RESUMEN
Regulatory genetic toxicology testing is essential for identifying potentially mutagenic hazards. Duplex Sequencing (DS) is an error-corrected next-generation sequencing technology that provides substantial advantages for mutation analysis over conventional mutagenicity assays including: improved accuracy of mutation detection, ability to measure changes in mutation spectrum, and applicability across diverse biological models. To apply DS for regulatory toxicology testing, power analyses are required to determine suitable sample sizes and study designs. In this study, we explored study designs to achieve sufficient power for various effect sizes in chemical mutagenicity assessment. We collected data from MutaMouse bone marrow and liver samples that were analyzed by DS using TwinStrand's Mouse Mutagenesis Panel. Average duplex reads achieved in two separates studies on liver and bone marrow were 8.4 × 108 (± 7.4 × 107) and 9.5 × 108 (± 1.0 × 108), respectively. Baseline mean mutation frequencies (MF) were 4.6 × 10-8 (± 6.7 × 10-9) and 4.6 × 10-8 (± 1.1 × 10-8), with estimated standard deviations for the animal-to-animal random effect of 0.15 and 0.20, for liver and bone marrow, respectively. We conducted simulation analyses based on these empirically derived parameters. We found that a sample size of four animals per group is sufficient to obtain over 80% power to detect a two-fold change in MF relative to baseline. In addition, we estimated the minimal total number of informative duplex bases sequenced with different sample sizes required to retain power for various effect sizes. Our work provides foundational data for establishing suitable study designs for mutagenicity testing using DS.
Asunto(s)
Médula Ósea , Secuenciación de Nucleótidos de Alto Rendimiento , Hígado , Pruebas de Mutagenicidad , Animales , Médula Ósea/efectos de los fármacos , Hígado/efectos de los fármacos , Pruebas de Mutagenicidad/métodos , Ratones , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Mutágenos/toxicidad , Mutación/efectos de los fármacos , Tasa de Mutación , Proyectos de Investigación , Mutagénesis/efectos de los fármacos , Mutagénesis/genética , MasculinoRESUMEN
Traditional therapy and extensive use of medications and intravaginal autolymphocyte therapy show different results of the treatment of vulvovaginal infections. The purpose of the article was to explore safe and highly effective methods to treat vulvovaginal infections and diseases of the pelvic organs. The standard clinical and laboratory screening of 70 patients of reproductive age was carried out to diagnose the diseases of the reproductive tract. The screening included the description of quantitative and qualitative characteristics of vaginal discharge, examining the mucous covering of the vulva and vagina, microscopic examination of Gram-stained vaginal swabs, endocervical cultures, and diagnosis of sexually transmitted infections using polymerase chain reaction. Intravaginal autolymphocyte therapy was used together with traditionally-accepted treatment schemes (etiotropic antibacterial and antifungal therapy) in the treatment of the main group (40 patients). Traditional treatment methods depending on the etiology of the development of infection were used in the control group (30 patients). The IgM, IgA, and IgG levels were also observed because of the possibility of causing embryo rejection. This study shows that in case of relapsing vulvovaginitis and mixed infections accompanied by disorders of the immune system at different levels, the use of intravaginal autolymphocyte therapy in a comprehensive therapy can be assessed as advisable and pathogenetically substantiated.
