Redox aspects of cytotoxicity and anti-neuroinflammatory profile of chloroquine and hydroxychloroquine in serum-starved BV-2 microglia.

Chloroquine (CQ) and hydroxychloroquine (HCQ) have long been used worldwide to treat and prevent human malarias. However, these 4-aminoquinolines have also shown promising potential in treating chronic illnesses with an inflammatory component, including neurological diseases. Given the current demand for serum avoidance during pharmacological testing and modeling of some pathologies, we compared cytotoxicities of CQ and HCQ in both serum-deprived and -fed murine BV-2 microglia. Furthermore, we assessed the anti-neuroinflammatory potential of both compounds in serum-deprived cells. Under both conditions, CQ showed higher cytotoxicity than HCQ. However, the comparable MTT-assay-derived data measured under different serum conditions were associated with disparate cytotoxic mechanisms of CQ and HCQ. In particular, under serum starvation, CQ mildly enhanced secondary ROS, mitochondrial hyperpolarization, and decreased phagocytosis. However, CQ promoted G1 phase cell cycle arrest and mitochondrial depolarization in serum-fed cells. Under both conditions, CQ fostered early apoptosis. Additionally, we confirmed that both compounds could exert anti-inflammatory effects in microglia through interference with MAPK signaling under nutrient-deprivation-related stress. Nevertheless, unlike HCQ, CQ is more likely to exaggerate intracellular prooxidant processes in activated starved microglia, which are inefficiently buffered by Nrf2/HO-1 signaling pathway activation. These outcomes also show HCQ as a promising anti-neuroinflammatory drug devoid of CQ-mediated cytotoxicity.

Skin Aging, Cellular Senescence and Natural Polyphenols.

The skin, being the barrier organ of the body, is constitutively exposed to various stimuli impacting its morphology and function. Senescent cells have been found to accumulate with age and may contribute to age-related skin changes and pathologies. Natural polyphenols exert many health benefits, including ameliorative effects on skin aging. By affecting molecular pathways of senescence, polyphenols are able to prevent or delay the senescence formation and, consequently, avoid or ameliorate aging and age-associated pathologies of the skin. This review aims to provide an overview of the current state of knowledge in skin aging and cellular senescence, and to summarize the recent in vitro studies related to the anti-senescent mechanisms of natural polyphenols carried out on keratinocytes, melanocytes and fibroblasts. Aged skin in the context of the COVID-19 pandemic will be also discussed.

Proteasome Biology: Chemistry and Bioengineering Insights.

Proteasomal degradation provides the crucial machinery for maintaining cellular proteostasis. The biological origins of modulation or impairment of the function of proteasomal complexes may include changes in gene expression of their subunits, ubiquitin mutation, or indirect mechanisms arising from the overall impairment of proteostasis. However, changes in the physico-chemical characteristics of the cellular environment might also meaningfully contribute to altered performance. This review summarizes the effects of physicochemical factors in the cell, such as pH, temperature fluctuations, and reactions with the products of oxidative metabolism, on the function of the proteasome. Furthermore, evidence of the direct interaction of proteasomal complexes with protein aggregates is compared against the knowledge obtained from immobilization biotechnologies. In this regard, factors such as the structures of the natural polymeric scaffolds in the cells, their content of reactive groups or the sequestration of metal ions, and processes at the interface, are discussed here with regard to their influences on proteasomal function.

Protoflavones in melanoma therapy: Prooxidant and pro-senescence effect of protoapigenone and its synthetic alkyl derivative in A375 cells.

AIMS: In our study, the anticancer effects of a semisynthetic p-quinol, protoapigenone 1'-O-butyl ether (PABut), were tested in human melanoma A375 cells also in comparison with natural congener, protoapigenone (PA). MAIN METHODS: The cytotoxic effect of PABut and PA was determined using MTT assay. Flow cytometry analysis was used to evaluate the influence of the compounds tested on ROS generation and cell cycle distribution in A375 cells. Moreover, apoptosis was evaluated by AO/EB dual staining as well as by flow cytometry. Markers of senescence were quantified by spectrofluorimetry and by Western blot analysis. KEY FINDINGS: Both PABut and PA showed significant cytotoxicity against melanoma A375 cells at sub-micromolar concentrations. Both protoflavones induced comparable cell cycle arrest in G2/M phase. However, a more profound upregulation of intracellular ROS levels was found following PABut treatment. An increased apoptosis in the cells following 48 h treatment with both protoflavones tested was also confirmed. Both compounds tested remarkably upregulated p21 protein levels in A375 cells. Unlike PA, PABut significantly decreased protein levels of NAD+-dependent deacetylase SirT1 and ß-actin accompanied by mild significant upregulation of mitochondrial SOD2 and senescence markers, p16 protein and SA-ß-Gal activity. However, a significant upregulation of p53 only following PA treatment was found. SIGNIFICANCE: These results suggest that PABut and PA confer high chemotherapeutic potential in melanoma cells and are suitable for further testing. Furthermore, modification of protoapigenone with 1'-O-butyl ether moiety can be associated with improved senescence-inducing effect and, thus, enhanced chemotherapeutic potency of PABut compared to the unmodified natural protoflavone.

Result of Prospective Validation of the Trisomy Test® for the Detection of Chromosomal Trisomies.

Noninvasive prenatal testing (NIPT) is one of the most common prenatal screening tests used worldwide. Trisomy Test® belongs to NIPT tests based on low-coverage whole-genome sequencing. In our prospective study, 7279 samples of pregnant women collected during approximately two years were analyzed. In this cohort, 117 positive cases for trisomies 21, 18, and 13 were reported. An in-house designed bioinformatic pipeline and proprietary biostatistical approach was used for the detection of trisomies. The pooled sensitivity and specificity of our test reached 99.12% and 99.94%, respectively. The proportion of repeatedly uninformative results after repeated blood draws was 1.11%. Based on the presented results, we can confirm that the Trisomy Test® is fully comparable with other commercial NIPT tests available worldwide.