Protective effect of antioxidant enzymes against drug cytotoxicity in MCF-7 cells

Aim: To evaluate protective effect of antioxidant enzymes against epirubicin-HCI (EPI) cytotoxicity in vitro. Materials and Methods: Viability of MCF-7 cells treated with EPI was measured using the MTT test. Glutathione (GSH), protein content and enzymatic activity were measured spectrophotometrically. NADPH — dependent cytochrome P-450 reductase (NADPH-CYP-450) and glutathione S-transferase pi (GST-pi) expression in MCF-7 cells were determined by Western blot analysis. Results: The IC50 values of EPI in MCF-7 cells were 1.0, 0.7 and 0.5 ng/ml respectively for 24, 48 and 72 h applications. Simultaneously enzymatic activity of glutathione S-transferase, glutathione peroxidase, GSH and expression of GST-pi, NADPH-CYP-450 reductase were increased in EPI (1 ng/ml) — treated cells at the end of the 24 h incubation. Addition of superoxide dismutase, catalase and GSH decreased cytotoxicity of EPI. Conclusion: We hypothesized that the production of reactive oxygen species and hydrogen peroxide as result of EPI treatment can cause cytotoxicity in MCF-7 cells and antioxidant enzymes protect the cells against this process.Цель: оценить защитный эффект антиоксидантных ферментов против цитотоксичности эпирубицина-HCI (ЕРІ) in vitro. Материалы и методы: жизнеспособность клеток линии MCF-7, обработанных ЕРІ, оценивали при помощи MTT-метода. Содержание глутатиона (GSH), концентрацию белка и ферментативную активность измеряли спектрофотометрически. Экспрессию NADPH-зависимой цитохром P-450 редуктазы (NADPH-CYP-450) и глутатион-S-трансферазы-pi в клетках линии MCF-7 определяли при помощи Вестерн блот-анализа. Результаты: значения IC50 для клеток MСF-7, обработанных EPI, составили 1,0, 0,7 и 0,5 нг/мл при 24, 48 и 72 ч инкубации соответственно. Через 24 ч инкубации клеток с 1 нг/мл ЕРІ отмечали повышение ферментативной активности глутатион-S-трансферазы и глутатион-пероксидазы, а также уровней экспрессии GST-рi и NADPH-CYP-450 редуктазы. Прединкубация клеток с супероксиддисмутазой, каталазой и GSH приводила к снижению цитотоксичности ЕРІ. Выводы: образование свободных радикалов кислорода и перекиси водорода при действии ЕРІ на клетки линии MCF-7 приводит к цитотоксическому эффекту, в снижении которого участвуют антиоксидантные ферменты

Media coverage and public understanding of sentencing policy in relation to crimes against children

This research examines how the media report on sentences given to those who commit serious crimes against children and how this impacts on public knowledge and attitudes. Three months of press and television coverage were analysed in order to establish the editorial lines that are taken in different sections of the media and how they are promoted by selective reporting of sentencing. Results indicate that a small number of very high profile crimes account for a significant proportion of reporting in this area and often, particularly in the tabloid press, important information regarding sentencing rationale is sidelined in favour of moral condemnation and criticism of the judiciary. Polling data indicate that public attitudes are highly critical of sentencing but also confused about the meaning of tariffs. The article concludes by discussing what can be done to promote a more informed public debate over penal policy in this area

Integrated analyses of single-cell atlases reveal age, gender, and smoking status associations with cell type-specific expression of mediators of SARS-CoV-2 viral entry and highlights inflammatory programs in putative target cells

The COVID-19 pandemic, caused by the novel coronavirus SARS-CoV-2, creates an urgent need for identifying molecular mechanisms that mediate viral entry, propagation, and tissue pathology. Cell membrane bound angiotensin-converting enzyme 2 (ACE2) and associated proteases, transmembrane protease serine 2 (TMPRSS2) and Cathepsin L (CTSL), were previously identified as mediators of SARS-CoV2 cellular entry. Here, we assess the cell type-specific RNA expression of ACE2, TMPRSS2, and CTSL through an integrated analysis of 107 single-cell and single-nucleus RNA-Seq studies, including 22 lung and airways datasets (16 unpublished), and 85 datasets from other diverse organs. Joint expression of ACE2 and the accessory proteases identifies specific subsets of respiratory epithelial cells as putative targets of viral infection in the nasal passages, airways, and alveoli. Cells that co-express ACE2 and proteases are also identified in cells from other organs, some of which have been associated with COVID-19 transmission or pathology, including gut enterocytes, corneal epithelial cells, cardiomyocytes, heart pericytes, olfactory sustentacular cells, and renal epithelial cells. Performing the first meta-analyses of scRNA-seq studies, we analyzed 1,176,683 cells from 282 nasal, airway, and lung parenchyma samples from 164 donors spanning fetal, childhood, adult, and elderly age groups, associate increased levels of ACE2, TMPRSS2, and CTSL in specific cell types with increasing age, male gender, and smoking, all of which are epidemiologically linked to COVID-19 susceptibility and outcomes. Notably, there was a particularly low expression of ACE2 in the few young pediatric samples in the analysis. Further analysis reveals a gene expression program shared by ACE2(+)TMPRSS2(+) cells in nasal, lung and gut tissues, including genes that may mediate viral entry, subtend key immune functions, and mediate epithelial-macrophage cross-talk. Amongst these are IL6, its receptor and co-receptor, IL1R, TNF response pathways, and complement genes. Cell type specificity in the lung and airways and smoking effects were conserved in mice. Our analyses suggest that differences in the cell type-specific expression of mediators of SARS-CoV-2 viral entry may be responsible for aspects of COVID-19 epidemiology and clinical course, and point to putative molecular pathways involved in disease susceptibility and pathogenesis

Investigation of protective effects of selenium and vitamin e against DNA oxidation, membrane damage and alteration of comt metabolism in smoke-exposed male mice (Mus Musculus, Balb/c)

In our study, the protective effects of vitamin E and Se (selenium) against cigarette smoke hazards on second-hand smoker (passive smoker) male mice (Balb/c) were investigated. Serum MDA levels in the smoke-exposed mice were found higher than serum MDA levels of control mice and Se- and vitamin E-treated mice. But, the MDA levels of smoke-exposed plus Se- and vitamin E-treated mice were found lower than MDA levels of smoke-exposed mice at the end of the three and five months. According to these results, application of vitamin E and Se, when given to smoke-exposed mice together, had an additive protective effect against cigarette smoke hazards (p<0.05). Vitamin E also had protective effect on formation of 8-OHdG in smoke-exposed mice. The serum 8-OHdG amounts of smoke-exposed plus vitamin E-treated mice were found low, but the serum 8-OHdG amounts of smoke-exposed mice were found high. Also 8-OHdG levels in the serum of the smoke-exposed mice were increased which occurs as a result of DNA oxidation (p<0.05). At the end of the three and five months, COMT (catechol-o-methyl transferase) activity of smoke-exposed mice livers were increased but, vitamin E and/or Se showed a significant protective effect on changing of COMT activity only at the end of the 5 months. Our results showed that MDA levels and 8-OHdG amounts were increased in the serum of smoke-exposed mice. On the other hand, vitamin E and Se had an additive protective effect against increasing MDA level. Also vitamin E had a protective effect against formation of 8-OHdG amounts and COMT activity alterations