OXYHEMOGLOBIN 
 
 ALLIED PRODUCTS. 
 
 FREDERICK STEARNS & CO. 
 
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 btate (QolUge of Agriculture 
 
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Cornell University Library 
 RS 163.09S8 
 
 Oxyhemoglobin and allied products. 
 
 '""l""™ 003 467 986 
 
|| Cornell University 
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 The original of this book is in 
 the Cornell University Library. 
 
 There are no known copyright restrictions in 
 the United States on the use of the text. 
 
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AN ILLUSTRATED MONOGRAPH 
 
 ON 
 
 OXYHEMOGLOBIN 
 
 ALLIED PRODUCTS. 
 
 PART I.— OXYHEMOGLOBIN, 
 
 ALBUMINATE OF IRON, AND 
 PEPTONATE OP IRON. 
 
 —Stewart. 
 
 PART II.— THE ABSORPTION OF 
 
 IRON IN THE ANIMAL BODY; 
 
 /V — MACALLUM. 
 
 PART III.— THE HAEMOGLOBINS AND RELATED 
 PRODUCTS OF THE MARKET.— Williams. 
 
 PUBLISHED UNDER THE DIRECTION OF 
 
 F. E. STEWART, M. D., Ph. G. 
 
 Director of Scientific Department, F. Stearns & Co. 
 
 Formerly Demonstrator and Lecturer on Materia 
 
 Medica and Pharmacy, Jefferson 
 
 Medical College, Etc. 
 
 PRESS OF 
 
 FREDERICK STEARNS & CO. 
 
 Detroit, Mich. 
 
 1895. 
 
09 S3 
 
 SCIENTIFIC DEPARTMENT. 
 
 < FREDERICK STEARNS & CO. 
 
 »«, P. E. STEWART, M. D., Ph. Q., DIRECTOR. 
 
 CHEMISTRY— C/C. SHERRARD, Ph. C. 
 Materia Meoica— L. H. (fARDNER, Ph. C. 
 PHARMAOOGNdsv — P. H. FRAZEE, Ph. C. 
 Botany— W. Q. RANKIN, Ph. C. 
 
 
 
 ■V 
 
 Therapy— F. E. STEWART, M. D., Ph. Q. 
 Pharmaceutical Assaying — 
 
 J. L. TEQARDEN, Ph. C 
 Microscopy — S. C. STEARNS. 
 
CONTENTS. 
 
 PAGE 
 
 Introduction. 
 Oxyhemoglobin : 
 
 Its administration in anaemia; action of gastric juice 
 upon, etc. The absorption of Iron; theories concerning 
 the same 1-3 
 
 Haemoglobin and Oxyhemoglobin. Discrimination be- 
 tween haemoglobin and oxyhaemoglobin. Preparation 
 of oxyhaemoglobin. Methaemoglobin 2-3 
 
 Composition of Haemoglobin. Composition not clearly 
 determined. Origin of the name "Haemoglobin." 
 Haemoglobin classed as a proteid food product. Chemi- 
 cal nature of haemoglobin. Effect of gastric digestion 
 on haemoglobin. Haematin. Description and solubili- 
 ties of hasmatin 3-4 
 
 Spectroscopic Assay. Examination of haemoglobin by 
 the spectroscope. Description of plate (frontispiece). 
 Absorption specta of oxyhaemoglobin 4-6 
 
 Therapeutics. Introduction of Sanguis Bovinus Exsic- 
 catus. Its preparation. Dr. Andrew H. Smith's for- 
 mula. Experiments at St. Luke's Hospital, New York. 
 Introduction of Haemoglobin Compound. "Haemofer- 
 rum." Dr. Campbell's theory. Experiments of Pon- 
 flck. Bunge's observations. Fost m^s explanation of 
 the action of living cells and tissues. Reports con- 
 cerning the administration of oxyhaemoglobin by Dr. 
 Desidero Varely, of Mexico. Contributions to the 
 literature of the subject by Drs. A. E. Sansom, Thos. 
 S. Butcher, E. O. Day, Guerder, of Paris, France, 
 Steibel and others. Editorial from Medical Record re- 
 lating to the work of Drs. Debove, Dujardin-Beaumetz 
 and Guerder 6-14 
 
 Oxyhaemoglobin as a Pharmaceutical Product. Ex- 
 periments with Oxyhaemoglobin (Haemoferrum brand). 
 Properties of Oxyhaemoglobin. Clinica 1 reports by Dr. 
 Chas. E. McCallum, Chas. "W. Ellis, and W. J. O'Reilly. 
 Dose of Oxyhaemoglobin. Best form of administration. 
 Iron not the only means for the cure of anaemia; do 
 not neglect those means which should accompany its 
 
 administration 14-17 
 
 Albuminate of Iron 17 
 
 Peptonate of Iron- 18 
 
Contents— Continued. 
 
 PAGE 
 
 The Absorption of Iron in the Animal Body. 
 
 Difficulty of determining chemico-physiological relations 
 of Iron. Masked Iron. Unsettling nature of Bunge's 
 researches. Haematogen 19-20 
 
 On the Absorption of Inorganic Iron Compounds. Con- 
 flicting opinions regarding. Micro-chemical method of 
 investigation. Experiments on guinea-pigs, with "pep- 
 tonate," albuminate, chloride, sulphate, and phosphate 
 of Iron. Experiments on kittens. Observation regard- 
 ing absorbed Iron in the villi. Iron -holding leucocytes. 
 Experiments on lake-lizards (Nectus lateralis). Con« 
 elusion : Inorganic and albuminate compounds of Iron 
 are absorbed. Mode of excretion of Iron 20-31 
 
 The Absorption of Organic Iron Compounds. Classes of 
 Organic Iron Compounds: (1) chromatins; (2) haemo- 
 globin, melanin, lardacein. Investigations limited to 
 representatives of chromatin class. Experiments on 
 lake-lizards (Nectus lateralis) with chromatin from 
 lamb's testicles and egg-yolk. Experiments on mice, 
 guinea-pigs and on Amblystromata. Leucocytes carry 
 Iron to the liver. Method of entrance of chromatin to 
 epithelial cells. Organic Iron Compounds are ab- 
 sorbed 31-44 
 
 Summary , 44-45 
 
 Explanation of Figures 45-47 
 
 The Hemoglobins and Belated Products of the Market. 
 
 Haemoglobins add to the elegance of medicinal ingestion. 
 Administration of Iron in form of Haemoglobin a 
 combination of rival ideas. Haematin formed by the 
 action of gastric juice on Oxyhemoglobin 51-52 
 
 Haemoglobin and Oxyhaemoglobin. Methaemoglobin. 
 
 . Spectrum. Absorption bands ... 52-53 
 
 Preparation of Haemoglobin. Spectroscopic examination 
 of Haemoglobin. Examination of the Haemoglobins of 
 the market, "a" Haemoglobin (Merck) ; "b" Hsemofer- 
 rum; "c" Ferroglobin ; "d" Ferrum Sanguinis (Chapo- 
 teaut); "e" Ferratin; "f" Hsemogallol; "g" Desiccated 
 Defibrinated Blood; "h" Haemoglobin Compound. 
 
 Table showing comparative tests. 
 
 LIST OP ILLUSTRATIONS. 
 
 Frontispiece. 
 
 Plates illustrating the counting of blood corpuscles. 
 
 Plates illustrating Macallum's article. 
 
INTRODUCTION. 
 
 This monograph emanates from our Scientific Department and 
 is in no sense of the term an advertisement. 
 
 Inasmuch as we have no exclusive control of the manufacture 
 and sale of any article herein mentioned, and as the treatise is 
 solely designed to advance the science of medicine and allied 
 beneficent arts, we believe that the most enthusiastic "altrurian" 
 can take no exception to it. 
 
 A word of explanation regarding our Scientific Department. 
 
 "We organized it upon the theory that, pharmacy being a branch 
 of medical science and practice, the standing of the manufacturing 
 pharmacist, like that of the physician, should be guaged rather by 
 his contributions to science, than by the extent of his business. All 
 commercial considerations have, therefore, been eliminated from 
 this department, which we maintain solely for the gratuitous dis- 
 semination of knowledge by the publication of the investigations of 
 modern scientists. It is, however, far from being a mere Bureau 
 of Publication, in that the outside investigator is supplied with 
 material for original laboratory or bedside experiments, and is, in 
 special cases, remunerated for his labors. 
 
 We make no false pretenses; we have a natural desire to aug- 
 ment the prestige of our house, and the conduct of this scientific 
 work brings us into a frequent contact with leaders in medicine and 
 pharmacy which necessarily encourages friendly relations and 
 ultimately swells our list of patrons. Again — the discussion of the 
 therapeutic value of drugs of which we, in common with other 
 houses, place new preparations upon the market, increases the 
 general demand and incidentally our own output. 
 
 Before a remedy is introduced to the profession at large, it 
 should be thoroughly tested in laboratory and hospital practice. 
 Private practitioners will not bring this about by merely ignoring 
 preparations which have not undergone such tests. They should 
 rather devote their energies to enforcing the proper performance of 
 duty in this regard by physicians connected with laboratories and 
 hospital dispensaries. 
 
 We hold that the manufacturing pharmacist is comparatively 
 little to blame for the present situation in regard to proprietary 
 medicines in that he has simply supplied what was called for, 
 leaving it to the profession to create a demand for something better. 
 This the latter can now do by co-operating in the work of our 
 
Scientific Department. As an evidence of liberal spirit, we offer to 
 remunerate them for such work as may be done under our control, 
 or in the laboratories of the universities, or in large hospitals. The 
 proper performance of the scientific work undertaken has been 
 assured by the careful selection of a staff of experts in the theory 
 and practice of medicine, pharmacy and chemistry. 
 
 The lofty standard set for the department will be persistently 
 maintained. It will not be permitted to degenerate into a mere 
 Advertising Bureau. Conducted as it is by high-grade physicians 
 and pharmacists •who can be held personally accountable for their 
 work, we feel justified in soliciting the active participation of the 
 profession in their labors. 
 
 TO PHYSICIANS. 
 
 The staff of the Scientific Department of Frederick Stearns 
 & Co. hereby extend a cordial and urgent invitation to practitioners 
 who may desire information on any subject pertaining to thera- 
 peutics or pharmacy, to address their inquiries to the laboratory. 
 If the problem submitted for solution is such as can only be passed 
 upon by a specialist, we shall without delay place it before one of 
 the numerous experts with whom we are in touch and the response 
 will be promptly forthcoming. 
 
PART I. 
 Oxyhemoglobin. 
 
 By F. E. STEWART, M. D., Ph. G., 
 
 Director of the Scientific Department of Frederick Stearns & Co. Formerly 
 Lecturer on and Demonstrator of Materia Medica and Pharmacy, Jeffer- 
 son Medical College, Medico-Chirurgical College, and Woman's Medi- 
 cal College, Philadelphia; Author of a Compend of Pharmacy; 
 Member of the American Medical Association, Ameri- 
 can Pharmaceutical Association, Judicial Council 
 Ninth International Medical Congress, etc 
 
Oxyhemoglobin. 
 
 Haemoglobin, the well known principle to which the blood owes 
 its color, is chiefly of interest to the physiologist as an oxygen car- 
 rier, and to the therapist because of the iron it contains. Its ad- 
 ministration in anaemia has been favored because of the avidity 
 with which, when merely exposed to the air, it absorbs and com- 
 bines loosely with oxygen, and second, because of the ease with 
 which, when brought into relation with the oxygen free tissues of 
 the body, it releases the oxygen for use in the respiratory processes. 
 Unfortunately, this theory does not take into consideration the ac- 
 tion of the gastric juice on haemoglobin. Bunge says, "Haemo- 
 globin 1 splits up rapidly under the action of the digestive ferments ; 
 the iron separating as haematin. It is not known whether a part of 
 the haematin becomes absorbed, as no quantitive experiments have 
 been made to decide this point." Haemoglobin would not act as an 
 oxygen-carrier, if injected directly into the blood, for experiments 
 have proven that 2 the haemoglobin thus injected appears in the 
 urine as urobilin, a product of the conversion of bilirubin. It is 
 almost certain that bilirubin arises from the haemoglobin. 
 
 The absorption of iron is being discussed by leading physiolo- 
 gists throughout the world. Bunge believes that no prepara- 
 tion of iron is absorbed, while Macallum has brought forward 
 evidence which is almost overwhelmingly in favor of its absorption. 
 The researches of the latter (published in the Journal of Physiology 
 for April, 1894), are so interesting and valuable that we have 
 included them in this Monograph. (Part II). Whether Iron is 
 absorbed or not, this cannot affect its position as a therapeutic 
 agent. Even Bunge himself, in his classical work of Physiologi- 
 cal and Pathological Chemistry, page 94., after using argu- 
 ments to prove that Iron is never absorbed, does not hesitate 
 to say that "It is nevertheless noteworthy, that there are few rem- 
 edies of the efficacy of which in cases of chlorosis, almost all physi- 
 
 1 Text Book of Physiological and Pathological Chemistry, G. Bunge; trans- 
 lated from the second German edition by the late L. C. Wooldridge, M. D., D. Sc. 
 London, Kegan-Paul Trench, Trubner & Co., 1890. 
 
 2 Ibid. p. 375. 
 
cians are so firmly convinced, as that of iron. It is not one of the 
 remedies which are recommended one' day, considered infallible 
 the next, and forgotten in a short time thereafter, for the adminis- 
 tration of Iron is as old as the history of medicine itself. Even 
 those skeptics who doubt the value of all other drugs will assure us 
 that chlorosis, which is frequently so obstinate, will almost invari- 
 ably yield in a few weeks to a vigorous treatment with iron." 
 
 The next question is, how much iron should be administered? 
 That, again, depends on whether it is absorbed or not. If it is 
 absorbed, a very small amount would suffice to supply the blood 
 with haemoglobin. While the amount of iron in haemoglobin is 
 but 0.42 per cent, this is ample for therapeutic purposes if it be ab- 
 sorbed. If however, it is not absorbed, much larger doses of iron 
 would have to be administered, in order to render the alkaline sul- 
 phides found in the intestine inert. 
 
 Another very interesting question bearing on the medicinal 
 value of haemoglobin is as to whether organic or inorganic prepara- 
 tions are the more readily absorbed. It has always been assumed 
 that vegetive intervention was necessary to raise inorganic matter 
 to the plane of animal life. It is because animals cannot subsist on 
 inorganic material that they feed on vegetables or the flesh of other 
 animals. Now, haemoglobin being an organic preparation, the 
 probability of its absorption seems to be greater than is the case with 
 inorganic preparations of iron. Being the form of iron found in the 
 blood, it can be cogently argued that it must be peculiarly adapted 
 for supplying the blood with iron. The comparative powers of 
 absorption of inorganic and organic iron are discussed at length in 
 Prof. Macallum's article. 
 
 Hsmoglobin and Oxyhemoglobin. 
 
 Physiological text books of recent issue have discriminated 
 more clearly between haemoglobin and oxyhaemoglobin. The 
 method for preparing haemoglobin given in Foster's Physiology 
 (3rd edition) furnishes oxyhaemoglobin rather than haemoglobin. 
 The crystals of haemoglobin have a dark red appearance, with a 
 strong purple or bluish tint, unlike the brighter color of oxyhaemo- 
 globin. The method referred to consists primarily of breaking up 
 the corpuscles by the addition of chloroform or bile salts or by 
 alternately freezing and thawing. The alkaline serum should be 
 first removed, either by allowing the blood to clot or by centrifug- 
 ally separating the corpuscles. The breaking up of the corpuscles 
 results in an aqueous solution of haemoglobin. The alkalinity of 
 the solution can, if necessary, be reduced by the cautious addition 
 
acetic acid, while the solvent power of the aqueous medium can 
 be diminished by the addition of one-fourth its bulk of alcohol. 
 When the blood of the dog, cat, horse, etc., is used, and the mixture 
 set aside in a temperature of 0°C. in. order to still further reduce 
 the solubility of the haemoglobin, it readily crystallizes. When 
 prepared from bullock's blood, however, crystals are very difficult 
 to obtain. The solution of haemoglobin should, therefore, be con- 
 centrated in vacuo and dried on glass plates. The resultant pro- 
 duct is pulverulent, or rather of a character resembling the scaly salts 
 of the pharmacopoeia. If this process is carefully conducted, the fin- 
 ished product is almost pure oxyhaemoglobin. The only impurity 
 present is a small amount of methaemoglobin, which in so far as it 
 affects the medicinal properties of haemoglobin, should hardly be 
 called an impurity, for it contains as much iron as haemoglobin it- 
 self.' Oxyhemoglobin and methaemoglobin have exactly the same 
 chemical composition, the only difference being that the oxygen is 
 more loosely combined in oxyhemoglobin than in methaemoglobin. 
 In other words, we may probably say, without being able to 
 state exactly what has taken place, that, under certain conditions, 
 the oxygen, loosely united to haemoglobin as oxyhemoglobin, 
 becomes more stably combined and is not removable either by means 
 of a vacuum, or of carbon-monoxide, or of a current of hydrogen ; and, 
 further, that the resulting substance (methaemoglobin) has the same 
 composition and crystalline forms as oxyhemoglobin, and may be 
 reconverted into the latter body by suitable means, such as reduc- 
 tion by ammonium sulphide and subsequent oxidation. 3 
 Composition of Hemoglobin. 
 
