Metformin/Leucine

Hypoxia stress has been demonstrated to impede animal embryonic development, spermatogenesis, and lactation, leading to decreased animal production performance. However, the impact of hypoxia-induced activation of hypoxia inducible factor-1 (HIF-1) signaling on milk protein and fat synthesis remains unclear. L-leucine, a branched-chain amino acid, is known to modulate milk protein and fat synthesis. Therefore, our study aimed to evaluate the effect of L-leucine on milk protein and fat synthesis under hypoxic conditions and shed light on the molecular mechanism using an in vitro model. The results indicated that hypoxia treatment significantly decreased the synthesis of α-casein and β-casein, as well as inhibited factors related to milk fat synthesis in bovine mammary epithelial cells (MAC-T). Additionally, hypoxia stress suppressed the activities of the mammalian target of rapamycin (mTOR) and protein kinase B (AKT). Interfering with HIF-1α significantly reversed the expression of AKT, mTOR and factors related to milk synthesis. Importantly, supplementation with L-leucine activated AKT/mTOR signaling, thereby enhancing milk protein and fat synthesis in MAC-T cells to some extent. In conclusion, these findings suggest that HIF-1 signaling plays an important role in milk synthesis and that L-leucine may stimulate the synthesis of milk protein and fat by activating the AKT/mTOR signaling pathway under hypoxic conditions, making it a potential additive for promoting milk synthesis inhibited by hypoxia.

Azole-resistant Aspergillus fumigatus (A. fumigatus) is an emerging worldwide pathogen. Pulmonary aspergillosis primarily affects severely immunocompromised patients and is also a particularly critical condition for cystic fibrosis (CF) patients. A recently designed gallium polypyridyl catecholate complex, GaS1, has previously demonstrated in vitro and in vivo antimicrobial activity against Gram-negative bacteria. In the present work GaS1 activity was assessed against A. fumigatus clinical isolates in a novel air-liquid-interface lung infection model, mimicking the conditions found in the CF airways. Furthermore, in this study both a solution for nebulisation and dry powders for inhalation were developed with a view to optimising GaS1 delivery to the lung. The solution for nebulisation was characterised for its osmolality and pH, while the dry powders were characterised by scanning electron microscopy, powder X-ray diffraction, thermal analysis and laser light scattering particle size analysis. The aerodynamic deposition profiles of all formulations were determined using a next generation impactor. GaS1, tested in a concentration range of 0.016-0.5 mg/mL, inhibited the growth of A. fumigatus lung isolates in a complex host-environment-mimicking medium at the non-toxic concentration of 0.063 mg/mL. A marked dose-dependent antifungal activity of GaS1 was also observed in the presence of differentiated human distal lung epithelial cells (NCI-H441) at the air liquid interface, with nearly no fungal growth detected at the macroscopic and microscopic level. A solution for nebulisation and three different dry powder inhaler formulations, prepared by spray-drying GaS1 with different concentrations of L-leucine, displayed suitable aerodynamic characteristics for GaS1 delivery to the lungs, while maintaining excellent antifungal activity. Overall, the results obtained highlight the potential of gallium-polypyridyl catecholate complexes for the management of difficult-to-treat A. fumigatus pulmonary infections.