Heat Shock Protein 70 Promotes Cell Survival by Inhibiting Lysosomal Membrane Permeabilization

Heat shock protein 70 (Hsp70) is a potent survival protein whose depletion triggers massive caspase-independent tumor cell death. Here, we show that Hsp70 exerts its prosurvival function by inhibiting lysosomal membrane permeabilization. The cell death induced by Hsp70 depletion was preceded by the release of lysosomal enzymes into the cytosol and inhibited by pharmacological inhibitors of lysosomal cysteine proteases. Accordingly, the Hsp70-mediated protection against various death stimuli in Hsp70-expressing human tumor cells as well as in immortalized Hsp70 transgenic murine fibroblasts occurred at the level of the lysosomal permeabilization. On the contrary, Hsp70 failed to inhibit the cytochrome c–induced, apoptosome-dependent caspase activation in vitro and Fas ligand–induced, caspase-dependent apoptosis in immortalized fibroblasts. Immunoelectron microscopy revealed that endosomal and lysosomal membranes of tumor cells contained Hsp70. Permeabilization of purified endo/lysosomes by digitonin failed to release Hsp70, suggesting that it is physically associated with the membranes. Finally, Hsp70 positive lysosomes displayed increased size and resistance against chemical and physical membrane destabilization. These data identify Hsp70 as the first survival protein that functions by inhibiting the death-associated permeabilization of lysosomes

Long Range Plan: Dense matter theory for heavy-ion collisions and neutron stars

Since the release of the 2015 Long Range Plan in Nuclear Physics, major events have occurred that reshaped our understanding of quantum chromodynamics (QCD) and nuclear matter at large densities, in and out of equilibrium. The US nuclear community has an opportunity to capitalize on advances in astrophysical observations and nuclear experiments and engage in an interdisciplinary effort in the theory of dense baryonic matter that connects low- and high-energy nuclear physics, astrophysics, gravitational waves physics, and data scienceComment: 70 pages, 3 figures, White Paper for the Long Range Plan for Nuclear Scienc

Activating methods in preschool education

Celem artykułu jest przedstawienie metod aktywizujących wykorzystywanych w nauczaniu przedszkolnym oraz wskazanie ich głównych zalet. Opracowanie artykułu poprzedziło studium literatury dotyczącej dydaktyki ogólnej i roli nauczyciela w rozwoju aktywności dziecka metod aktywizujących i ich wykorzystania na poziomie edukacji przedszkolnej. W artykule odniesiono się do przeprowadzonych wśród nauczycieli wychowania przedszkolnego z województwa warmińsko-mazurskiego wyników badań własnych, których celem było poznanie opinii nauczycieli przedszkoli na temat przydatności i stopnia wykorzystania metod aktywizujących w procesie edukacyjnym, a także zalet i częstotliwości ich stosowania w pracy z dziećmi w wieku przedszkolnym.The aim of the article is to present the activating methods used in preschool education and to indicate their main advantages. The preparation of the article was preceded by a literature study on general didactics and the role of a teacher in the development of a child’s activity of activating methods and their use at the pre-school level. The article refers to the results of own research conducted among teachers of pre-school education from the Warmian-Masurian Voivodeship, the aim of which was to find out the opinions of kindergarten teachers on the usefulness and degree of use of activating methods in the educational process, as well as the advantages and frequency of their use in working with children in preschool age

The comparison of the motivation systems of secondary school and higher education teachers in the existing legislation

Human capital is the driving force of the company functioning and it has a significant impact on its competitiveness on the market. The behavior and performance of human capital are affected by the way of management which includes the motivation responsible for the increase in work efficiency. The existing legal acts regulate the financial as well as non-financial issues motivating school employees, which are inadequate and they often act as a demotivating stimulus. The teachers employed in educational institutions and universities are more often exposed to job burnout, which partially might be due to the lack of the appropriate motivation systems, the implementation of which would bring the expected results. This article is to compare the motivation factors resulting from the legal acts governing both types of school.Kapitał ludzki stanowi siłę napędową funkcjonowania firmy oraz ma istotny wpływ na jej konkurencyjność na rynku. Na zachowania i działanie kapitału ludzkiego wpływ ma sposób zarządzania, który obejmuje motywację odpowiedzialną za wzrost efektywności pracy. Obowiązujące akty prawne regulują kwestie finansowe, jak i pozafinansowe motywujące pracowników szkół, które są niewystarczające i często działają jako bodziec demotywujący. Nauczyciele zatrudnieni w placówkach oświatowych oraz szkołach wyższych znacznie częściej narażeni są wypalenie zawodowe, które częściowo może być związane z brakiem odpowiednich systemów motywacyjnych, których wdrożenie przyniosłoby oczekiwane efekty. Niniejszy artykuł ma na celu porównanie wykorzystywanych czynników motywacyjnych wynikających z obowiązujących w obu typach szkół aktów prawnych

The role of a headmaster in motivating teachers

Celem artykułu jest wskazanie najważniejszych narzędzi, jakimi dysponuje dyrektor szkoły w aspekcie motywowania pracowników oraz podkreślenie jego roli w tym procesie. Opracowanie artykułu poprzedziło studium literatury dotyczącej motywacji nauczycieli, roli dyrektora oraz pełnienia funkcji kierowniczej w szkole, które jednoznacznie wskazują na istnienie dużej ilości narzędzi gotowych do wykorzystania po uprzednim dostosowaniu ich do specyfiki grupy.The aim of this article is to show important incentive tools a principal has which and underlining his role in the process of motivation. Drawing the article up preceded the literature review concerning motivation of teachers, the role of the director and performing the managerial function at school, which are pointing out to the existence of a lot of ready tools for using the group after the adaptation of them to the specificity

