Chloroquine-induced accumulation of autophagosomes and lipids in the endothelium

Chloroquine (CQ) is an antimalarial drug known to inhibit autophagy flux by impairing autophagosome–lysosome fusion. We hypothesized that autophagy flux altered by CQ has a considerable influence on the lipid composition of endothelial cells. Thus, we investigated endothelial responses induced by CQ on human microvascular endothelial cells (HMEC-1). HMEC-1 cells after CQ exposure were measured using a combined methodology based on label-free Raman and fluorescence imaging. Raman spectroscopy was applied to characterize subtle chemical changes in lipid contents and their distribution in the cells, while the fluorescence staining (LipidTox, LysoTracker and LC3) was used as a reference method. The results showed that CQ was not toxic to endothelial cells and did not result in the endothelial inflammation at concentrations of 1–30 µM. Notwithstanding, it yielded an increased intensity of LipidTox, LysoTracker, and LC3 staining, suggesting changes in the content of neutral lipids, lysosomotropism, and autophagy inhibition, respectively. The CQ-induced endothelial response was associated with lipid accumulation and was characterized by Raman spectroscopy. CQ-induced autophagosome accumulation in the endothelium is featured by a pronounced alteration in the lipid profile, but not in the endothelial inflammation. Raman-based assessment of CQ-induced biochemical changes offers a better understanding of the autophagy mechanism in the endothelial cells

Obraz turystyki w łódzkich mediach i jego odbiór przez mieszkańców w pierwszym roku pandemii COVID-19

Celem opracowania jest analiza przekazu medialnego (przedstawionego w Internecie, lokalnej prasie, telewizji, radiu) dotyczącego turystyki i jego odbioru głównie wśród mieszkańców Łodzi (145 e-ankiet) w pierwszym roku pandemii COVID-19. Turystyka dla Łodzi jest istotną działalnością społeczno-gospodarczą. Pandemia COVID-19 w znacznym stopniu ograniczyła tę działalność. Znalazło to swoje odzwierciedlenie w informacjach przekazywanych przez lokalne media, które szczególnie akcentowały trudności w funkcjonowaniu łódzkich instytucji i przedsiębiorstw turystycznych w 2020 r., spowodowane wprowadzonymi ogólnokrajowymi ograniczeniami w związku z pandemią COVID-19. Opinię tę, przekazywaną przez lokalne media, podzielali również respondenci. Świadczy to o znaczącym wpływie lokalnych mediów na kształtowanie opinii o trudnej sytuacji instytucji i przedsiębiorstw turystycznych w Łodzi, będącej wynikiem pandemii COVID-19.The aim of this study is to analyse the media coverage (presented on the internet, local press, television and radio) concerning tourism and its reception mainly among the inhabitants of Łódź (145 e-questionnaires) during the first year of the COVID-19 pandemic. Tourism is an important socio-economic activity for Łódź and the COVID-19 pandemic significantly reduced this activity. This was reflected in the information provided by the local media, which particularly emphasized the difficulties in the functioning of Łódź institutions and tourism businesses in 2020 caused by the nationwide restrictions imposed due to the pandemic. This position, provided by the local media, was also shared by the respondents and indicates the significant influence of the local media on the formation of opinions about the difficult situation of tourist institutions and businesses in Łódź caused by the pandemic

Raman microscopy reveals how cell inflammation activates glucose and lipid metabolism

Metabolism of endothelial cells (ECs) depends on the availability of the energy substrates. Since the endothelium is the first line of defence against inflammation in the cardiovascular system and its dysfunction can lead to the development of cardiovascular diseases, it is important to understand how glucose metabolism changes during inflammation. In this work, glucose uptake was studied in human microvascular endothelial cells (HMEC-1) in high glucose (HG), and additionally in an inflammatory state, using Raman imaging. HG state was induced by incubation of ECs with a deuterated glucose analogue, while the EC inflammation was caused by TNF-α pre-treatment. Spontaneous and stimulated Raman scattering spectroscopy provided comprehensive information on biochemical changes, including lipids and the extent of unsaturation induced by excess glucose in ECs., induced by excess glucose in ECs. In this work, we indicated spectroscopic markers of metabolic changes in ECs as a strong increase in the ratio of the intensity of lipids / (proteins + lipids) bands and an increase in the level of lipid unsaturation and mitochondrial changes. Inflamed ECs treated with HG, revealed enhanced glucose uptake, and intensified lipid production i.a. of unsaturated lipids. Additionally, increased cytochrome c signal in the mitochondrial region indicated higher mitochondrial activity and biogenesis. Raman spectroscopy is a powerful method for determining the metabolic markers of ED which will better inform understanding of disease onset, development, and treatment

Tunicamycin induced endoplasmatic reticulum changes in endothelial cells investigated in vitro by confocal Raman imaging

This paper describes how tunicamycin (Tu), the most widely used pharmacological agent for inducing endoplasmic reticulum (ER) stress, interacts with endothelial cells. Our results show that tunicamycin enters the cells and accumulates within the ER area. ER stress takes place when improperly folded or damaged proteins begin to accumulate; however, spectroscopic markers of these changes have not been identified as yet. In this work, Raman spectroscopy and scanning electron microscopy imaging of individual endothelial cells treated with Tu were performed. The changes in the biochemical composition of endothelial cells induced by Tu attributed to ER stress were studied in detail. A main feature of the Tu impact on the cells was a decrease of the phospholipid content in the area of ER, and the most abundant lipid with phosphorus groups found there, was identified as sphingomyelin

Fixed versus live endothelial cells : the effect of glutaraldehyde fixation manifested by characteristic bands on the Raman spectra of cells

This work shows an impact of glutaraldehyde (GA) fixation on endothelial cells. Raman spectroscopy imaging was used as a method to monitor biochemical content of the cells due to GA fixation since this is an approach frequently used for studying cells by means of Raman imaging. To get a deeper insight into the changes and to understand them better the measurements of live and fixed cells were performed using two lasers, i.e. 488 and 532 nm. It has been demonstrated that GA fixation affects lipids, proteins, nucleic acid and carbohydrates to small extent. The application of 488 nm laser line seems to be more efficient for live cells due to the small impact of cytochrome resonance on Raman spectra, however 532 nm line is more beneficial for fixed cells due to higher quantum efficiency of the detector, thus leading to higher intensity of Raman bands. Generally, the changes due to fixation are not pronounced but cannot be ignored and the knowledge about them can help in a proper interpretation of data collected for fixed versus live cells