PKC-θ is a negative regulator of TRAIL-induced and FADD-mediated apoptotic spectrin aggregation

Introduction. During studies on chemotherapy-induced apoptosis in lymphoid cells, we noted that aggregation of spectrin occurred early in apoptosis, i.e. before activation of initiator caspase(s) and prior to exposure of phosphatidylserine (PS). We also found that protein kinase C theta (PKC-θ) co-localized with spectrin in these aggregates. Our previously published studies indicated that in formation of early apoptotic spectrin aggregates, either PKC-θ or other apoptosis-related proteins are involved. Taking into consideration above data, we decided to test the effect of PKC-θ and Fas-associated death domain protein (FADD) on spectrin aggregation in these cells during tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. Material and methods. For PKC-θ gene (PRKCQ) or FADD gene expression silencing in Jurkat T cells we used lentiviral particles containing shRNA and scrambled shRNA, respectively. Spectrin aggregates were detected by Western blotting after Triton-X 100 extraction in pellet and soluble fractions or by confocal imaging. Results. TRAIL-induced apoptosis results in spectrin aggregation and leads to translocation and aggregation of PKC-θ. We found that phorbol-myristate acetate, a PKC activator and translocation inducer, has only a small effect on spectrin aggregation. To further confirm this, we have also shown that knock down of PRKCQ in Jurkat T cells accelerates the formation of TRAIL-induced spectrin aggregates. Transient overexpression of the β-spectrin C-terminal fragment, containing multiple S/T phosphorylation sites, potential substrate sites for PKC-θ, accelerated the formation of spectrin aggregates. Silencing of downstream TRAIL receptor effector gene, FADD, delayed aggregation of spectrin, but did not reduce PKC-θ localization to the plasma membrane. Conclusions. In summary, our results show for the first time involvement of spectrin aggregation in TRAIL receptor-FADD apoptotic pathway and indicate that TRAIL-induced spectrin aggregate formation is mediated by FADD and negatively regulated by PKC-θ

Epidermal T Cell Dendrites Serve as Conduits for Bidirectional Trafficking of Granular Cargo

Dendritic epidermal T cells (DETCs) represent a prototypical lineage of intraepithelial γδ T cells that participate in the maintenance of body barrier homeostasis. Unlike classical T cells, DETCs do not recirculate and they remain persistently activated through their T cell receptors (TCR) at steady state, i.e., in absence of infection or tissue wounding. The steady state TCR signals sustain the formation of immunological synapse-like phosphotyrosine-rich aggregates located on projections (PALPs) which act to anchor and polarize DETC’s long cellular projections toward the apical epidermis while the cell bodies reside in the basal layers. The PALPs are known to contain pre-synaptic accumulations of TCR-containing and lysosomal granules, but how this cargo accumulates there remains unclear. Here, we combined anti-Vγ5 TCR, cholera toxin subunit B (CTB), and LysoTracker (LT)-based intravital labeling of intracellular granules, with high resolution dynamic microscopy and fluorescence recovery after photobleaching (FRAP) to characterize the steady state composition and transport of DETC granules in steady state epidermis. Intradermal fluorescent Vγ5 antibody decorated DETCs without causing cellular depletion, dendrite mobilization or rounding up and became slowly internalized over 48 h into intracellular granules that, after 6 days, colocalized with LAMP-1 and less so with LT or early endosomal antigen-1. Intradermal CTB was likewise internalized predominantly by DETCs in epidermis, labeling a partly overlapping set of largely LAMP-1+ intracellular granules. These as well as LT-labeled granules readily moved into newly forming dendrites and accumulated at the apical endings. FRAP and spatiotemporal tracking showed that the inside tubular lengths of DETC cellular projections supported dynamic trafficking of lysosomal cargo toward and away from the PALPs, including internalized TCR and lipid raft component ganglioside GM1 (labeled with CTB). By contrast, the rate of GM1 granules transport through comparable dendrites of non-DETCs was twice slower. Our observations suggest that DETCs use chronic TCR activation to establish a polarized conduit system for long-range trans-epithelial transport aimed to accumulate mature lysosomes at the barrier-forming apical epidermis. The biological strategy behind the steady state lysosome polarization by DETCs remains to be uncovered