Domestic hot water consumption in multi-apartment buildings

The article presents the analysis of the hot water consumption in apartment buildings. This study was conducted basing on the data gathered from 626 apartments during two following years. By statistical analysis of the collected data, the correlation between the DHW consumption and both the size of the apartments and the number of occupants in the dwellings was evaluated. The dependencies between the hot water consumption and these two parameters were calculated. The results were compared to the data available in the literature. The article lists the areas of the greatest inaccuracies in the calculations basing on the available indicators and proposes an alternative method that corrects them. The results were used to develop a calculation model for daily hot water usage in multi-apartment buildings

Dielectrophoresis-Based SERS Sensors for the Detection of Cancer Cells in Microfluidic Chips

The detection of freely circulating cancer cells (CTCs) is one of the greatest challenges of modern medical diagnostics. For several years, there has been increased attention on the use of surface-enhanced Raman spectroscopy (SERS) for the detection of CTCs. SERS is a non-destructive, accurate and precise technique, and the use of special SERS platforms even enables the amplification of weak signals from biological objects. In the current study, we demonstrate the unique arrangement of the SERS technique combined with the deposition of CTCs cells on the surface of the SERS platform via a dielectrophoretic effect. The appropriate frequencies of an alternating electric field and a selected shape of the electric field can result in the efficient deposition of CTCs on the SERS platform. The geometry of the microfluidic chip, the type of the cancer cells and the positive dielectrophoretic phenomenon resulted in the trapping of CTCs on the surface of the SERS platform. We presented results for two type of breast cancer cells, MCF-7 and MDA-MB-231, deposited from the 0.1 PBS solution. The limit of detection (LOD) is 20 cells/mL, which reflects the clinical potential and usefulness of the developed approach. We also provide a proof-of-concept for these CTCs deposited on the SERS platform from blood plasma

Detection of Circulating Tumor Cells Using Membrane-Based SERS Platform: A New Diagnostic Approach for ‘Liquid Biopsy’

The detection and monitoring of circulating tumor cells (CTCs) in blood is an important strategy for early cancer evidence, analysis, monitoring of therapeutic response, and optimization of cancer therapy treatments. In this work, tailor-made membranes (MBSP) for surface-enhanced Raman spectroscopy (SERS)-based analysis, which permitted the separation and enrichment of CTCs from blood samples, were developed. A thin layer of SERS-active metals deposited on polymer mat enhanced the Raman signals of CTCs and provided further insight into CTCs molecular and biochemical composition. The SERS spectra of all studied cells—prostate cancer (PC3), cervical carcinoma (HeLa), and leucocytes as an example of healthy (normal) cell—revealed significant differences in both the band positions and/or their relative intensities. The multivariate statistical technique based on principal component analysis (PCA) was applied to identify the most significant differences (marker bands) in SERS data among the analyzed cells and to perform quantitative analysis of SERS data. Based on a developed PCA algorithm, the studied cell types were classified with an accuracy of 95% in 2D PCA to 98% in 3D PCA. These results clearly indicate the diagnostic efficiency for the discrimination between cancer and normal cells. In our approach, we exploited the one-step technology that exceeds most of the multi-stage CTCs analysis methods used and enables simultaneous filtration, enrichment, and identification of the tumor cells from blood specimens

Detection of circulating tumor cells (CTCs) by SERS-based immunomagnetic optofluidic device

https://kent-islandora.s3.us-east-2.amazonaws.com/node/14388/83895-thumbnail.jpgThe isolation and characterization of circulating tumor cells (CTCs) has great potential for non-invasive biopsy. In this study, a surface-enhanced Raman spectroscopy (SERS) method was developed using magnetic nanoparticles and a solid SERS-active substrate integrated with an external field-assisted microfluidic device to efficiently isolate CTCs from blood samples. A new SERS substrate was used, developed by physically modifying the surface with a femtosecond laser, sputtering the active SERS layer and chemically modifying the surface with anti-EpCAM antibodies. Magnetic nanoparticles (Fe2O3) were coated with SERS active metal and then modified with para-mercaptobenzoic acid (p-MBA), which acts simultaneously as a Raman reporter and a linker with anti-EpCAM antibodies. The sensitive immune recognition of tumor cells is aided by the introduction of a controlled external magnetic field into the microfluidic chip. The integration of the SERS-active platform and p-MBA labeled immuno-Ag@Fe2O3 nanostructures with the microfluidic device ensures lower demand for samples and analytes, precise operation, increases the reproducibility of spectral responses and enables miniaturization and portability of the presented approach. We used four target tumor cell lines with relatively large (human prostate metastatic adenocarcinoma cells (LNCaP)), medium (adenocarcinomic human alveolar basal epithelial cells (A549)), weak (human prostate tumor line (PC3)) and no expression of EpCAM (tumor cells) cervical cancer (HeLa)) to estimate the detection limits on the basis of constructed calibration curves blood samples from lung cancer patients were used to validate the developed method.[1] M. Czaplicka, K. Niciński, A. Nowicka, T. Szymborski i A. Kamińska, Cancers, 2020, 12 (3315), 1-21. </ol