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Biomedical subjects

M Brune

Publications and source records attributed to M Brune.

At least 91 records · Page 5Linked to original sources

Inhibition of haem-iron absorption in man by calcium.

The inhibiting effect of Ca on non-haem-Fe absorption is well established. Present studies showed that Ca inhibited haem-Fe absorption to the same extent when the same amount of Ca (165 mg Ca as CaCl2) was added to a meal. Attempts were made to examine the mechanism for this inhibition in the present studies. Meat is the only known dietary factor influencing haem-Fe absorption. The present studies were designed to examine whether Ca interfered with the enhancing effect of meat on haem-Fe absorption. We found that the inhibition was the same whether biosynthetically radio-Fe-labelled haemoglobin was given in meals with or without meat. The haem-Fe absorption ratio with:without added Ca was 0.59 (SE 0.07) when Ca was added to a hamburger meal, and 0.52 (SE 0.03) when added to a wheat roll. These values were not significantly different (t 0.95; P = 0.35). The inhibition of haem-Fe absorption by Ca is, thus, a direct effect on the absorption of haem-Fe and not an indirect counteracting effect of the well-known enhancing effect of meat on haem-Fe absorption. Control studies were conducted to ensure that haem-Fe had not been degraded to non-haem-Fe during preparation of the foods. Since Ca inhibits the absorption of haem- and non-haem-Fe to the same extent, the present results strongly suggest that Ca interferes with the transport of Fe through the mucosal cell, and at a late stage, is common for haem- and non-haem-Fe transport.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Iron absorption from bread in humans: inhibiting effects of cereal fiber, phytate and inositol phosphates with different numbers of phosphate groups.

Iron absorption was measured from five kinds of bread made from various types of flour and fermented in different ways in order to obtain a wide variation in the content of fiber, phytate (inositol hexaphosphate) and its degradation products, inorganic phosphate and inositol phosphates with fewer numbers of phosphate groups (inositol pentaphosphate through monophosphate). Each experiment had 9-10 subjects and, in each subject, iron absorption was measured from control rolls made from low extraction wheat flour and one kind of test roll using two different radioiron tracers: 55Fe and 59Fe. The inhibition of iron absorption was closely related to the content of phytate-phosphorous as determined using the AOAC method, and to the sum of the tri- through hexaphosphate groups as determined using the HPLC method. As an example, prolonged fermentation of whole-rye bread reduced total inositol phosphates to the same amount as in the control rolls and increased fractional iron absorption to the same high level, in spite of a fiber content five times as great. The results strongly suggest that the inhibitory effect of bran on iron absorption is due to its content of phytate and other inositol phosphates present after fermentation, rather than to its content of fiber or other constituents. Thus, effective fermentation will increase the bioavailability of iron in whole-meal bread.

Absorption↗

Bioavailability in man of iron in human milk and cow's milk in relation to their calcium contents.

Iron absorption from human milk and cow's milk was compared in the same subjects using two radio-iron tracers and extrinsic labeling of the iron. Previously observed higher iron absorption from human milk was confirmed as 19.5 +/- 17.3% (mean +/- SD) in cow's milk versus 48.0 +/- 25.5% in human milk; mean individual absorption ratio was 0.39 +/- 0.18 (p less than 0.0001). Cow's milk has a several times higher content of calcium, recently established to be a potent inhibitor of iron absorption. Adding calcium chloride to human milk in amounts to equate the calcium content of cow's milk significantly reduced iron absorption [39.1 +/- 17.8% (mean +/- SD) versus 21.3 +/- 10.6%]; mean individual absorption ratio with or without added calcium was 0.58 +/- 0.12 (mean +/- SD) (p less than 0.0001). Differences in calcium content could explain at least 70% of the difference in iron bioavailability between milks. It is suggested that the remaining 30% difference may mainly be an artifact related to inhomogeneous radioiron-labeling of milk iron compounds, especially in human milk, and that the main difference in true iron absorption between human and cow's milk is related to their different calcium contents. The rather high fractional iron absorption from human milk can be explained by its low iron content. There is nothing to indicate that human milk contains any enhancer of iron absorption. The marked inhibiting effect of calcium on iron absorption should be considered in the design of infant formulas to achieve an optimal balance in the contents of calcium and iron.

Adult↗

Calcium and iron absorption: mechanism of action and nutritional importance.

We investigated the inhibitory effect of calcium on iron absorption in 57 human subjects. Three studies suggested that the effect is not located in the gastrointestinal tract. The presence of phytate in a meal and formation of calcium-iron-phytate complexes is not a prerequisite for the inhibition. The relative increase in iron absorption by ascorbic acid was the same in meals with and without calcium, suggesting that calcium did not influence the balance between enhancing and inhibiting ligands in the gastrointestinal lumen. No inhibiting effect on iron absorption was seen when adding 3 mg calcium to 0.01 mg iron (molar ratio Ca/Fe = 420). Previous studies showing a marked inhibition by calcium had a lower molar ratio, but greater amounts of calcium were given. This suggests that a minimal concentration of calcium is needed to achieve an effect. The present results indirectly support our original hypothesis that the inhibitory effect of calcium on iron absorption is situated within the intestinal mucosal cells. The practical nutritional implications of the inhibitory effect of calcium are considerable since addition of milk, milkshake or cheese to common meals such as pizza or hamburger meals reduced iron absorption by 50-60%. It is recommended to reduce the intake of dairy products with the main meals providing most of the dietary iron, especially for those having the highest iron requirements i.e. children, teenagers and women at childbearing age.

