have revealed overlapping roles for Notch3 with other Notch proteins, which widen the range of developmental functions. In the adult, Notch3, in collaboration with other Notch proteins, is involved in stem cell regulation in different tissues in stem cell regulation in different tissues, and it also controls the plasticity of the vascular smooth muscle phenotype involved in arterial vessel remodelling. Overexpression, gene amplification and mis-activation of Notch3 are associated with different cancers, in particular triple negative breast cancer and ovarian cancer. Mutations of Notch3 are associated with a dominantly inherited disease CADASIL (cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy), and there is further evidence linking Notch3 misregulation to hypertensive disease. Here we discuss the distinctive roles of Notch3 in development, health and disease, different views as to the underlying mechanisms of its activation and misregulation in different contexts and potential for therapeutic intervention.

The use of music therapy practices in the prevention of occupational diseases pianists

For 10 years the author has been dealing with the theme of the relationship between music and medicine; in her research and publications she focuses on the health and developmental functions of music; touches upon the little-studied topic - the need for interaction between specialists of musical art and medicine; and actively develops methods for the prevention of occupational diseases of pianists.

Trim45 is essential to the development of the diencephalon and eye in zebrafish embryos

ABSTRACT Trim45 is one of the RING (really interesting new gene) finger containing E3 ligase, which belongs to TRIM (Tripartite motif) protein family. Its molecular biological functions have been well characterized but not in light of developmental aspects. Here, we are reporting its expression patterns and developmental functions in zebrafish embryos. First, maternal transcripts of trim45 were found at one cell stage while its zygotic messages appeared at 30% epiboly. trim45 transcripts were restricted to the optical tectum, hypothalamus, hindbrain, and pharyngeal endoderm at 24 hpf (hour post-fertilization), and further to the retinal ganglion cell layer and cranial ganglion at 36 hpf. Second, ectopic expression of trim45 by injecting its mRNAs into embryos at one cell stage caused significant expansion of the diencephalon and eye fields at 24 hpf. In contrast, knock-down of trim45 with anti-sense trim45 morpholinos reduced the size of the two tissues at 24 hpf. Finally, the spatial distribution of the transcripts from olig2 and rx1/rx3, markers for the midbrain and eye respectively, were significantly decreased in the thalamus and eye fields respectively at 24 hpf. Based upon these observations, we proposed possible roles of Trim45 in the development of the diencephalon and eye in zebrafish embryos.

Intracellular Enhancement of BMP Signaling by LIM-Domain Protein FHL3 Controls Spatiotemporal Emergence of the Neural Crest Driven by WNT Signaling

How multiple morphogens are coordinated in space and time to position key embryonic tissues remains elusive. During neural crest formation, bone morphogenetic protein (BMP), fibroblast growth factor (FGF) and WNT signaling cooperate by acting either on the paraxial mesoderm or directly on the neural border ectoderm, but how each tissue interprets this complex information remains poorly understood. Here we show that Fhl3, a scaffold LIM domain protein of previously unknown developmental function, is essential for neural crest formation by linking BMP and WNT signaling thereby positioning the neural crest-inducing signaling center in the paraxial mesoderm. During gastrulation, Fhl3 promotes Smad phosphorylation and Smad-dependent wnt8 activation in the paraxial mesoderm, thus enhancing the response of mesoderm compared to ectoderm although both germ layers are subjected to a common extracellular BMP gradient. This promotes a non-cell autonomous WNT signaling ensuring neural border ectoderm specification by the underlying mesoderm. During neurulation, in turn, neural crest inducers activate fhl3 in the ectoderm, promoting BMP/Smad-dependent WNT activity required for neural crest specification. We show that Fhl3 binds Smads downstream of BMP signals and promotes Smad binding to wnt8 regulatory elements. Our findings highlight how Fhl3, acting cell-autonomously, ensures a fine spatial and temporal coordination of BMP and WNT signaling at several steps of neural crest development.

The relationship between chronic paediatric feeding disorders and caregiver stress.

Paediatric feeding disorders (PFDs) are common. Earlier studies have shown an association between PFD and caregiver stress, but these studies have been hampered by insufficient power. This study reports stress for caregivers of children diagnosed with PFD. These caregivers were then compared to community norms of the Parental Stress Index-Short Form. Caregivers also completed the Mealtime Behaviour Questionnaire, Child Behaviour Checklist and Child Development Inventory. Linear regression and hierarchical regression analyses assessed the relationship among the variables. Caregivers of 840 children with PFD responded. Negative child behaviours and lower child developmental function predicted higher levels of total parenting stress, parental distress, parent-child dysfunctional interaction and caregiver perceptions of their child as difficult. Higher rates of child internalizing and externalizing problems and child mealtime aggression were associated with parent perceptions of their child as difficult and contributed to total parental stress. Parents of older children reported higher levels of stress, whereas parents of younger children were more likely to be defensive responders. Overall, parents of children with PFD have more caregiver-related stress. These caregivers may benefit from consultations with mental health professionals to provide the most appropriate care to affected families.