 As Foster has pointed out, " 3 The exact nature of the 
 proteid constituent of haemoglobin has not as yet been clearly 
 determined. It was supposed to be globulin (hence the name 
 haemetoglobulin, contracted into haemoglobin), but though belonging 
 to the globulin family, it has characteristics of its own ; it is possibly 
 a mixture of two or more distinct proteids. It has been provision- 
 ally named globin and is said to be free from ash." Haemoglobin 
 therefore must be classed as a food product which probably plays 
 a similar part in nutrition to that of other proteid substances. 
 Another interesting feature in relation to this product is that its 
 alkaline solution is capable of being reduced by reducing agents; 
 the spectrum changing at the same time. The reduced solution 
 
 1 The Chemical Basis of the Animal Body; an appendix to Foster's Text 
 Book of Physiology (sixth edition) by A. Sheridan Lea, M. A., D. Sc, F. B. S. 
 p. 229. 
 
 2 Ibid, p. 230. 
 
 3 Ibid. p. 868. 
 
 3 
 
will, like the haemoglobin, take up oxygen again on being brought 
 into contact with the air or oxygen. This would seem to indicate 
 that the oxygen-holding power of haemoglobin is connected exclu- 
 sively with its hsematin constituent. 
 
 '"By the action of strong sulphuric acid, hsematin may be 
 robbed of all its iron. It still retains the feature of possessing 
 color, the solution of iron-free haematin being a dark rich brownish 
 red ; but is no longer capable of combining loosely with oxygen. 
 This indicates that the iron is in some way associated with the 
 peculiar respiratory functions of haemoglobin ; though it is obviously 
 an error to suppose, as was once supposed, that the change from 
 venous to arterial blood consists essentially in a change from a fer- 
 rous to a ferric salt. 
 
 As already stated, when administered as a medicine, haemo- 
 globin is split up by the gastric juice; its globin being converted into 
 a soluble peptone (which is absorbed) while its hsematin is set free. 
 Haematin is a remarkably stable substance, strongly resembling 
 iodine ; or, if powdered, appearing dark or light brown according to 
 the fineness of the powder. It may be heated to 180° without decompo- 
 sition ; but, by stronger heating, it is finally decomposed, liberating 
 an odor of hydrocyanic acid and leaving a residue (12.5 p. c.) of pure 
 oxide of iron. It is quite insoluble in water, alcohol, ether, chloro- 
 form or benzol ; but is somewhat soluble in strong acetic add, espec- 
 ially if warm. It is also soluble in alcohol, (not water) to which 
 some acid has been added ; and readily soluble in alkaline solutions, 
 or in alcohol containing alkalies. 8 
 
 Spectroscopic Assay. 
 
 As Oxyhemoglobin prepared from bullock's blood does not 
 crystallize readily, if at all, it appears on the market in the form of 
 a more or less scaly powder, resulting from the method of drying 
 it. The microscope, therefore, does not furnish a reliable method 
 for determining its purity. This must be ascertained by means of 
 the spectroscope, for a tolerably dilute solution is found to absorb 
 certain rays of light in a peculiar and characteristic manner. The 
 spectrum of oxyhemoglobin may be seen by referring to the frontis- 
 piece of this monograph. Here it will be observed that " 3 a portion 
 of the red end of the spectrum is absorbed, as is also a much larger 
 portion of the blue end; but what is most striking is the presence of 
 two strongly marked absorption bands, lying between the solar 
 
 ilbid. p. B68. 
 
 3 Text Book of Physiology by M. Foster, M.A., M.D., L.L.D., K.R.S. (Sixth 
 edition). London, Macmillan & Co., and New York, 1891. Part II., p. 561. 
 
lines D and E. Of these, the one towards the red side, sometimes 
 spoken of as the band (a), is the thinnest, (and, in extremely dilute 
 solutions, the only one visible) but the mo3t intense. Its middle lies 
 at some little distance to the blue side of D. Its position may be more 
 exactly denned by expressing it in wave-lengths. As is well known, 
 the rays of light which make up the spectrum differ in the length 
 of their rays, diminishing from the red end where the rays are 
 longest, to the blue end where they are shortest. Thus Frauen- 
 hofer's line D corresponds to rays having a wave-length of 589.4 
 millionths of a millimetre. Using the same unit, the centre of this 
 absorption band (a) of haemoglobin corresponds to the wave-length 
 578. The other, sometimes called 6, much broader, lies a little to 
 the red side of E, its blueward edge, even in moderately dilute solu- 
 tions, coming close up to that line. Its centre corresponds to about 
 wave-length 539. Each band is thickest in the middle, and gradu- 
 ally thins away at the edges. These two absorption bands are ex- 
 tremely characteristic of haemoglobin. Even in very dilute solu- 
 tions, both bands are visible ; in fact they may be seen in a thickness 
 of one cm. in a solution containing one grm. of haemoglobin in ten 
 litres of water. This is the case when scarcely any of the extreme 
 red end, and very little of the blue end, is cut off. They then ap- 
 pear not only faint but narrow. As the strength of the solution is 
 increased, the bands broaden, and become more intense; at the same 
 time both the red end, and still more the blue end, of the whole 
 spectrum, are encroached upon. This may go on until the two ab- 
 sorption bands become fused together into one broad band. The 
 only rays of light which then pass through the haemoglobin solution 
 are those in the green between the blueward edge of the united 
 bands and the general absorption which is now rapidly advancing 
 f rom the blue end, and those in the red between the united bands 
 and the general absorption at the red end. If the solution be still 
 further increased in strength, the interval on the blue side 
 of the united bands becomes absorbed also, so that the only rays 
 which pass through are the red rays lying to the red side of d; these 
 are the last to disappear, and hence the natural color of the solution 
 as seen by transmitted light. Exactly the same appearances are 
 seen when crystals of haemoglobin are examined with a microspec- 
 troscope. They are also seen when arterial blood itself (diluted with 
 saline solutions so that the corpuscles remain in as natural a condi- 
 tion as possible) is examined with the spectroscope, as well as when 
 a drop of blood, which from the necessary exposure to air is always 
 arterial, is examined with the microspectroscope. In fact, the 
 spectrum of haemoglobin is the spectrum of normal arterial blood. 
 
When oxyhemoglobin loses its oxygen and becomes reduced to 
 haemoglobin, either by submitting the solution to the air pump or 
 adding reducing agents, such as a few drops of ammonium sulphide 
 or of an alkaline solution of ferrous sulphate, kept from precipita- 
 tion by the presence of tartaric acid, or even if, to the solution of 
 oxyhemoglobin, be added an unpurified solution of blood corpuscles, 
 such as is afforded by the washings of a clot, for example, the oxy- 
 gen in loose combination is immediately seized upon by the reduc- 
 ing agent. The two absorption bands now disappear, and, in their 
 place, there is seen a single, much broader, but at the same time, 
 much fainter band whose middle occupies a position about midway 
 between the two absorption bands of the unreduced solution, though 
 the redward edge of the band shades away rather farther towards 
 the red than does the other edge towards the blue; its centre cor- 
 responds to about wave-length 555. At the same time the general 
 absorption of the spectrum is different from that of the unreduced 
 solution; Jess of the blue end is absorbed. Even when the solutions 
 become tolerably concentrated, many of the bluish-green rays to 
 the blue side of the single band still pass through. Hence the dif- 
 ference in color between haemoglobin which retains the loosely 
 combined oxygen, and haemoglobin which has lost its oxygen and 
 become reduced. In tolerably concentrated solutions, or tolerably 
 thick layers, the former lets through the red and the orange-yellow 
 rays ; the latter the red and the bluish-green rays. Accordingly, 
 the one appears scarlet, the other purple. In dilute solutions, or in 
 a thin layer, the reduced haemoglobin lets through so much of the 
 green rays that they preponderate over the red, and the resulting 
 impression is one of green. In the unreduced haemoglobin or oxy- 
 hemoglobin, the potent yellow which is blocked out in the reduced 
 haemoglobin, makes itself felt, so that a very thin layer of oxyhem- 
 oglobin, as in a single corpuscle seen under the microscope, appears 
 yellow rather than red. 
 
 Therapeutics. 
 
 Attracted by the remarkable results attained by the use of 
 defibrinated blood quaffed at the butcher's shambles, I took up an 
 investigation of this practice in 1879, and shortly 'afterwards intro- 
 duced Sanguis Bovinus Exsieoatus (dessicated blood). This was pre- 
 pared in the following manner: Fresh bullock's blood was defibrin- 
 ated in the ordinary way by stirring and then dried on iron plates 
 in the form of scales. If done properly, an odorless product with 
 an albuminous taste resulted. It dissolved readily in cold water 
 
 lANew Method of Rectal Alimentation by P. E. Stewart, M. D., Ph. G., 
 Medical Record, Jan. 3, 1880. 
 
forming a perfectly clear blood-red solution. After standing for 
 some time, however, the scales lost their ready solubility and could 
 be dissolved only with difficulty. Again, in the Medical Record, a 
 few months later, I recommended defibrinated Mood as a substitute 
 for extract of beef. In the Therapeutic Gazette for January 1884, 
 I again called attention to the use of this article for use as a thera- 
 peutic agent, under the caption "Haemoglobin." 
 
 While I was conducting the experiments in my New York 
 laboratory which led to the introduction of desiccated blood, Dr. 
 Andrew H. Smith of that city was experimenting with defibrinated 
 blood as a means of supplementary rectal alimentation at St. Luke's 
 Hospital. When I called Dr. Smith's attention to the desiccated 
 product, he immediately put it into use. During his experiments 
 with it he sent me the following formula on a postal card : 
 
 1$. Sanguinis Bovini Exsiccati 3 viij 
 
 Aquae § vj. 
 
 Spiritus Frumenti 
 
 Glycerine aa § j 
 
 Misce. Sig. Tablespoonf ul three or four times a day. 
 
 This formula has been variously modified by the different man- 
 ufacturing houses and we have in consequence several so-called 
 "fluid" foods. These have had a very large sale in spite of their 
 disagreeable nature. While open to many objections, they have 
 certainly done good work in a great many cases. 
 
 In 1890, I attempted to improve on these fluid preparations by 
 another modification of Dr. Smith's formula, using fresh 
 defibrinated bullock's blood in place of the dried article. 
 This I introduced under the name of haemoglobin compound; 
 the formula for the preparation containing three fluidounces each 
 of defibrinated blood and extract of malt, and an ounce each of 
 whiskey and glycerine. Since my introduction of this preparation, 
 Frederick Stearns & Co. have brought forward an oxyhaemoglobin to 
 which they have given the trade name "Hsemoferrum." 
 
 The dry preparations of haemoglobin on the market certainly 
 possess manifest advantages over those furnished in fluid form, al- 
 though they lack the stimulating properties of the latter. This ob- 
 jection, is, however, easily overcome by administering small quanti- 
 ties of some stimulant. 
 
 The question as to the power of the various tissues of the body to 
 absorb albuminous substances is one of great interest. Albumen, 
 being a colloidal substance, will not dialyze through animal mem- 
 branes outside the body, and it has, therefore, been urged that the 
 action of the gastric juice is necessary to fit such substances for 
 
 7 
 
absorption. The evidence against this theory, however, is strong. 
 Eggs and other albuminous substances are successfully employed 
 for rectal alimentatipn, and physiologists claim that the lower 
 bowel contains no digestive ferment. Dr. Campbell, of Georgia,, 
 answered this objection by saying that a retro-staltio action was 
 set up which carried the injected substance above the ileo-coecal 
 valve and that it was digested in the fluid of the small intestines. 
 This suggestion, however, has not been generally accepted by the 
 medical profession. 
 
 Some years ago, Dr. Andrew H. Smith, of New York, had under 
 his care in St. Luke's Hospital, a man in the last stage of pulmon- 
 ary phthsis, to whom an injection of 120 grammes of blood was 
 given every evening. The patient died just after an enema had 
 been administered. At the autopsy, the large intestine was found 
 to be very evenly coated with thickened blood for a distance of 
 nearly three feet. While this is an argument in favor of retograde 
 peristalsis, there is no proof that the injected material was carried 
 above the ileo-coecal valve. 
 
 A more striking argument in favor of the power possessed by 
 certain living tissues to absorb colloidal substances is furnished by 
 the experiments of Ponfick, who published in "L'Union Medicale" 
 in 1879 or '80 reports of his investigations in regard to intra-periton- 
 eal injection of defribrinated blood. He reports five cases of pro- 
 found anemia which were greatly improved by injection of from 15 
 to 20 ounces of defribrinated blood into the peritoneal cavity. 
 While we are all aware that extra vasated blood is absorbed in cases 
 of surgical operations, it has also been claimed that the surface of 
 the skin itself is capable of absorbing oils and fats applied by in- 
 unction. Cod liver oil is frequently used in this manner in the 
 treatment of debility where the stomach is in an enfeebled condition. 
 It is, however, doubtful whether the good effects should not be at- 
 tributed to the massage accompanying the inunction. 
 
 Bunge has pointed out that ' "We have been satisfied to account 
 for the absorption of food from the alimentary canal by the laws of 
 diffusion and osmosis, but we now know that, as regards osmosis, 
 the wall of the intestine does not act as would a dead membrane. 
 We know that the intestinal wall is covered with epithelium and 
 that every epithelial cell is in itself an organism, a living being with 
 the most complex functions. We know that it takes up food by the 
 active contraction of protoplasm in the same way as observed in 
 independent naked animal cells, such as amoebae and rhizopods. 
 Observations on the intestinal epithelium of cold-blooded animals 
 
 i Text Book of Physiological and Pathological Chemistry, p. 3. 
 
 8 
 
have made it obvious that the cells grasp the particles of fat con- 
 tained in the food by means of their protoplasmic processes ; and that 
 they incorporate the fat globules with the protoplasm of the cell, 
 which finally passes them on to the commencement of the chyle- 
 vessels. So long as this active intervention of cells was unknown, 
 it was impossible to understand the remarkable fact that, although 
 the minute drops of fat were able to pass through the intestinal 
 wall, yet finely divided pigments, intentionally introduced into the 
 intestine, remained quite unabsorbed. At the present time, we know 
 that all unicellular organisms possess the power of selecting their 
 food, of taking up the useful and rejecting the useless substances." 
 How much these facts may account for the apparent power of 
 absorption possessed by the mucous and serous membranes it 
 is impossible to say. 
 
 Foster, in his classical work on 'physiology, explains the 
 structure and functions of the body in a comprehensive manner, 
 using the simple organisms known to biologists as amoebae as a type 
 of the more complex organisms of the system. The amoebae are al- 
 most wholly composed of undifferentiated protoplasm in the midst 
 of which lies a nucleus (though this is sometimes absent). He calls 
 attention to the fact that this protoplasm possesses certain funda- 
 mental vital properties. It is contractile, irritable and automatic, 
 receptive and assimilative, metabolic and secretory ; also respiratory 
 and reproductive. The tissues of higher animals may be regarded 
 as groups of peculiarly associated amoebae. Their accompanying 
 physiological phenomena depend upon the combination of the 
 fundamental properties possessed by the original protoplasm. In 
 the evolution of living beings, it has finally come to pass that, in 
 higher animals and plants certain groups of the constituent amcebaed- 
 iform units or cells have, in company with the change in their 
 structure, been set apart for the manifestation of some of the funda- 
 mental properties of protoplasm, to the exclusion or at least to the 
 complete subordination of the other properties. While one group 
 of cells constituting the nervous tissue manifests the properties of 
 irritability and automatism, with an almost complete absence of 
 contractility and a great restriction of the other qualities, another 
 group is characterized by the activity of its protoplasm in the 
 secreting of digestive fluids, and in the promoting of the peculiar 
 chemical and physiological changes known as digestion and assimi- 
 lation. Is it not possible that other tissues in which these properties 
 are only latent may become stimulated temporarily? Admitting 
 
 i Text Book of Physiology by M. Foster, A. M., M. D. etc., Phila. Henry 
 C. Lee's Sons & Co. 1881. 
 
this possibility, can we not account for the absorption and assimi- 
 lation of food by mucous and serous membrane. 
 
 'Dr. Desiderio Varela, of Mexico, reports an interesting instance 
 of the efficacy of this agent in anaemia. The patient was a young 
 woman, 20 years of age, who had become excessively debilitated. 
 
 He says that every known preparation of iron and tonics had 
 been tried in vain, and at last the situation became such that it was 
 determined to adopt the procedure recommended in like cases by 
 Drs. F. E. Stewart and A. H. Smith, of New York, viz. : Enemata 
 of defribrinated blood. On the third day, after using four or five 
 ounces per diem in this manner, a favorable change was noticed, 
 and, about the fifth day, the stomach became tolerant of light foods, 
 and a favorable prognosis was ventured upon. On the twelfth day 
 all rectal alimentation ceased, and at the end of a month, she re- 
 turned to her household duties. 
 