Adult↗

Structural and catalytic properties of a deletion derivative (delta 133-157) of Escherichia coli adenylate kinase.

Escherichia coli adenylate kinase (AKe) as well as the enzyme from yeast and mitochondria differs from the muscle cytosolic variant (AK1) by an insertion of 25 amino acid residues that are missing in AK1. The extra sequence, highly homologous in "large" size variants, is situated between residues 133 and 157 in AKe. Removal of 25 codons in the corresponding adk gene resulted in expression of a modified form of adenylate kinase (delta 133-157 AKe) which still conserved 7% of the maximal activity of the wild-type protein. The apparent Km for nucleotide substrates was increased by a factor of 4.6 (ADP), 23 (ATP) or 43 (AMP) in delta 133-157 AKe when compared with the wild-type enzyme. The secondary structure of delta 133-157 AKe, as well as its thermal stability were very similar to the parent protein. However, the deleted protein was much more sensitive than the wild-type enzyme to inactivation by trypsin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of trypsin digested delta 133-157 AKe revealed accumulation of several well defined fragments which were not observed in the case of wild-type enzyme. We conclude that the additional sequence, although necessary for expression of full activity in AKe, is not critical for catalysis. It is perhaps responsible for interaction of enzyme with other cellular components although a different mechanism of water shielding for large and small size variants of AK can be also envisaged.

Adenylate Kinase↗

Identification of valine/leucine/isoleucine and threonine/alanine/glycine proton-spin systems of Escherichia coli adenylate kinase by selective deuteration and selective protonation.

Adenylate kinase from two types of Escherichia coli strains, a wild-type and a leucine-auxotrophic strain, was purified. On the one hand, growing the leucine-auxotrophic bacteria on a medium containing deuterated leucine yielded E. coli adenylate kinase with all leucine residues deuterated. On the other hand, by growing the wild-type bacteria on deuterated medium with phenylalanine, threonine and isoleucine present as protonated specimens, 80% randomly deuterated enzyme with protonated phenylalanine, threonine and isoleucine residues could be prepared. Use of these proteins enabled identification of the spin systems of these amino acid residues in the n.m.r. spectra of the protein.

Adenylate Kinase↗

Calcium: effect of different amounts on nonheme- and heme-iron absorption in humans.

We investigated the effect of calcium on iron absorption in 126 human subjects. Addition of calcium chloride to wheat rolls significantly reduced iron absorption. Doses between 40 and 600 mg Ca were studied. The inhibition was clearly dose related up to 300 mg Ca. Calcium added to the dough when making the rolls reduced phytate degradation during fermentation and baking. As little as 40 mg Ca added to 80 g flour reduced phytate degradation by 50%, thus increasing the phytate content of the rolls to levels interfering with iron absorption. Calcium also had a direct dose-related inhibiting effect on iron absorption, noted by adding calcium to the rolls after they had been baked instead of to the dough. Iron absorption was reduced by 50-60% at doses of 300-600 mg Ca. Giving 165 mg Ca as milk, cheese, or calcium chloride reduced absorption by 50-60%. The same amount of calcium also significantly reduced heme-iron absorption, suggesting that the effect of calcium is related to the mucosal transfer of iron. The observed marked inhibitory effect on iron absorption of calcium in amounts frequently encountered in normal meals has important nutritional implications.

Absorption↗

Dose-dependent inhibitory effect of phenolic compounds in foods on nonheme-iron absorption in men.

Nonheme-iron absorption from a typical Southeast Asian meal was studied to examine the effect of a common vegetable, Yod Kratin, which contains a considerable amount of iron-binding phenolic groups. Yod Kratin (leaves of the lead tree) is a very popular vegetable in Thailand. It is consumed at least once a week year round, sometimes every day, together with the main meal. With a common portion size of the vegetable (20 g), iron absorption was reduced by almost 90%. As little as 5 g inhibited iron absorption by 75%. Addition of ascorbic acid partly counteracted inhibition. Adding 100 mg ascorbic acid reduced inhibition of iron absorption from 5 g Yod Kratin by half and the inhibition from 10 g Yod Kratin by a quarter. The study illustrates the marked effect of iron-binding phenolic compounds on iron nutrition and, thus, the importance of acquiring knowledge of the content of such compounds in different foods.

Absorption↗

Competitive inhibition of iron absorption by manganese and zinc in humans.

