Bin Sun

Mitochondrial amidoxime reducing component (mARC) proteins are molybdopterin-containing enzymes of unclear physiological function. Both human isoforms mARC-1 and mARC-2 are able to catalyze the reduction of nitrite when they are in the reduced form. Moreover, our results indicate that mARC can generate nitric oxide (NO) from nitrite when forming an electron transfer chain with NADH, cytochrome b5, and NADH-dependent cytochrome b5 reductase. The rate of NO formation increases almost 3-fold when pH was lowered from 7.5 to 6.5. To determine if nitrite reduction is catalyzed by molybdenum in the active site of mARC-1, we mutated the putative active site cysteine residue (Cys-273), known to coordinate molybdenum binding. NO formation was abolished by the C273A mutation in mARC-1. Supplementation of transformed Escherichia coli with tungsten facilitated the replacement of molybdenum in recombinant mARC-1 and abolished NO formation. Therefore, we conclude that human mARC-1 and mARC-2 are capable of catalyzing reduction of nitrite to NO through reaction with its molybdenum cofactor. Finally, expression of mARC-1 in HEK cells using a lentivirus vector was used to confirm cellular nitrite reduction to NO. A comparison of NO formation profiles between mARC and xanthine oxidase reveals similar Kcat and Vmax values but more sustained NO formation from mARC, possibly because it is not vulnerable to autoinhibition via molybdenum desulfuration. The reduction of nitrite by mARC in the mitochondria may represent a new signaling pathway for NADH-dependent hypoxic NO production.

Brown adipose tissue uncoupling protein-1 (UCP1) plays a major role in the control of energy balance in rodents. It has long been thought, however, that there is no physiologically relevant UCP1 expression in adult humans. In this study we show, using an original approach consisting of sorting cells from various tissues and differentiating them in an adipogenic medium, that a stationary population of skeletal muscle cells expressing the CD34 surface protein can differentiate in vitro into genuine brown adipocytes with a high level of UCP1 expression and uncoupled respiration. These cells can be expanded in culture, and their UCP1 mRNA expression is strongly increased by cell-permeating cAMP derivatives and a peroxisome-proliferator-activated receptor-gamma (PPARgamma) agonist. Furthermore, UCP1 mRNA was detected in the skeletal muscle of adult humans, and its expression was increased in vivo by PPARgamma agonist treatment. All the studies concerning UCP1 expression in adult humans have until now been focused on the white adipose tissue. Here we show for the first time the existence in human skeletal muscle and the prospective isolation of progenitor cells with a high potential for UCP1 expression. The discovery of this reservoir generates a new hope of treating obesity by acting on energy dissipation.

Beef contains functional fatty acids such as conjugated linoleic acid and long-chain fatty acids. This review summarizes results from studies comparing the fatty acid composition of beef from cattle fed either grass or grain-based feed. Since functional lipid components are contributed through dietary consumption of beef, the fatty acid composition is reported on mg/100 g of meat basis rather than on a percentage of total fat basis. Beef from grass-fed contains lesser total fat than that from grain-fed in all breeds of cattle. Reduced total fat content also influences the fatty acid composition of beef. A 100 g beef meat from grass-fed cattle contained 2,773 mg less total saturated fatty acids (SFA) than that from the same amount of grain-fed. Grass-fed also showed a more favorable SFA lipid profile containing less cholesterol-raising fatty acids (C12:0 to C16:0) but contained a lesser amount of cholesterol-lowering C18:0 than grain-fed beef. In terms of essential fatty acids, grass-fed beef showed greater levels of trans-vaccenic acid and long-chain n-3 polyunsaturated fatty acids (PUFA; EPA, DPA, DHA) than grain-fed beef. Grass-fed beef also contains an increased level of total n-3 PUFA which reduced the n-6 to n-3 ratio thus can offer more health benefits than grain-fed. The findings signify that grass-fed beef could exert protective effects against a number of diseases ranging from cancer to cardiovascular disease (CVD) as evidenced by the increased functional omega-3 PUFA and decreased undesirable SFA. Although grain-fed beef showed lesser EPA, DPA, and DHA, consumers should be aware that greater portions of grain-fed beef could also achieve a similar dietary intake of long-chain omega-3 fatty acids. Noteworthy, grain-fed beef contained higher total monounsaturated fatty acid that have beneficial roles in the amelioration of CVD risks than grass-fed beef. In Hanwoo beef, grain-fed showed higher EPA and DHA than grass-fed beef.