H. E. Parker

The effects of copper on collagen cross-linking.

Abstract Evidence in this report indicates that copper is directly involved in the process of collagen cross-linking. The bones from chicks fed copper-deficient diets became brittle and fragile. The collagen in copper-deficient bone was more easily extracted and contained less total aldehyde than corresponding controls. After chromatography on CM-cellulose columns and acrylamide gel electrophoresis, monomer or α-collagen and the cross-linked dimer, β-collagens, could be separated and quantitated. The α to β ratio of collagen from copper-deficient bone was approximately twice that of collagen from control bone, indicating a decrease in the cross-linking.

The Effect of Magnesium and Fluoride on Bone Pyrophosphatase Activity

Summary The addition of Mg ions to bone homogenates stimulated pyrophosphatase activity in the absence of F ions, but inhibited pyrophosphatase activity in the presence of F ions. However, femur pyrophosphatase levels were increased in chicks fed diets containing high levels of both Mg and F for 3 or 6 days as compared with chicks fed diets containing identical levels of either element alone.

Observations on a magnesium-fluoride interrelationship in chicks.

The addition of high levels of both magnesium and fluoride to either practical or semipurified diets complete in all known nutrients produced the following results in young chicks: (1) greater reduction in growth than with either magnesium or fluoride alone; (2) the development of a characteristic leg weakness; (3) decreased calcification of the bone. Increasing the calcium level in the diets from 1.0 to 1.5% reduced but did not prevent the magnesium-fluoride relationship. The growth reduction, leg weakness and reduced bone ash caused by high dietary magnesium and fluoride were obtained only when one-day-old chicks were fed the experimental diets; these results were not observed when chicks at two weeks of age were fed the same diets, nor could the interaction be shown in weanling rats. Plasma alkaline phosphatase activity in chicks was elevated about twofold by added fluoride regardless of the dietary magnesium or calcium levels. The level of alkaline phosphatase in the plasma decreased with age in all groups at approximately the same rate. Dietary magnesium or fluoride levels did not influence the conversion of 3-phosphoglycerate to phosphoenolpyruvate by muscle homogenates. 11 references, 4 tables.

The partial characterization of an amine oxidase in bone tissue.

Summary Evidence is presented which demonstrates that chick bone contains an amine oxidase which could function in the oxidative deamination of collagen lysyl residues to α-amino adipic acid-Δ-semialdehyde residues. This function would be directly related to the process of collagen cross-linking. The enzyme was slightly soluble, had an apparent molecular weight from gel filtration studies of 250,000, and could be partially purified by ammonium sulfate precipitation. When benzylamine was used as the substrate in kinetic studies, a K mapp of 0.15 mM was obtained. The enzyme required copper and pyridoxal phosphate and was competitively inhibited by beta-amino propionitrile, butylamine, and the peptide, lysine-vasopressin. Benzylamine oxidation was noncompetively inhibited by ascorbic acid, penicillamine, and isoniazid. The lysine in lysine-vasopressin was oxidized to the semial-dehyde. The pH optimum for oxidations was approximately pH 7.7.

Effects of dietary magnesium and fluoride on the magnesium content of tissues from growing chicks.

Two factorial experiments were conducted in which the effect of supplemental magnesium and fluoride on the magnesuim content of plasma (total and diffusible), bone, muscle, heart, liver and kidney were determined. Bone, liver and total and diffusible plasma magnesium were greater in birds fed diets containing high levels of both magnesium and fluoride than in birds fed diets high in either magnesium or fluoride alone. Muscle magnesium did not vary greatly with the dietary treatments and the magnesium content of the heart was remarkably constant. Previous work in this laboratory has shown that the addition of high levels of both magnesium and fluoride to the diets of growing chicks caused a greater reduc tion in growth rate than the addition of high levels of either magnesium or fluoride alone (1, 2). Leg weakness and reduced bone ash were observed in the birds fed high magnesium-high fluoride diets but not in birds fed diets high in either ele ment alone. The lack of a rapid and accurate method of analysis for magne sium limited the amount of information obtained on magnesium distribution in the earlier studies. The acquisition of an atomic absorption spectrophotometer which facilitates rapid, precise and accu rate analysis made it possible to deter mine the influence of dietary magnesium and fluoride levels on the distribution of magnesium in various tissues. Two experiments were conducted to determine: 1), the normal magnesium content of chick plasma, bone, liver, kid ney, muscle and heart; 2), the effects of excess dietary magnesium and fluoride on the magnesium content of the above tis sues; and 3), the effects of excess dietary magnesium and fluoride on the distribu tion of total and diffusible magnesium in the blood plasma. EXPERIMENTAL

Effect of dietary magnesium and fluoride on citric acid content of chick bones.

Summary Addition of both magnesium and fluoride to the diet of growing chicks caused a greater reduction in the citric acid content of bone than that caused by addition of either element alone. Bones from chicks maintained on either the basal diet or diets supplemented with 0.25% magnesium contained approximately 0.42% citric acid and those from chicks maintained on high fluoride diets contained approximately 0.29% citrate. Bones from chicks fed diets high in both magnesium and fluoride contained only 0.16% citric acid.

Some Comparisons of Thyroid Glands of Bobwhites and Japanese Quail

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Influence of parental dietary iodine level on thyroid activity of eighteen day chick embryos.

Summary Thyroid activity in 18-day embryos from hens fed 3 levels of dietary iodine was studied with the use of I131. The thyroid glands of embryos from hens on the lowest level of iodine synthesized and released thyroxine at a much faster rate than embryos from hens on higher levels of iodine. This rapid synthesis and release of thyroxine in more active thyroid glands can confound measusements of thyroid activity by I131 “uptake” procedures if care is not taken in selecting proper time intervals between injection and autopsy.