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J Hegenauer

Publications and source records attributed to J Hegenauer.

At least 19 recordsLinked to original sources
PubMed1993

Intrinsic stoichiometric equilibrium constants for the binding of zinc(II) and copper(II) to the high affinity site of serum albumin.

Intrinsic stoichiometric equilibrium constants were determined for zinc(II) and copper(II) binding to bovine and human serum albumin. Data were obtained from equilibrium dialysis experiments. Metals were presented to apoprotein as metal chelates in order to avoid metal hydrolysis and to minimize nonspecific metal-protein interactions. Scatchard analysis of the binding data indicated that the high affinity class for both zinc and copper was comprised of one site. Results of binding experiments done at several pH values suggested that while both histidyl and carboxyl groups appear to be involved in copper binding, histidyl residues alone were sufficient for zinc binding. These amino acid residues were used in combination to model several binding sites used in the formulation of equilibria expressions from which stoichiometric constants were calculated. The log10K for bovine serum albumin were calculated to be 7.28 for Zn(II) and 11.12 for Cu(II). Those for human serum albumin were determined to be 7.53 and 11.18 for Zn(II) and Cu(II), respectively. These constants were used in equilibria to simulate speciation of metal-albumin and metal-chelator and to illustrate relative binding affinities. This comparison of binding strengths was possible only through the calculation of an intrinsic stoichiometric binding constant.

Animals↗
PubMed1992

Metal-ion-directed site-specificity of hydroxyl radical detection.

A wide variety of .OH detectors are in use for determination of biological .OH production. The chemical generation of .OH is site-specific with respect to the metal-binding site, and thus .OH detectors with metal-binding properties may affect the biological damage and bias .OH detection. The present study shows that both salicylate and phenylalanine, added as low molecular weight .OH indicators, decreased Cu(II) binding to erythrocyte ghosts. In a cell-free system, Cu(II) complexed to both salicylate and phenylalanine. Phenylalanine is a stronger Cu(II) chelator than salicylate, both when competing for Cu(II) bound to ghosts and when competing directly with each other. When OH radicals were generated by ascorbate and Cu(II), the amount of .OH detected as dihydroxybenzoates was proportional to the amount of .OH produced. However, when phenylalanine was added to this system, the efficiency of .OH detection by salicylate strongly decreased, concomitant with the transfer of Cu(II) binding from salicylate to the amino acid. This decrease was larger than that predicted by calculations for random competition of the two detectors for .OH. Deoxyribose and mannitol, which do not bind copper appreciably, competed poorly with salicylate for the .OH. Hydroxylation of phenylalanine, on the other hand, was only slightly affected by the presence of salicylate and unaffected by deoxyribose and mannitol. These results suggest that the detection of .OH by low molecular weight .OH indicators was related to the relative affinity of the detectors for the catalyzing metal, and thus partially site-specific. Furthermore, glutamate, which does not contain an aromatic ring but binds Cu(II) with considerable affinity, competed strongly with salicylate for the .OH, indicating that metal-binding properties rather than the presence of an aromatic ring were the cause of the deviation from random competition. The results indicate that .OH indicators with metal-binding properties affect the distribution of catalytic metal ions in a biological system, causing a shift of free radical damage and localizing a site-specific reaction of .OH on these detectors, with a resulting positive bias in the apparent .OH production.

Ascorbic Acid↗
PubMed1987

Isolation and characterization of a new zinc-binding protein from albacore tuna plasma.

The protein responsible for sequestering high levels of zinc in the plasma of the albacore tuna (Thunnus alalunga) has been isolated by sequential chromatography. The glycoprotein has a molecular weight of 66,000. Approximately 8.2% of its amino acid residues are histidines. Equilibrium dialysis experiments show it to bind 3 mol of zinc/mol of protein. The stoichiometric constant for the association of zinc with a binding site containing three histidines was determined to be 10(9.4). This protein is different from albumin and represents a previously uncharacterized zinc transport protein.

Amino Acids↗
PubMed1986

Age-related differences in the bone mineralization pattern of rats following exercise.

The effect of 12 weeks of treadmill exercise on the mineralization of trabecular and cortical bone was studied in rats 7, 14, and 19 months of age. Bone mineralization was evaluated by measuring concentrations of Ca, Mg, and hydroxyproline as well as uptake of 45Ca concentration in the femur, humerus, rib and calvaria. The 7- and 14-month-old rats increased mineralization in those cortical bones directly involved in exercise. The 19-month animal responded to exercise by increasing mineralization in all bones examined, including the nonweight bearing trabecular calvaria and cortical rib. From these data, it is apparent that the older animals undergo a total skeletal mineralization in response to exercise compared with local adaptation in the younger animal. Further, we provide evidence to support the use of the rat as a model in which to study mammalian bone physiology during the aging process.

