Ferroptosis, an iron-dependent form of cell death, and dysregulated microRNA (miRNA) expression correlate with colorectal cancer (CRC) development and progression. The tumor suppressor ability of miR-148a-3p has been reported for several cancers. Nevertheless, the role of miR-148a-3p in CRC remains largely undetermined. Here, we aim at investigating the molecular mechanisms and regulatory targets of miR-148a-3p in the CRC cell death mechanism(s). To this end, miR-148a-3p expression was evaluated in SW480 and SW620 cells and normal colon epithelial CCD 841 CoN cells with quantitative real-time polymerase chain reaction (qRT-PCR). Data reported a reduction of miR-148a-3p expression in SW480 and SW620 cells compared to non-tumor cells (p < 0.05). Overexpression of miR-148a selectively inhibited CRC cell viability (p < 0.001), while weakly affecting normal CCD 841 CoN cell survival (p < 0.05). At the cellular level, miR-148a-3p mimics promoted apoptotic cell death via caspase-3 activation (p < 0.001), accumulation of mitochondrial reactive oxygen species (ROS) (p < 0.001), and membrane depolarization (p < 0.001). Moreover, miR-148a-3p overexpression induced lipid peroxidation (p < 0.01), GPX4 downregulation (p < 0.01), and ferroptosis (p < 0.01), as revealed by intracellular and mitochondrial iron accumulation and ACSL4/TFRC/Ferritin modulation. In addition, levels of SLC7A11 mRNA and protein, the cellular targets of miR-148a-3p predicted by bioinformatic tools, were suppressed by miR-148a-3p’s overexpression. On the contrary, the downregulation of miR-148a-3p boosted SLC7A11 gene expression and suppressed ferroptosis. Together, these in vitro findings reveal that miR-148a-3p can function as a tumor suppressor in CRC by targeting SLC7A11 and activating ferroptosis, opening new perspectives for the rationale of therapeutic strategies through targeting the miR-148a-3p/SLC7A11 pathway.

Buffalo (Bubalus bubalis) is the 5th species bred in the world after cows, sheep, goats and pigs with 203,939,158 heads (FAO 2023). Buffaloes are mainly reared for milk production and, globally, in...

Canine mammary tumours (CMTs) are the most common cancer in intact female dogs. In addition to surgery, additional targeted and non-targeted therapies may offer survival benefits to these patients. Therefore, exploring new treatments for CMT is a promising area in veterinary oncology. CMT cells have an altered lipid metabolism and use the oxidation of fatty acids for their energy needs. Here we investigated the tumoricidal effects of teglicar, a reversible inhibitor of carnitine palmitoyl transferase 1A (CPT1A), the rate-limiting enzyme for fatty acid import into mitochondria, on two CMT cells, P114 and CMT-U229. Viability and apoptosis were examined in CMT cells using the crystal violet assay, trypan blue assay, and flow cytometry analysis. The expression of mediators of apoptosis signalling (e.g., caspase-9, caspase-8, and caspase-3) was assessed by quantitative real-time polymerase chain reaction and western blot analyses. Teglicar was able to decrease cell viability and induce apoptosis in P114 and CMT-U229 cells. At the molecular level, the effect of teglicar was associated with an upregulation of the mRNA expression levels of caspase-9, caspase-8, and caspase-3 and an increase in their protein levels. In summary, our results show that teglicar has a potential effect against CMTs through the induction of apoptotic cell death, making it a promising therapeutic agent against CMTs.

