A novel expression cassette for the efficient visual selection of transformed tissues in florists' chrysanthemum (Chrysanthemum morifolium Ramat.).

Constructs carrying visual reporter genes coupled with efficient promoters could facilitate the process of identification and selection of stable transformants in recalcitrant crops. Here, a novel construct utilizing a ribulose-1,5-bisphosphate carboxylase (RbcS) promoter combined with the green fluorescent protein (GFP) reporter gene to initiate very high expression of GFP in florist's chrysanthemum (Chrysanthemum morifolium Ramat.) was described. Based on this expression cassette, a new regeneration protocol using leaf discs as explants was developed for the Agrobacterium-mediated transformation of Chrysanthemum genotype ‘1581’, and a transformation efficiency of 7% was obtained. The expression of two different GFP constructs targeted to either cytosol or plastids was compared in transgenic lines. Both GFP constructs were expressed at such a high level that the green fluorescence dominated red fluorescence in the leaf tissues, allowing easy observation and microdissection of transformed tissues even without a GFP filter. Under normal light, plants with GFP targeted to plastids had a light green phenotype deriving from the high GFP expression. Quantitative reverse transcriptional PCR analysis showed that the plastid targeted construct with intron had significantly higher steady state transcript levels of GFP mRNA. This novel expression cassette may allow direct visual selection of transformed tissues independent of antibiotic selection in a wide range of plant specie

A novel expression cassette for the efficient visual selection of transformed tissues in florist's chrysanthemum (Chrysanthemum morifolium Ramat.)

Constructs carrying visual reporter genes coupled with efficient promoters could facilitate the process of identification and selection of stable transformants in recalcitrant crops. Here, a novel construct utilizing a ribulose-1,5-bisphosphate carboxylase (RbcS) promoter combined with the green fluorescent protein (GFP) reporter gene to initiate very high expression of GFP in florist's chrysanthemum (Chrysanthemum morifolium Ramat.) was described. Based on this expression cassette, a new regeneration protocol using leaf discs as explants was developed for the Agrobacterium-mediated transformation of Chrysanthemum genotype ‘1581’, and a transformation efficiency of 7% was obtained. The expression of two different GFP constructs targeted to either cytosol or plastids was compared in transgenic lines. Both GFP constructs were expressed at such a high level that the green fluorescence dominated red fluorescence in the leaf tissues, allowing easy observation and microdissection of transformed tissues even without a GFP filter. Under normal light, plants with GFP targeted to plastids had a light green phenotype deriving from the high GFP expression. Quantitative reverse transcriptional PCR analysis showed that the plastid targeted construct with intron had significantly higher steady state transcript levels of GFP mRNA. This novel expression cassette may allow direct visual selection of transformed tissues independent of antibiotic selection in a wide range of plant species.Key words: Florist's chrysanthemum (Chrysanthemum morifolium), genetic transformation, GFP gene, RbcS promoter, RbcS targeting signal

Functional Implications of Neuroendocrine Differentiated Cells in Prostate Cancer

This thesis focuses on NE differentiation in prostate cancer, especially in prostate cancer models. We studied the effects of androgen depletion on the NE differentiated status of in vivo and in vitro prostatic tumor models. Knowledge concerning the function of NE cells in the normal human prostate and in prostate cancer is limited. A number of groups have studied NE differentiation in different epithelial systems. NE cells were found to be present in most prostatic adenocarcinomas (DiSant'Agnese, 1994). There is accumulating evidence that NE cells and tumors with NE cells are related to the androgen independent and poorly differentiated types of prostate cancer. There are indications that at least some secretion products of prostatic NE cells, like GRP and 5-HT, affect both prostate cancer growth and possibly also tumor differentiation. However, the growth modulating effects of neuropeptides and their relation to circulating androgen levels and androgen sensitivity of prostate cancer need to be further assessed. Studies directed towards identifying the role of NE cells in hormonally treated prostate cancer may contribute to the understanding of the transition of androgen dependent to androgen independent prostate cancer. There are still unanswered questions that need to be answered: How is the process of neuroendocrine differentiation regulated and can this process be influenced? Are the NE cells necessary for the regulation of growth and differentiation of the prostate or do they play a role in the maintenance of prostatic homeostasis? Can these cells induce (androgen independent) growth of surrounding cells, for instance by ligand independent activation of the androgen receptor (AR) through neuropeptides secreted by NE cells

The effectiveness of psychological interventions for post-traumatic stress disorder in children, adolescents and young adults: A systematic review and meta-analysis