 Dr. A. Ernest Sansom contributed a valuable article to the 
 Lancet (April, 1882), in which he recorded two very interesting 
 cases which rapidly improved after the injection of defribrinated 
 blood. One suffered with dilated stomach, intense anaemia and 
 purpura, together with severe pain and vomiting; the other was one 
 of probable renal calculi, though the diagnosis was less clear. 
 
 Dr. Thos. S. Butcher, in the Medical Bulletin for November, 
 1882, makes a strong plea in behalf of the administration of bul- 
 lock's blood as a remedy in ansemia. He prefers to use the fresh 
 blood (not defribrinated) as a drink immediately after the animal 
 has been slaughtered, without waiting for it to become cooL He 
 says that, while there is something repulsive to a delicate and fine 
 lady in the idea of drinking the warm, fresh blood from a dying 
 animal, yet these persons finally become accustomed to it, and do 
 not object to the taste of the fluid. The treatment is resorted to 
 three times per week ; the amount first administered being about 
 two fluidounces. This is gradually increased until a full glass is 
 drank each time. The only addition allowed is a small quantity of 
 salt, which makes it more palatable to some patients. Dr. Butcher 
 says that, where the directions have been carefully followed out 
 and the treatment persisted in for a sufficient length of time, the 
 average gain in weight has been fully one pound per week, and the 
 general results have been such as to warrant his urging upon the 
 profession this most effectual line of treatment. He enumerates 
 six cases of anaemia in which this method was employed; one patient 
 gaining 36 pounds in 36 weeks, another 12 pounds in 12 weeks, 
 
 l Beprinted from the El Genio Medico-Cmrurgico, Jan. 22, 1883, in La Es- 
 cuela de Medicina, Mexico, March 1882, and in London Medical Eecord, April 15, 
 1882. 
 
 10 
 
another 16 pounds in 16 weeks. Two others gained from 1^ to 1J£ 
 pounds per week during 14 or 15 weeks, and another about 5 pounds 
 a month. Some of these cases were far gone in the downward 
 course of their disease, but in all health was perfectly restored. 
 
 As the result of five years experience at the Royal Hospital for 
 Women and Children, Dr. E. O. Day reports favorably in regard to 
 the use of bullock's blood in the treatment of delicate children. His 
 method of preparing the blood is to prevent its coagulation by dilu- 
 tion with water; drying it by means of water bath and powdering 
 in a mortar. The dose for a child is a teaspoonf ul during the day. 
 
 Dr. Guerder, of Paris, France, has been very successful with 
 defribrinated blood in the treatment of various diseases. He gave 
 the powder to 8 convalescents, all of whom had been affected with 
 typhoid fever in a grava form. Three out of the eight were unable 
 to retain it, but the other five convalesced much more rapidly than 
 their companions. He reports equally good results in the treatment 
 of convalescents from adynamic fever, anaemia and other debilitated 
 conditions of the system. Of the 24 cases affected with chlorosis, 
 in which he gave this remedy, two vomited it frequently and di- 
 gested it with difficulty; two others, who were dyspeptic, suffered 
 oppression of the stomach in consequence and disagreeable eructa- 
 tions which caused them to discontinue its use. In the remaining 
 cases, the results were very satisfactory. One case of the number 
 that he has reported will serve as an illustration of his successful 
 application of this remedy. 
 
 "Case 3: — Mademoiselle Emilie M., aet. 17 years, in easy circum- 
 stances, is one of those chlorotic girls that are the despair of a physi- 
 cian. She has been treated by several physicians and by myself for 
 two years past. Tonics and chalybeates of all sorts, as well as fer- 
 ruginous mineral waters, have been successively administered with- 
 out great results. Menstruation regular but scanty. For some 
 time past the young girl, naturally tall and of stout build, was los-' 
 ing flesh very fast ; complexion sallow; lips pale; could not bear 
 fatigue. The least emotion, the slightest exertion, brought on vio- 
 lent palpitations of the heart. The appetite was quite gone, and 
 the sight of food provoked nausea. If forced to eat, she was taken 
 with vomiting immediately after; short dry cough, but nothing 
 about the lungs to excite apprehension. I prescribed the powder of 
 meat. The patient took it with some repugnance, and vomited it 
 at once. After trying the meat powder unsuccessfully for two 
 days (in spite of the willingness of the patient to submit to any 
 treatment), I was compelled to give it up. I proposed "gavage"' 
 (artificial feeding), but she would not consent to it. I then had re- 
 
 11 
 
course to the powder of blood. The treatment was commenced 
 October 6th, at first in the dose of 3, then 6 teaspoonfuls a day. 
 The first effect, singularly enough, after three or four days' treat- 
 ment, was to cause the nausea to disappear, and the patient began 
 to cat a little. At the end of a fortnight the gain was manifest, 
 the appetite was good and the digestion normal, the palpitations 
 were gone and she was able to walk out every day. The treatment 
 has been continued ever since, with occasional interruptions, and 
 the young girl is now quite well." 
 
 Dr. Guerder says that the therapeutic indications of this agent 
 are extremely numerous. In a general way it may be said that it 
 is always indicated when there exists impediment of the organism, 
 whatever its cause and nature. In simple anaemia, a better recon- 
 stituent cannot be found. The perfect tolerance for this product 
 manifested by delicate stomachs make it a precious resource. On 
 account of its iron, and the ease with which it can be borne, it is an 
 excellent remedy in chlorosis. It has been found of great value in 
 the treatment of dyspepsia and diseases of the alimentary canal; it 
 is a preventive and curative in the treatment of infantile diarrhoea; 
 it has shown itself of value as a tonic and reconstituent in pulmon- 
 ary phthisis, and, as Dr. Steibel says in his recent treatise, "In my 
 experience the powder of defibrinated blood rapidly stimulates the 
 appetite and the digestive functions. It advantageously replaces 
 iron, especially in the sufficiently numerous cases in which the lat- 
 ter is not well borne. With this action as a chalybeate is combined 
 another no less valuable. It is an excellent aliment, reduced to the 
 smallest volume possible, and of perfect digestibility. * * * 
 As a chalybeate, the blood powder presents the advantage of not 
 provoking constipation. In all debilitated states of the organism it 
 will prove a valuable therapeutic adjuvant and nutrient. 
 
 In my article on "Haemoglobin as a Therapeutic Agent" (Thera- 
 peutic Gazette, 1884. p. 55), I fell into the same error that other 
 writers on this subject have been guilty of. Referring to the most 
 important functions of the red corpuscles, viz., that of carrying 
 oxygen, I said: "But it is to the power of haemoglobin to hold a 
 large portion of oxygen in its embrace that this function of the red 
 corpuscles is due, and haemoglobin has this property to an equal ex- 
 tent wben separated from the corpuscles. Its administration, there- 
 fore, places in the blood an agent capable of restoring its lost func- 
 tion of carrying oxygen. Respiration is again established, and the 
 slumbering vital fires are again aroused into activity." The error 
 was, however, quite natural under the circumstances. Before ad- 
 vancing the theory, I consulted with several leading physiologists 
 
 12 
 
who supported my view. That is, we unanimously accepted the 
 theory as being reasonable, but the article referred to contains the 
 following admission: "I have not yet demonstrated my theory 
 with scientific exactness." I was particularly impressed even at 
 that time, by the fact that the effects of haemoglobin administration 
 were entirely out of proportion to what might be expected from the 
 amount given; i. e. if its virtues were to be attributed to the value 
 of blood as a nutrient alone. I had seen such remarkable results 
 from the administration of desiccated blood in 20 grain doses, three 
 times a day, that I began to suspect that it was sometimes absorbed 
 without being acted upon by the gastric juice, and that its effects 
 were due to its going directly into circulation. 
 
 The Medical Record (1884) says, "our ideas of providing 
 nourishment for patients with weak and irritable digestive 
 or ns have been somewhat revolutionized of late years by the re- 
 markable results obtained by Drs. Debove and Dujardin-Beaumetz 
 with their methods of forced feeding. They have clinically demon- 
 strated that proper nourishment plays no unimportant part in the 
 therapeutics of certain chronic and convalescent states." The 
 editor calls attention to the fact that indications for forced feeding 
 are presented frequently in practice ; much more frequently indeed, 
 than many will admit. He says that "the splendid results obtained 
 by S. "Weir Mitchell, and L. Playfair from systematic feeding, show 
 that its application is not limited to that class. Many cases of 
 hysteria, anaemia, chlorosis, and convalescence from acute diseases 
 and organic affections are accompanied by loss of appetite and even 
 disgust for food, due in great part to the fact that the stomach has 
 become so unaccustomed to the presence of food that it has par- 
 tially lost its digestive power, the best restorative of which is food 
 in small quantities frequently repeated." The Record then refers 
 to the various meat powders and to the excellent results obtained 
 by Guerder in the use of dried blood, suggesting the positive proba- 
 bility that the iron in the latter constitutes an important factor in 
 its usefulness ; its proportion of 0.30 part per hundred being suffi- 
 ciently large to represent additional doses of iron. De'bove and , 
 Dujardin-Beaumetz had indifferent results from the use of dried 
 blood. That Guerder attributes to the large quantities which they 
 administered and their faulty methods of preparing it. Indeed, 
 their results with his preparations were highly satisfactory. Guer- 
 der had administered it in 51 cases. Of this number 44 took it well 
 and without inconvenience for several weeks. The convalescents 
 from typhoid fever were unable to retain it at all, while to the re- 
 maining 4 chlorotics it was disagreeable, producing unpleasant sen- 
 
 13 
 
sations in the stomach and was sometimes vomited unchanged after 
 several hours. Guerder not only obtained excellent ' results in. 
 chlorosis, convalescence from acute diseases etc., but the effects up- 
 on phthisical patients were most remarkable. When the disease 
 was not far advanced, there was invariably an early increase of 
 strength, weight and appetite. 
 
 Oxyhemoglobin as a Pharmaceutical Product. 
 
 If the favorable results attending the administration of bullock's 
 blood can be properly attributed to oxyhaemoglobin we have, with- 
 in reach, a method of securing these results without the unpleasant 
 associations attending the use of the blood itself as a therapeutic 
 agent. By way of experiment, oxyhaemoglobin, ("Hasmoferrum" 
 brand) prepared in the manner already described, was administered 
 in three grain doses, twice in 24 hours to 72 persons, in order to deter- 
 mine whether or not it would produce unpleasant effects. This test, 
 which was prolonged for one week, demonstrated that such doses 
 will not, in the large majority of cases, disturb the stomach or bowels. 
 Furthermore, that, being neutral in reaction, it can be administered 
 indefinitely without injuring the teeth; that, being non-astringent, 
 it will not produce constipation, and that, unlike other preparations 
 of iron, it will not cause headache. It is true that the week's 
 experiment developed a few cases of slight gastric disturbance, 
 headache, diarrhoea, and constipation ; but these could not be fairly 
 attributed to the haemoglobin. It would be very remarkable to find 
 72 persons, among whom there were no cases of dyspepsia or con- 
 stipation. Twenty-three persons of the 72 reported that it increased 
 the appetite, and, of these, 18 slept better while taking the medicine. 
 As these people were all in comparatively good health and eating 
 three square meals a day, the effects were probably due to imagina- 
 tion. We all know that, if sugar and water were given to a hun- 
 dred patients with the statement that they might expect certain ef- 
 fects, a large portion would be confident that they experienced 
 these effects. On the other hand, what physician has not been ex- 
 asperated by the powerful imagination of patients, who ascribe 
 every unpleasant symptom to his medicines? As the general results 
 from these preliminary tests were favorable, the preparation was 
 sent to a number of physicians in Detroit for clinical test. Dr. Hal 
 C. Wyman tested it in the Detroit Emergency Hospital and reported 
 that in his hands the preparation proved a "splendid" tonic. Dr. 
 E. B. Smith reported that he was pleased with the good results ob- 
 tained in his private practice, and at the East Market Dispensary. 
 Dr. Oscar Le Seure used it in a number of cases of anaemia with 
 
 14 
 
satisfactory results. He reports "The preparation is well borne by 
 the stomach and improvement is rapid." Dr Dayton Parker used 
 it in the wards of Emergency Hospital with gratifying results, par- 
 ticularly so in a case of anaemia following ovariotomy. Dr. John 
 Lee reported equally good results in the outdoor clinic of St. Mary's 
 Hospital. Dr. P. M. Hickey reported the case of Miss D, aged 18, 
 who was referred to him for treatment Oct. 31, 1893 in a chlorotic 
 condition. Upon physical examination, an anaemic murmur was 
 discovered. She was placed upon oxyhemoglobin for some 
 weeks, at the end of which time the anaemic murmur disappeared, 
 and her condition was so improved that she reported herself well 
 and resumed her work. Dr. Hamilton E. Smith states that he used 
 the preparation with satisfactory results. Dr. W. E. Chittick, in 
 his experiments at St. Mary's Hospital, found oxyhaemoglobin to be 
 the best possible form in which to administer iron, and as valuable 
 in point of assimilation as any other form he had ever tried. His 
 experience demonstrated that it was acceptable to the most delicate 
 stomach. 
 
 The preparation was subsequently sent to a number of physi- 
 cians in different parts of the country in order to verify the obser- 
 vations made in Detroit. Dr. J. C. Martin, of Kansas City, Kas., 
 found it of value in the treatment of nervous exhaustion. Dr. J. M. 
 Shaffer, of Keokuk, la., employed it with satisfactory results in 
 cases where the usual iron preparations offended the patient. Dr. 
 P. G. Gober, of Alma, Wis., reports good results in anaemia and 
 recommends the preparation on account of its easy assimilation by 
 weak stomachs. 
 
 Among the many physicians who have tested the "Hsemofer- 
 rum' brand of oxyhemoglobin with equally good results may be 
 mentioned Drs. P. S. Stuart, Buffalo, N. Y., J. F. O'Keefe, Mt. Clem • 
 ens, Mich., L. B. "Weathers, San Antonio, Texas, Wm. A. Dixon, 
 Alpena, Mich., D. "W. Byers, Reading, Mass.,' J. "W. "Wiggenton, 
 Beebe, Ark., W. L. "Wilson, Midland, Mich., T. A. Pearson, Hope- 
 well, N. J., W. F. Reed, Cheboygan, Mich., T. C. Giroux, Brooklyn, 
 N. Y., Geo. W. Stewart, Saginaw, Mich., A. C. Tuchler, San Fran- 
 cisco, Cal., J. W. Hauxhurst, West Bay City, Mich., H. Schoenfield, 
 Trenton, O., M. C. Kelly, Mt. Clemens, Mich., E. K. Thompson, 
 McPherson, Kas., M. C. L. Kitchen, Saginaw, Mich., Frances Mam- 
 mell, McPherson, Kas., B. H. Beckwith, Saginaw, Mich., Lewis 
 Rochet, Fremont, O., C. H. Sample, Saginaw, Mich., E. S. Blair, 
 Correctionville, Iowa, C. D. Cope, Ionia, Mich., J. O. Chisholm, 
 New Concord, Ohio, and many others. The preparation has been 
 tested by several hundred physicians with equally good results. 
 
 15 
 
Occasionally untoward effects are reported ; for instance, in a small 
 proportion of cases, the oxyhemoglobin in pill form did not agree 
 with the stomach and could not be taken ; others again did not find 
 the rapid improvement from its use which has been claimed by a 
 great majority. 
 
 The bulk of the evidence is, however, entirely in favor of oxy- 
 hsemoglobin, because, while easy of assimilation, it is free from the 
 disagreeable effects usually attending the administration of inor- 
 ganic iron compounds. 
 
 For instance, Dr. Chas. E. McCallum, of Midland, Mich., makes 
 the following interesting report: 
 
 "Miss. L., Aged 17. Had never menstruated. Weight 85 pounds. Tall and 
 slender. Complexion almost transparent, with a greenish cast of countenance. 
 Had rheumatism of the knee joints. Both were considerably swollen, and al- 
 though she did not suffer any very severe pain, the joints were so much stiff- 
 ened as to make locomotion very difficult. Had considerable palpitation of the 
 heart and vertigo, also insomnia. Pulse very small and weak. 
 
 "Sept. 1st, 1894, was placed on oxyhemoglobin ("Hsemoferrum" brand). 
 Dose- One pilloid three times daily before meals and one at bed-time. 
 
 "Sept. 21st. Had gained 5 pounds. Appetite improved. Slept better. 
 Did not complain of knee joints so much. Swelling had almost disappeared! n 
 one. Color was slightly improved. Countenance brighter. Dose increased to 
 six pilloids per day. 
 
 "Oct. 30th. Had gained 12 pounds and was an entirely different looking 
 individual. She had also menstruated two days. Menstrual blood quite bright 
 in color. Knee joints almost entirely well and she was taking outdoor exercise 
 daily.' 
 