Aging↗
PubMed1986

Effects of long-term dietary manganese and copper deficiency on rat skeleton.

Experiments were designed to produce biochemical changes in both serum and bone in rats by long-term dietary deficiencies of manganese (Mn) and copper (Cu). Weanling rats were fed one of three diets: 1) normal Mn and Cu (N), 2) low Mn and Cu (L) and 3) depleted Mn and normal Cu (D). After 12 mo, serum Ca and P were significantly greater in the D rats than the N rats (134, 50 mg/L and 103, 39 mg/L, respectively). Serum and femur Mn levels were lower in D rats than in normal rats. Serum and femur Cu levels were lower in L rats than N rats. Femur Ca concentrations were significantly lower in D rats than in N rats (180 and 272 mg/g, respectively). Radiographic differences could be detected in some rats fed the L and D diets. In the two deficient groups, increased serum Ca was negatively correlated with bone Ca (r = -0.69; P less than 0.01). This biochemical association may represent alterations in regulatory control of Ca at the level of the bone (decreased mineralization) combined with an increase in bone resorption. The effect of long-term dietary deficiencies in Mn and Cu should be considered in human bone metabolism.

Animal Nutritional Physiological Phenomena↗
PubMed1984

Effects of iron deficiency and exercise on myoglobin in rats.

The effects of iron deficiency and endurance training on muscle myoglobin (Mb), body weights, and blood lactic acid concentration were studied in rats. Fifty animals were divided into four groups: anemic trained (AT), normal trained (NT), anemic sedentary (AS), and normal sedentary (NS). Following 5 weeks of dietary control, the mean hemoglobin values for the AT and AS rats were 0.013 +/- 0.002 mmol X l-1 (8.7 +/- 1.4 g X dl-1) and 0.014 +/- 0.003 mmol X l-1 (9.2 +/- 1.7 g X dl-1) respectively, and did not significantly change throughout the study. AT and NT rats were run on a motor driven treadmill 4 days/week for 6 weeks up to a pre-established time of 90 min. Following the training, body weights of the AT (157 +/- 13 g) and NT (153 +/- 13 g) rats were lower than their respective sedentary groups AS (172 +/- 9 g) and NS (176 +/- 15 g). Resting blood lactic acid concentration following training was lower in both trained groups, AT (3.3 +/- 2.0 mM) and NT (2.3 +/- 1.9 mM) compared to AS (8.2 +/- 2.6 mM) and NS (3.8 +/- 1.6 mM). Training increased Mb concentration in hearts of both the anemic and normal trained groups (AT, 0.66 +/- 0.13 mg X g-1; NT, 0.95 +/- 0.08 mg X g-1) compared to the sedentary groups (AS, 0.44 +/- 0.08 mg X g-1; NS, 0.70 +/- 0.13 mg X g-1). Only the AT rats showed an increase in skeletal muscle Mb. This study provides evidence that myoglobin may limit aerobic metabolism.

Anemia, Hypochromic↗
PubMed1984

Retention and distribution of iron added to cow's milk and human milk as various salts and chelates.

Iron supplementation of infant formulas is recommended by most national and international organizations, but the optimal form of supplementation has not been determined. We have compared the bioavailability and tissue distribution of iron from four iron chelates and two commonly used iron salts. Weanling C57BL/6J mice were fed for 1 week an evaporated cow's milk diet supplemented with vitamins and minerals (except for iron). Following the adjustment period, mice were divided into 12 groups of 20 each. Six groups continued to receive the cow's milk diet for 18 hours, while the other six groups were fed a similar diet based on human milk. Individual groups received a single dose of milk radioactively labeled with Fe(II)Cl2, Fe(II)SO4, Fe(III)NTA, Fe(III)EDTA, Fe(III)citrate or Fe(III)lactobionate. Wholebody retention was measured after 4 days; animals were then killed and individual tissues were counted for radioactivity. Iron from FeCl2, FeSO4 and FeNTA were the best retained from both milk diets. Fe citrate had a significantly lower iron retention than all other groups in either diet and is probably not an effective chelate for delivering iron to milk diets. Iron bioavailability was higher from the human milk diets than from the cow's milk diets from all vehicles used except citrate and lactobionate. Absorption of Fe citrate was similar from the two milk diets, while percent retention from Fe lactobionate was higher from cow's milk than from human milk. Tissue distribution of retained iron was similar for the milk diets and among the groups, indicating that, once absorbed, iron from the different vehicles is metabolized in a similar manner.

