Excerpt PART I. A CASE WITH HEMOLYTIC ANEMIA AND MARKED THROMBOPENIA The subject of Gaucher's disease continues to occupy a fairly prominent position in the medical literature of recent years. The ...

Our invention relates to the isolation and sep aration from impurities of Organic cyclic come pounds, with a hydroxyl group attached to the cyclic nucleus, such as cholesterol, ergosterol, isocholesterol, phytosterols, borneol, homologues and isomers of cholesterol, eugenol, and vanillin. Heretofore, the most common sources for the isolation of cholesterol have been spinal cords and brain tissue because the process of extrac tion by solvents and crystallization was rela tively simple in that no impurities occurred which interfered with the process of crystallization. The disadvantages of these sources is that they are, first, expensive and secondly, the amount of cholesterol in these sources is not greater than that which can be obtained from more common sources, such as lanolin and fish liver oils. For a long time it was known that sources such as lanolin and fish liver oils contained large quan tities of cholesterol but the method of extraction by solvents was ineffectual in that the impurities made crystallization difficult.

Journal Article The Journal of Biological Chemistry Get access Albert E. Sobel, Albert E. Sobel Pediatric Research Laboratory and the Division of Biochemistry, The Jewish Hospital of Brooklyn, New York Search for other works by this author on: Oxford Academic PubMed Google Scholar Henry Yuska, Henry Yuska Pediatric Research Laboratory and the Division of Biochemistry, The Jewish Hospital of Brooklyn, New York Search for other works by this author on: Oxford Academic PubMed Google Scholar David D. Peters, David D. Peters Pediatric Research Laboratory and the Division of Biochemistry, The Jewish Hospital of Brooklyn, New York Search for other works by this author on: Oxford Academic PubMed Google Scholar Benjamin Kramer Benjamin Kramer Pediatric Research Laboratory and the Division of Biochemistry, The Jewish Hospital of Brooklyn, New York Search for other works by this author on: Oxford Academic PubMed Google Scholar Nutrition Reviews, Volume 39, Issue 10, October 1981, Pages 374–377, https://doi.org/10.1111/j.1753-4887.1981.tb06717.x Published: 01 October 1981 Article history Received: 28 October 1939 Published: 01 October 1981

Es wird gezeigt, dass bei Entfernung eines Teiles der wasserlöslichen Phase einer meta-stabilen, verkalkenden Lösung, Kalk- und Phosphorionen so angereichert werden, dass «kristallisierte Kerne» von Hydroxyapatit auftreten.

Received for publication June 15, 1964. tion of beryllium ion which would interfere with crystal growth of bone and tooth mineral. In our test system, during the pretreatment stage, 10 mg. of tertiary calcium phosphate were shaken in flasks with 100 ml. of solutions of beryllium chloride in water, varying in concentration from 10-5 to 10-l mM/1. Control samples were either pretreated with 10-1 mM/l calcium chloride or received no pretreatment. The solid phase was separated by centrifugation, washed with distilled water, and dried at 105' C. An aliquot of 2.5 mg. was then added to 50 ml. of in vitro calcifying solution. Ca (10.0 mg. per cent) X P (5.0 mg. per cent) = 50, adjusted to a pH of 7.3, and incubated for 20 hours. Following incubation, the calcium and phosphate contents of the supernatant solution were determined. The removal of calcium and phosphate from solution and the appearance of additional white precipitate in the flask were taken as indications of crystal growth of the calcium phosphate. As seen in the table, incubation of control samples of calcium phosphate receiving no treatment or shaken previously with 10-1 mM/1 calcium chloride resulted in the removal of calcium and phosphorus from solution and crystal growth of calcium phosphate. However, prior treatment of the calcium phosphate with as little as 10-3 mM/l beryllium resulted in partial inhibition of crystal growth, while 10-2 mM/l beryllium totally inhibits crystal growth. No inhibition is seen when the beryllium concentration is reduced to 10-4 mM/l. The results of this study indicate that extremely low concentrations of beryllium ion (1 part per 100,000,000) can limit crystal growth. This function may be performed in vivo by critical concentrations of naturally occurring ions, such as citrate, polyphosphate, or some unknown substance(s). The ability to limit crystal growth of hydroxyapatite has theoretical implications for the dissolution of such crystals. These studies call attention to the possibility that some unknown ion(s) or compound(s) in trace amounts may be responsible for controlling the size of hydroxyapatite in vivo and in vitro.