 Dr. Chas. W. Ellis, of Saginaw, Mich., reports as follows. 
 
 "B. P., female, sixteen years of age, has been suffering from chlorosis ac- 
 companied with irregular heart action, defective appetite and general feeling 
 of lassitude and despondency. She was placed upon oxyhemoglobin ("Haemo- 
 lerrum" brand) with a satisfactory result. This preparation has been fairly 
 tested in this case and I can cheerfully and heartily bear testimony to its ex- 
 cellence as a remedy when other forms of iron cannot be taken/ 1 
 
 Dr. W. J. O'Reilly, attending physician, St. Mary's Hospital, 
 Saginaw, Mich., reports: 
 
 "In September 1894, I was called to see Miss. M. B., who had spent nearly 
 the whole previous twelve months in bed under treatment by gynecologists 
 for uterine displacement and ovaritis. The constant retention of a pessary 
 had produced pelvic cellulitis and an abscess which had not been entirely 
 cured. 
 
 "I found her in an anaemic weakened condition and suffering from gastric 
 catarrh, obstinate constipation etc. She had been obliged to use a laxative 
 daily for twelve months and had been kept on a milk diet for several months 
 bj direction of previous medical attendants. 
 
 "The condition of her stomach was such that ordinary preparations of iron 
 bid fair to aggravate the gastric catarrh and constipation. I therefore pre- 
 scribed oxyhemoglobin ("Ha?moferrum" brand) and solid food. She at once 
 showed a marked improvement which had continued up to date (November 1st). . 
 She now walks from her home to my office for consultation. 
 
 16 
 
"A remarkable result attending' this treatmeut is that, since taking oxy- 
 hemoglobin, she has been able to dispense with laxatives or cathartics and her 
 anaemia has entirely disappeared." 
 
 It seems to be conclusively proven that all the good results 
 attending the employment of defibrinated blood are obtained when 
 oxyhemoglobin is administered in doses of from three to five grains, 
 three or four times a day. Oxyhemoglobin, therefore, may be con- 
 sidered as occupying a relation to defibrinated blood similar to that 
 which quinine bears to cinchona. Oxyhemoglobin may, pharma- 
 ceutically speaking, be termed the "active principle" of blood. 
 The use of this ' 'active principle" does away with the large amount 
 of inert material represented by the stroma of the corpuscles, serum, 
 and other constituents of blood, and also with the disagreeable 
 associations connected with blood administration. 
 
 The best form for administering oxyhsemoglobin seems to be 
 that of pills, coated with a soluble preparation of gelatin. I would 
 like to caution the profession, however, not to demand more from 
 oxyhemoglobin as a therapeutic agent in anaemia than can reason- 
 ably be expected of iron. It is folly to look for a specific for anae- 
 mia in any one agent, and iron is only one of the remedies to be used 
 in this pathological condition. The cause should be sought for 
 and removed when possible. The patient ought to be supplied with 
 nutritious food largely of meats. Outdoor air, hydro-therapy, 
 change of occupation and scene, improved hygienic conditions, etc. , 
 etc . should be employed together with iron in the treatment of 
 anaemia and other debilitated conditions of the system. The physi- 
 cian should not rely on iron alone, no matter how much the partic- 
 ular preparation thereof has to recommend it. The question is not, 
 "Is oxyhsemoglobin a specific in anaemia?" but "Is oxyhemoglobin 
 a better form of iron than the inorganic and organic preparations 
 usually employed?" The mass of evidence accumulated in the 
 manner hereinbefore indicated will, we believe, justify an affirma- 
 tive response to this query. 
 
 Albuminate of Iron. — Most compounds of iron possess a decided 
 astringency and respond to the characteristic test with ferricyanide 
 of potassium. The styptic property, together with the excessive 
 acidity of the most popular of these iron preparations, renders their 
 universal use impossible. Efforts have been made from time to time 
 to produce an iron preparation which would have all the therapeutic 
 properties of iron and still be free from its most objectional features. 
 
 Dieterich and Barthel 1 found that iron could be made to unite 
 with such organic substances as lactose, mannit, inulin, dextrose 
 
 1 Pharm. Rundschau. 1888, p. 39. 
 
 17 
 
gelatine, albumin, and peptone, and further that these compounds 
 would not respond to the usual iron tests. This reason, together with 
 the fact that they were not astringent, led these chemists to designate 
 these preparations and others of a like nature as "indifferent" iron 
 compounds. Of these, the albuminate and peptonate of iron have 
 become very popular in America, as well as in Europe where they 
 were originated. 
 
 The albuminates of different manufacturers have been found 
 to contain valuable percentages of iron, and some of these were not 
 even permanent preparations. Quite recently a new artificial album- 
 inate of iron has appeared, which is claimed to contain a constant 
 percentage of iron. While this is apparently true, this compound 
 possesses the same property as the ordinary albuminates in that it is 
 altered by the digestive juice into ferric and ferrous chloride. The 
 most constant and definite albuminate of iron is doubtless the natu- 
 ral compound of the blood (haemoglobin). Until recently it has not 
 been possible to obtain it pure on a large scale. The action of gas- 
 tric juice converts the iron into a compound called haematin, only a 
 small portion of ferric chloride being formed. 
 
 Peptonate of Iron. — By gradually adding a solution of chemi- 
 cally neutral ferric chloride to a solution of beef peptone, the liquid 
 will be seen to assume a dark red color, and finally a point will be 
 reached when no response to the well known iron test with potas- 
 sium ferricyanide can be obtained. Not until a mineral acid has 
 been added can this test be obtained. Since, with a definite strength 
 solution of a given sample of peptone, the combining proportions of 
 iron and peptone are constant, we are justified in saying that a 
 chemical compound and not a mere mixture has been produced. 
 As evidence of this, we may mention that this soluble compound is 
 entirely devoid of astringency, and does not corrode the teeth, as is 
 the case with ferric chloride and most other iron compounds. 
 
 Peptone in the free state dialyzes readily through animal mem- 
 branes, but when combined with iron does not. Free peptone re- 
 sponds to the well-known biuret reaction, but when combined with 
 iron it does not; yet the least excess beyond that required to satisfy 
 the affinity of iron immediately responds to the characteristic test. 
 These facts establish beyond doubt that the products of proteolytic 
 digestion unite chemically with iron. 
 
 Objection has quite recently been offered to the use of the term 
 "Peptonate of Iron" since it does not represent a simple, definite, 
 chemical substance. Strictly speaking, this expression would indi- 
 cate a compound formed by the union of iron and peptonic acid, 
 an acid which does not exist. Nevertheless the name "Peptonate 
 
 18 
 
of Iron" is sanctioned by common usuage among scientific men and 
 can therefore not be misleading. The absurdity of the terms "Pep- 
 tonized Iron" and "Peptonated Iron" which have been suggested as 
 more proper, becomes evident when they are associated with "Pep- 
 tonized" or "Peptonated Milk." Peptonization is a process of diges- 
 tion and no one would think of applying it to iron. Moreover, in 
 using the expression "Peptonate of Iron," we are only following the 
 example set by chemists the world over who invariably speak of the 
 compounds produced by adding iron, and mercury to egg albumen, 
 as albuminate of iron, and albuminate of murcury respectively. In- 
 stances are not wanting in the U. S. Pharmacopoeia to sustain this 
 view. 
 
 The composition of the peptonates, like that of the albuminates 
 is probably not constant, for the reason that the so-called peptone 
 of the market is a mixture of several products of proteid digestion. 
 Popularly speaking the name "peptone" is applied to all the pro- 
 ducts of the action of pepsin and hydrochloric acid on albumin. 
 But this digested proteid matter is a mixture of variable quantities 
 of parapeptone, albumoses, hemipeptone, etc., of which only the true 
 peptones are readily dialyzable. Probably each one of these pro- 
 ducts is capable of uniting with iron to form a definite compound. 
 But peptone (so-called) being, as has already been stated, a variable 
 mixture of all these proteid bodies, it can be readily seen why "Pep- 
 tonate of Iron" is not an absolutely definite chemical compound. 
 Doubtless if all conditions in the manufacture are equal, an appar- 
 ently constant "Peptonate of Iron" can be made. 
 
 Whatever may be said of the absorption and assimilation of 
 iron, "Peptonate of Iron" because of its peculiar properties, is quite 
 generally admitted to be an exceptional form of iron ingestion. Be- 
 ing neutral, it does not derange the digestive organs and is therefore 
 indicated where there is anaemia, chlorosis, or wherever prolonged 
 use of iron is necessary. 
 
Method of Counting Red Blood Corpuscles. 
 
 The average number of red corpuscles in human blood may be 
 probably put down at about 5 millions in a cubic millimetre (the 
 range in different mammals is said to be from 3 to 18 millions), but 
 the relation of corpuscles to plasma varies a good deal even in health, 
 and very much in disease. Obviously the relation may be affected (1) 
 by an increase or decrease of the plasma, (2) by an actual decrease or 
 increase of red corpuscles. Now, tho former must frequently take 
 place. The blood, as we have already urged, is always being acted 
 upon by changes in the tissues and, indeed, is an index of those 
 changes; hence the plasma must be continually changing, though 
 
 Human red blood corpuscles 1000 diameters, 
 always striving to return to the normal condition. Thus when a 
 large quantity of water is discharged by the kidney, the skin, or the 
 bowels, that water comes really from the blood, and the drain of 
 water must tend to diminish the bulk of the plasma, and so to in- 
 crease the relative number of red corpuscles, though the effect is 
 probably soon remedied by the passage of water from the tissues 
 into the blood. So again when a large quantity of water is drunk, 
 this passes into the blood and tends temporarily to dilute the plasma 
 (and so to diminish the relative number of red corpuscles), though 
 this condition is in turn soon remedied by the passage of the super- 
 fluous fluid to the tissues and excretory organs. The greater or 
 
HEMACYTOMETER OF GOWERS. 
 
 A, Pipette for measuring the diluting solution; B, capillary 
 
 tube for measuring blood; C, divisions on slide, 
 
 cover-glass and springs; D, vessel to mix 
 
 solutions; E, mixer; F, pointed 
 
 needle for sticking finger. 
 
less number of red corpuscles, then, in a given bulk of blood may 
 be simply due to more or less plasma, but we have reason to think 
 that the actual number of the corpuscles in the blood does vary from 
 time to time, This is especially seen in certain forms of disease 
 which may be spoken of under the general term of anaemia (there 
 being several kinds of anaemia), in which the number of red cor- 
 puscles is distinctly diminished. 
 
 The redness of blood is due to the presence of haemoglobin. 
 The degree of redness, may be influenced however, not only by the 
 number of red corpuscles in each cubic millimeter of blood, but also 
 by the amount of haemoglobin in each corpuscle, and to a less degree 
 by the size of the corpuscles. If we compare with a common 
 standard, the redness of two specimens of blood unequally red, and 
 then determine the relative number of corpuscles in each, we may 
 find that the less red specimen has as many corpuscles as the redder 
 
 Crystals of Haemoglobin from human blood. 
 
 one, or at least the deficiency in redness is greater than can 
 be accounted for by the paucity of red corpuscles. Obviously in 
 such a case the red corpuscles have too little haemoglobin. In some 
 cases of anaemia the deficiency of haemoglobin in each corpuscle is 
 more striking than the scantiness of red corpuscles. 
 
 The number of corpuscles in a specimen of blood is determined 
 by mixing a small but carefully measured quantity of the blood 
 with a large quantity of some indifferent fluid, — e. g. a 5 p. c. solu- 
 tion of sodium sulphate, — and then actually counting the corpuscles 
 in a known minimal bulk of the mixture. 
 
 This perhaps may be most conveniently done by means of las- 
 sez. A glass slide in a metal frame, is ruled into minute rectangles, 
 — e. g. \ mm. by J mm., — so as to give a convenient area of ^th of 
 
of a square mm. Three small screws in the frame permit a cover- 
 slip to be brought to a fixed distance,— e.g. £ mm. from the surface 
 of the slide. The blood having been diluted, — e.g. to 100 times its 
 volume, — a small quantity of the diluted (and thoroughly mixed) 
 blood, sufficient to occupy fully the space between the coverslip and 
 the glass slide when the former is brought to its proper position, is 
 placed on the slide, and the coverslip brought down. The volume 
 of diluted blood now lying over each of the rectangles will be f^th 
 (At x I) of a cubic mm. ; and if, when the corpuscles have subsided, 
 the number of corpuscles lying within a rectangle be counted, the 
 result will give the number of corpuscles previously distributed 
 through ^th of a cubic mm. of the diluted blood. This multiplied 
 by 100 will give the number of corpuscles in 1 cubic mm. of the 
 diluted blood and again multiplied by 100 the number in 1 cubic 
 mm. of the entire blood. It is advisable to count the number of 
 corpuscles in several of the rectangles, and to take the average. 
 For the convenience of counting, each rectangle is subdivided into 
 a number of very small squares, — e.g. into 20, — each with a side of 
 ■& mm., and so an area of 3-foth of a square mm. 
 
PART II. 
 
 The Absorption of Iron in the Animal 
 Body. 
 
 By A. B. MACALLUM, M. D., Ph. D. 
 
 Associate Professor of Physiology, University of Toronto. 
 
 Containing investigations as to the absorption of both 
 
 inorganic and organic iron compounds. 
 
 (Illustrated). 
 
 Taken from the Journal of Physiology, Vol. XVI, Nos. 3 and i, April 19, 1894, 
 Dy special permission of the editors. 
 
THE ABSORPTION OF IRON IN THE ANIMAL BODY. 
 
 By A. B. MACALLUM, M.D., Ph.D., 
 Associate-Professor of Physiology, University of Torontoi. (Plate XI.) 
 
 The chemico-physiological relations of iron have been the sub- 
 ject of much speculation during the last fifty years. The chief 
 difficulty in investigating the subject experimentally is the fact 
 that, when iron enters into the composition of organic structures, 
 it becomes "masked" and can no longer be detected by its ordinary 
 chemical reactions. 
 
 The question is one of great practical interest, and Bunge's 
 researches have in recent years done much to unsettle the medical 
 dogma that the iron contained in drugs enters directly into combi- 
 nation with the red corpuscles to form hEemoglobin, 
 
 In 1890, an investigation on the formation of blood corpuscles 
 in larval amphibia led me to conclude that haemoglobin is formed 
 from nuclein (the chromatin of histologists), and the iron-holding 
 character of other nucleins has been demonstrated by Bunge 2 , and 
 ZalesM 3 . The discovery of micro-chemical methods 4 for detecting 
 iron in cells has aided me in establishing the generalization that the 
 most important of all elements in the life of every cell is an iron- 
 holding compound. The prozymogens, if not the zymogens them- 
 selves, also contain iron. 
 
 I regard Bunge's theory of the direct conversion of iron-con- 
 taining nucleins in the food (hcematogen) into haemoglobin as 
 extremely doubtful, but the whole question of the synthesis of 
 organic iron compounds is in an uncertain state, chiefly owing to 
 our ignorance of the constitution of the nuclein molecule. There 
 are, however, certain allied questions which can be more readily 
 answered, and I have attempted in the following pages to determine 
 first whether or not inorganic compounds of iron are absorbed, and 
 secondly whether certain organic compounds of iron are absorbed. 
 
 iThe expense attending the investigation of the absorption of organic 
 compounds of iron (chromatins) was generously defrayed by a grant from the 
 Elizabeth Thompson Science Pund. 
 
 2Zett. /. Phyglol. Chem. Vol. IX., p. 49, 1885. 
 
 3 Ibid. Vol. X., p. 453, 1886. 
 
 « Macallum. Proc. Bay. Soc. Vol. L., 1891. 
 
 19 
 
The second part of my paper must however be regarded as a prelim- 
 inary communication, as my work in this direction is still incom- 
 plete. 
 
 I. ON THE ABSORPTION OF INORGANIC IRON 
 COMPOUNDS. 
 
 The literature of this subject is very abundant but it consists 
 largely of clinical records. The only papers, however, which I will 
 mention are those which embrace observations on the quantitative 
 estimation of iron ingested and excreted. In these the difference 
 between the amounts is relied on to show whether iron salts are 
 absorbed or not. The principal researches of this nature are those 
 of Kletzinsky 1 , Hamburger 2 , Gottlieb 3 , Kunkel 4 , Kumberg 5 , Busch 6 , 
 Marfori', Coppola 8 , and Stender 9 . 
 
 A careful perusal of these papers will convince the enquirer 
 that the methods adopted are not calculated to answer the question, 
 and that the various observers obtained different results. 
 
 In none of the series of experiments referred to, however, was 
 the micro-chemical method employed to any extent except in those 
 of Stender, although Kunkel's experiments on the livers of mice fed 
 with iron indicate how valuable such a method would be in results, 
 and it seemed to me that it would be easy to determine with it 
 whether there is absorption of iron salts, and if so, in what manner 
 and through what physiological agents it takes place. 
 