Background/Objectives: Colorectal cancer (CRC) remains one of the most common and deadly malignancies worldwide, driven by metabolic reprogramming and mitochondrial dysfunction, which support tumor growth and progression. Several studies showed that nutrition is a contributing factor in the prevention and management of CRC. In this context, carnitines, amino acid derivatives abundant in food of animal origin, such as meat and milk, are crucial for mitochondrial function. Recently, l-carnitine and acetyl-l-carnitine have received particular attention due to their antioxidant, anti-inflammatory, and antitumor properties. However, to date, there is no conclusive evidence on the effects of l-carnitine and acetyl-l-carnitine in CRC or the underlying molecular mechanism. Methods: In this study, we investigated in HCT 116 and HT-29 CRC cells the effects of l-carnitine and acetyl-l-carnitine on mitochondrial homeostasis by XF HS Seahorse Bioanalyzer and cell death pathways by flow cytometry and western blot assays. Results: Data showed that l-carnitine and acetyl-l-carnitine reduced cell viability (p < 0.001), modulated cellular bioenergetics, and induced oxidative stress (p < 0.001). These phenomena promoted autophagic flux and the mitophagy process via PINK1 and Parkin modulation after 72 h of treatment. Of note, the combined treatment with l-carnitine and acetyl-l-carnitine showed a synergistic effect and enhanced the effect of single carnitines on tumor cell growth and metabolic dysfunction (p < 0.05). Moreover, exposure to l-carnitine and acetyl-l-carnitine promoted CRC cell apoptosis, suggesting a mechanism involving mitophagy-related cell death. These data were associated with increased SIRT4 expression levels (p < 0.01) and the activation of AMPK signaling (p < 0.01). Conclusions: Overall, the results, by supporting the importance of nutritional factors in CRC management, highlight l-carnitine and acetyl-l-carnitine as promising agents to target CRC metabolic vulnerabilities.

This study investigated the use of K-means and hierarchical clustering, to group Italian Mediterranean buffalo using routinely collected test-day records. The analysis was first conducted on a combined dataset comprising three buffalo herds and subsequently on each herd individually. The main objective was to determine whether data-driven groupings could be implemented to support improvements in general herd management strategies. Results indicated that K-means consistently outperformed hierarchical clustering across all datasets, as reflected by average silhouette scores (0.17–0.18 vs. 0.10–0.12 for K-means and hierarchical, respectively), favorable Davies–Bouldin Index (DBI; 2.05–2.16 vs. 2.11–2.5 for K-means and hierarchical, respectively) and Calinski–Harabasz Index values (CHI; 1034–3877 vs. 729–2109 for K-means and hierarchical, respectively). K-means identified two clusters in the combined dataset and in two of the three herds, while three clusters were identified in the remaining herd. Cluster composition analysis revealed that days in milk and milk yield were the main discriminating factors when two clusters were formed. When three clusters emerged, K-means also identified a subgroup of animals that differed from the others in both age and lactation stage. These findings were supported by the analysis of variance (ANOVA), which showed statistically significant differences among most of the evaluated variables.

Cortisol and DHEA(S) are markers of allostatic load and resilience; as such, they may be added to the definition of performance measures alongside immunocompetence. We aimed at studying the potential carryover effects of management practices on growth performances and the interrelationship between the hypothalamic–pituitary–adrenal (HPA) axis, immunocompetence and performances during the preweaning period in dairy calves. This study consisted of first a retrospective cohort study and second a prospective study. Calves had their growth performances and disease occurrence recorded, and their hair steroids (T1 at 20.03 ± 0.39 and T2 at 50.83 ± 0.41 days from birth) and plasmatic and colostral immunoglobulin G (IgG) was measured by immunoassays. The occurrence of preweaning diseases and the duration of individual housing negatively correlated to body weight (BW) at 60 days and the hair cortisol-to-DHEA(S) ratio (HC/HDHEA(S)) (T2) was negatively correlated to the daily weight gain (DWG) (−0.36; p < 0.05), plasmatic IgG (plaIgG) and the apparent efficiency of absorption (AEA) (−0.41; p < 0.01). Our results confirmed that the occurrence of diseases during the preweaning period and individual housing duration are pivotal for calves’ growth. This prospective study highlighted a detrimental effect of allostatic load on immunocompetence and described a link between resilience and growth performances.