ABSTRACT
The fatty acid (FA) and phospholipid composition of dietary lecithin may influence FA digestibility and milk production in cattle. Eight multiparous Holstein cows (99.4 ± 9.2 d in milk [DIM]; 48.9 ± 3.8 kg milk/d) were enrolled in a 3 × 3 incomplete Latin square design with 3 treatments provided as continuous abomasal infusates spanning 14-d experimental periods: water (CON), soybean phospholipids (SOY; 74.5 g of deoiled soy lecithin), or sunflower phospholipids (SUN; 133.5 g of hydrolyzed sunflower lecithin). Cows were fed the same diet, which contained (% dry matter) 27.0% neutral detergent fiber (NDF), 15.6% crude protein (CP), 26.2% starch, and 5.87% FA. Treatments did not modify body weight, milk fat, protein, or lactose contents, or the efficiency of producing energy-corrected milk. Cows infused with SUN had greater milk yields than those receiving SOY or CON treatments. Cows infused with SUN had higher total solids, protein, and lactose yields than cows receiving the SOY or CON treatments. Sunflower phospholipids enhanced feed efficiency (milk yield/dry matter intake) relative to SOY or CON. Treatment did not affect intakes or apparent total-tract digestibilities for NDF, CP, starch, or 16-carbon (16C) FA. Cows receiving SUN had greater total FA and 18-carbon (18C) FA intakes than SOY or CON, but treatments did not impact their digestibility. Milk FA composition was modified by treatment. Cows receiving SUN had a greater concentration of polyunsaturated FA and lower concentrations of saturated FA and monounsaturated FA in milk relative to SOY or CON. In conclusion, the abomasal infusion of SUN improved milk production and milk FA composition, indicating potential benefits for dairy cow nutrition and milk quality.
ABSTRACT
BACKGROUND: The immune system plays a crucial role in initiating, progressing, and disseminating HNSCC. This study aims to investigate the differences in immune microenvironments between 2D-4-culture and 3D-4-culture models of head and neck squamous cell carcinoma (HNSCC) cells (FaDu), human fibroblasts (HF), human monocytes (THP-1), and human endothelial cells (HUVEC). METHODS: For the 3D-4-culture model, FaDu:HF:THP-1 (2:1:1) were inoculated in an ultra-low attachment culture plate, while HUVECs were placed in a transwell chamber. The ordinary culture plate was used for the 2D-4-culture model. Tumor-associated macrophage markers (CD163), tumor-associated fibroblast markers (FAP), and epithelial-mesenchymal transition (EMT) were detected by western blot. Inflammatory cytokines (IL-4, IL-2, CXCL 10, IL-1 ß, TNF-α, CCL 2, IL-17 A, IL-6, IL-10, IFN-γ, IL-12 p 70, CXCL 8, TGFß1) in the supernatant were measured by flow cytometry. HUVEC migration was observed under a microscope. The 3D spheres were stained and observed with a confocal microscope. CCK8 assay was used to detect the resistance of mixed cells to cisplatin in both 2D-4-culture and 3D-4-culture. RESULTS: After three days of co-culture, the 3D-4-culture model showed increased expression levels of CD163 and FAP proteins (both P < 0.001), increased expression of E-cadherin protein and N-cadherin protein expression (P < 0.001), decreased expression of vimentin (P < 0.01) and Twist protein (P < 0.001). HUVEC migration significantly increased (P < 0.001), as did the concentrations of IP-10, MCP-1, IL-6, and IL-8 (all P < 0.001). Confocal microscopy showed that 3D-4-culture formed loose cell clusters on day 1, which gradually became a dense sphere surrounded by FaDu cells invading the inside. After co-culturing for 24 h, 48 h, and 72 h, the resistance of mix cells to cisplatin in 3D-4-culture was significantly higher than in 2D-4-culture (P < 0.01 for all). CONCLUSION: Compared to 2D-4-culture, 3D-4-culture better simulates the in vivo immune microenvironment of HNSCC by promoting fibroblast transformation into tumor-associated fibroblasts, monocyte transformation into tumor-associated macrophages, enhancing endothelial cell migration ability, partial EMT formation in HNSCC cells, and is more suitable for studying the immunosuppressive microenvironment of HNSCC.