Board characteristics and sustainability in higher education institutions: The case of the United Kingdom

We explored the relationship between board characteristics and sustainability of higher education institutions in the United Kingdom (UK). We analysed 153 UK universities using data for the year 2019. Our analysis revealed that board size, the number of students on the board, and the number of academic members on the board were found to have significant and positive relationships with sustain-ability. Also, the composition of the sustainability commit-tee was shown to have a significant and positive impact on sustainability score. However, the relationships between board gender diversity, the number of external members on the board, and the number of board meetings held during the year with sustainability score were not significant. The results provide guidance to universities for developing their sustainability practices

Shrek in Polish and French dubbing - analysis of the transfer of cultural elements

Celem naszej pracy magisterskiej jest przedstawienie i analiza elementów kulturowych zawartych w polskim i francuskim dubbingu filmu animowanego „Shrek”. Nasza analiza będzie oparta na różnych technikach i procedurach tłumaczeniowych według Jean-Paula Vinay i Jean Darbelnet, Teresy Tomaszkiewicz, Arkadiusza Belczyka i Marzeny Chrobak. Analiza dotyczyć będzie dwóch dubbingów: polskiego i francuskiego w reżyserii Bartosza Wierzbiety i Nathalie Raimbault oraz oryginalnej wersji amerykańskiej.W pierwszym rozdziale przedstawimy film „Shrek” i jego fenomen.W następnym rozdziale skupimy się na ogólnych informacjach o tłumaczeniu audiowizualnym. Przedstawimy trzy rodzaje tego tłumaczenia. Bardziej szczegółowo omówimy również dubbing w Polsce i Francji. Wymienimy również strategie, którymi tłumacze mogą się kierować podczas wykonywania swojego zadania. Porozmawiamy także o nazewnictwie procesu tłumaczenia filmu.W rozdziale trzecim zajmiemy się zagadnieniami, które pojawiają się w przekładzie audiowizualnym, wymienimy i krótko opiszemy siedem procedur Jeana Paula Vinay i Jeana Darbelneta. Skoncentrujemy się na strategiach stosowanych podczas transferu elementów kulturowych. Będziemy bazować na tych przedstawionych przez Teresę Tomaszkiewicz, Arkadiusza Belczyka i Marzenę Chrobak. W dalszej części przedstawimy i wyjaśnimy również pojęcie intertekstualności, ponieważ często pojawia się ono w filmach animowanych, zwłaszcza w „Shreku”. Poruszymy również kwestię komizmu, który jest ściśle związana z tym filmem. Omówimy podział komizmu i gier słownych według Wojciecha Kalagi i Patricka Zabalbeascoa. Zaprezentowane zostaną także techniki tłumaczenia humoru według Tomaszkiewicza i Dirka Delabastitia.W rozdziale czwartym skupiamy się na tłumaczu audiowizualnym. Zobaczymy kim jest tłumacz i jakie cechy powinien posiadać. Zaprezentujemy również dialogi dubbingu francuskiego i polskiego.W ostatnim, piątym rozdziale zostanie przedstawiona analiza elementów kulturowych występujących w polskim i francuskim dubbingu pierwszej części „Shreka”. Analiza ta obejmie również techniki stosowane przez każdego autora dialogów. Podczas analizy staraliśmy się scharakteryzować i nazwać strategie i techniki tłumaczeniowe. Mogliśmy też zobaczyć powtórki strategii i teorii. Obie wersje zawierają wiele odniesień do kultury docelowej (np. jedzenie, postacie). Z analizy można wywnioskować, że obie wersje były zorientowane głównie na teorię naturalizacji. Ważne było dostosowanie tekstu do odbiorców z danego kraju, aby mogli go jak najlepiej zrozumieć.The objective of our master thesis is to present and analyze the cultural elements included in the Polish and French dubbing of the animated film "Shrek". Our analysis will be based on various translation techniques and procedures according to Jean-Paul Vinay and Jean Darbelnet, Teresa Tomaszkiewicz, Arkadiusz Belczyk and Marzena Chrobak. The analysis will concern two dubbing: Polish and French directed by Bartosz Wierzbięta and Nathalie Raimbault, as well as the original American version.In the first chapter we will present the movie "Shrek" and its phenomenon.In the next chapter, we will focus on general information about audiovisual translation. We will present three types of this translation. We will also discuss dubbing in Poland and France in more detail. We will also list the strategies that translators can follow during their task. We will also talk about the naming of the film's translation process.In the third chapter, we will address the issues that appear in