CaMKIIδ Met281/282 oxidation is not required for recovery of calcium transients during acidosis.

CaMKII is needed for the recovery of Ca2+ transients during acidosis but also mediates postacidic arrhythmias. CaMKIIδ can sustain its activity following Met281/282 oxidation. Increasing cytosolic Na+ during acidosis as well as postacidic pH normalization should result in prooxidant conditions within the cell favoring oxidative CaMKIIδ activation. We tested whether CaMKIIδ activation through Met281/282 oxidation is involved in recovery of Ca2+ transients during acidosis and promotes cellular arrhythmias post-acidosis. Single cardiac myocytes were isolated from a well-established mouse model in which CaMKIIδ was made resistant to oxidative activation by knock-in replacement of two oxidant-sensitive methionines (Met281/282) with valines (MM-VV). MM-VV myocytes were exposed to extracellular acidosis (pHo 6.5) and compared to wild type (WT) control cells. Full recovery of Ca2+ transients was observed in both WT and MM-VV cardiac myocytes during late-phase acidosis. This was associated with comparably enhanced sarcoplasmic reticulum Ca2+ load and preserved CaMKII specific phosphorylation of phospholamban at Thr17 in MM-VV myocytes. CaMKII was phosphorylated at Thr287, but not Met281/282 oxidized. In line with this, postacidic cellular arrhythmias occurred to a similar extent in WT and MM-VV cells, whereas inhibition of CaMKII using AIP completely prevented recovery of Ca2+ transients during acidosis and attenuated postacidic arrhythmias in MM-VV cells. Using genetically altered cardiomyocytes with cytosolic expression of redox-sensitive green fluorescent protein-2 coupled to glutaredoxin 1, we found that acidosis has a reductive effect within the cytosol of cardiac myocytes despite a significant acidosis-related increase in cytosolic Na+. Our study shows that activation of CaMKIIδ through Met281/282 oxidation is neither required for recovery of Ca2+ transients during acidosis nor relevant for postacidic arrhythmogenesis in isolated cardiac myocytes. Acidosis reduces the cytosolic glutathione redox state of isolated cardiac myocytes despite a significant increase in cytosolic Na+. Pharmacological inhibition of global CaMKII activity completely prevents recovery of Ca2+ transients and protects from postacidic arrhythmias in MM-VV myocytes, which confirms the relevance of CaMKII in the context of acidosis.NEW & NOTEWORTHY The current study shows that activation of CaMKIIδ through Met281/282 oxidation is neither required for CaMKII-dependent recovery of Ca2+ transients during acidosis nor relevant for the occurrence of postacidic cellular arrhythmias. Despite a usually prooxidant increase in cytosolic Na+, acidosis reduces the cytosolic glutathione redox state within cardiac myocytes. This novel finding suggests that oxidation of cytosolic proteins is less likely to occur during acidosis.

Effect of dietary aluminum on mineral metabolism of adult males.

During a 40-day balance study, eight adult males were fed two levels of aluminum: 5 mg daily (control diet) and 125 mg daily (test diet). These two levels of dietary aluminum are representative of the upper and lower limits of aluminum that are present in the diets of Americans. Initially subjects excreted significantly more phosphorus in their feces when fed the test diet rather than the control diet. However, subjects excreted similar amounts of phosphorus in their feces when fed the test and control diets for more than 12 days. Subjects excreted significantly less fluoride in their urine and less, but not significantly less, phosphorus and cyclic AMP in their urine when fed the test diet rather than the control diet. The dietary aluminum levels had no overall effect on the retention of phosphorus, calcium, magnesium, iron, zinc, or copper by these subjects.

Excretion and retention of low or moderate levels of aluminium by human subjects.

During a 40-day balance study, eight adult males were fed two levels of aluminium: 5 mg/day for 20 days (control diet) and 125 mg/day for 20 days (test diet). Every subject excreted more than 96% and more than 74% of his aluminium intake in his faeces when fed the test and control diets, respectively. Subjects excreted two- to five-fold more aluminium in their urine and had significantly higher levels of aluminium in their sera when fed the test diet rather than the control diet. No retention of aluminum was detected when faecal and urinary losses of aluminium were compared with intakes.

Effects of dietary tin on zinc, copper, iron, manganese, and magnesium metabolism of adult males.

The effects of dietary tin on zinc, copper, iron, manganese, and magnesium metabolism were determined in eight adult males. Subjects were fed mixed diets containing 0.11 mg tin daily (control diet) and 49.67 tin daily (test diet) in this 40-day study. The level of tin in the control diet was typical of the levels of tin found in diets that contain only fresh and frozen foods; the level of tin in the test diet was typical of the amount of tin in diets that contain 2 cups of certain canned foods. When subjects were fed the test diet they lost significantly more zinc (p less than 0.01) in their feces and significantly less zinc (p less than 0.05) in their urine. Subjects retained significantly less zinc (p less than 0.01) when fed the test diet rather than the control diet. The fecal and urinary losses of copper, iron, manganese, and magnesium were not significantly affected by the dietary treatments.

Effects of dietary tin and aluminum on selenium utilization by adult males.

The main purpose of these studies was to determine whether the amounts of tin and aluminum that can enter foods during processing and storage are sufficient to affect the utilization of selenium by human subjects. Two 40-day balance studies were conducted. The eight adult males who participated in the first study lost significantly more selenium in their feces when fed a test diet containing 50 mg tin daily than when fed the control diet containing 0.1 mg tin daily. During the first study subjects tended to excrete less selenium in the urine when fed the test diet rather than the control diet. In the second study, the dietary treatments (5 and 125 mg aluminum daily) had no effect on the excretion and apparent retention of selenium by eight adult males.