Background: The mitogenomes of vascular plants are one of the most structurally diverse molecules. In the present study we characterize mitogenomes of a rare and endangered speciesPulsatilla patens. We investigated the gene content and its RNA editing potential, repeats distribution and plastid derived sequences.Results: The mitogenome structure of early divergent eudicot, endangeredPulsatilla patensdoes not support the master chromosome hypothesis, revealing the presence of three linear chromosomes of total length 986 613 bp. The molecules are shaped by the presence of extremely long, exceeding 87 kbp repeats and multiple chloroplast-derived regions including nearly complete inverted repeat. Since the plastid IR content of Ranunculales is very characteristic, the incorporation into mitogenome could be explained rather by intracellular transfer than mitochondrial HGT. The mitogenome contains almost a complete set of genes known from other vascular plants with exception ofrps10 andsdh3, the latter being present but pseudogenized. Analysis of long ORFs enabled the identification of genes which are rarely present in plant mitogenomes, including RNA and DNA polymerases, albeit their presence even at species level is variable. Mitochondrial transcripts ofP. patenswere edited with a high frequency, which exceeded the level known in other analyzed angiosperms, despite the strict qualification criteria of counting the editing events and taking into analysis generally less frequently edited leaf transcriptome. The total number of edited sites was 902 andnad4 was identified as the most heavily edited gene with 65 C to U changes. Non-canonical, reverse U to C editing was not detected. Comparative analysis of mitochondrial genes of threePulsatillaspecies revealed a level of variation comparable to chloroplast CDS dataset and much higher infrageneric differentiation than in other known angiosperm genera. The variation found in CDS of mitochondrial genes is comparable to values found amongPulsatillaplastomes. Despite the complicated mitogenome structure, 14 single copy regions of 329 kbp, not splitted by repeats or plastid-derived sequences (MTPT), revealed the potential for phylogenetic, phylogeographic and population genetics studies by revealing intra- and interspecific collinearity.Conclusions: This study provides valuable new information about mitochondrial genome of early divergent eudicots,Pulsatilla patens, revealed multi-chromosomal structure and shed new light on mitogenomics of early eudicots.

An active form of vitamin D₃ (1,25-dihydroxyvitamin D₃) acts through vitamin D receptor (VDR) initiating genomic response, but several studies described also non-genomic actions of 1,25-dihydroxyvitamin D₃, implying the role of PDIA3 in the process. PDIA3 is a membrane-associated disulfide isomerase involved in disulfide bond formation, protein folding, and remodeling. Here, we used a transcriptome-based approach to identify changes in expression profiles in PDIA3-deficient squamous cell carcinoma line A431 after 1,25-dihydroxyvitamin D₃ treatment. PDIA3 knockout led to changes in the expression of more than 2000 genes and modulated proliferation, cell cycle, and mobility of cells; suggesting an important regulatory role of PDIA3. PDIA3-deficient cells showed increased sensitivity to 1,25-dihydroxyvitamin D₃, which led to decrease migration. 1,25-dihydroxyvitamin D₃ treatment altered also genes expression profile of A431ΔPDIA3 in comparison to A431WT cells, indicating the existence of PDIA3-dependent genes. Interestingly, classic targets of VDR, including CAMP (Cathelicidin Antimicrobial Peptide), TRPV6 (Transient Receptor Potential Cation Channel Subfamily V Member 6), were regulated differently by 1,25-dihydroxyvitamin D₃, in A431ΔPDIA3. Deletion of PDIA3 impaired 1,25-dihydroxyvitamin D₃-response of genes, such as PTGS2, MMP12, and FOCAD, which were identified as PDIA3-dependent. Additionally, response to 1,25-dihydroxyvitamin D₃ in cancerous A431 cells differed from immortalized HaCaT keratinocytes, used as non-cancerous control. Finally, silencing of PDIA3 and 1,25-dihydroxyvitamin D₃, at least partially reverse the expression of cancer-related genes in A431 cells, thus targeting PDIA3 and use of 1,25-dihydroxyvitamin D₃ could be considered in a prevention and therapy of the skin cancer. Taken together, PDIA3 has a strong impact on gene expression and physiology, including genomic response to 1,25-dihydroxyvitamin D₃.