PHYSICO-CHEMICAL HYPOTHESES OF CARIES SUSCEPTIBILITY AND THE ROLE OF DIETARY PHOSPHATE.-Over twenty years ago we postulated that the addition of soluble phosphate to the diet would reduce the incidence of dental caries. This hypothesis was based on physicochemical reasoning and on studies interrelating the composition of the diet to that of blood, bone, and teeth. In 1942 studies were presented relating the composition of blood serum to that of growing bones;' later we related it to that of growing teeth." 3 In these studies the composition of the blood serum was regulated by the diet. The addition of soluble phosphate (as Na2HP04) to the diet reduced the COa:P04 ratio of both the serum and the mineral of bones and growing teeth.>" We reasoned that teeth low in carbonate content would be less susceptible to caries since carbonate is preferentially dissolved.v-" In 1952, in collaboration with James H. Shaw of the Harvard School of Dental Medicine, we presented a paper" in which it was shown, by means of a controlled study in Cotton rats that teeth of animals on a high-phosphate diet were less susceptible to caries than the teeth of animals on a high-calcium diet. The carbonate content of the teeth was lower in animals who received a high-phosphate diet. This appeared to confirm our working hypothesis. A definite relationship was shown between the phosphate content of the diet and caries susceptibility, and at that time we assumed that the reduction of caries-susceptibility was due to the reduction of the carbonate content of the teeth. Since that time, it has been confirmed by others that caries-suceptibility is reduced by the addition of soluble phosphates to the diet. But the mechanism of this effect is still a subject for further study. The principal questions appear to be: (1) Is the effect systemic? (2) Is it a direct local effect? (3) Is it a combination of both? SYSTEMIC CHANGES OF BLOOD SERUM COMPOSITION.-Some of the systemic effects of adding phosphate to the diet, on blood composition, are given in Table 1. The relationship between serum and extracellular fluid was not established in this study, but it is generally accepted that the electrolyte composition of lymph closely resembles that of the ultrafiltrate of blood serum. It must be held in mind that the composition of both blood and lymph can be changed by means other than the diet. For example, in metabolic acidosis the carbonate is lowered, whereas in metabolic alkalosis the carbonate is elevated. In starvation there is a tendency for the carbonate to be lowered and the inorganic phosphate to be elevated. Parathormone'" and other hormones'! also exert an influence on the calcium and phosphate serum levels as well as on the citrate levels (see Figs. 1-4).

Using rachitic epiphyseal cantilage from rats, a study was made of strontification and calcification in vitro. The formation of a crystal from a solution takes place in 3 stages, (1) the formation of nuclei of crystallization (nucleation); (2) growth of crystals upon these nuclei; and (3) the cessation of crystal growth. These studies with rachitic rat cartilage indicate that in solutions of Ca and phosphate, nucleation takes place with relative ease. Nucleation takes place with Sr and phosphate, but is more difficult to achieve. Small amounts of Ca inhibit nucleation with Sr but once Sr phosphate nuclei have been produced, Ca ion does not inhibit the crystal growth of Ca phosphate. X-ray diffraction studies indicate that the crystals are Sr hydroxyapatite and Ca hydroxyapatite. (H.H.D.)

1. A study has been made of the repair of bony defects in the calvaria of albino rats. 2. An accelerated rate of bone repair was observed in experimental defects into which chondroitin sulphate-treated demineralised bone was implanted. 3. Acid-soluble collagen reconstituted with chondroitin sulphate was also more effective as an implant than was acid-soluble collagen reconstituted with sodium chloride. 4. It is concluded from these studies that chondroitin sulphate treatment accelerated the rate of new bone formation induced by demineralised bone, by reconstituted acid-soluble collagen, and to a lesser extent by Gelfoam. It was also found that demineralised bone and fresh homogenous bone promoted bone repair, but that chondroitin sulphate-treated demineralised bone promoted the most rapid rate of bone repair among the substances tested. 5. The possible role of chondroitin sulphate in bone formation is discussed.

More than a decade has passed since the thought was presented that there is an inverse relationship between dietary phosphate and caries susceptibility.' This was based in part on animal studies,2 in which diets varying from high to low in phosphate content resulted in teeth varying from low to high in carbonate concentration, respectively. It was suggested that the low-carbonate teeth were less soluble in dilute acids and therefore more resistant to at least one type of caries, whereas high-carbonate teeth were more acid soluble and less caries-resistant. By 19523 this concept led to the first trial experiments on a series of cotton rats in which low-carbonate and high-carbonate teeth were produced by feeding high-phosphate and low-phosphate diets, respectively, prior to being placed on a cariogenic diet.4 Two diets with greatly differing Ca/P ratios were selected in an attempt to highlight the effect on tooth carbonate composition. These experiments demonstrated that a high phosphate diet is cariostatic. The present report is an extension of these findings.