Background: Children and adolescents display different symptoms of post-traumatic stress disorder (PTSD) than adults. Whilst evidence for the effectiveness of psychological interventions has been synthesised for adults, this is not directly applicable to younger people. Therefore, this systematic review and meta-analysis synthesised studies investigating the effectiveness of psychological interventions for PTSD in children, adolescents and young adults. It provides an update to previous reviews investigating interventions in children and adolescents, whilst investigating young adults for the first time. / Methods: We searched published and grey literature to obtain randomised control trials assessing psychological interventions for PTSD in young people published between 2011 and 2019. Quality of studies was assessed using the Cochrane Risk of Bias tool. Data were analysed using univariate random-effects meta-analysis. / Results: From 15 373 records, 27 met criteria for inclusion, and 16 were eligible for meta-analysis. There was a medium pooled effect size for all psychological interventions (d = −0.44, 95% CI −0.68 to −0.20), as well as for Trauma-Focused Cognitive Behavioural Therapy (TF-CBT) and Eye Movement Desensitisation and Reprocessing (EMDR) (d = −0.30, 95% CI −0.58 to −0.02); d = −0.46, 95% CI −0.81 to −0.12). / Conclusions: Some, but not all, psychological interventions commonly used to treat PTSD in adults were effective in children, adolescents and young adults. Interventions specifically adapted for younger people were also effective. Our results support the National Institute for Health and Care Excellence guidelines which suggest children and adolescents be offered TF-CBT as a first-line treatment because of a larger evidence base, despite EMDR being more effective

Playing hide and seek: tumor cells in control of MHC class I antigen presentation

MHC class I (MHC-I) molecules present a blueprint of the intracellular proteome to T cells allowing them to control infection or malignant transformation. As a response, pathogens and tumor cells often downmodulate MHC-I mediated antigen presentation to escape from immune surveillance. Although the fundamental rules of antigen presentation are known in detail, the players in this system are not saturated and new modules of regulation have recently been uncovered. Here, we update the understanding of antigen presentation by MHC-I molecules and how this can be exploited by tumors to prevent exposure of the intracellular proteome. This knowledge can provide new ways to improve immune responses against tumors and pathogens.Chemical Immunolog

Segmentation of NKX2.5 Signal in Human Pluripotent Stem Cell-Derived Cardiomyocytes

Human pluripotent stem cell-derived Cardiomyocytes (hPSC-CMs) become increasingly popular in recent years for disease modeling and drug screening. NKX2.5 gene is a key transcription factor that regulates cardiomyocyte differentiation. A human embryonic stem cell (hESC) reporter line with NKX2.5 in GFP signal allows us to monitor the specificity and efficiency of human cardiac differentiation. We intend to develop an automatic analysis pipeline for the NKX2.5 signal. However, the NKX2.5 signal captured from fluorescence microscopy is highly heterogeneous. It is not possible to be properly segmented using traditional thresholding methods. Therefore, in this paper, one machine learning method: enhanced Fuzzy C-Means clustering (EnFCM) and two deep learning models: U-Net and DeepLabV3+, are evaluated on the segmentation performance. Parameters have been tuned for each method so as to reach to the optimal segmentation performance. The results show that EnFCM reaches the performance of 0.85. U-Net and DeepLabV3+ have a superior performance. Their performances are 0.86 and 0.89 respectively.</p

How Smart are Smart Materials?:A Conceptual and Ethical Analysis of Smart Lifelike Materials for the Design of Regenerative Valve Implants

It may soon become possible not just to replace, but to re-grow healthy tissues after injury or disease, because of innovations in the field of Regenerative Medicine. One particularly promising innovation is a regenerative valve implant to treat people with heart valve disease. These implants are fabricated from so-called 'smart', 'lifelike' materials. Implanted inside a heart, these implants stimulate re-growth of a healthy, living heart valve. While the technological development advances, the ethical implications of this new technology are still unclear and a clear conceptual understanding of the notions 'smart' and 'lifelike' is currently lacking. In this paper, we explore the conceptual and ethical implications of the development of smart lifelike materials for the design of regenerative implants, by analysing heart valve implants as a showcase. In our conceptual analysis, we show that the materials are considered 'smart' because they can communicate with human tissues, and 'lifelike' because they are structurally similar to these tissues. This shows that regenerative valve implants become intimately integrated in the living tissues of the human body. As such, they manifest the ontological entanglement of body and technology. In our ethical analysis, we argue this is ethically significant in at least two ways: It exacerbates the irreversibility of the implantation procedure, and it might affect the embodied experience of the implant recipient. With our conceptual and ethical analysis, we aim to contribute to responsible development of smart lifelike materials and regenerative implants.</p

Self-expanding stents and aortoiliac occlusive disease: A review of the literature

The treatment of symptomatic aortoiliac occlusive disease has shifted from open to endovascular repair. Both short- and long-term outcomes after percutaneous angioplasty and stenting rival those after open repair and justify an endovascular-first approach. In this article, we review the current endovascular treatment strategies in patients with aortoiliac occlusive disease, indications for primary and selective stenting in the iliac artery, and physical properties and future perspectives of self-expanding stents