audiovisual translation, we will enumerate and briefly describe seven processes by Jean Paul Vinay and Jean Darbelnet. We will focus on the strategies that are applied during the transfer of cultural elements. We will build on those introduced by Teresa Tomaszkiewicz, Arkadiusz Belczyk and Marzena Chrobak. In what follows, we will also present and explain the concept of intertextuality because it often appears in animated films, especially in "Shrek". We will also address the issue of comedy, which is closely related to this film. We will discuss the division of comedy and word games according to Wojciech Kalaga and Patrick Zabalbeascoa. The humor translation techniques according to Tomaszkiewicz and Dirk Delabastitia will also be presented.In the fourth chapter, we focus on the audiovisual translator. We will see who the translator is and what characteristics he should have. We will also present the French and Polish translators of those dubbings.The last, fifth chapter, will present an analysis of the cultural elements which appear in the Polish and French dubbing of the first part of "Shrek". This analysis will also encompass the techniques used by each dialogue writer. During the analysis, we tried to characterize and name the translation strategies. We could also see a repetition of strategies and theories. Both versions contain many references to the target culture (eg food, characters). From the analysis it can be concluded that both versions were mainly oriented towards the theory of naturalization. It was important to adapt the text to the recipients of a given country so that they could understand it as well as possible.L'objectif de notre mémoire de master est de présenter et d'analyser les éléments culturels inclus dans le doublage polonais et français du film d'animation "Shrek". Notre analyse sera basée sur diverses techniques et procédures de traduction selon Jean-Paul Vinay et Jean Darbelnet, Teresa Tomaszkiewicz, Arkadiusz Belczyk et Marzena Chrobak. L'analyse concernera deux doublages : polonais et français réalisés par Bartosz Wierzbięta et Nathalie Raimbault, ainsi que la version originale américaine.Dans le premier chapitre nous présenterons le film "Shrek" et son phénomène.Dans le chapitre suivant, nous nous concentrerons sur les informations générales de la traduction audiovisuelle. Nous présenterons trois types de cette traduction. Nous aborderons également plus en détail le doublage en Pologne et en France. Nous allons également lister les stratégies que les traducteurs peuvent suivre au cours de leur tâche. Nous parlerons également de la dénomination du processus de traduction du film. Dans le troisième chapitre, nous aborderons les enjeux qui apparaissent dans la traduction audiovisuelle, nous allons énumérer et décrire brièvement sept procédés de Jean Paul Vinay et Jean Darbelnet. Nous nous concentrerons sur les stratégies qui sont appliquées lors du transfert des éléments culturels. Nous nous appuierons sur celles introduites par Teresa Tomaszkiewicz, Arkadiusz Belczyk et Marzena Chrobak. Dans ce qui suit, nous présenterons et expliquerons églament le concept d'intertextualité car il apparaît souvent dans les films d'animation, notamment dans "Shrek". Nous aborderons également la question du comique, qui est étroitement liée à ce film. Nous évoquerons la division du comique et de jeux de mots selon Wojciech Kalaga et Patrick Zabalbeascoa. Les techniques de traduction de l'humour selon Tomaszkiewicz et Dirk Delabastitia seront également présentées.Dans le quatrième chapitre, nous nous concentrons sur le traducteur audiovisuel. Nous verrons qui est le traducteur et quelles caractéristiques il doit avoir. Nous présenterons également les dialoguistes des doublages français et polonais.Le dernier et cinquième chapitre, présentera une analyse des éléments culturels qui apparaissent dans le doublage polonais et français du premier volet de "Shrek". Cette analyse englobera également les techniques utilisées par chaque dialoguiste. Au cours de l'analyse, nous avons essayé de caractériser et de nommer les stratégies de traductions. Nous pourrions également voir une répétition des stratégies et des théories. Les deux versions contiennent de nombreuses références à la culture cible (par exemple culinaire, personnages). De l'analyse, on peut conclure que les deux versions étaient principalement orientées vers la théorie de la naturalisation. Il était important d'adapter le texte aux destinataires d'un pays donné afin qu'ils puissent le comprendre le mieux possible