 Influenced by these views as to the value of the micro-chemical 
 method, I made a number of experiments with guinea-pigs, kittens, 
 lake-lizards and Amblystomata, using different preparations of iron 
 and administering them either with or without food in various 
 doses. At different intervals, after administering a preparation, or 
 during the course of feeding with the preparation, the animal was 
 killed, the abdomen opened, parts of the intestine, liver, spleen 
 and kidney were removed, put directly into 95 per cent, alcohol or, 
 after hardening for ten minutes in a saturated solution of corrosive 
 sublimate, into alcohols successively of 50, 70, and 95 per cent, 
 strengths. For control purposes teased-out portions of the fresh 
 mucous membrane of the intestine were treated with ammonium 
 sulphide and examined under the microscope. After the hardening 
 was completed in alcohol, sections were made of the various parts 
 
 i Zeitsch. d. GeseUsch. der Aerzte zu Wien, 1854, il. 281. 
 
 2 Zeitsch. f. Physiol. Chem. "Vol. II. 1878,p. 191. 
 
 3 Arch. f. Exp. Path, u. Pharm. Vol. XXVI. p. 139, 1889; Zeitsch. f. physiol. 
 Chem. Vol. XV. p. 371, 1891. 
 
 * Arch. f. d. gee. Physiol. Vol. L. p. 1, 1891. 
 
 5 and 6 Kobert. Arbeiten des Pharm. Inst, zu Dorpat. Vol. VII. 1891. 
 
 i Arch.f. exp. Pathol, u. Pharm. Vol. XXIX p. 212, 1891. 
 
 8 Rendiconti deUa R. Acad, dei JAncei. Vol. VI. p. 362, 1890. 
 
 9 Kobert. hoc. cit. 
 
 20 
 
either by the free hand (liver and kidney), or by the paraffin or eel- 
 loidin methods. Sections made by the paraffin method from tissues 
 hardened in corrosive sublimate were fixed on the cover slip by 
 Gaule's method, and the paraffin having been removed by the usual 
 process, they were immersed in a mixture of equal parts of solutions 
 of hydrochloric acid and potassic ferrocyanide of 0.5 and 1.5 per 
 cent, strengths respectively. Here they were allowed to lie for 
 about ten minutes, then they were washed in distilled water, dehy- 
 drated, cleared in cedar oil and mounted in benzole balsam. Sec- 
 tions made with the free hand, or with the celloidin method, were 
 either mounted on the slide in a mixture of glycerine and ammonium 
 sulphide and examined under the microscope, or placed v a ferro- 
 cyanide mixture for ten minutes, then carefully washed in distilled 
 water, dehydrated in alcohol and, after clearing in oil of cedar, 
 mounted in benzole balsam. Portions of the intestine hardened in 
 alcohol were also put in a mixture of alcohol and ammonium sul • 
 phide in the proportion of 5 to 1. Material hardened in corrosive 
 sublimate, on account of the presence of this reagent in the tissues, 
 gave no useful preparations with ammonium sulphide, but they fur- 
 nished, nevertheless, instructive ones when treated with the ferro- 
 cyanide mixture. In studying the preparations made with ammon- 
 ium sulphide, I teased out portions of the mucosa and mounted 
 them in a mixture of glycerine and ammonium sulphide. The gly- 
 cerine retards the evaporation of the sulphide, and such prep- 
 arations may be kept weeks and even months. In the case of 
 the liver, spleen and kidney teased-out preparations yielded nothing 
 of value, but sections of alcohol material, made sufficiently thin 
 with a little practice by the free hand, were useful when mounted 
 in the glycerine sulphide mixture. I have found that material 
 hardened in alcohol furnished the best preparations, and that it gave 
 results not equalled by those given by material fixed with corrosive 
 sublimate, or those obtained by the use of ammonium sulphide on 
 fresh material. The latter method is apt to lead to error, since the 
 living or non-hardened tissues are slowly penetrated by ammonium 
 sulphide, and when the penetration does take place, the cellular 
 elements are more or less altered, giving confusing results. Alcohol 
 has the advantage that it hardens rapidly and does not extract the 
 salts of iron which are in the tissues in an albuminate form. In 
 order to prevent diffusion of iron salts from without into the mu- 
 cosa of the intestine, the latter, laid open immediately after removal, 
 was in every case quickly washed free from adherent food matters 
 and then dropped into alcohol. 
 
 In guinea-pigs there is absorption of iron in the intestinal 
 
 21 
 
mucosa. This is readily seen in well-fed animals, less readily so in 
 those whose stomachs and intestines are almost empty. The 
 intestinal mucosa, after treatment with alcohol and when tested 
 with ammonium sulphide, acquires a more or less dark color, due 
 to the formation of sulphide of iron, which, under the microscope, 
 is seen to be limited to the sub-epithelial portions of the tips of the 
 villi. On closer examination the iron is found to be deposited in 
 leucocytes which, in their disposition, form together a cap, as it 
 were, for the extreme end of the lacteal vessel. The dark green re- 
 ■ action is not uniformly diffused through each cell, the nucleus be- 
 ing free from it, while, in addition to that present in the cytoplasm 
 as a whole, there are masses in it which yield a greater intensity of 
 color. The leucocytes are not as numerous immediately under the 
 membrane on which the epithelium rests. They may occasionally 
 be found between the epithelial cells of the tips of the villi; not 
 however, as much loaded with iron as those are which are found 
 about the end of the lacteal. The iron of these cells originates from 
 the food in great part, for, if the animal be kept without food for a 
 week, the tips of the villi give but a feeble reaction. What is pre- 
 sent in such preparations is derived from the bile, and this was 
 shown by the results obtained from feeding guinea-pigs with egg- 
 yolk. The latter, according to Bunge, contains but a trace of inor- 
 ganic iron — and when it is fed to the animal but a feeble reaction 
 for inorganic iron is obtainable in the villi. When, however, a 
 trace of ether is added to the yolk given, the amount of bile poured 
 into the intestine becomes greater, the absorption more vigorous, 
 and then one finds the tips of the villa give a marked iron reaction 
 almost as distinct as that present in the animal fed on its ordinary 
 diet. This iron must therefore be derived from the bile. In all 
 well-fed animals the iron reaction obtained is the more marked the 
 nearer the part examined is to the pyloric opening, and at a distance 
 of from seven to ten inches from it, it is usually absent altogether. 
 In ordinary guinea-pigs it is indeed very seldom that one finds 
 inorganic or albuminate iron in the epithelial cells themselves. In 
 order to meet the objection that possibly the iron in the lymph cells 
 is that in the process of excretion, and also to determine how the 
 iron compounds reach these elements, I fed several animals, which 
 had fasted for about four days, with various preparations of iron, 
 but using for this purpose specially some of the commercial "pep- 
 tonate" of iron of which about 100 — 200 mgrm. were administered 
 per os daily to each animal. The intestinal mucosa of animals so 
 fed for three days became black in the ammonium sulphide solution, 
 and on examination with the microscope the reaction, as before,. 
 
 22 
 
was almost confined to the tips of the villi, which were similarly 
 affected down to the distal end of the small intestine. It was fur- 
 ther found that the epithelial cells themselves covering the tips 
 were loaded with iron; the leucocytes were massed below in great 
 numbers and a large number had wandered between the epithelial 
 cells in such a way as, in many of the villi, to displace and distort 
 the cells. In very thin sections treated with the f errocyanide mix- 
 ture and mounted in balsam, the distribution of the iron was more 
 clearly seen. Sometimes, in the epithelial cells, the blue reaction 
 was a diffuse one with blue granules collected in groups here and 
 there in the cell. In some instances it was found in the inner end of 
 the cell chiefly, while again the protoplasmic processes in the hya- 
 line border gave an intense reaction. Fig. 4 shows some of these 
 details distinctly. In this are represented three cells, in two of 
 which the inner ends appear loaded with iron and they were fixed 
 in the act of transferring it to the underlying tissue. This involves 
 an internal secretion, a process that plays an extensive part in ab- 
 sorption. The iron compound appears to be secreted in a soluble 
 form, for I found that the underlying elements, the connective tis- 
 sue fibres, yielded frequently a deep homogeneous blue reaction 
 when the lower ends of the epithelial cells gave the same. When 
 the amount of. iron in the epithelial cell is small, that part of it in 
 the lower end is dissolved in the protoplasm, but when the amount 
 is large, it appears to be precipitated in a granular form. The dis- 
 solved form of the iron compound is possibly an albuminate, but 
 the character of the granular form is difficult to determine, and it 
 is equally difficult to do so in the matter of the deposit in the form 
 of granular masses in the leucocytes below, although on the ground 
 that some of these cells win their way with their iron back into the 
 blood vessels, it is allowable to believe that the iron-holding masses 
 in them, in order not to impede their movements, must be of al- 
 buminate composition and not inorganic with fixed shape. 
 
 The further course of the absorbed iron it is difficult to demon- 
 strate, unless iron salts are given for some time. It is easy, of course, 
 to determine that the leucocytes take up some of it. That fact is 
 the one the most prominently seen even when the iron is given in 
 large doses and for a long time, but it is only in the latter case that 
 one is able by micro-chemical means to show that another and 
 probably more important method of transfer exists. When the ex- 
 cised serosa of the upper part of the small intestine from a guinea- 
 pig, fed for some' time with either the phosphate, chloride or "pep- 
 tonate," was treated, after hardening in alcohol, with the acid 
 ferrocyanide mixture and after the usual course of treatment 
 
 23 
 
mounted in balsam, the venules appeared blue while the arterioles 
 were unaffected. In all these cases the blue color was found in both 
 the contents and the intima of the vessels, and where the venule 
 was empty, in the intima alone. In the contents there was a very- 
 light blue in the red corpuscles, a white corpuscle here and there 
 contained iron and the plasma was shown to carry it by a color 
 deeper than that given by the red discs. When the dose was very 
 great indeed the iron in the plasma took a granular form, at least in 
 alcohol preparations, but in this condition the muscular as well as 
 inner coat of the vessel was blue. These venules are radicles of the 
 portal vein. In the liver inorganic iron was found in all the tissues 
 of the peripheral zone of the lobules, in part in a granular form in 
 the peripheral hepatic cells, but diffused mainly through both cell 
 and nucleus of each element. When the dose of iron given was not 
 great, then the iron was mainly if not wholly confined to the peri- 
 pheral zone. With large doses a greater portion of the lobule was 
 impregnated with iron, and other elements came prominently to 
 view, especially in the "peptonate" preparations. There were leuc- 
 ocytes in the angles of the capillaries in all parts of the lobule but 
 very frequently in the central portion, and their occurrence was 
 manifested under the low power by the strong reaction which they 
 gave for iron (fig. 5). Sometimes each cell was a mass of blue 
 material, or it contained large blue masses. In others again, the cyto- 
 plasm had a diffuse blue tint with one or more clumps of iron-hold- 
 ing substance. Did these leucocytes come from the villi of the 
 intestine? Some of them undoubtedly must have done so, for, as 
 already stated, in the contents of the venules in the serosa of the 
 upper portion of the small intestine there were a few iron-holding 
 leucocytes. It is probable, however, from a difference in the 
 arrangement and deposition of the iron in them, that a large number 
 have taken up the iron from the plasma after they became entan- 
 gled in the capillaries. Some of them appear to be so large as to 
 occlude the capillary channels. What they do with the iron which 
 they contain is a matter of inference only. They probably transfer 
 it to the liver cells through the capillary wall, or, if they again 
 become free, they pass with it on to the general circulation. They 
 are apparently not much discommoded by the amount of iron which 
 they contain, for I found similar iron-holding leucocytes of all 
 shapes and sizes both in the spleen pulp and in the venules leading 
 off from it. 
 
 The iron-holding leucocytes of the villi, when they leave the 
 latter, probably pass into the transverse branches of the capillary 
 network or into the collecting venule of the villus. Outside of the 
 
 24 
 
■villi the occurrence of iron-holding leucocytes, except in vessels, is 
 extremely rare, although when excessive doses, as, for example, 
 0-5 grm. daily of ferric phosphate, were given, they were increased 
 in number, owing to the saturation of all the tissues of the intestinal 
 wall with iron. 
 
 Whatever iron salt was administered, whether the "peptonate," 
 phosphate, chloride or sulphate, when the dose was small, i.e. under 
 50 mgrms., the evidence of its absorption was veiy plain in the villi 
 of the upper end of the small intestine. "When the dose was larger, 
 as in the case of the phosphate and "peptonate," its presence was 
 observed in the villi far down in the intestine, but the reaction was 
 the less distinct the more remote the villus examined was from the 
 pylorus. When either of the two was given in very large doses, 
 (0-5 grm. daily of the phosphate for example), the villi near the 
 ccecum gave an intense reaction. 
 
 These results can be explained readily. The iron salt of the 
 chyme", when the latter is thoroughly mixed with the biliary and 
 pancreatic fluids, becomes wholly precipitated if the alkalinity of 
 the two latter fluids is sufficiently great. The alkali present may 
 be completely destroyed by a large quantity of iron salt in solution, 
 and when this occurs the excess of the iron salt not precipitated and 
 remaining in solution is absorbed. When the quantity in excess of 
 that necessary to destroy the alkalinity is very great, all the villi of 
 the intestine are in a position to absorb some of it. If on the other 
 hand the dose is small, absorption in the upper end of the intestine 
 is favored by the circumstance that the three fluids, chyme, bile, 
 and pancreatic juice, do not immediately and intimately mingle 
 and, therefore, the iron is not at once precipitated, some of it being 
 absorbed before that occurs. The quantity of acid in the chyme is 
 a factor of some importance, and when the iron given is in the salt 
 form and not the oxide, the acidity of the chyme is not decreased, 
 the acid of the salt displaced taking the place of the hydrochloric 
 acid. When the oxide or the reduced metal is administered, their 
 solution takes up a portion or all of the acid without contributing 
 in turn to the acidity of the chyme, and, therefore, in the intestine 
 the alkalinity of the bile and pancreatic juices goes farther in the 
 precipitation of the salts of iron in solution in the out-poured chyme. 
 The larger the amount of free acid in the latter, the greater must be 
 the quantity of iron absorbed. 
 
 I have made some experiments also upon the absorption of iron 
 in kittens. One of two from the same litter was given, through a 
 pipette by the mouth, 120 mgrms., in solution, of tartrate of iron 
 and ammonia and both were killed four hours after with chloro- 
 
 25 
 
form. The villi of the second kitten gave absolutely no reaction 
 for inorganic iron while in the other fed with the iron salt, the 
 sub-epithelial portion of the tips of the villi gave a marked one. In 
 both the amount of milk in the stomach was the same, the gastric 
 contents in one still holding apparently the greater part of the iron 
 administered. 
 
 It was noted in this and in other cases where the dose adminis- 
 tered was comparatively small, how free the epithelial cells them- 
 selves were from inorganic iron. Although in the sub-epithelial 
 portions of the villi in the kitten the amount of iron was large, the 
 epithelial cells when fixed did not contain the slightest trace of it. 
 The possible explanation is that the cells transfer with great rapid- 
 ity the iron which they absorb, and that it is only when the cells 
 are fatigued by overwork in this transference, that some of the iron 
 absorbed is seen in them. 
 
 I have already referred to the collection of the leucocytes in 
 the tips of the villi and to the invasion of the epithelial layer by 
 leucocytes in the "peptone" preparations. This is shown in Fig. 2, 
 representing an optical section of a villus treated with ammonium 
 sulphide and glycerine. The migration into the epithelial layer 
 does not occur in every villus, while in some exhibiting this appear- 
 ance it may be more marked than in others. The epithelial cells 
 were very often greatly affected by the invasion, for not only were 
 they considerably displaced, but they were in such a condition of 
 disintegration that in some stained sections the extreme tips of some 
 of the villi appeared denuded of epithelium. Whether the intra- 
 epithelial leucocytes were the cause of this disintegration or not, 
 they contained in addition to iron a great part of the disintegrated 
 material. An invasion of the epithelial layer by leucocytes was 
 also obtained in the villi when a quantity of albuminate of iron, 
 made according to Marf ori's 1 method, was given to a guinea-pig. 
 The animal had been fasting for three days when the ppwdered al- 
 buminate, suspended in water, was given and five hours afterwards 
 it was killed. As this compound is insoluble in weak acids, but 
 readily soluble in weak alkaline solutions, it was not surprising that 
 the villi from the opening of the pancreatic duct to the coecum 
 gave clear evidence of its absorption, while the villi for half an inch 
 beyond the pyloric valve contained no traces of it. The villi near 
 the distal end of the small intestine contained as much iron, judg- 
 ing by the reaction, as those which were situated near the opening 
 of the pancreatic duct. The number of iron-holding leucocytes was 
 not great, yet they carried a full complement of iron, and in many 
 
 l Arch. fur exper. Path, unci Pharm. Vol. X?XX., p. 212, 1891. 
 
 26 
 
of the villi a majority of them were intra-epithelial. Their presence 
 in the latter situation was not, as in the "peptone" preparations, ac- 
 companied by a disintegration of epithelial cells, although the lat- 
 ter were frequently much displaced. 
 
