Multiple respiratory chain deficiencies represent a common cause of mitochondrial diseases and are associated with a wide range of clinical symptoms. We report a subject, born to consanguineous parents, with growth retardation and neurological deterioration. Multiple respiratory chain deficiency was found in muscle and fibroblasts of the subject as well as abnormal assembly of complexes I and IV. A microsatellite genotyping of the family members detected only one region of homozygosity on chromosome 17q24.2-q25.3 in which we focused our attention to genes involved in mitochondrial translation. We sequenced MRPL12, encoding the mitochondrial ribosomal protein L12 and identified a c.542C>T transition in exon 5 changing a highly conserved alanine into a valine (p.Ala181Val). This mutation resulted in a decreased steady-state level of MRPL12 protein, with altered integration into the large ribosomal subunit. Moreover, an overall mitochondrial translation defect was observed in the subject's fibroblasts with a significant reduction of synthesis of COXI, COXII and COXIII subunits. Modeling of MRPL12 shows Ala181 positioned in a helix potentially involved in an interface of interaction suggesting that the p.Ala181Val change might be predicted to alter interactions with the elongation factors. These results contrast with the eubacterial orthologues of human MRPL12, where L7/L12 proteins do not appear to have a selective effect on translation. Therefore, analysis of the mutated version found in the subject presented here suggests that the mammalian protein does not function in an entirely analogous manner to the eubacterial L7/L12 equivalent.

Astragalus L. species are recognized for their prominent biological activities, making them useful in the treatment of several diseases. Astragalus spruneri (AS), a perennial prostrate herbaceous wild plant endemic to Southeast Europe and Turkey, remains insufficiently explored to date. The present study investigated total phenolic content, DPPH radical-scavenging activity, and in vitro cytotoxic activity of extracts from aerial parts of A. spruneri. We analyzed the potential of methanolic (ASM) and ethanolic (ASE) extracts to induce cell death in MCF-7 cells using the Incucyte® Cytotox NIR dye and bright-field microscopy. ASE of the leaf was richer in flavonoids, whereas ASM of the leaf displayed higher antioxidant activity than the stem. Live-cell analysis showed that ASE of the leaf exhibited the highest cytotoxic potency (EC50 = 125 μg/mL after 48 hours) and induced a time- and concentration-dependent increase in Cytotox NIR signal. Compared with the vehicle control, ASE of the leaf produced an earlier onset and more significant sustained cytotoxic activity in MCF-7 cells. Western blot analysis further demonstrated that AS extract modulated PUMA protein expression. The Cytotox NIR assay provided new insights into distinct extract- and tissue-dependent cytotoxic responses against MCF-7 cancer cells, with ethanolic leaf extracts exhibiting the most prominent effect. Thus, it can be concluded that A. spruneri serves as a valuable source of natural bioactive compounds with promising cytotoxic activity. Hence, further research is needed to confirm its anticancer potential and assess its safety profile.