Zinc and manganese may interfere with iron absorption because of similar physicochemical properties and shared absorptive pathways. The effects of zinc and manganese on iron absorption were studied in human subjects by using paired observations and a dual-radioisotope method (55Fe and 49Fe). Manganese inhibited iron absorption both in solutions and in a hamburger meal. Fractional iron absorption is strongly dose dependent. Adding 2.99 mg Mn to 0.01 mg Fe reduced iron absorption to the same extent as increasing the iron dose 300-fold to 3 mg, strongly indicating a direct competitive inhibition of manganese on iron absorption. In the same experiment with zinc, no inhibitory effect was observed, suggesting different pathways for the absorption of zinc and iron. An intraluminal interaction may occur, because a fivefold excess of zinc to iron (15 mgZn/3 mg Fe) reduced iron absorption by 56% when given in a water solution but not when given with a hamburger meal.

Absorption↗

Rice and iron absorption in man.

Several studies suggest that iron absorption is low from rice-based meals. Comparisons of the effect of rice starch and wheat starch on iron absorption showed that rice had no effect on absorption. Observed differences in effects on iron absorption between different batches of rice starch could be explained by differences in the phytate content. Phytate is always present in rice grains. The content varies markedly depending on the method of milling. In polished rice, the content of phytate-P varied between 11.5 and 66 mg/100 g rice (data from 45 rice mills in Thailand). The bioavailability of iron in a meal composed of meat, a vegetable and rice varied about threefold (22.1-7.5 per cent) depending on the different phytate content of the rice (4 levels between 30 and 175 mg phytate-P were studied). The inhibitory effect of the phytate in rice was overcome by adding different amounts of an ascorbic acid-rich vegetable (green collard) to the meals.

Biological Availability↗

Adenylate kinases from thermosensitive Escherichia coli strains.

The adk genes from several thermosensitive (ts) mutants of Escherichia coli were cloned and sequenced. The mutations responsible for the thermolability of the gene product, the enzyme adenylate kinase, were established. From five independently isolated strains analysed, two contain a CCG to TCG transition changing proline 87 to serine (P87S), another two have a TCT to TTT transition that mutates serine 129 to phenylalanine (S129F), and the last one was found not to contain a mutation in the adk gene. Overproducing strains were constructed that contain ts genes in the genome as well as in the plasmids. These strains grow at high temperature, although much slower than wild-type. Most probably, the high rate of synthesis of adenylate kinase compensates for the destruction of the thermolabile protein by the elevated temperature. Mutated proteins were purified. The P87S but not the S129F mutation was found to cause thermosensitivity of the adenylate kinase reaction. Revertants of thermosensitivity were isolated and the nature of the mutation was determined by the RNase digestion method of RNA-DNA hybrids and by DNA sequencing. The revertants of the P87S mutation regained the wild-type sequence, whereas the revertants of the S129F strain retained the original mutation in the adenylate kinase gene. These results are discussed in the light of the three-dimensional structure of the enzyme and the possible role of adenylate kinase in phospholipid synthesis.

Adenylate Kinase↗

Iron absorption in man: ascorbic acid and dose-dependent inhibition by phytate.

The dose-dependent inhibitory effect of sodium phytate on iron absorption was studied in man by serving wheat rolls containing no phytates and rolls to which various amounts (seven dose levels between 2 and 250 mg expressed as phytate phosphorus) were added just before serving. Fe in the two kinds of rolls was labeled with two radioisotopes of Fe (55Fe, 59Fe) and the rolls were served on alternate days. The inhibition of Fe absorption was strongly related to the amount of phytate added; 2 mg inhibited absorption by 18%, (p less than 0.001), 25 mg by 64% (p less than 0.001), and 250 mg by 82% (p less than 0.001). The addition of ascorbic acid significantly counteracted the inhibition whereas the corresponding effect of meat was less well defined and only seen at the highest phytate level. The marked inhibition of Fe absorption by phytates and the significant counteracting effect of ascorbic acid have wide nutritional implications.

Adult↗

Iron absorption: no intestinal adaptation to a high-phytate diet.

The present study examined the possibility that a high bran and phytate intake over a long period would induce changes in the intestines or its microflora leading to a reduction of the inhibitory effect of dietary phytates on iron absorption. Such changes might occur in vegetarians and populations in developing countries with a usually very high dietary intake of phytates. The inhibitory effect of bran on Fe absorption was compared between a group of strict vegetarians with a regular high phytate intake and a control group by use of wheat rolls with and without bran labeled with 55Fe and 59Fe. The average individual decrease of the Fe absorption from adding the bran was 92 and 93% in the two groups, respectively. No intestinal adaptation to a high phytate intake could be observed. This finding has wide nutritional implications.

Adaptation, Physiological↗

The role of vitamin C in iron absorption.

Iron requirements remain the same despite the current lower energy requirement. This means that more iron must be absorbed per unit energy. A higher bioavailability of the dietary iron can be achieved by increasing the content of food components enhancing iron absorption (ascorbic acid, meat/fish) or by decreasing the content of inhibitors (e.g., phytates, tannins). The latter is not feasible considering the recent and reasonable trend toward increasing the intake of dietary fibre. The key role of ascorbic acid for the absorption of dietary nonheme iron is generally accepted. The reasons for its action are twofold: (1) the prevention of the formation of insoluble and unabsorbable iron compounds and (2) the reduction of ferric to ferrous iron, which seems to be a requirement for the uptake of iron into the mucosal cells.

Ascorbic Acid↗