Fatty acid oxidation is the major energy pathway used by the kidney, although glycolysis becomes more important in the low oxygen environment of the medulla. Fatty acid oxidation appears to be reduced in renal fibrosis, and drugs that reverse this improve fibrosis. Expression of glycolytic genes is more variable, but some studies have shown that inhibiting glycolysis reduces renal fibrosis. To address the role of glycolysis in renal fibrosis, we have used a genetic approach. The crucial control point in the rate of glycolysis is 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase. Phosphorylation of the kidney isoform, PFKFB2, on residues Ser(468) and Ser(485) stimulates glycolysis and is the most important mechanism regulating glycolysis. We generated transgenic mice with inactivating mutations of Ser(468) and Ser(485) in PFKFB2 (PFKFB2 KI mice). These mutations were associated with a reduced ability to increase glycolysis in primary cultures of renal tubular cells from PFKFB2 KI mice compared to WT cells. This was associated in PFKFB2 KI mice with increased renal fibrosis, which was more severe in the unilaternal ureteric obstruction (UUO) model compared with the folic acid nephropathy (FAN) model. These studies show that phosphorylation of PFKFB2 is important in limiting renal fibrosis after injury, indicating that the ability to regulate and maintain adequate glycolysis in the kidney is crucial for renal homeostasis. The changes were most marked in the UUO model, probably reflecting a greater effect on distal renal tubules and the greater importance of glycolysis in the distal nephron.

The current study explored the effect of intramuscular injection of vitamin B-12 (VB12) in early-lactation dairy cows on subsequent low-moisture part-skim Mozzarella cheese quality and VB12 levels during cheese processing and storage. Twenty-four peripartum dairy cows were blocked based on parity and milk yield and randomly assigned into 2 treatments: basal diet (CON) and basal diet with an intramuscular injection of 10 mg of VB12 per cow per week (VB12). Raw milk was collected to determine VB12 content and then used to make low-moisture part-skim Mozzarella cheese 8 wk after injection. The VB12 content of raw milk and cheese was determined using ultra-performance liquid chromatography coupled with tandem mass spectrometry. We found that VB12 content was significantly increased in milk (15.43 vs. 3.30 ng/mL) and fresh cheese (3.72 ng/g vs. undetectable) from the VB12 group compared with the CON group. However, approximately 70% of VB12 was lost in the whey during cheese making, and no VB12 was detectable in either cheese treatment after 8 wk of storage. Furthermore, no significant differences were observed in fat and protein contents in the cheese between the 2 groups. For cheese color, the b* value increased and the a* value decreased slightly in fresh VB12 cheese. Functional properties of stretchability, flowability, and meltability of VB12 cheese were initially comparable to that of CON cheese, but higher flowability and meltability was observed in VB12 cheese after 8 wk of storage. In summary, intramuscular injection of VB12 in early-lactation dairy cows increases the content of VB12 in milk and fresh cheese with no adverse effect on cheese quality, but substantial VB12 is lost during cheesemaking and declines rapidly during storage.