Animals↗
PubMed1983

Transitory hematologic effects of moderate exercise are not influenced by iron supplementation.

A young women's exercise/fitness class tested the idea that administration of supplemental iron would prevent "sports anemia" that may develop during exercise and training and improve iron status of exercising females of menstrual age. Fifteen women (aged 18-37) were selected for each of three treatment groups: (1) no supplemental iron; (2) 9 mg X d-1 of Fe; and (3) 18 mg X d-1 of Fe (1 US Recommended Daily Allowance). Women exercised at approximately 85% of maximal heartrate for progressively increasing lengths of time in a jogging program and worked up to 45 min of exercise 4 d X week-1 for 8 weeks. Hematologic analysis was performed in weeks 1, 5, and 8. A significant decline in hemoglobin (Hb) concentration and hematocrit (Hct) was observed at week 5 when all data were examined without regard for iron intake; these red cell indices returned to pre-exercise levels by week 8. Reduction of mean cell hemoglobin concentration (MCHC) indicated that the midpoint decline was not caused by simple hemodilution during exercise. Serum ferritin (SF) concentration changed in parallel with Hb and Hct. Although the midpoint decline in SF was not statistically significant, it ruled out the possibility that turnover of red cell iron was directed to storage. Lowered MCHC and SF suggested lower availability of iron during the synthesis of a new generation of red cells. Few iron treatment effects of magnitude were observed. Iron did not prevent the midpoint decline in Hb concentration. Iron intake did not affect SF, serum iron, transferrin saturation, or final Hb, and Hct.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1983

Adaptations of lactate metabolism in iron-deficient rats.

Effects of dietary iron deficiency on lactate metabolism were studied in weanling female rats. Following an iron-deficient diet for 5 weeks, mean hemoglobin concentration was lowered to 6.4 g/dl relative to 12.2 in the control group. Mean plasma iron levels were 58 and 162 micrograms/dl, respectively. Significantly elevated resting lactate levels were observed in whole blood and plasma from iron-deficient anemic (relative to control) rats. Total activity of lactate dehydrogenase (LDH) was elevated in soleus and gastrocnemius muscles in response to iron deficiency from 269 +/- 51 to 364 +/- 60 (Mean +/- SD) and from 265 +/- 65 to 372 +/- 61 IU . 10(-3) . g-1, respectively. The LDH activity in heart was lowered from 700 +/- 61 to 593 +/- 45 IU . 10(-3) . g-1. The M3H and M2H2 isozymes in soleus were increased from 12.7 +/- 2.8 to 20.4 +/- 5.8% and from 19.4 +/- 6.1 to 28.2 +/- 3.6%, respectively. Similar increase was observed in M2H2 and MH3 in gastrocnemius from 9.8 +/- 0.9 to 14.8 +/- 2.0% and from 17.4 +/- 2.0 to 20.5 +/- 2.3%, respectively. The H4 isozyme was significantly reduced in soleus, gastrocnemius, and plantaris muscles from 27.7 +/- 4.7 to 12.4 +/- 4.4, from 15.8 +/- 1.9 to 7.2 +/- 2.9, and from 10.5 +/- 2.9 to 3.9 +/- 2.1%, respectively. It was suggested that iron-deficiency anemia induces an elevation of lactate production following an increase in total LDH activity and change in LDH isozyme patterns.

Anemia, Hypochromic↗
PubMed1982

Bioavailability of iron- and copper-supplemented milk for Mexican school children.

Fortification of dairy products with trace metals requires use of assimilable compounds that do not catalyze off-flavors due to lipid peroxidation but show good biological availability. The Fe(III) and Cu(II) chelates of the promising chelator, lactobionic acid, have been compared to Fe(II) and Cu(II) salts for their ability to improve hematological status in a mildly anemic population. Fe- and Cu-fortified cow milk was administered to children (aged 6 to 15) in the Durango, Mexico, "school lunch" program. Children drank milk providing 20 mg Fe and 3 mg Cu as ferric/cupric lactobionate ("chelate") or ferrous/cupric chloride ("salt") for 5 of 7 days/wk for 3 months. Supplementation with "salt" and "chelate" raised Hb significantly by 1 and 0.3 g/dl, respectively, above the control (unsupplemented) group. No significant change was observed in incremental serum ferritin, serum Fe, or transferrin saturation, or in final serum Cu. Ferric lactobionate shows poorer bioavailability than ferrous ion in the presence of Cu, but milk can be an excellent vehicle for Fe or Cu supplementation.

Adolescent↗
PubMed1982

Mitochondrial NADH dehydrogenase in iron-deficient and iron-repleted rat muscle: an EPR and work performance study.

