Display omitted
We recently constructed a multicellular spheroid model of pancreatic tumor based on a triple co-culture of cancer cells, fibroblasts and endothelial cells and characterized by the ...presence of fibronectin, an important component of the tumor extracellular matrix. By combining cancer cells and stromal components, this model recreates in vitro the three-dimensional (3D) architecture of solid tumors. In this study, we used these hetero-type spheroids as a tool to assess the penetration of doxorubicin (used as a model drug) through the whole tumor mass either in a free form or loaded into polymer nanoparticles (NPs), and we investigated whether microscopy images, acquired by Confocal Laser Scanning Microscopy (CLSM) and Light Sheet Fluorescence Microscopy (LSFM), would be best to provide reliable information on this process. Results clearly demonstrated that CLSM was not suitable to accurately monitor the diffusion of small molecules such as the doxorubicin. Indeed, it only allowed to scan a layer of 100 µm depth and no information on deeper layers could be available because of a progressive loss of the fluorescence signal. On the contrary, a complete 3D tomography of the hetero-type multicellular tumor spheroids (MCTS) was obtained by LSFM and multi-view image fusion which revealed that the fluorescent molecule was able to reach the core of spheroids as large as 1 mm in diameter. However, no doxorubicin-loaded polymer nanoparticles were detected in the spheroids, highlighting the challenge of nanomedicine delivery through biological barriers. Overall, the combination of hetero-type MCTS and LSFM allowed to carry out a highly informative microscopic assessment and represents a suitable approach to precisely follow up the drug penetration in tumors. Accordingly, it could provide useful support in the preclinical investigation and optimization of nanoscale systems for drug delivery to solid tumors.
The present study examined the psychometric properties of the Chinese version of the Trait Emotional Intelligence Questionnaire Short Form (TEIQue-SF). Analyses were performed using a sample of ...undergraduates (
= 585) recruited from four universities across China. Confirmatory factor analysis of the Chinese TEIQue-SF supported the one-factor structure of trait emotional intelligence. Measurement invariance analyses were conducted across the Chinese sample and a sample of Canadian undergraduate students (
= 638). Although the two samples demonstrated configural and partial metric invariance, scalar invariance was not found. Cross-cultural implications and explanations of the present findings, as well as suggestions for future research are discussed.
The Sleeping Beauty (SB) transposon is an advanced tool for genetic engineering and a useful model to investigate cut-and-paste DNA transposition in vertebrate cells. Here, we identify novel SB ...transposase mutants that display efficient and canonical excision but practically unmeasurable genomic re-integration. Based on phylogenetic analyses, we establish compensating amino acid replacements that fully rescue the integration defect of these mutants, suggesting epistasis between these amino acid residues. We further show that the transposons excised by the exc+/int- transposase mutants form extrachromosomal circles that cannot undergo a further round of transposition, thereby representing dead-end products of the excision reaction. Finally, we demonstrate the utility of the exc+/int- transposase in cassette removal for the generation of reprogramming factor-free induced pluripotent stem cells. Lack of genomic integration and formation of transposon circles following excision is reminiscent of signal sequence removal during V(D)J recombination, and implies that cut-and-paste DNA transposition can be converted to a unidirectional process by a single amino acid change.
Tagging of endogenous stress response genes can provide valuable in vitro models for chemical safety assessment. Here, we present the generation and application of a fluorescent human induced ...pluripotent stem cell (hiPSC) reporter line for Heme oxygenase-1 (HMOX1), which is considered a sensitive and reliable biomarker for the oxidative stress response. CRISPR/Cas9 technology was used to insert an enhanced green fluorescent protein (eGFP) at the C-terminal end of the endogenous
HMOX1
gene. Individual clones were selected and extensively characterized to confirm precise editing and retained stem cell properties. Bardoxolone-methyl (CDDO-Me) induced oxidative stress caused similarly increased expression of both the wild-type and eGFP-tagged HMOX1 at the mRNA and protein level. Fluorescently tagged hiPSC-derived proximal tubule-like, hepatocyte-like, cardiomyocyte-like and neuron-like progenies were treated with CDDO-Me (5.62–1000 nM) or diethyl maleate (5.62–1000 µM) for 24 h and 72 h. Multi-lineage oxidative stress responses were assessed through transcriptomics analysis, and HMOX1-eGFP reporter expression was carefully monitored using live-cell confocal imaging. We found that eGFP intensity increased in a dose-dependent manner with dynamics varying amongst lineages and stressors. Point of departure modelling further captured the specific lineage sensitivities towards oxidative stress. We anticipate that the newly developed HMOX1 hiPSC reporter will become a valuable tool in understanding and quantifying critical target organ cell-specific oxidative stress responses induced by (newly developed) chemical entities.
