VITAMINS 



372 



VITAMINS 



These include colloidal suspensions of 

 various benzidine dyes (trypan blue, 

 isamin blue, pyrrhol blue, trypan red, 

 etc.), of silver, Higgins ink, lamp black 

 etc.; and of simple suspensions of India 

 ink, carmine, graphite and so on. They 

 are injected intravenously, intraperi- 

 toneally or subcutaneously. The litera- 

 ture is enormous. Consult latest issue 

 of the Quarterly Cumulative Index 

 Medicus. For chemistry of Benzidine 

 dyes see Evans, H. M. and Schulemann, 

 W., Science, 1914, 39, 443. 



The following experiment is suggested. 

 Give each of a dozen or more white 

 mice 1 cc. of 0.5% trypan blue in sterile 

 aq. dest. intraperitoneally and in the 

 course of a few minutes the beginning of 

 deposition of the dye in the ears will be 

 noted. Give similar doses every sec- 

 ond day for 8 days. A few hours after 

 the last draw a little blood from the 

 tail and observe that some of the mono- 

 cytes have taken up the dye. Then 

 autopsy the mice and study the dis- 

 tribution of the dye in the tissues. 

 The skin, kidneys, adrenals, liver, 

 spleen ana bone marrow will be found 

 quite deeply colored while the nervous 

 system has escaped. The heaviest 

 accumulation will be in the peritoneal 

 cavity near the sites of injection and in 

 the loose connective tissue everywhere. 

 Examination of fresh mounts in physio- 

 logical salt solution will reveal that the 

 dye is concentrated within (1) the 

 epithelial cells of the convoluted tubules 

 of the kidney, of the adrenal and choroid 

 plexus; (2) certain cells of the ovary 

 and testicle; (3) the macrophages of 

 loose connective tissue and especially 

 of the spleen, liver, bone marrow, ad- 

 renals and lymph nodes — fibroblasts 

 are colored less deeply; and (4) the 

 "specific endothelia" of the five organs 

 mentioned. If permanent preparations 

 are desired fix in 10% formalin and im- 

 bed in paraffin. 



Vital staining in the narrow sense is 

 used for many purposes. (1) To iden- 

 tify phagocytic cells of the reticulo- 

 endothelial system and to see how they 

 behave in normal and pathological 

 conditions. (2) To locate injured cells 

 because some cells that do not ordi- 

 narily stain take up the dye when 

 injured. (3) To influence the activity 

 of R. E. cells by blocking them with 

 particulate matter. This has not been 

 very successful. See R. E. Blockade 

 (Victor, J., Van Buren, J. R. and 

 Smith, H. P., J. Exper. Med., 1930, 51, 

 531-548). (4) To measure the absorp- 

 tion by membranes of particulate matter 

 (Wislocki, G. B., Anat. Rec, 1921, 21, 

 29-33). (5) To distinguish between 



malignant and non-malignant cells (Lud- 

 ford, R. J., Arch. f. exp. Zellf., 1933, 

 14, 42-55). (6) To determine pH of 

 different organs and tissues by injec- 

 tion with phthalein indicators (Rous, 

 P., J. Kxper. Med., 1925, 41, 739-759). 

 (7) To identify calcium salts laid down 

 (Alizarin Red S and Madder). See 

 method for Reticulo-endothelial system. 

 It is sometimes very worthwhile to 

 inject simultaneously three materials, 

 for example Higgins' Ink intravenously, 

 trypan blue or Niagara blue intraperi- 

 toneally, and lithium carmine intra- 

 pleurally (Foot, McClung, p. 116). 

 An interesting experiment is to feed 

 Sudan III or Scharlach (scarlet = 

 Sudan IV) colored lipids. Make solu- 

 tion in olive oil (about 20%). Intro- 

 duce by stomach tube into a cat. There 

 is slight staining of fatty tissue within 

 24 hrs. and maximum in 3-7 days 

 (Hadjioloff, A., Bull. d'Hist. AppL, 

 1938, 15, 81-98). Try also inducing 

 cat to drink large amount of milk or 

 cream colored with Sudan III or Sudan 

 black, see colored illustrations of Gage 

 and Fish (S. H. and P. A., Am. J. Anat., 

 1924-25, 34, 1-81). History of vital 

 staining (Conn, H. J. and Cunningham, 

 R. S., Stain Techn., 1932, 7, 81-90, 

 115-119). See Chorioallantoic Mem- 

 brane, Carmine, Indigo-Carmine, 

 Manganese Dioxide, Higgins' Ink, 

 Protargol (silver), Lampblack, Leuco- 

 Dyes, Nuclei, Titanium Dioxide, Tho- 

 rium Dioxide, Copper, Platinum, Iron, 

 Mercury, Lymphatic Vessels. 

 Vitamins — Written bj^ C. Carruthers, Wash- 

 ington University School of Medicine, 

 St. Louis. February 15, 1951 — Only a 

 few vitamins are susceptible of micro- 

 scopic localization. Deficiencies in most 

 of them leave structural imprints 

 in the tissues. A list may be therefore 

 useful giving briefly required tech- 

 niques. Current information is sup- 

 plied in Annual Reviews of Biochem- 

 istry and Physiology. 



A useful background is provided by 

 Sherman, H. C. (Chemistry of Food and 

 Nutrition, New York: Macmillan, 1941, 

 611 pp.). For a summary of tissue 

 changes in vitamin deficiencies see 

 Wollach, S. B. and Bessev, O. H. (Phvs- 

 iol. Rev., 1942, 22, 233-^290). For the 

 biochemistry and chemical determina- 

 tion of the vitamins the following are 

 recommended: Gyorgy, P., Vitamin 

 Methods, New York, Academic Press. 

 1950, 571 pp.; Williams, R. J., Eakerj 

 R. E., Beerstecker, E., and Shive, W., 

 The Biochemistry of B. Vitamins, New 

 York, Reinhold Publishing Co., 1950, 

 741 pp.; Harrow, B., One Family. 

 Vitamins, Enzymes, Hormones, Minne- 