Zinc-Substituted Pheophorbide A Is a Safe and Efficient Antivascular Photodynamic Agent

The present study focuses on the photodynamic activity of zinc-substituted pheophorbide a against human endothelial cells. Previously, zinc pheophorbide a has been shown to be a very potent photosensitizer but also a strong albumin binder. Binding to albumin significantly reduces its availability to cancer cells, which may necessitate the use of relatively high doses. Here we show that zinc pheophorbide a is very effective against vascular endothelial cells, even in its albumin-complexed form. Albumin complexation increases the lysosomal accumulation of the drug, thus enhancing its efficiency. Zinc pheophorbide a at nanomolar concentrations induces endothelial cell death via apoptosis, which in many cases is considered a desirable cell death mode because of its anti-inflammatory effect. Additionally, we demonstrate that in comparison to tumor cells, endothelial cells are much more susceptible to photodynamic treatment with the use of the investigated compound. Our findings demonstrate that zinc pheophorbide a is a very promising photosensitizer for use in vascular-targeted photodynamic therapy against solid tumors, acting as a vascular shutdown inducer. It can also possibly find application in the treatment of a range of vascular disorders. Numerous properties of zinc pheophorbide a are comparable or even more favorable than those of the well-known photosensitizer of a similar structure, palladium bacteriopheophorbide (TOOKAD®)

Toward a magic or imaginary bullet? Ligands for drug targeting to cancer cells: principles, hopes, and challenges

Monika Toporkiewicz, Justyna Meissner, Lucyna Matusewicz, Aleksander Czogalla, Aleksander F SikorskiLaboratory of Cytobiochemistry, Faculty of Biotechnology, University of Wrocław, Wrocław, PolandAbstract: There are many problems directly correlated with the systemic administration of drugs and how they reach their target site. Targeting promises to be a hopeful strategy as an improved means of drug delivery, with reduced toxicity and minimal adverse side effects. Targeting exploits the high affinity of cell-surface-targeted ligands, either directly or as carriers for a drug, for specific retention and uptake by the targeted diseased cells. One of the most important parameters which should be taken into consideration in the selection of an appropriate ligand for targeting is the binding affinity (KD). In this review we focus on the importance of binding affinities of monoclonal antibodies, antibody derivatives, peptides, aptamers, DARPins, and small targeting molecules in the process of selection of the most suitable ligand for targeting of nanoparticles. In order to provide a critical comparison between these various options, we have also assessed each technology format across a range of parameters such as molecular size, immunogenicity, costs of production, clinical profiles, and examples of the level of selectivity and toxicity of each. Wherever possible, we have also assessed how incorporating such a targeted approach compares with, or is superior to, original treatments.Keywords: targeting, drug delivery, tumor, monoclonal antibody, EGFR, cance

How organisational board compositions lead to a higher job satisfaction: an empirical analysis of US and UK companies

The relationship between board characteristics and micro-level organizational factors is an area that has been significantly under-researched, and there is a lack of understanding of how these two elements interact with each other. Hence, we aim to explore how board characteristics could potentially have an impact on individual-level job satisfaction. The dataset used for this study encompasses a total of 4020 observations gathered from 804 companies listed in the FTSE 350 and S&P 500 indices, and it covers the period spanning from 2016 to 2021. The results of the adopted multiple regression analysis showed significant positive relationships between board gender diversity, diversity of specific skills, board independence, board meeting attendance, board size, and average board tenure and employees’ job satisfaction of the companies under analysis. However, cultural diversity was not found to have a significant impact on employees’ satisfaction. We draw out the theoretical implications of these findings and provide practical recommendations regarding companies’ boards composition and structure that help them to enhance the level of their employees’ job satisfaction

video_6_Epidermal T Cell Dendrites Serve as Conduits for Bidirectional Trafficking of Granular Cargo.avi