 These results seem to indicate that combinations of iron and 
 proteid influence in some way the activity of the leucocytes, and, in 
 order to obtain further evidence on this point, observations were 
 made on the lake-lizards (Neatus lateralis) in the laboratory. The 
 animals used had been for over thirty months without food, and 
 the intestinal cavities of two examined contained nothing more 
 than small masses of inspissated mucus impregnated with bile, but 
 the mucosa of the same gave preparations of which that represented 
 in Fig. 3 is typical. There was iron present in small quantities in 
 the outer ends of the epithelial cells, but it was in the leucocytes 
 that it was most abundant, the cytoplasm of these containing, in 
 addition to what was diffused through it, large granular masses im- 
 pregnated with iron. Leucocytes giving -such a reaction were 
 found scattered all through the mucosa. In the liver and spleen 
 iron-holding leucocytes were found, in the former in the angles of 
 the capillaries and in the latter in the adenoid tissue. To determine 
 if in these animals the leucocytes are affected as they are in guinea- 
 pigs by the albuminate of iron, a quantity of the dried albuminate 
 of iron was dissolved in a very weak solution of bicarbonate of soda. 
 and injected through the vent into the intestine of one. Eight 
 hours afterward the animal was killed, the liver, spleen and intestine 
 removed and put into alcohol. The latter organ was only partially 
 opened. In the liver and spleen the amount of iron in the leucocy- 
 tes was greatly increased and the liver cells gave a deeper diffuse- 
 reaction. In the sections from the intestine the leucocytes wero 
 found loaded with an excess of iron, so much so that the ammonium 
 sulphide gave them the appearance of huge collections of greenish 
 black granules. In some parts of the intestine they were collected 
 in large numbers under the epithelium with a few situated in the 
 latter layer, but in other parts, especially near the vent, where 
 naturally the bulk of the injected fluid collected, they were present 
 in very large numbers in the epithelial layer, many of them fixed 
 in all shapes and while migrating with their load of iron compound 
 into the cavity. The best preparations which I obtained, however, 
 were those from the unopened portions of the intestine, for here the 
 contents were retained in the sections. A very large number of 
 leucocytes excessively charged with iron were found in the cavity 
 and others were fixed in the act of passing into it, while in the sub- 
 epithelial leucocytes the iron was abundant. In all the preparations 
 
 27 
 
the epithelial cells themselves -were comparatively free from the iron 
 compound. 
 
 In guinea-pigs fed with the chloride, phosphate, or sulphate of 
 iron, I have not found that the leucocytes were similarly affected, 
 even when these preparations were given without food, nor does the 
 iron present in the ordinary food of the animals produce a like effect, 
 although in all these cases the leucocytes gave unmistakable 
 evidence of the absorption of these compounds. 
 
 It follows from the results of those experiments that inorganic 
 and albuminate compounds of iron are absorbed, at least in the por- 
 tions of the intestine near the pylorus, and in all parts of the small 
 intestine when the iron compound is not precipitable on m i xture 
 with the bile and the pancreatic juice. Undoubtedly also, sulphides 
 in the bowel must remove from solution a quantity of iron propor- 
 tionate to their abundance. In ordinary diet the extent of the 
 mucosa which absorbs iron must be, in proportion to that which 
 does not, remarkably small. In may possibly be that in the human 
 subject when the iron is specially increased the extent of the absorb- 
 ing surface is increased, the more so when there is a diminution in 
 the amount of pancreatic and biliary fluids, a condition possible in 
 anaemias. The view held by some that iron salts are not absorbed 
 and that these exercise their effect by stimulating the mucosa to 
 greater physiological activity, is opposed by the results of these 
 experiments. If iron salts stimulate, it is because they penetrate 
 the epithelium of the mucosa and in doing so are transferred to the 
 underlying elements. In other words, they are absorbed. But the 
 extent of the absorbing area is limited; so likewise must be the 
 extent of the area supposedly stimulated and, therefore, the bene- 
 ficial effects of the stimulation of the mucosa alone must be small. 
 What then is the purpose served by the absorption of iron salts? 
 Leaving out of consideration the possible answer that the iron of 
 combinations becomes assimilated, that is, is united in the animal 
 cell with other constituents to form what the histologist calls 
 chromatin, we may discuss some explanations of its effects. The 
 experiments with the feeding of "peptonate'' and of albuminate of 
 iron are, of course, too few to enable one to infer anything concern- 
 ing the action of iron-holding proteids of this character but all the 
 experiments without exception indicate that the leucocytes have a 
 special affinity for inorganic and albuminate compounds of iron, 
 and it is not too much to infer that this affinity involves a stimulating 
 or chemiotactic effect upon the leucocytes, and that iron salts exer- 
 cise an effect on other cells corresponding to their function. 
 
 The number of leucocytes which are engaged at any one time 
 
 28 
 
in the absorption of iron is comparatively small, owing to the small 
 extent of the intestinal mucosa bathed by a solution of an iron salt, 
 and, therefore, the stimulating effect would appear to be small, but 
 it must be remembered that, with the constant stream of iron-hold- 
 ing plasma and iron-holding leucocytes from the villi in the upper 
 portion of the small intestine to the liver, spleen, etc. , the other 
 cells of the body, including leucocytes with little or no inorganic 
 iron, are put in position to obtain some of that absorbed. There is 
 also another way in which the question may be viewed. Inorganic 
 compounds of iron, like those of calcium, potassium, and sodium, 
 have been, since the dawn of animal life on the globe, constituents 
 of its media, and of its food, and it is possible that the animal cell 
 has, in acquiring a tolerance for them, accommodated its functions to 
 their presence and has established with them a physiological equili- 
 brium which it may be possible to maintain in the absence of such 
 compounds. The view that the iron of inorganic combinations goes 
 directly into combination with nuclein and albumin to form, 
 chromatin, is one that may in the future be proved correct, but 
 whether it will happen so or not, it is not now, nor apparently will 
 it be then, incompatible with the explanations of other possible 
 functions of iron salts like those just referred to. 
 
 In investigating the absorption of iron I had opportunities for 
 determining the mode of excretion, when the iron absorbed is in 
 excess of the needs of the organism. The material illustrating this 
 was obtained from two guinea-pigs, one of which was given a very 
 large quantity of the "peptonate" preparation of iron, while the 
 other received a correspondingly large dose of ferrous sulphate. 
 The secretion of the Lieberkuhnian glands of the animal fed with 
 the ' 'peptonate" gave an intense iron reaction, and this was the case 
 not only with the small intestine but also with the coecum and 
 upper portion of the colon. In the superficial epithelium of the two 
 latter parts there was no evidence of either absorption or secretion 
 of iron compounds. In the cells of the Lieberkuhnian glands no 
 iron reaction could be obtained, nor was it obtained in the same 
 structures in the preparations from the animal fed with ferrous sul- 
 phate, although in the latter case the secreted material in the lumina 
 of the glands appeared to consist, in large part, of an iron compound. 
 Possibly the explanation for this is that the cells rapidly transfer 
 with the secretion the iron compound to the lumen and in excess- 
 ively small quantities at any one moment. The kidneys in both 
 animals gave a very slight diffuse reaction for iron not confined to 
 any part of the organs. It was only in the animal fed with ferrous 
 sulphate that the liver yielded more than ordinary evidence of the 
 
 29 
 
excretion of iron. The periphery of the lobules gave an intense iron 
 reaction partly diffused through the hepatic cells, and partly local- 
 ized in granules situated in that part of each cell bordering on the 
 bile capillary. The latter gave a feeble reaction for iron, especially 
 at the periphery of the lobules. The epithelium of some of the bile 
 duclets contained free iron, probably absorbed from the bile. 
 
 In man the liver and kidneys are the most active organs in the 
 excretion of iron. This has been shown by the observations of 
 Hunter 1 , Mott 2 , and others in cases of pernicious anaemia. In the 
 liver and kidney from a patient who had died from a complication 
 of troubles in which pernicious anaemia was supposed by some to be 
 a factor, I found this excretion illustrated to a remarkable degree. 
 The bile capillaries were, in some parts of the sections, filled and 
 distended with secreted iron compounds. Where this was the case 
 the cells surrounding a capillaiy were almost free from jnorganic 
 iron compounds, but where the capillary was feebly or not at all 
 injected, then the portion of each cell touching the capillary was 
 loaded with granules of a ferric compound. The sections of the 
 cortical portions of the kidneys gave an exceedingly intense reaction 
 for iron, which, on examination under the microscope, was found 
 chiefly in the convoluted tubules, the cells of these exhibiting both 
 a diffused and a localized reaction, the latter given by an abundant 
 collection of granules distributed in that half of the cell adjacent 
 to the lumen. The intestine in this case yielded no evidence what- 
 ever of the excretion of iron. 
 
 The results of experiments on guinea-pigs and of observations 
 on cases of pernicious anasmia show that, in different animals, the 
 organ for the excretion of iron may not be the same when this is 
 greatly in excess. This fact has, to a certain extent, been illustrated 
 also in a rutting bitch, in the intestine, liver and kidney of which I 
 could find no evidence of excretion of iron; but sections of its 
 uterus, after being hardened in alcohol, gave in a few seconds a 
 marked reaction for iron confined to the mucosa. Closer examina- 
 tion of these resulted in showing that the iron was deposited in three 
 situations : in leucocytes scattered in large numbers throughout the 
 inter-glandular elements, in the cells of the long convoluted glands, 
 and in the secretion found in the lumina of these. The leucocytes 
 were very much enlarged, the enlargement apparently being due to 
 the brownish masses of iron which they contained. The iron 
 appeared to be in the phosphate form. The cytoplasm gave a reac- 
 tion also for iron. In the gland cells the iron wpj3 both diffused 
 
 i The Practmmer, Vol. XLIII. 1889. 
 a Wd. Vol. XLV. 1890. 
 
 30 
 
through, the cytoplasm and localized in the form of a row of granules 
 immediately adjacent to the lumen of the gland. The diffuse reac- 
 tion obtained was slight, but that obtained with the granules was an 
 intense one, and the arrangement of the granules seemed to suggest 
 that they were caused by precipitation, in an inert portion of the 
 cell, of iron which had been in solution in the more active parts of 
 the cytoplasm. The substance in the lumen of each gland gave a 
 weak, but still distinct reaction for iron. Whether the leucocytes 
 in the inter-glandular tissue carried the iron to the mucosa from 
 other parts, or obtained it by absorption from the lymph bathing 
 them, it was not possible to determine definitely. I could find no 
 leucocytes in the blood vessels which contained inorganic iron com- 
 pounds, and this, considered in connection with the large size of the 
 iron masses in the inter-glandular cells, suggests that the latter 
 derived their iron from the lymph of the inter-glandular tissue. In 
 this case the lymph must have been also the source of the iron in 
 the gland cells. 
 
 Why different organs in different animals should serve for the 
 excretion of a great excess of iron, it is difficult to say. Possibly 
 this depends on differences in the degree of activity which the 
 organs present in different animals. It is quite as difficult to say- 
 why the kidney in some cases of pernicious anaemia and not in 
 others should give such marked evidence, in the convoluted tubules, 
 of the excretion of iron. 
 
 II. THE ABSORPTION OF ORGANIC IRON 
 COMPOUNDS. 
 
 The expression "organic iron compounds" includes two chief 
 classes of substances : — 
 
 1. The assimilated compounds of iron; that is, nuclein and re- 
 lated compounds, including Bunge's haematogen. These may be 
 conveniently termed the "chromatins." 
 
 2. Compounds produced by a degenerative process from the 
 first class. This includes haemoglobin, melanin, and lardacein. 
 
 I have limited my investigation to representatives of the 
 chromatin class. 
 
 Bunge believes that haematogens are formed only in vegetable 
 life, and constitute the only compounds of iron which are absorbed 
 by animals. 
 
 Feeding experiments with these substances have been made by 
 Socin 1 and Busch 2 . In one experiment Socin found the amount of 
 ingested iron exceeded that in the excreta. In the other two experi- 
 
 1 ZettscKf. Physiol. Chem. Vol. XV., p. 93, 1891. 
 
 2 Robert. XjOC. cit. 
 
 31 
 
ments the reverse was the case. These experiments were made on 
 dogs. "With mice he found that iron-free food mixed with organic 
 or inorganic iron compounds did not sustain life any longer than 
 food free from iron, whereas control animals fed on a more natural 
 food (coagulated egg-yolk) lived healthily. Busch from analyses 
 of the iron in the urine after feeding on hsematogen, haemoglobin 
 and hsematin, concludes that of these hsematogen is least absorbable. 
 Bokay 1 found that nuclein is not digested by artificial pancreatic 
 juice, that it leaves the body chiefly in the faeces, and causes but a 
 slight increase in the urinary phosphates 4 . 
 
 I began my own observation by feeding lake-lizards (Necturus 
 lateralis) on chromatin isolated by artificial gastric digestion from 
 lambs' testicles. The lizards were killed, and the intestines investi- 
 gated by the micro-chemical methods already described. The re- 
 sults were negative. I then fed mice on chromatins mixed with 
 starch and lard ; they died in from six to fifteen days with symptoms 
 of diarrhoea. They also died in a few days when the chromatin 
 was mixed with coagulated egg-yolk, but thrived well on the egg- 
 yolk without such admixture. I also gave to a lake-lizard the fluid 
 from an artificial pancreatic digestion of nuclein; this fluid, how 
 ever, contained little if any nuclein in solution, and micro-chemical 
 investigation of the lizard's intestine gave negative results. 
 
 The failure of all these experiments led me to use a less abnor- 
 mal kind of food; and, since, according to Miescher 3 , egg-yolk itself 
 contains 1 to 1*5 per cent, of nuclein (haematogen), that food sub- 
 stance appeared likely to yield the best results. 
 
 I used unboiled egg-yolk, for when egg-yolk is hard-boiled the 
 yolk spherules become thereby fixed in form, and the chromatin- 
 holding particles are set free only when the spherules are digested, 
 but when the yolk is administered fresh the spherules readily under- 
 go fragmentation and the chromatin-holding particles are liberated 
 and put in a form in which the epithelial cells, if they possess the 
 power, can invaginate them. In the spherules the chromatin is 
 partly in a granular form 4 and, apparently, partly as envelope 
 material to its fat globules, the latter varying in size and shading 
 
 1 Zeitsch. f. Physiol. Chem. Vol. I., p. 157, 1887-8. 
 
 2 In Gumllch's experiments, a description of which was recently published 
 [Ze.it. filr Physiol. Chemie, "Vol. XVIII., parts 6 and 6, p. 508, 1894), the absorption 
 of nucleinic acid in the dog appeared to be shown by the increase in the amount 
 of phosphorus excreted in the urine after the administration of a quantity of 
 compound.— P. M. Popoff (ibid. p. 533) found that nuclein compounds enter into 
 solution only in very small quantities in gastric digestion but in greater quan- 
 tities as the result of the action of artificial pancreatic juice, and ne points out 
 that the occurrence of such compounds in a dissolved form is aE that is neces- 
 sary for their absorption in the intestinal tract. 
 
 3 Miescher. Hoppe-Seyler's med. chem. Unters. Pt. 4, p. 502, 1871. 
 
 i Miescher. Qoc. cit.j localized the nuclein which he discovered in egg-yolk 
 in the granules of the yellow yolk spherules. 
 
 33 
 
off into the small granules in such a way as to suggest that the lat- 
 ter are also fat globules of almost infinitesimal size surrounded by 
 chromatin. Fig. 14 gives a representation of two yolk spherules 
 ■which were fixed by heat and in which the iron, set free by sul- 
 phuric acid alcohol, was converted by ammonium sulphide into the 
 sulphide. In it can be seen smaller and larger fat globules sur- 
 rounded by an iron-holding envelope. The fat is, therefore, closely 
 associated with the chromatin, and as we know the former is in 
 some way absorbed by the intestinal epithelium, the conclusion did. 
 not appear to be a strained one that both constituents are absorbed 
 together. 
 
 The first experiments ■were made with mice. The animals were 
 isolated and fed for periods ranging from five to fifteen days with 
 fresh egg-yolk and preparations of the small intestine were made in 
 various ways to determine the effect. Owing to the liquid or semi- 
 liquid form ot the food the animals partook but lightly of it, and 
 consequently there were found in the intestinal epithelium no very 
 marked indications of absorption, even of fat. In a series of prep- 
 arations from one ot the animals ■which was allowed to live for the 
 longer period, I found that, in the internal or lower half of each 
 epithelial cell, the protoplasm possessed a greater capacity for stain- 
 ing matters like eosin than is ordinarily exercised by the same cells 
 and, further, that this same protoplasm was denser and more finely 
 granular. Owing to the smallness of the sections, manipulation 
 with acid alcohols to set free the masked iron 1 to determine if this 
 staining capacity was due to chromatin, did not meet with much 
 success, although the teased-out cells after hardening in alcohol and 
 long treatment with warm ammonium hydrogen sulphide and 
 glycerine gave results whicn seemed to support that view. These 
 experiments, however, indicated the direction in which my succeed- 
 ing observations were to be conducted. 
 
