No. 4553 February 2, 1957 NATURE 261 Effect of Ethisterone, [3-<Estradiol and two weeks, four showed vital staining appearances Activity similar to the controls, while the remaining two Progesterone on the Phagocytic es. Reticulo-Endothelial System animals showed reduced activity of the macrophag of the The results, therefore, suggest that, in the male THE reticulo-endothelial cells are known to form guinea pig, ethisterone, [3-restradiol and progesterone an important part of the defence mechanism of the have little or no effect on the activity of the reticulo­ body against infection. At the site of a cute infections, endothelial system and in this respect differ greatly neutrophil leucocytes attack the invading organisms from the cestrogens. while the reticulo-endothelial macrophages phago­ The substances used in these investigations were cytose the dead bacteria and dead tissue cells. provided by Dr. Tindall, of Organon Laboratories, In chronic infections such as tuberculosis, the Ltd., to whom we offer our grateful thanks. phagocytic activity of the macrophages forms the T. NICOL main line of defence against the invading organisms. R. s. SNELL Further, there is much circumstantial evidence in Department of Anatomy, both acute and chronic infections to show that the King's College, 1·eticulo-endothelial cells raise the humoral resistance London, W.C.2. Nov. 9. patient by the production of antibodies. It of the 1 Nicol, T., Helmy, I. D., and Abou-Z1kry, A., Brit. J. Sura., 40, 166 has already been demonstra ted that the phagocytic (1952). activity of the reticulo-endotholial system is stimu­ • Nicol, T., and Snell, R. S., Nature, 174, 554 (1954): 177, 430 (1956). 2 [178, 1405 (1956) J. lated by restrogens1 ; that cortisone , testosterone, ' Snell, R. S., and Nicol, T., Nature androstene-3[3-l 71X-diol3 depress the phagocytio • Nicol, T., and Snell, R. S., Nature, 175, 995 (1965). activity ; and that deoxyoortioosterone• has no appreciablo effect. The present communication deals with the e ffects Isolation of Nigerose from Floridean of ethisterone, [3 -restradiol and progesterone. Forty­ Starch three male guinea pigs were used. The reticulo­ IT is generally agreed that the iodine-staining by giving all endothelial macrophages were studied polysaccharide isolated from many red algae is a blue the animals one daily injection of trypan glucan of the amylopectin type in which the cha.in­ being subcutaneously for the last six days prior to forming links are IX-1 : 4 and the branch links, was killed by chloroform. The dosage of the dye 01:-l : 6-glucosidic. The floridean starch from Dilsea per calculated on the basis of O ·8 ml. of a 1 edulis appears to differ, in finer details of structure, body­ cent solution in distilled water per 100 gm. from one sample to another. This difference turns only weight. Seven of the animals were given dye m ainly on the presence or the absence of a third type as controls. The hormone treatment 1 and were used of link, namely, the 1: 3-glucosidic linkage • The shown given to the remaining thirty-six animals is m ajority of investigations favour the view that in Table 1. 1 : 3-glucosidic linkages are in fact not present in Dilsea starch•. We therefore think it desirable to Table 1. HORMONE TREATMENT GIVEN TO THmTY•SIX ANIMALS record some observations made in the course of a Dally dose Duration comprehensive linkage analysis of £1.oridean starch No.of Hormone used intra- of which establish that a small proportion of 1 : 3-links animals muscularly treatment is an integral part of the starch structure and, more­ ---6 Ethisterone (Organon) dis- 0 ·5 mgm. 2 weeks over, that these links have the IX-configuration. solved in ethyl oleate Floridean starch was extracted from Dilsea edulis 6 Ethisterone (Organon) dis- 0·5 mgm. 4 weeks by the method of Barry et al. 1 and was purified by solved in ethyl oleate precipitation with iodine• and by use of cetyl tri­ 6 /1-CE•tradl ol (Organon) dis- 0·5mgm. 2 weeks methyl ammonium bromide ('Cetavlon') to precipitate solved In ethyl oleate a galactan sulphate impurity•. The starch (92·5 per 6 /1-CEstradlol ~rganon) dis- 0 ·5 mgm. 4 weeks cent polyglucose) had [1X]~ + 187° (water) and blue in et yl oleate solved value, 0·065. The starch (12·5 gm.) was hydrolysed 6 Progesterone (Organon) dis- 6·0 mgm. 2 weeks by 0·33N sulphuric acid (1·25 litres) at 100° In ethyl oleate solved to the stage of an apparent conversion into glucose 6 Progesterone (Organon) dis- 3 ·0mgm. 3 weeks of 53·4 per cent (2·15 hr.). After being cooled, ethyl oleate solved In neutralized and concentrated, the hydrolysate was chromatographed on a column of charcoal-'Celite'•. but Specimens were taken from the splean, liver and Glucose was the only monosaccharide obtained, into two lymph nodes and fixed in Heidenhain's Susa fluid. the disaccharides were readily separated entirely of Sections were cut at 10µ thick and stained with weak fractions, the first consisting almost as its [3-octa­ eosin, dilute carbol fuchsin or alum carmine. The isomaltose (289 mgm.), characterized [1XJt 96 ·9° in activity of the reticulo-endothelial system in the acetate (melting point 142 ·3° ; + organs studied was assessed by the number of dye­ chloroform). The second disaccharide fraction was bearing cells and the intensity of the vital staining. mainly maltose but contained a small amount of In the animals which received ethisterone and another disaccharide, which was separated from the [3-restradiol, a moderate number of dye granules was maltose by taking advantage of its greater electro­ present in the macrophages of the spleen, liver and phoretic mobility in borate buffer•. The amorphous lymph nodes and the vital staining appearances solid obtained (34·6 mgm.) migrated as a single closely resembled those in the control animals. In substance during both paper chromatography and the six animals which received 3 mgm. of progesterone electrophoresis and had mobilities in each system daily for three weeks the vital staining appearances identical with those of authentic nigerose. It showed were similar to those of the controls. In the group [1Xn + 136·9° (water) and its f,-octa-acetate had which received 5 mgm. of progesterone daily for melting point and mixed melting point 147-48° and © 1957 Nature Publishing Group.