Transactions of the New York Academy of SciencesVolume 22, Issue 4 Series II p. 233-243 SECTION OF CHEMICAL SCIENCES: NUCLEI FORMATION AND CRYSTAL GROWTH IN MINERALIZING TISSUES* Albert E. Sobel, Albert E. Sobel Department of Biochemistry, The Jewish Hospital of Brooklyn, Brooklyn, N.Y.Search for more papers by this authorPenni A. Laurence, Penni A. Laurence Department of Biochemistry, The Jewish Hospital of Brooklyn, Brooklyn, N.Y.Search for more papers by this authorMartin Burger, Martin Burger Department of Biochemistry, The Jewish Hospital of Brooklyn, Brooklyn, N.Y.Search for more papers by this author Albert E. Sobel, Albert E. Sobel Department of Biochemistry, The Jewish Hospital of Brooklyn, Brooklyn, N.Y.Search for more papers by this authorPenni A. Laurence, Penni A. Laurence Department of Biochemistry, The Jewish Hospital of Brooklyn, Brooklyn, N.Y.Search for more papers by this authorMartin Burger, Martin Burger Department of Biochemistry, The Jewish Hospital of Brooklyn, Brooklyn, N.Y.Search for more papers by this author First published: February 1960 https://doi.org/10.1111/j.2164-0947.1960.tb00686.xCitations: 36 † This paper, illustrated with slides, was presented at a meeting of the Section on January 5, 1960. The work described was supported in part by the United States Air Force under contract AF 41(657)-229 monitored by the School of Aviation Medicine, USAF, Randolph AFB, Texas; the National Institute of Dental Research, Public Health Service, Grant D-S5, Bethesda, Md.; and the Office of Naval Research Project NR 105–025, Washington, D.C. Reproduction in whole or in part is permitted for any purpose of the United States government AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume22, Issue4 Series IIFebruary 1960Pages 233-243 RelatedInformation

The growth of nuclei into crystals requires a metastable solution phase with respect to the growing particles (supersaturation), and the presence of impurities. The rate of growth is proportional to the diameter of the particles, and is related to the surface to volume ratio of the crystal. Trace components can modify cyrstal habit and may be factors in determining the characteristics of bone mineral. Orientation of bone cystals parallel to the collagen fibers implies that the organic matrix acts as a habit modifier. The small size of bone hydroxyapatite crystals (640 Å or less), in contrast to the synthetic hydroxyapatite crystals prepared in the laboratory (2 · 107 Å or 2.0 mm), suggests that factors in vivo operate to limit their size. Cessation of crystal growth in a supersaturated solution is likely to be caused by surface adsorption of inhibiting ions or compounds. A mechanical barrier imposed by the collagen fibrils, and decreased renewal of the fluids surrounding the crystal may also be involved. The inorganic portion of mineralized tissues in two phases: a crystalline core of hydroxyapatite of [Ca10(PO4)6(OH2)] and an adsorbed outer shell which is of an amorphous or gel-like nature, containing anions such as carbonate, citrate, fluorides, and cations such as Na, Mg, Pb, and Sr. This does not preclude entry of a few such ions into the lattice. The inorganic composition of bones and teeth depends on the composition of the surrounding fluids (which can be regulated by dietar constituents). For example, the CO3/PO4 ratio of minerals in bone and teeth is related to the CO3/PO4 ratio of blood serum.

The alkaline phosphatase activity of pre-osseous tibial cartilage of rachitic bone stored in the deep freeze for two weeks at -25 degrees centigrade was only slightly less than that of fresh controls from the same animals. The deep frozen pre-osseous tissue did not calcify in in vitro calcifying media containing either inorganic phosphorus or organic phosphate ester. The fresh controls calcified equally well in both media. In addition, after deep-freeze storage the tissue hydrolysed the organic phosphate to the same degree as did the fresh tissue. Bones heated at 65 degrees centigrade will calcify in vitro after calcium chloride treatment despite the destruction of phosphatase activity. It appears unlikely that a relationship exists between alkaline phosphatase and the minimal system required for calcification of pre-osseous cartilage in vitro. These findings do not exclude the possibility that alkaline phosphatase plays some critical role in vivo.

1) Fibers of acid-soluble collagen reconstituted with chondroitin sulfate showed sheet-like structures without indication of spacings. After treatment for 2 days with basal salt solution at pH 7.3 and 37°C, containing neither calcium nor phosphate ions, non-striated fibers were formed of 300-1500 Å thickness. If either calcium or phosphate ions were present partial transformation to the form with 640 Å spacings took place. If both of these ions were present this transformation occurred more rapidly and more completely. 2) These findings suggest that calcium and phosphate ions interact with sites on the collagen fiber where the breaking of electrostatic bonds catalyzes transformation to the 640 Å spaced fiber.