Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused considerable disruption worldwide. For efficient SARS-CoV-2 detection, new methods of rapid, non-invasive sampling are needed. This study aimed to investigate the stability of SARS-CoV-2 in a novel medium for gargle-lavage (GL) self-sampling and to compare the performance of SARS-CoV-2 detection in paired self-collected GL and clinician-obtained nasopharyngeal swab (NPS) samples. The stability study for SARS-CoV-2 preservation in a novel medium was performed over 14 days (4 °C, 24-27 °C, and 37 °C). In total, 494 paired GL and NPS samples were obtained at the University Hospital in Olomouc in April 2021. SARS-CoV-2 detection in paired samples was performed with a SARS-CoV-2 Nucleic Acid Detection Kit (Zybio, Chongqing Municipality, Chongqing, China), an Elecsys® SARS-CoV-2 Antigen assay (Roche Diagnostics, Mannheim, Germany), and a SARS-CoV-2 Antigen ELISA (EUROIMMUN, Lübeck, Germany). The stability study demonstrated excellent SARS-CoV-2 preservation in the novel medium for 14 days. SARS-CoV-2 was detected in 55.7% of NPS samples and 55.7% of GL samples using rRT-PCR, with an overall agreement of 91.9%. The positive percent agreement (PPA) of the rRT-PCR in the GL samples was 92.7%, and the negative percent agreement (NPA) was 90.9%, compared with the NPS samples. The PPA of the rRT-PCR in the NPS and GL samples was 93.2% when all positive tests were used as the reference standard. Both antigen detection assays showed poor sensitivity compared to rRT-PCR (33.2% and 36.0%). rRT-PCR SARS-CoV-2 detection in self-collected GL samples had a similar PPA and NPA to that of NPSs. GL self-sampling offers a suitable and more comfortable alternative for SARS-CoV-2 detection.
In this work, a large set of betulinic acid derivatives modified with various aromatic substituents at the position C-3 were prepared via Suzuki-Myiaura cross-coupling. All compounds were tested for their in vitro cytotoxic activity in 8 cancer and 2 healthy cell lines. Derivatives 6h, 6i, and 6o had the lowest IC50 in the CCRF-CEM cell line (0.69-4.0 μM) and had high selectivity. In addition, 6h and 6i also showed significant activity in daunorubicin-resistant CEM and taxol-resistant K562 cell lines; therefore, they were selected for the evaluation of the mechanism of action. First, the effect of 6h, 6i, and 6o on cell death induction was studied. To our surprise, we have not detected almost any apoptotic cells, even following a long-time exposure of CCRF-CEM cells to the compounds. On the other hand, a dramatic cell number decrease was observed, proportional to the time of the compound's exposure. Based on this data it was concluded that the effect of compounds is cytostatic rather than cytotoxic, which was further confirmed by subsequent studies of the impact of 6h, 6i, and 6o on the cell cycle. Detailed cell cycle analysis revealed a block in the G1 phase accompanied by reduced expression of phosphorylated forms of the RB protein as well as cyclin A protein. Evaluation of the pharmacological properties of the most promising compounds revealed their high stability in the presence of phosphate buffer, human plasma, and microsomes and limited permeability determined using permeability through artificial membrane (PAMPA) and cell permeability assay: Caco-2 and MDCK-MDR1 cell lines. Compounds 6h, 6i, and 6o were selected for further drug development; their cytostatic effect may be advantageous in this process since we expect fewer non-specific interactions and toxicity than in highly cytotoxic compounds. In addition, the activity of 6h and 6i against resistant CEM-DNR and K562-TAX leukemic cell lines makes them promising as a possible future alternative to currently used therapies.
- MeSH
- antitumorózní látky * farmakologie MeSH
- apoptóza MeSH
- Caco-2 buňky MeSH
- cytostatické látky * farmakologie MeSH
- fenotyp MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory * MeSH
- pentacyklické triterpeny farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- databáze jako téma MeSH
- financování organizované MeSH
- lékařská onkologie metody organizace a řízení trendy MeSH
- lidé MeSH
- nádory podle lokalizace MeSH
- registrace klasifikace normy statistika a číselné údaje MeSH
- sběr dat využití MeSH
- software MeSH
- statistika jako téma MeSH
- vládní organizace využití MeSH
- výsledky a postupy - zhodnocení (zdravotní péče) ekonomika metody trendy MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
- MeSH
- chorobopisy statistika a číselné údaje MeSH
- databáze jako téma MeSH
- epidemiologické faktory MeSH
- financování organizované MeSH
- incidence MeSH
- internet statistika a číselné údaje využití MeSH
- lékařská onkologie metody organizace a řízení trendy MeSH
- lidé MeSH
- nádory epidemiologie klasifikace mortalita MeSH
- on-line systémy využití MeSH
- registrace klasifikace normy statistika a číselné údaje MeSH
- software MeSH
- statistika jako téma MeSH
- věkové faktory MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
- MeSH
- databáze jako téma přístrojové vybavení využití MeSH
- internet přístrojové vybavení trendy využití MeSH
- lékařská onkologie metody organizace a řízení trendy MeSH
- lidé MeSH
- nemocniční informační systémy využití MeSH
- registrace normy statistika a číselné údaje MeSH
- sběr dat metody přístrojové vybavení využití MeSH
- statistika jako téma MeSH
- využití lékařské informatiky MeSH
- zobrazování dat trendy využití MeSH
- Check Tag
- lidé MeSH
- MeSH
- databáze faktografické MeSH
- lidé MeSH
- nádory * epidemiologie klasifikace mortalita MeSH
- software MeSH
- statistické modely MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- chemické látky znečišťující vodu farmakokinetika toxicita MeSH
- finanční podpora výzkumu jako téma MeSH
- kosterní svaly metabolismus MeSH
- monitorování životního prostředí metody statistika a číselné údaje MeSH
- řeky MeSH
- rtuť farmakokinetika terapeutické užití MeSH
- ryby MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- Geografické názvy
- Česká republika MeSH
- MeSH
- morbidita MeSH
- mortalita MeSH
- nádory epidemiologie klasifikace prevence a kontrola MeSH
- registrace MeSH
- Geografické názvy
- Česká republika MeSH