 A large number of guinea-pigs were then fed for different 
 periods with undiluted yolk in excess. Each animal was given 
 about 10 c.c., three times a day, administered by the mouth by 
 means of a glass pipette, and when it was killed the small intestine 
 ■was removed and hardened, one part in alcohol and another in cor- 
 rosive sublimate. The sections from the sublimate preparations 
 made with paraffin and fixed on the cover glass by Gaule's method, 
 were either stained with hsematoxylin and eosin, or put through 
 nitric acid alcohol for 8 — 10 hours at 35" C. and then treated with 
 
 I The method here referred to will be described at full length, In a forth- 
 coming paper on the distribution of assimilated iron in animal and vegetable 
 cells. 
 
 33 
 
the acid ferrocyanide mixture to determine the distribution of 
 "masked" iron. The alcohol material was also treated in the same 
 ■way, with this exception, that the sections were fastened to the 
 cover slip with collodion before the paraffin was removed, and when 
 the sections were made by the celloidin process they were passed 
 directly through nitric or sulphuric acid alcohol. The results of 
 these experiments and methods were very interesting. 
 
 In the guinea-pig, as ordinarily fed, the "masked" iron exhibits, 
 in its distribution in the epithelium of the intestine very little differ- 
 ence from that represented in Fig. 6, in which the iron is shown in 
 the chromatin of the nuclei and in a narrow zone immediately about 
 some of the nuclei, but in preparations from animals fed with yolk 
 for two or three days, the epithelial cells situated on the sides of the 
 villi and below the tips of the same have the iron distributed as 
 represented in Fig. 7, in which the whole of the protoplasm in the 
 lower half of each cell and in the leucocytes below give a uniformly 
 diffuse Prussian blue reaction. The epithelial cells at the tips of 
 the villi are so much distorted by the fat present in them, that a 
 division of each into an internal and external part is impossible, 
 except in some cases where the absorption of fat has ceased to take 
 place. In preparations stained with haematoxylin and eosin, the 
 cells immediately below the tip give usually the appearance repre- 
 sented in Fig. 8, but with this exception, that the bodies enclosed in 
 cavities of the protoplasm in the external half of each cell shown in 
 the figure are not present in the preparations from all the animals 
 fed with egg-yolk. The internal half of each cell is loaded with 
 finely granular, eosinophilous material in which vacuoles are dis- 
 cernible, the outer half having its protoplasm arranged in a coarse 
 meshwork, the cavities of which were probably occupied by fat 
 globules. The nuclei in the majority of these cells contain an eosin- 
 ophilous substance filling the spaces left by the chromatin network. 
 In some villi the absorption of fat had occurred to such an extent 
 that the epithelium was separated from the underlying tissue, as 
 represented in the figure, and in these cases one sees sometimes a 
 condition which explains the destiny of the finely granular, eosin- 
 ophilous material in the inner ends of the cells. The corrosive sub- 
 limate has coagulated a proteid material extending from each cell 
 and containing some eosinophilous granules. This condition I 
 regard as due to internal secretion on the part of the epithelial cells, 
 and it has its parallel in the same epithelial cells when they are 
 transferring iron salts from themselves to the internal elements, a 
 process which I have already described. There is a difference, how- 
 ever, in the two in one respect, a difference which may only be 
 
 84 
 
apparent; in the internal secretion of iron, the latter appears to be- 
 come dissolved before passing from the epithelial cell internally. 
 In the yolk preparations, the eosinophilous material is in a granular 
 form, and could one be certain that it was not due to irregular pre- 
 cipitation by the corrosive sublimate, it would be an indication that 
 the epithelial cell secretes internally matters in a solid or semi-solid 
 form. The irregular form taken by the secreted matter may be due 
 to conditions governing the diffusion of the hardening reagent, but, 
 from the way in which each shred of the hardened proteid secretion 
 Is arranged in its attachment to an epithelial cell, one is disposed . 
 to regard it as fixed in the condition which it occupied in the living 
 tissue. Whether this secretion contains ' 'masked" iron chromatin 
 I cannot say. 
 
 Now, as in the secreting cells of all sorts, and especially in those 
 of the Lieberktlhnian glands, secretory activity is associated with 
 the presence of a chromatin in the part of the cell remote from the 
 lumen (Fig. 9), it might be urged that the increase of the "masked" 
 iron in the inner ends of the superficial epithelial cells of a villus 
 was due not to an iron compound absorbed, but to secretory activity 
 bringing about an increase of the substance governing that process. 
 This objection has some force, for so constantly is the presence of a 
 chromatin (prozymogen) connected with the processes of secretion, 
 especially in those glands which furnish a ferment, that one hesi- 
 tates to deny the existence of a prozymogen in another phase of cell 
 activity but little, if at all, different from that of secretion. The 
 only evidence on which one can rely to show that some of the chro- 
 matin of the yolk was absorbed in this case is, that the iron reaction 
 of the liver from an animal fed for some time with yolk is of a dif- 
 ferent character from that given by the liver of the animal after its 
 ordinary diet. In two guinea-pigs, one fed for seven days, the other 
 for six, when it died in a comatose condition, there was undoubted 
 evidence of the absorption of the chromatin, for, in sections of the 
 livers hardened in alcohol, the application of warm ammonium 
 hydrogen sulphide for some hours gave a reaction which was most 
 marked at the periphery of each lobule, but present also in the cen- 
 tral zone of the same. The reaction in the central portion was more 
 difficult to get and was best obtained when the sections were very 
 thin. In these preparations one can determine that the iron reac- 
 tion gradually becomes less intense as one follows the tissues of the 
 lobules from the periphery to the centre. In the liver of the ani- 
 mal after ordinary diet the reaction with ammonium hydrogen sul- 
 phide is obtained only in the outer zone and faintly in the outer 
 portion of the middle zone. In these, further, iron in a coarsely 
 
 35 
 
granular form is almost always found in the cells at the periphery 
 of the lobule, but such granules are absent in preparations from the 
 animals fed with yolk for some time. In the latter the iron reac- 
 tion in the liver cell is a diffuse one and it is present in the leucocy- 
 tes of the capillaries as well. What seems to show that this reac- 
 tion is not due to inorganic or albuminate iron, but to that in the- 
 organic form, is the length of time required to bring it out. These 
 results have been obtained also in experiments on Amblystomata. 
 In Amblystomata the cellular elements, although not of the size 
 of those in Necturus, are very large, and as it was possible to obtain 
 these animals in numbers sufficient for experients in this line, I 
 used them for this purpose. The yolk was given through a glass 
 pipette, introduced through the mouth into the stomach, and in 
 quantities corresponding to the size of the animal. Owing to the 
 ease with which they may be fed, and because of the retention on 
 the part of the stomach of what is so given, these animals are very 
 suitable objects for experiments of this sort. Now, in those recent- 
 ly captured or kept in captivity for a week or so without being fed, 
 there is comparatively little inorganic iron in the liver, no more, in 
 fact,, than there is in the liver of Necturus after long captivity, and 
 in sections of the liver of such, when hardened in alcohol and 
 treated with nitric acid alcohol, or sulphuric acid alcohol, to set 
 free the "masked" iron, the latter is wholly confined to the nuclei 
 of the cells. When, however, yolk is given daily in such quantities 
 as are retained to these animals for a period of four days or more, 
 the liver cells, which are loaded with fat, present a different reac- 
 tion. When hardened in alcohol, sections of the livers give, after 
 treatment with sulphuric acid alcohol or nitric acid alcohol for 
 sixty hours, a reaction for iron like that shown in Fig. 13. The 
 cytoplasm of the hepatic cell, arranged in a coarse meshwork en- 
 closing the fat droplets, carries in its trabeculae irregularly shaped 
 masses of iron-holding substance which are sometimes extended 
 along and in the trabeculae. The nucleus gives the usual reaction 
 for "masked" iron. The iron in the cytoplasm is of the "masked" 
 character, for isolated cells mounted with a mixture of ammonium 
 hydrogen sulphide and glycerine, and kept, at a temperature of 
 60"C, gave an iron reaction in the masses in the cytoplastic trabe- 
 cules only after three days, which became more marked on the fifth 
 day. The nuclei in these cells gave but a feeble reaction for iron, 
 owing, apparently, to the fact that the "masked" iron in the cyto- 
 plasm had used up all the decomposing energy of the sulphide and 
 therefore, apparently, little of the active reagent reached the nuclei. 
 That the iron in the masses of the cytoplasm is not due to the dif- 
 
 86 
 
fusion from the nucleus, was shown by the non-oocurrence of such 
 iron compounds in the cytoplasm of hepatic cells from Ambly- 
 stomata recently captured, or kept in captivity without food. In 
 > those Ambtystornata, in the intestine of which on capture, food 
 ', matters, such as mollusca, worms and insects, were found, the cyto- 
 plasm of the hepatic cells gave no reaction for "masked" iron. The 
 difference in this respect between the liver cells of the recently 
 captured or fasting animal, and those of the animal which had been 
 fed for several days with egg-yolk, was readily shown by putting 
 thin sections of the organs, hardened in alcohol, taken from animals 
 in the two conditions, into the same dish of sulphuric acid alcohol 
 for three days. At the end of this time, treatment of the two kinds 
 of sections with the acid f errocyanide mixture gave different results 
 in both cases. In both the nuclei exhibited an equally intense iron 
 reaction, but in the cytoplasm of the cells from the fed animals, the 
 masses of the trabecules were as distinctly iron-holding as in the 
 cells treated with ammonium hydrogen sulphide, while the cells 
 from the livers of the other animals exhibited no such reaction. 
 
 This subjection of sections to treatment under exactly the same 
 conditions, considered in connection with the results obtained by 
 using ammonium hydrogen sulphide, shows that the iron found in 
 the masses situated in the cytoplasmic trabeculae was due to a 
 "masked" combination there placed. That it was a chromatin was 
 shown by its capacity for taking up staining matters. In sections 
 of the liver in this case, when hardened either by the alcohol or 
 corrosive sublimate method and stained with haematoxylin, saf ranin, 
 or eosin, the cytoplasmic masses were colored, though not as deeply 
 and intensely as the nuclear elements, yet as distinctly as the lat- 
 ter. In sections from the liver of the unfed or recently captured 
 animal, when similarly treated, there was a complete absence of 
 such stainable material in the cytoplasm. When sections of the 
 liver from the fed animal were treated with sulphuric acid alcohol 
 to set the "masked" iron free and subjected to the Prussian blue re- 
 action, subsequent staining with saf ranin gave a violet-like combi 
 nation of the blue and the saf ranin red in the chromatin of both the 
 cell and the nucleus. 
 
 That the 'Cytoplasmic chromatin in this case comes from the 
 chromatin of the yolk, and is not due to a prozymogen called into 
 Existence by the abnormal quantity of food matter in the hepatic 
 cell, I tried to show by feeding Ambtystornata with olive oil. In 
 those fed with yolk the hepatic cells were very greatly loaded with 
 fat. A similar excess of fat in the liver cells was obtained by feed- 
 ing with oil, but in these there was no cytoplasmic chromatin. 
 
 37 
 
In thin sections from the intestine of the Ambly&toma fed on 
 egg-yolk, the presence of "masked" iron compounds in the cyto- 
 plasm of the epithelial cells was not indicated readily, owing to the 
 abundance of fat in the same. The more active the absorption of 
 the yolk elements had been, the less, therefore, was it possible to 
 demonstrate the existence of cytoplasmic chromatin. I have found, 
 however, as in the intestinal epithelium of guinea-pigs after they 
 are fed on yolk for some time, that the inner portion of each epith- 
 elial cell not overloaded with fat, was filled with an eosinophilous, 
 finely granular material from which sulphuric acid alcohol sets free 
 iron. I found also that if an Amblystoma is fed for several days 
 with yolk and then allowed to fast for several days, in its intestinal 
 epithelium now freed from fat there is, as shown by sulphuric acid 
 alcohol, a slight amount of "masked" iron diffused through the 
 cytoplasm. Whether this is derived from the yolk, or is that of a 
 prozymogen, it is as difficult to decide as in the case of the intesti- 
 nal cells of the guinea-pig under similar conditions. 
 
 There is a difference between the chromatin of the yolk and 
 that of the hepatic cytoplasm of an Amblystoma fed with yolk, 
 which is probably to be explained as due to a chemical transforma- 
 tion effected by the absorbing cells or by the liver cells. According 
 to Bunge 1 the chromatin of egg-yolk, or haematogen, as he terms it, 
 when isolated, gives no immediate reaction with ammonium sul- 
 phide, but after the lapse of half-an-hour, a slight green color ap- 
 pears, which, in the course of several hours, deepens to an intense 
 dark green, and several days after becomes black and opaque, the 
 various appearances being hastened by increasing the volume of 
 ammonium sulphide added. I find that the addition of ammonium 
 hydrogen sulphide to fresh yolk, calls forth in two or three minutes 
 at most the fullest iron reaction of which it is capable. The differ- 
 ence in this respect between isolated and unisolated yolk chromatin, 
 is to be explained by the finer division and distribution of the latter 
 in the spherules which the reagent disintegrates and partially dis- 
 solves before attacking the chromatin, and by the consequently 
 readier action of the sulphide. "When, on the other hand, the 
 spherules have been fixed by heat or by alcohol before subjection to 
 ammonium hydrogen sulphide, the iron reaction is almost as slow- 
 ly obtained as in the case of the isolated yolk chromatin, the slow- 
 ness of the reaction being partly caused by the difficulty with 
 which the sulphide penetrates the spherules. In the livers of the 
 Amblystomata fed with yolk the cytoplasm chromatin is readily 
 
 i Loo. ett. I have elsewhere criticised Bunge's description of this com- 
 pound and have pointed out some mistakes into which he has fallen. 
 
 38 
 
reached by the ammonium hydrogen sulphide, but it takes several 
 days to give the iron reaction, this indicating that the iron there is 
 more firmly held than it is in the yolk. This difference corresponds 
 to that found in the yolk as it is being assimilated in the developing 
 larvae of Amblystoma. In these the yolk chromatin is in form of 
 homogenous spherules 'which, when fresh or hardened in alcohol, 
 give, in a minute or two after treatment with ammonium hydrogen 
 sulphide, a dark green reaction for iron, also shown, though less 
 readily, by the nuclei of the cells containing the spherules. Until 
 the spherules are completely dissolved by the containing cells the 
 nuclei give this reaction, but, when they have disappeared, then the 
 iron reaction in the nuclei is obtained with as great a difficulty as 
 it is in those of the adult animals. In other words, the yolk chro- 
 matin, as it is assimilated, undergoes a change whereby the iron in 
 it is more firmly fixed or more "masked." It is extremely probable 
 that the difference between chromatin of hen's egg and that of the 
 hepatic cytoplasm of Arribtystomata fed with it, is to be explained 
 in the same way 1 . 
 
 The yolk of old or stale eggs gives, with ammonium sulphide, 
 the iron reaction almost immediately, and is, in this respect, more 
 suitable than the fresh yolk to determine if the absorption of yolk 
 chromatin obtains. I made two experiments with it, one with a 
 guinea-pig kept without food for seven days, the other with a lake- 
 lizard which had been kept without food for over two years. 
 
 The guinea-pig was killed five hours after it was given ISc.c. 
 of stale yolk, and parts of the small intestine and the liver were 
 removed and put into alcohol. Pieces of both organs were, in the 
 fresh condition, teased out on the slide in a drop of ammonium 
 hydrogen sulphide and immediately examined under the micro- 
 scope. The preparations from the liver yielded nothing distinctly 
 demonstrating that absorption had taken place, but in the tips of 
 the villi in which fat absorption was prominently shown, there was 
 a dull green color apparently confined to the sub-epithelial elements. 
 The fat-holding epithelial cells of the fresh villi were easily removed 
 in some instances by teasing, and in some of these denuded villi 
 
 i Some change takes place in the iron-holding substance in the cytoplasm 
 of the hepatic cell of Amblystoma, as shown by its staining reaction. In the 
 animals fed with egg-yolk for some time, the hepatic cells, when hardened with 
 alcohol or corrosive sublimate and stained with haematoxylin and eosin, were 
 found to contain, in that part next the blood capillary, eosinophilous masses 
 and in that part adjacent to the bile capillary masses which had a violet stain. 
 Both species of contents were formed of an organic iron compound and be- 
 tween both there were transition elements in which the affinity for haematoxy- 
 lin was as great as that for eosin. It would appear as if the eosinophilous sub- 
 stance which, judging from its position, is possibly derived by the cell from the 
 blood, becomes converted by the cell into that substance which manifests a 
 greater affinity for haematoxylin. 
 
appearances were obtained with ammonium hydrogen sulphide 
 which resembled those represented in Fig. 10. Here greenish 
 granules were found in the adenoid tissue and the leucocytes which, 
 in some instances, were collected in groups immediately under the 
 epithelial layer. Halfway down the villus the greenish reaction 
 became indistinct. In sections of the villi hardened in alcohol, the 
 ammonium hydrogen sulphide gave a green reaction most fre- 
 quently confined to a zone immediately below the epithelium of the 
 tips of the villi, but sometimes extending into the lower ends of the 
 epithelial cells themselves. The reaction of the sub-epithelial zone 
 was chiefly in the leucocytes, and in these the iron was rarely in the 
 granular form observed in the fasting animal or in one fed on 
 ordinary diet. This difference indicates that the iron in these cases 
 is derived from different sources. The difference between the dis- 
 position of the iron in the fresh villus, and that found in the villus 
 hardened in alcohol, is to be attributed to the ammonium hydrogen 
 sulphide altering the arrangement and character of the sub-epithelial 
 elements when these were fresh, and to the extraction by the alco- 
 hol of the fat droplets and globules in the leucocytes which, conse- 
 quently, were more uniformly colored by the reagent than was the 
 case with the fresh elements. 
 