<p>Dendritic epidermal T cells (DETCs) represent a prototypical lineage of intraepithelial γδ T cells that participate in the maintenance of body barrier homeostasis. Unlike classical T cells, DETCs do not recirculate and they remain persistently activated through their T cell receptors (TCR) at steady state, i.e., in absence of infection or tissue wounding. The steady state TCR signals sustain the formation of immunological synapse-like phosphotyrosine-rich aggregates located on projections (PALPs) which act to anchor and polarize DETC’s long cellular projections toward the apical epidermis while the cell bodies reside in the basal layers. The PALPs are known to contain pre-synaptic accumulations of TCR-containing and lysosomal granules, but how this cargo accumulates there remains unclear. Here, we combined anti-Vγ5 TCR, cholera toxin subunit B (CTB), and LysoTracker (LT)-based intravital labeling of intracellular granules, with high resolution dynamic microscopy and fluorescence recovery after photobleaching (FRAP) to characterize the steady state composition and transport of DETC granules in steady state epidermis. Intradermal fluorescent Vγ5 antibody decorated DETCs without causing cellular depletion, dendrite mobilization or rounding up and became slowly internalized over 48 h into intracellular granules that, after 6 days, colocalized with LAMP-1 and less so with LT or early endosomal antigen-1. Intradermal CTB was likewise internalized predominantly by DETCs in epidermis, labeling a partly overlapping set of largely LAMP-1⁺ intracellular granules. These as well as LT-labeled granules readily moved into newly forming dendrites and accumulated at the apical endings. FRAP and spatiotemporal tracking showed that the inside tubular lengths of DETC cellular projections supported dynamic trafficking of lysosomal cargo toward and away from the PALPs, including internalized TCR and lipid raft component ganglioside GM1 (labeled with CTB). By contrast, the rate of GM1 granules transport through comparable dendrites of non-DETCs was twice slower. Our observations suggest that DETCs use chronic TCR activation to establish a polarized conduit system for long-range trans-epithelial transport aimed to accumulate mature lysosomes at the barrier-forming apical epidermis. The biological strategy behind the steady state lysosome polarization by DETCs remains to be uncovered.</p

video_1_Epidermal T Cell Dendrites Serve as Conduits for Bidirectional Trafficking of Granular Cargo.avi

<p>Dendritic epidermal T cells (DETCs) represent a prototypical lineage of intraepithelial γδ T cells that participate in the maintenance of body barrier homeostasis. Unlike classical T cells, DETCs do not recirculate and they remain persistently activated through their T cell receptors (TCR) at steady state, i.e., in absence of infection or tissue wounding. The steady state TCR signals sustain the formation of immunological synapse-like phosphotyrosine-rich aggregates located on projections (PALPs) which act to anchor and polarize DETC’s long cellular projections toward the apical epidermis while the cell bodies reside in the basal layers. The PALPs are known to contain pre-synaptic accumulations of TCR-containing and lysosomal granules, but how this cargo accumulates there remains unclear. Here, we combined anti-Vγ5 TCR, cholera toxin subunit B (CTB), and LysoTracker (LT)-based intravital labeling of intracellular granules, with high resolution dynamic microscopy and fluorescence recovery after photobleaching (FRAP) to characterize the steady state composition and transport of DETC granules in steady state epidermis. Intradermal fluorescent Vγ5 antibody decorated DETCs without causing cellular depletion, dendrite mobilization or rounding up and became slowly internalized over 48 h into intracellular granules that, after 6 days, colocalized with LAMP-1 and less so with LT or early endosomal antigen-1. Intradermal CTB was likewise internalized predominantly by DETCs in epidermis, labeling a partly overlapping set of largely LAMP-1⁺ intracellular granules. These as well as LT-labeled granules readily moved into newly forming dendrites and accumulated at the apical endings. FRAP and spatiotemporal tracking showed that the inside tubular lengths of DETC cellular projections supported dynamic trafficking of lysosomal cargo toward and away from the PALPs, including internalized TCR and lipid raft component ganglioside GM1 (labeled with CTB). By contrast, the rate of GM1 granules transport through comparable dendrites of non-DETCs was twice slower. Our observations suggest that DETCs use chronic TCR activation to establish a polarized conduit system for long-range trans-epithelial transport aimed to accumulate mature lysosomes at the barrier-forming apical epidermis. The biological strategy behind the steady state lysosome polarization by DETCs remains to be uncovered.</p