Iron may affect both respiratory O2 transport and mitochondrial electron transport in the performance of muscle work. This study was designed to elucidate the molecular defect of iron-deficient work performance by identifying heretofore unmeasurable mitochondrial enzymes that are diminished by iron deficiency and may be restored by iron repletion. Female rats were made iron-deficient by dietary control and were repleted by oral iron. Iron deficiency reduced physical work capacity (treadmill running time), haemoglobin (Hb), and mitochondrial iron-sulphur (Fe-S) centres in heart and skeletal muscles; mitochondrial number was unaffected. Oral iron supplementation restored work capacity and Hb within 4 d to normal or near-normal levels, but in general Fe-S centres of mitochondria due to NADH dehydrogenase remained at iron-deficient levels. Subnormal concentrations of mitochondrial iron-dependent NADH dehydrogenase in muscle are not by themselves rate-limiting in work performance.

Anemia, Hypochromic↗
PubMed1979

Iron(III)--phosphoprotein chelates: stoichiometric equilibrium constant for interation of iron(III) and phosphorylserine residues of phosvitin and casein.

Estimates of the strength of iron binding to model phosphoproteins were obtained from equilibrium dialysis experiments. Iron-free phosvitin (chicken and frog) or alpha sl-casein (cow) was dialyzed against the iron(III) chelates of nitrilotriacetate (NTA), )ethylenedinitrilo)tetraacetate (EDTA), or citrate. Protein-bound metal was measured at equilibrium; competition of chelator and phosphoprotein for iron(III) was determined by reference to comprehensive equilibrium equations presented in the Appendix. Analysis of the iron-binding data for phosvitin suggested that clusters of di-O-phosphorylserine residues (SerP.SerP) were the most probable iron-binding sites. A stoichiometric equilibrium constant of 10(18.0) was calculated for the formation of the Fe3+(SerP.SerP) chelate. When comared on the basis of phosphate content, casein bound iron more weakly than phosvitin. However, if the stoichiometric equilibrium constant for the formation of the casein Fe3+(SerP.SerP) chelate (10(17.5) was adjusted to account for the fact that a smaller percentage of casein phosphoserines occurs in di-O-phosphorylserine clusters, the affinity of casein and phosvitin for iron was very similar. A theoretical comparison showed that the "strengths" of the ferric chelates can be ranked: EDTA greater than phosphoprotein di-O-phosphorylserine greater than citrate greater than NTA.

Animals↗
PubMed1979

Removal of cadmium(II) from crystallized ferritin.

The cadmium content of crystallized horse spleen ferritin, usually about 2% by weight without special treatment, can be substantially decreased by prolonged dialysis against certain chelating agents, chaotropic ions, or weakly reducing anions. For example, neutral bisulphite buffer (2M) removed 95% of the bound cadmium of crystallization without affecting the iron content, and may thus be valuable for preparing "metal-free" holoferritin for physical-chemical studies.

Cadmium↗
PubMed1979

Degradation of ascorbic acid (vitamin C) in iron-supplemented cows' milk.

The fate of [6-carbon-14] ascorbic acid in iron-supplemented and unsupplemented raw milk was studied by anion-exchange chromatography, which permitted quantitative analysis of the conversion of ascorbate to dehydroascorbate and diketogulonate as a function of time. Iron catalyzed an increase in the rate of autoxidation of ascorbate to dehydroascorbate but did not alter the equilibrium concentrations of ascorbate, dehydroascorbate, and diketogulonate. The conversion of ascorbate to dehydroascorbate and of dehydroascorbate to diketogulonate occurred rapidly even in unsupplemented milk. Thus, trace metal supplementation may not affect materially the vitamin C content of stored milk.

2,3-Diketogulonic Acid↗
PubMed1977

Correlation of serum ferritin and liver ferritin iron in the anemic, normal, iron-loaded rat.

We developed a two-site immunoradiometric assay for rat serum ferritin that uses antibody immobilized on agarose. Individual serum ferritin values were significantly correlated with iron stores as determined chemically by liver ferritin iron content. This group correlation was not sufficiently great, however, to allow confident prediction of iron stores in a given animal on the basis of serum ferritin alone. Significant differences in mean liver ferritin iron concentration between groups of rats raised on diets of differing iron content were not always reflected by differences in mean serum ferritin values. Data correlating the serum levels of ferritin and a liver-specific transaminase suggested that hepatocellular death may sometimes contribute ferritin to the serum. Strong postitive correlations between serum ferritin and iron stores in the rat were not observed when serum transaminase levels were in the normal range.

Alanine Transaminase↗