Background: Sarcopenia is linked with high rates of adverse surgical outcomes, and computed tomography angiography (CTA)-based psoas measurements are used as imaging sarcopenia surrogates. Their prognostic value in patients with chronic limb-threatening ischemia (CLTI) undergoing revascularization remains uncertain. Objectives: To evaluate whether CTA-derived psoas muscle indices predict complications and mortality after lower-limb revascularization for CLTI. Methods: We performed a retrospective cohort study of consecutive adults who underwent open, hybrid, or endovascular revascularization for CLTI at a single tertiary center (March 2018–December 2021). Psoas muscle area (PMA) and density (PMD) were measured preoperatively on CTA at the mid-L3 vertebral level. Psoas muscle index (PMI) was calculated as PMA/height². Patients were stratified by tertiles for each index (lowest tertile = “sarcopenic” vs. upper two tertiles). Outcomes included early in-hospital complications, late complications, overall complications, late mortality, and overall mortality. Group comparisons used χ²/Fisher tests with false discovery rate (FDR) adjustment; multivariable logistic regression with AIC-guided selection assessed independent predictors. Results: A total of 234 patients were included (median age 68 years; 65.4% men). Early complications occurred in 15.8%; late complications in 70.3%; overall mortality during follow-up was 26.6% (38/143 within follow-up data). In tertile analyses, none of the psoas-derived measures were significantly associated with early complications, late complications, overall complications, or mortality after FDR correction. Lower PMD showed consistent but non-significant trends toward higher late complications (84% vs. 64%), overall complications (87% vs. 72%), overall mortality (38% vs. 21%), and late mortality (37% vs. 20%) (all p < 0.05 unadjusted; all p_adj ≥ 0.139). In multivariable models, PMA, PMD, and PMI were not independent predictors of any outcome. Conclusions: In this retrospective cohort study, preoperative CTA-derived psoas indices were not independent predictors of early, late, or overall complications, nor of in-hospital or follow-up mortality after revascularization for chronic limb-threatening ischemia. Although lower psoas muscle density showed consistent trends toward higher risk, these associations did not reach statistical significance after adjustment. Taken together, our findings suggest that psoas-based measures have limited prognostic value in this setting and should be interpreted cautiously, while their potential role warrants confirmation in larger, prospective studies.

Background: Chronic limb-threatening ischemia (CLTI) is associated with high morbidity and substantial healthcare utilization. Length of hospital stay (LOS) after lower limb revascularization is influenced by procedural complexity, but patient physiological reserve may also play a role. We evaluated whether CT-derived lean psoas muscle area (LPMA) is independently associated with LOS in patients undergoing revascularization for CLTI. Methods: We retrospectively analyzed 234 consecutive patients treated with endovascular, hybrid, or open revascularization for CLTI (Rutherford 4–5) between 2018 and 2021. Sarcopenia markers were derived from preoperative CT at the L3 level, including psoas muscle area (PMA), muscle density (PMD), and LPMA. Multivariable linear regression models with log-transformed LOS were used to estimate relative effects on hospitalization duration. Results: Median age was 68 years and 65.4% were male; 76.5% of admissions were urgent. Median LOS was 6 days (IQR 4–9). Procedure type was the strongest determinant of LOS: hybrid (β = 0.69, p < 0.001) and open surgery (β = 0.73, p < 0.001) were associated with approximately 99% and 108% longer LOS compared with endovascular treatment. Higher LPMA was independently associated with shorter LOS (β = −0.00049, p = 0.004). Smoking (β = −0.21, p = 0.003) and history of myocardial infarction (β = −0.19, p = 0.030) were associated with shorter LOS, whereas dialysis showed a non-significant trend toward longer hospitalization (β = 0.36, p = 0.056). Conclusions: In patients undergoing lower limb revascularization for CLTI, CT-derived LPMA demonstrated a modest but independent association with hospital stay duration after adjustment for procedural and clinical factors. Given the exploratory nature of this study, these hypothesis-generating findings support further evaluation of imaging-based muscle assessment as an adjunct marker of physiological reserve in this high-risk population.