T. H. Jukes

Further Observations on the “Animal Protein Factor”

Summary and Addendum 1. Fermentation products of Streptomyces aureofaciens were found to promote growth in depleted chicks on various diets which were adequately supplied with vitamin B12. 2. Growth responses in chicks on a corn-soybean diet were also produced by crystalline aureomycin hydrochloride and by cultures in which the aureomycin, as measured by antibiotic potency, was destroyed by alkaline hydrolysis. 3. Responses were also obtained with succinylsulfathiazole, streptomycin and 3-nitro-4-hydroxyphenylarsonic acid, but these substances appeared less potent than aureomycin. No responses were obtained with dried whey or dried brewers yeast. Following the submission of this manuscript, Dr. Paul Gyorgy kindly showed us a manuscript (Gyorgy, P., Stokes, J., Jr., Smith, W. H., and Goldblatt, H., Transactions of Conference on Biological Antioxidants, sponsored by the Macy Foundation, November, 1949) which described a beneficial exerted by aureomycin on growth and the prevention of dietary hepatic necrosis in rats. The assumption was made that aureomycin might act through the suppression of the intestinal flora. The assistance of Vincent Stryeski, Alfred Plattner and Joseph Lang in the care of the animals is gratefully acknowledged.

Assay of Anti-Pernicious Anemia Factor with Euglena.:

Summary The algal flagellate Euglena gracilis var. bacillaris was shown to exhibit a quantitative growth response to crystalline antipernicious anemia factor, using a chemically defined medium. Thymidine was inactive.

Antithyroid Effects of Aminotriazole

Aminotriazole, Like other antithyroid compounds and low-iodine diets, produces adenomatous changes in the thyroid glands of rats when fed continuously for long periods. Such changes are reversible if the antithyroid regime is discontinued. The antithyroid compounds that are naturally present in food are not regarded as causes of cancer.

Observations on the Effect of 4-Amino-Pteroylglutamic Acid on Mice

Summary The 4-amino analogue of pteroylglutamic acid was fed to mice as a dietary supplement. There was some indication of reversal by pteroylglutamic acid of the effect on mice when the level of the antagonist was 0.3 part per million of diet. Death was found to occur within a few days with levels of 1 part per million of diet or higher. The effect was not reversed by feeding high levels of pteroylglutamic acid. This finding may be contrasted with the inhibitory effect of the 4-amino compound on S. jecalis R which is reversed by pteroylglutamic acid.

Prevention of Chemotherapeutic Effects of 4-Amino-N10-Methyl-Pteroyl-glutamic Acid on Mouse Leukemia by Citrovorum Factor.

Summary The effect of 4-amino-N10-methyl-PGA in prolonging the survival time of mice with transplanted leukemia AK4 can be blocked almost completely by prior administration of 1/3 to 2/3 as much by dry weight of a preparation of citrovorum factor. Thus, citrovorum factor is at least 12 to 24 times as active as pteroylglutamic acid in preventing the antileukemic effect of 4-amino-N10-methyl-PGA.

Acceleration of Pteroylglutamic Acid Deficiency in Mice and Chicks by a Chemical Antagonist

Summary Mice on a purified diet with added succinylsulfathiazole developed no signs of pteroylglutamic acid deficiency within 6 weeks. When a crude synthetic preparation of a pteroylglutamic acid antagonist was added to the diet a syndrome appeared which was characterized by slow growth, anemia and leucopenia. Pteroylglutamic acid prevented the appearance of the syndrome. The development of pteroylglutamic acid deficiency in chicks on a purified diet was aggravated by adding the antagonist to the diet, and the effects of the antagonist were reversed by pteroylglutamic acid.

Vitamin B12b Some Properties and its Therapeutic Use.

Summary A pink fraction obtained from liver and from Streptomyces aureofaciens has been separated from vitamin B12 by silicic acid chromatography and has been shown to have different ultraviolet and visible absorption spectra. This new substance, vitamin B12b, has been prepared in crystalline form and has been found to be effective parenterally in the treatment of patients with Addisonian pernicious anemia in amounts of 1 to 2 μg daily. We are indebted to Mrs. Helen Jakubowski for performing the blood examination.

The role of vitamin B12 in metabolic processes.

Publisher Summary This chapter analyzes the role of vitamin B 12 in metabolic processes. The fundamental source of vitamin B 12 in animal nutrition appears to be in nonphotosynthetic organisms. A microbiological synthesis of vitamin B 12 may take place in the rumen and elsewhere in the digestive tract of cattle, providing a supply of the vitamin that is accumulated in the liver, thus accounting for the effectiveness of extracts of this organ in the treatment of pernicious anemia. The growth of certain thermobacteria is stimulated by thymidine or vitamin B 12 . The requirement for vitamin B 12 in certain lactic acid organisms can be obviated by the addition of ascorbic acid. Microorganisms that respond to vitamin B 12 or thymidine for growth on a medium containing pteroylglutamic acid may be divided into five groups depending on their response to various desoxyribosides, vitamin B 12 and ascorbic acid. Studies also show that vitamin B 12 is concerned in reactions involving a single carbon transfer in the synthesis of methionine, serine, thymine and purines in E. coli . Studies also show that vitamin B 12 is involved in the metabolism of methionine in sulfanilamide-inhibited E. coli .

Effect of Various Levels of Vitamin B12 Upon Growth Response Produced by Aureomycin in Chicks

Summary 1. The vit. B12 requirement for growth of chicks was studied on a deficient diet with and without the addition of aureomycin. In some experiments, a “sparing effect” of aureomycin upon vit. B12 requirement was noted but in others no effect was observed. A growth-promoting effect for aureomycin was noted both in the presence and absence of added vit. B12. 2. The mortality of deficient chicks on the diet containing no added vit. B12 was markedly reduced by aureomycin. 3. The addition of sulfamethazine to the diet did not appear to effect the quantitative requirement of chicks for vit. B12.

Some Observations on Growth Factors Required by Leuconostoc citrovorum.

Summary 1. Leaconostoc citrovorum was found to respond to a growth-promoting factor in the concentrated liver extract, but this organism did not respond to vitamin B12. The alkali-stable nature of the “citrovorum factor” further contrasts it with vitamin B12. 2. Two fractions were separated from liver extract by paper strip chromatography; one of these fractions was presumably thymidine and promoted growth of Lactobacillus leichmannii and Leuconostoc citrovorum. The other fraction was inactive for L. leichmannii but active for L. citrovorum. 3. Although thymidine or high levels of pteroylglutamic acid (PGA) when tested singly were only partially effective in promoting growth of L. citrovorum, the simultaneous addition of thymidine plus PGA produced marked growth of the organism. This finding suggests a functional relationship between thymidine, PGA and the “citrovorum factor.”