Efficiency and accuracy of visual search develop at different rates from early childhood through early adulthood

Most studies of visual search across the life span have focused on classic feature and conjunction searches in which observers search for a fixed, simple shape target among relatively homogeneous distractors over a block of multiple trials. In the present study, we examine a more realistic task in which participants (4 to 25 years-old) look for images of real objects, presented amongst a heterogeneous array of other objects. The target is unique on every trial, unlike in previous developmental studies of visual search. Our new touchscreen-based “Pirate-Treasure” search also allows the testing of younger children within a video-game-like task. With this method, we tested a large sample (n = 293) of typically developing children and young adults. We assessed the developmental course of different search metrics like search efficiency, motor response differences, and accuracy (misses and false-alarm errors). Results show the most rapid time courses in development for accuracy. Search slopes reach the young adult level most slowly. The intercepts of the Reaction Time (RT) × Set Size function are often attributed to nonsearch perceptual and motor components of the task. The intercept time course is intermediate between accuracy and slope. Interestingly, these developmental functions follow time courses proposed in neuropsychological models of executive function development. This suggests that a single, video-game-like search task could be useful in routine assessments of cognitive development.

Bookmarking by Non-pioneer Transcription Factors during Liver Development Establishes Competence for Future Gene Activation

SummaryTranscription factor binding to enhancer and promoter regions critical for homeostatic adult gene activation is established during development. To understand how cell-specific gene expression patterns are generated, we study the developmental timing of association of two prominent hepatic transcription factors with gene regulatory regions. Most individual binding events display extraordinarily high temporal variations during liver development. Early and persistent binding is necessary, but not sufficient, for gene activation. Stable gene expression patterns are the result of combinatorial activity of multiple transcription factors, which mark regulatory regions long before activation and promote progressive broadening of active chromatin domains. Both temporally stable and dynamic, short-lived binding events contribute to the developmental maturation of active promoter configurations. The results reveal a developmental bookmarking function of master regulators and illuminate remarkable parallels between the principles employed for gene activation during development, during evolution, and upon mitotic exit.

The impact of entrustment assessments on feedback and learning: Trainee perspectives.

Assessment for and of learning in workplace settings is at the heart of competency-based medical education. In postgraduate medical education (PGME), entrustable professional activities (EPAs) and entrustment scales are increasingly used to assess competence. However, the educational impacts of these assessment approaches remain unknown. Therefore, this study aimed to explore trainee perceptions regarding the impacts of EPAs and entrustment scales on feedback and learning processes in the clinical setting. Four focus groups were conducted with postgraduate trainees in anaesthesia, emergency medicine, general internal medicine and nephrology at McMaster University in Hamilton, Ontario, Canada. Data collection and analysis were informed by principles of constructivist grounded theory. Entrustable professional activities representing well-defined tasks are perceived as potentially effective drivers for feedback and learning. Use of EPAs and entrustment scales, however, may augment existing tensions between developmental (for learning) and decision-making (of learning) assessment functions. Three key dilemmas seem to influence the impact of EPA-based assessment approaches on residents' learning: (a) standardisation of outcomes versus flexibility in assessment to align with individual learning experiences; (b) assessment tasks focusing on performance standards versus opportunities for learning, and (c) feedback focusing on numeric entrustment scores versus narrative and dialogue. Use of entrustment as an assessment outcome may impact trainees' motivation and feelings of self-efficacy, further enhancing tensions between learning and performance. Entrustable professional activities and entrustment scales may support assessment for learning in PGME. However, their successful implementation requires the careful management of dilemmas that arise in EPA-based assessment in order to support competence development.

Women’s Developmental Networks and Career Satisfaction: Developmental Functions as a Mediator

The current research empirically examined the developmental networks of female workers and how the network configurations influence their subjective career success, with particular attention to the gender and status composition of the network along with other network characteristics. Data from 427 female workers with a college degree or higher in the Republic of Korea were analyzed using egocentric network analysis and structural equation modeling methods. The findings suggested that (a) the network characteristics of intimacy and network size showed a direct positive effect on career satisfaction, while contact frequency and network range showed a direct negative effect; (b) intimacy, network size, and developers’ status indirectly affected career satisfaction mediated by the function of role modeling, while career support and psychosocial support played no mediating role between network characteristics and career satisfaction; and (c) women benefited from relationships with higher status developers, regardless of their gender, mainly through enhanced role modeling. Based on these, practical implications and recommendations for future research were presented.

Indole-3-lactic acid, a metabolite of tryptophan, secreted by Bifidobacterium longum subspecies infantis is anti-inflammatory in the immature intestine.

Background:Necrotizing enterocolitis (NEC), a necrotic inflammation of the intestine, represents a major health problem in the very premature infant. Although prevention is difficult, the combination of ingestion of maternal-expressed breastmilk in conjunction with a probiotic provides the best protection. In this study, we establish a mechanism for breastmilk/probiotic protection.Methods:Ultra-high-performance liquid chromatography-tandem mass spectrometry of B. infantis secretions was used to identify an anti-inflammatory molecule. Indole-3-lactic acid (ILA) was then tested in an established human immature small intestinal cell line, necrotizing colitis enterocytes and other immature human enteroids for anti-inflammatory effects and to establish developmental function. ILA was also examined in immature and mature enterocytes.Results:We have identified indole-3-lactic acid, a metabolite of breastmilk tryptophan, as the anti-inflammatory molecule. This molecule is developmentally functional in immature but not mature intestinal enterocytes, ILA reduces the IL-8 response after IL-1β stimulus. It interacts with the transcription factor aryl hydrocarbon receptor (AHR) and prevents transcription of the inflammatory cytokine IL-8.Conclusions:This molecule produced by B. infantis (ATCC No. 15697) interaction with ingested breastmilk functions in a complementary manner and could become useful in the treatment of all at risk premature infants for necrotizing enterocolitis if safety and clinical studies are performed.