Effects on mammalian cell transcriptome reveal a plant extract's medicinal properties: Potential anti-COVID-19 and anti-cancer properties of Sarcopoterium spinosum

Plants with medicinal properties are usually identified based on traditional medicine knowledge or using low-throughput screens for specific pharmacological activities. Here, we suggest a different approach to uncover a range of pharmacological activities of a chosen plant extract without the need for functional screening. This tactic predicts biological activities of a plant extract based on pathway analysis of transcriptome changes caused by the extract in mammalian cell culture. In this work, we identified transcriptome changes after exposure of cultured cells to an extract of the medicinal plant Sarcopoterium spinosum. Gene Set Enrichment Analysis (GSEA) confirmed known anti-inflammatory and anti-cancer activities of the extract and predicted novel biological effects on oxidative phosphorylation and interferon pathways. Experimental validation of these pathways uncovered strong activation of autophagy, including mitophagy, and astounding protection from SARS-CoV-2 infection. Our study shows that gene expression analysis alone is insufficient for predicting biological effects since some of the changes reflect compensatory effects, and additional biochemical tests provide necessary corrections. In conclusion, this study defines the advantages and limitations of an approach towards predicting the biological and medicinal effects of plant extracts based on transcriptome changes caused by these extracts in mammalian cells.

Influence of Potassium Ions on Act of Amphotericin B to the DPPC/Chol Mixed Monolayer at Different Surface Pressures

Amphotericin B (AmB) is an antifungal drug that rarely develops resistance. It has an affinity with the cholesterol on mammalian cell membranes, disrupting the structure and function of the membranes, which are also affected by potassium ions. However, the mechanism is unclear. In this paper, the Langmuir monolayer method was used to study the effects of potassium ions on the surface pressure–mean molecular area of isotherms, elastic modulus and the surface pressure–time curves of a 1,2-dipalmitoyl-sn-glycero-3-phosphocholine/cholesterol (DPPC/Chol) monolayer and a DPPC/Chol/AmB monolayer. The morphology and thickness of the Langmuir–Blodgett films were studied via atomic force microscopy. The results showed that AmB can increase the mean molecular area of the DPPC/Chol mixed monolayer at low pressures (15 mN/m) but reduces it at high pressures (30 mN/m). The potassium ions may interfere with the effect of AmB in different ways. The potassium ions can enhance the influence of AmB on the stability of monolayer at low surface pressures, but weaken it at high surface pressures. The potassium ions showed significant interference with the interaction between AmB and the cholesterol-enriched region. The results are helpful for us to understand how the effect of amphotericin B on the phospholipid membrane is interfered with by potassium ions when amphotericin B enters mammalian cell membrane.

Differentiating Trypanosoma cruzi in a Host Mammalian Cell Imaged in Aqueous Liquid by Atmospheric Scanning Electron Microscopy

Using Atmospheric Scanning Electron Microscopy (ASEM), we visualized interaction between infectious stage of Trypanosoma cruzi and completely intact host mammalian cell. Plasma membrane appears translucent under ASEM, which not only enables direct observation of T. cruzi within its host cell, but also reveals internal structures of the parasite itself.

Macromolecular cryoprotectants for the preservation of mammalian cell culture: lessons from crowding, overview and perspectives

Cryopreservation is a process used for the storage of mammalian cells at a very low temperature, in a state of ‘suspended animation’.

Assessing Phototoxicity in a Mammalian Cell Line: How Low Levels of Blue Light Affect Motility in PC3 Cells

Phototoxicity is a significant constraint for live cell fluorescence microscopy. Excessive excitation light intensities change the homeostasis of the observed cells. Erroneous and misleading conclusions may be the problematic consequence of observing such light-induced pathophysiology. In this study, we assess the effect of blue light, as commonly used for GFP and YFP excitation, on a motile mammalian cell line. Tracking PC3 cells at different light doses and intensities, we show how motility can be used to reliably assess subtle positive and negative effects of illumination. We further show that the effects are a factor of intensity rather than light dose. Mitotic delay was not a sensitive indicator of phototoxicity. For early detection of the effect of blue light, we analysed the expression of genes involved in oxidative stress. This study addresses the need for relatively simple and sensitive methods to establish a dose-response curve for phototoxicity in mammalian cell line models. We conclude with a working model for phototoxicity and recommendations for its assessment.

Analyses of Safety Profile and Homologous Antibody Responses to a Mammalian Cell-Based, MF59-Adjuvanted, A/H5N1, Pandemic Influenza Vaccine across Four Phase II/III Clinical Trials in Healthy Children, Adults, and Older Adults

Modern cell culture-based technology eliminates vaccine manufactures reliance on embryonated chicken eggs, which may become compromised during an avian influenza pandemic. Four studies (total N = 6230) assessed the immunogenicity and safety of mammalian cell-based, MF59®-adjuvanted, A/H5N1 vaccine (aH5N1c; AUDENZ™) as two doses administered on Days 1 and 22 in children (NCT01776554), adults (NCT01776541; NCT02839330), and older adults (NCT01766921; NCT02839330). Immunogenicity of formulations at 7.5 μg and 3.75 μg antigen per dose were assessed by hemagglutination inhibition and microneutralization assays on Days 1, 22, 43, and 183 or 387. Solicited local and systemic adverse events (AEs) were recorded for 7 days after each vaccination. Unsolicited AEs were collected for 21 days after each vaccination, and serious and other selected AEs were recorded for one year. Antibody responses after two 7.5 μg doses met CBER licensure criteria in all age groups. Overall, an age-related response was evident, with the highest responses observed in children <3 years old. In children, antibody titers met seroconversion criteria 12 months after vaccination. MF59 allowed for antigen dose sparing. Solicited AEs were mild to moderate in nature, of short duration, and less frequent after the second dose than the first, demonstrating a favorable risk-benefit profile.