During early mammalian development, transient pools of pluripotent cells emerge that can be immortalised upon stem cell derivation. The pluripotent state, 'naïve' or 'primed', depends on the ...embryonic stage and derivation conditions used. Here we analyse the temporal gene expression patterns of mouse, cattle and porcine embryos at stages that harbour different types of pluripotent cells. We document conserved and divergent traits in gene expression, and identify predictor genes shared across the species that are associated with pluripotent states
and
Amongst these are the pluripotency-linked genes
and
The novel genes discovered include naïve- (
and
) and primed-associated (
and
) genes as well as naïve to primed transition genes (
and
). Both
and
play a role in pluripotency since their knockdown results in differentiation and downregulation of key pluripotency genes. Our interspecies comparison revealed new insights of pluripotency, pluripotent stem cell identity and a new molecular criterion for distinguishing between pluripotent states in various species, including human.
For decades, the Big 5 model of personality has dominated as one of the most popular and widely used models of personality. The current article reviews the development and prevalence of the Big 5 ...model and explores the position of other personality variables not (or not fully) encompassed by this model, to see what lies ‘beyond the Big 5’. We start by comparing the Big 5 to the alternative six-factor HEXACO model, the Supernumerary Personality Traits, and a psychobiological model of personality. We then discuss the Dark Tetrad traits, self-defeating interpersonal styles, and trait emotional intelligence as examples of narrower personality traits which predict outcomes over and above the Big 5. Comparisons between these different personality models and the extent to which other personality traits can predict different psychological outcomes will be used to address the discussion of whether the Big 5 is a sufficient measure of personality, or whether other personality variables need to be also considered for a more comprehensive model of personality. Areas of consideration for researchers investigating Big 5 personality will also be discussed.
Peripheral blood was collected from a 1-year-old male patient with an X-linked recessive mutation of Iduronate 2-sulfatase (IDS) gene (NM_000202.7(IDS):c.85C>T) causing MPS II (OMIM 309900). ...Peripheral blood mononuclear cells (PBMCs) were reprogrammed by lentiviral delivery of a self-silencing hOKSM polycistronic vector. The pluripotency of the iPSC line was confirmed by the expression of pluripotency-associated markers and in vitro spontaneous differentiation towards the 3 germ layers. The iPSC line showed normal karyotype. The cell line offers a good platform to study MPS II pathophysiology, for drug testing, early biomarker discovery and gene therapy studies.
Peripheral blood was collected from a 3-year-old male patient with an X-linked recessive mutation of Iduronate 2-sulfatase (IDS) gene (NM_000202.7(IDS):c.85C>T) causing MPS II (OMIM 309900). ...Peripheral blood mononuclear cells (PBMCs) were reprogrammed by lentiviral delivery of a self-silencing hOKSM polycistronic vector. The pluripotency of the iPSC line was confirmed by the expression of pluripotency-associated markers and in vitro spontaneous differentiation towards the 3 germ layers. The iPSC line showed normal karyotype. The cell line offers a good platform to study MPS II pathophysiology, for drug testing, early biomarker discovery and gene therapy studies.
Traumatic brain injury (TBI) occurs more frequently in homeless persons than the general public. Both homelessness and TBI have been linked to experiences of violence (e.g., aggression and ...victimization). This study aimed to understand the temporal occurrences of events over the life course that contribute to vulnerabilities to TBI, victimization, aggression, and homelessness. A life-course perspective was used in this thematic analysis of in-person interviews with homeless persons. A total of 33 homeless persons met the inclusion criteria. Twenty-five of 33 (76%) participants had a self-reported history of TBI. Seventy-six percent of TBI events occurred before the onset of homelessness. Assault was the most common mechanism of TBI. During childhood, TBI was a frequently reported event, and parent- or guardian-related physical and sexual abuse were also accentuated with peer abuse, which may have contributed to a unique developmental trajectory. Aggressive behaviors were reported more commonly in persons who previously endured physical, sexual, and emotional victimization early in childhood. The cumulative effect of early adverse events, including TBI and other forms of victimization, subsequent aggression, and further TBI occurring later in life, may create an “at-risk” or vulnerable state preceding homelessness. Precipitating events during adulthood may contribute to a state of homelessness. Homelessness itself may facilitate the context for recurring physical and emotional injury, some of which may be preventable. Future studies should examine the temporality of events related to victimization by physical trauma, such as TBI, aggression, and homelessness.
Peripheral blood was collected from a 7-year-old male patient with an X-linked recessive mutation of Iduronate 2-sulfatase (IDS) gene (NM_000202.7(IDS):c.182C>T) causing MPS II (OMIM 309900). ...Peripheral blood mononuclear cells (PBMCs) were reprogrammed by lentiviral delivery of a self-silencing hOKSM polycistronic vector. The pluripotency of the iPSC line was confirmed by the expression of pluripotency-associated markers and in vitro spontaneous differentiation towards the 3 germ layers. The iPSC line showed normal karyotype. The cell line offers a good platform to study MPS II pathophysiology, for drug testing, early biomarker discovery and gene therapy studies.
PDF