 There was evidence of the transference of yolk chromatin from 
 the epithelial cells to the underlying elements, and this was found 
 in those villi in which the process of fat absorption had not distorted 
 the cells. Sections of these villi, obtained from material hardened 
 in alcohol, when treated with ammonium hydrogen sulphide, gave 
 preparations like that of which Fig. 12 is an illustration. The inner 
 portions of some of the cells at the extreme tip of a villus gave a 
 faint greenish reaction immediately after the reagent was added, 
 but in the corresponding portions of other epithelial cells the reac- 
 tion was given also by granular elements lying between and among 
 the fat droplets. When the cell was loaded with fat the greenish 
 reaction was not obtainable. The underlying leucocytes appeared, 
 frequently, placed against the lower ends of the epithelial cells, and 
 when this was the case the greenish granules in the latter were 
 accompanied by similar elements in the more superficially placed 
 leucocytes. The iron compound in the lower portions of the- epith- 
 elial cells differed, in the readiness with which it reacts on the addi- 
 tion of the sulphide, from that found in the same structures when 
 fresh yolk was fed, as in the earlier experiments, 
 
 The lake-lizard, five hours after 4 c.c. of stale yolk was injected 
 into its intestine, was killed and the intestine removed, cut open 
 ajid put in alcohol. Sections, as well as teased-out portions of the 
 
 40 
 
hardened mucosa, were examined when mounted in a mixture of 
 the sulphide and glycerine, both before and after the application of 
 warmth. The results of this experiment were not as decisive as 
 those of the last described, for the reason, apparently, that as the 
 digestive and absorptive powers had been so long unused, a longer 
 time than five hours for the absorption of the yolk should have been 
 allowed. The yolk had mingled with mucus and in many places 
 formed with it a coagulum which, even in the fresh state, came 
 away leaving the surface of the mucosa perfectly clean. In other 
 parts, chiefly at the tips of the longitudinal folds, fat absorption 
 had occurred, causing an extraordinary distortion of the epithelial 
 cells and amongst these, shaped and moulded by the fat-loaded cells, 
 were leucocytes which also held fat and at the same time gave a 
 very strong iron reaction with ammonium hydrogen sulphide. Be- 
 low these points there were large collections of leucocytes with 
 scarcely less iron than had those amongst the fat-holding epithelial 
 cells. Leucocytes were also found in the tips of other folds project- 
 ing beyond the free border between fat-holding epithelial cells, the 
 part projecting being surrounded by yolk and containing, both fat 
 and iron compounds. That a portion of this iron was of the organic 
 form was shown by the fact that the reaction first obtained with 
 ammonium hydrogen sulphide became deeper.when the preparation 
 was kept for an hour at 60°C. It of course does not follow that this 
 was derived from the yolk, but that they received a part of it from 
 this source, appeared to be indicated by the greater abundance of 
 iron-holding leucocytes under those tips of which the epithelial cells 
 were loaded with fat. 
 
 How the chromatin is transferred to the liver I cannot say 
 definitely, but the capacity of the leucocytes for carrying inorganic 
 compounds of iron and the occurrence of iron-holding leucocytes in 
 the capillaries of the liver of an Amblystoma fed for a time with 
 yolk, gives a basis for the opinion that they are the chromatin car- 
 riers. Furthermore, we know that in the performance of their 
 function as phagocytes they invaginate and dissolve the remains of 
 disintegrated cells, and there is very little apparent difference be- 
 tween this and that of taking up particles, holding chromatin, which 
 may reach them through the process of internal secretion already 
 described. This view would explain the occurrence of leucocytes 
 which gave the diffuse iron reaction in the villi of guinea-pigs fed 
 with stale yolk. 
 
 There remains another question : How does the chromatin of 
 the yolk gain access to the interior of the epithelial cells? Upon 
 this point I have not much to offer, although I have often en- 
 
 41 
 
deavored to determine the mode of entrance. The intimate rela- 
 tions of the fat and the chromatin in the yolk suggest that the 
 method of absorption of both is the same, that is, that when the 
 epithelial cell takes in fat, it also receives the chromatin which ap- 
 pears to form envelopes for the most minute fat particles in the yel- 
 low yolk spherule. The chromatin of yolk would, when deprived 
 of its intimate association with fat, therefore, be unabsorbable, even 
 though mechanically mixed with a quantity of the same. This 
 would explain some of the results of Sucin's experiments already 
 quoted. That investigator fed mice with pure haematogen (of yolk) 
 mixed with the proteids of serum, hog's fat, and starch, and found 
 that the animals did not live as long as those fed on absolutely iron- 
 free food, while those fed on hard-boiled egg-yolk mixed with starch 
 and cellulose lived and throve on the diet. If the fat of yolk assists 
 in the absorption of its chromatin, then yolk from which the fat 
 has been extracted with ether, mixed with hog's fat and starch, 
 ought to give the same result when fed to mice as was obtained in 
 Socin's experiments with pure haemetogen mixed with other food 
 elements. The result of such an experiment would indeed be in- 
 teresting. 
 
 As already mentioned, I have made frequent attempts to de- 
 termine how the particles of chromatin enter the epithelial cells. I 
 have in all of these used alcohol material, which was teased out on 
 tl^e slide and mounted in glycerine and ammonium hydrogen sul- 
 phide. The idea with which I made these attempts was that the 
 chromatin particles, rendered greenish by their iron reaction, ought 
 to be easily seen with careful adjustment of light and the best opti 
 cal appliances obtainable. Of all these endeavors but one seemed 
 to give good results. This was furnished by the epithelium of an 
 Ambiy stoma which had, 24 hours before being killed, been artificial- 
 ly fed with yolk. The appearances to be described were found in 
 isolated spots only on the tips of the longitudinal folds in the in 
 testine. The free margin of the cell was covered with granular 
 matter from the yolk, in which one could see very minute fat 
 globules intermingled with particles having a greenish color. The 
 line of separation of this material from the striated border was, in 
 the majority of the few cases observed, not distinct. The striated 
 border did not give the usual appearance, the strise being apparent- 
 ly replaced, each by a row of oval vesicles with greenish envelopes. 
 "With i - 5 mm. apochromatic immersion (Zeiss) and compensation 
 ocular 4, the vesicles could be seen connected by a grayish line. 
 On the 'protoplasmic side of the margin were also minute vesicles 
 with greenish envelopes, apparently of the same character as those 
 
 42 
 
in the striated border and the reticulated protoplasm itself had a 
 slightly greenish tinge (Fig. 15). "Whether the oval vesicles in the 
 margin were in the rodlets ("stabchen" of the Germans) or between 
 the same it is not possible to say, for in this case one deals with 
 structures which, in their size and optical properties, approach the 
 limits of microscopic vision. It was also impossible to determine 
 whether these vesicles were being invaginated by the epithelial cells 
 or were mechanically entangled by the rodlets. It may, in fact, ,be 
 that the oval vesicles were not connected with absorption at all and 
 that they were merely appearances in the rodlets, although, on this 
 view, the green reaction in their envelopes would be difficult to ex- 
 plain. That the rodlets are not always simple structureless ele- 
 ments, I have, several times, found to be the case in preparations 
 from the guinea-pig (Fig. 11), in which each rodlet appeared to be a 
 series of beadlets or granules. These may have been produced by the 
 reagent used, but their occurrence indicates that the rodlets are not 
 homogeneous elements. Heidenhain 1 observed in the salamander, 
 the dog and the cat, a thickening of the lower end of each rodlet. 
 In my preparations the basal granule of each rodlet was most distinct. 
 It does not appear that the absorption of chromatin and fat 
 goes on in the proportion in which they occur in yolk. In sections, 
 made by the celloidin method, of the intestinal mucosa and its 
 adherent yolk, from an Amblystoma fed with yolk, I found above 
 and resting upon the epithelium, here and there, homogenous 
 clumps of chromatin which are not found in the yolk in its ordinary 
 condition. When these sections are for time kept in contact with 
 ammonium hydrogen sulphide at 60° C. , the clumps give a decided 
 iron reaction. The occurrence may be explained by the existence, 
 on the part of the epithelial cell, of a capacity for the absorption of 
 fat greater than that for the absorption of chromatin. 
 
 1 have already referred to the occurrence of bodies in the outer 
 halves of the intestinal epithelial cells in preparations from a guinea- 
 pig which had been fed with yolk for thirty-six hours. These 
 elements are not iron-holding, as they do not yield any iron reac- 
 tion after treatment for days with ammonium hydrogen sulphide, 
 or after treatment with acid alcohols. They possess, however, a 
 greater affinity for dyes like eosin and aurantia than the surround- 
 ing cytoplasm, and solutions of Ehrlich's heematoxylin give them a 
 feeble violet color. Like nuclear chromatin they absorb and tena- 
 ciously hold mineral reagents, as, for example, ammonium molyb- 
 date 2 and this fact, considered in connection with their capacity for 
 
 i Archiv fOr d. ges. Physiol. Vol. LXIII. Supplement, 1888. 
 
 2 As employed by Lilienfeld and Monti to determine micro-chemically the 
 presence of phosphorus (Zett. fttir Physiol. Chemie. Vol. XVII. p..4i0); 
 
 43 
 
absorbing dyes, would seem to suggest that they belong to the 
 nuolein class of compounds. Somewhat similar bodies have been 
 observed by Heidenhain 1 in the epithelial cells of the intestinal villi 
 in a new-born pup which had suckled, and he has figures also, in 
 one of the plates accompanying his paper, of a portion of a villus 
 from a rabbit after being fed with milk, in the epithelial of which 
 are shown a number of similar bodies. He found their size and 
 number to vary in different cells, while they were absent in the 
 foetus and in a pup twelve days old. He regarded them as albumin- 
 ous excretions from the protoplasm which appear at the commence- 
 ment of the absorptive process, but gradually vanish. If the bodies 
 which I find in my preparations are of the same nature as those 
 described by Heidenhain, a different explanation of their origin 
 appears necessary. It is remarkable that they should occur after 
 the commencement of milk or yolk feeding, and as both foods con- 
 tain proteids intimately associated with the fat globules, the absorp- 
 tion of the proteids, as well as of the fats, must entail upon the cells 
 the disposal of some of the former which are at first physiologically 
 cumbersome as it were, and which, possibly, are disposed of tem- 
 porarily in the form of these protoplasmic masses. The casein of 
 milk and the vitellin of yolk, both phosphorus-holding compounds, 
 may thus contribute to their formation. The only difficulty experi- 
 enced in accepting this view of their origin is that it does not ex- 
 plain, in the results obtained with a yolk diet, why these elements 
 are absent where the yolk absorption goes on most vigorously, that 
 is, in the cells at the extreme tips of the villi, while they are present 
 in those cells in which yolk absorption is less active, and why also 
 these elements are absent after two days of a yolk diet. Further 
 investigation is necessary upon all these points. 
 
 SUMMARY. 
 1. The experiments on the administration of inorganic com- 
 pounds of iron to guinea-pigs and other animals have resulted in 
 showing that the intestinal mucosa absorbs these to an extent which 
 varies with the nature of the compound and with the quantity of it 
 given. When the dose is small, absorption occurs only in that part 
 of the intestine adjacent to the pylorus and measuring only a few 
 inches in length, yet when the quantity given at any one time is 
 large, the absorptive area may embrace the whole of the small intes- 
 tine. In the former case the result appears to depend on the com- 
 plete precipitation, as hydroxide, of the iron of the salt unabsorbed, 
 in the thoroughly mixed bile, chyme, and pancreatic juice ; and in 
 
 i Lou. cit. 
 
 44 
 
the latter case the large amount of the iron salt, apparently, first 
 destroys the alkalinity of these fluids, the excess of the salt un- 
 affected and remaining in solution then undergoing absorption. 
 
 2. The intestinal epithelial cells transfer the absorbed iron at 
 once to the underlying elements when the quantity absorbed is 
 small, but, with a large amount absorbed, the epithelial cells are 
 found to contain some of it. 
 
 3. Though some of the sub-epithelial leucocytes of the villi ap- 
 pear to carry part of the absorbed iron into the general blood circu- 
 lation, probably the more important agent in the transference of the 
 inorganic iron from the villi to other parts of the body is the blood- 
 plasma. 
 
 4. Marfori's albuminate and the commercial "peptone" of iron, 
 when administered to guinea-pigs, seem to stimulate the leucocytes 
 to invade the epithelial layer of the intestinal villi. 
 
 5. Of the organic iron compounds belonging to the ' 'chromatin" 
 class, that present in egg-yolk (hsematogen of Bunge) undergoes ab- 
 sorption in the intestine of the guinea-pig and of Amblystoma. In 
 these, but more especially in the latter, after they are fed with egg- 
 yolk for several days, the cytoplasm of the liver cells yields marked 
 evidence of the presence of an organic iron compound belonging to 
 the "chromatin" class and derived from the yolk administered. 
 
 6. The mode of absorption of yolk "chromatin" is obscure, but 
 the process appears, in some way, to be connected with the absorp- 
 tion of the fat with which the iron compound is closely associated in 
 yolk. 
 
 EXPLANATION OF FIGURES*. 
 
 PLATE XI. 
 
 Note. In the preparation of all the figures (except 15) Abbe's 
 camera was employed, and all, with the exception of two, are illus- 
 trated as they were seen with an immersion apochromatic objective 
 (Zeiss 3 mm., 2 mm. or 1.5 mm. focus). The exceptions are Figs. 1 
 and 2, in the drawing of which Zeiss D. was used. 
 
 Fig. 1. Section of a villus from the pyloric end of the small 
 intestines of a guinea-pig kept on ordinary diet. Alcohol, acid fer- 
 rocyanide mixture, balsam, x 305. 
 
 Fig. 3. A portion of the mucosa of the intestine in a lake- 
 lizard, "e," epithelial cells, "1," iron-carrying leucocytes, "r," red 
 blood corpuscles, also shown to contain inorganic iron. Alcohol, acid 
 ferrocyanide mixture, balsam, x 620. 
 
 Fig. 4. A portion of the epithelium and underlying elements 
 * Taken from the Journal of Physiology, April, 1894; p. 295. 
 
 45 
 
of an intestinal villus of a guinea-pig after the administration of 
 "peptonate" of iron. "L," leucocyte, "be," blood capillary. Alco- 
 hol acid ferrocyanide mixture, balsam, x 1340. Drawn with the 
 diaphragm of Abbe's condenser removed from the microscope. 
 
 Fig. 5. Portion of the section of the liver of the guinea-pig fed 
 with "peptonate of iron. "L," leucocytes, "he," hepatic cells, "bc,'> 
 blood capillary, x 1240. Drawn with the diaphragm of the conden- 
 ser removed. 
 
 Fig. 6. A portion of the tip of an intestinal villus oi a guinea- 
 pig kept on its ordinary diet, to show the distribution in the cells of 
 the organic iron compounds (chromatins). "E," epithelial cells, "1," 
 leucocytes, "a,'' neuclei of adenoid elements. In the cytoplasm of 
 two of the leucocytes are found granules of an inorganic (?) iron com 
 pound. Alcohol, nitric acid alcohol, acid ferrocyanide mixture, 
 balsam, x 1240. Drawn with the diaphragm of the condenser re- 
 moved. 
 
 Fig: 7. Epithelium and underlying elements from the side of a 
 villus of a guinea-pig on the second day of the course of yolk-feed- 
 ing, to show the distribution of organic iron compounds (chromatins) 
 "L," leucocytes, "a," the sub-epithelial "membrane." Alcohol, nit- 
 ric acid alcohol, acid ferrocyanide mix ture, balsam, x 1240. Drawn 
 with the diaphragm of the condenser removed. 
 
 Fig. 8. Epithelium and underlying elements of a villus of the 
 same animal. "A,'' sub-epithelial "membrane," "s," the secretion 
 from epithelial cells. Corrosive sublimate, hsematoxylin, eosin, 
 balsam, x 1240. 
 
 Fig. 9. Portion of a Lieberkiihnian gland of a guinea-pig to 
 show the distribution of organic compounds of iron (Chromatins) 
 and especially of those connected with secretion. Alcohol, nitric 
 acid alcohol, acid ferrocyanide mixture, balsam, x 620. Drawn 
 with the diaphragm of the condenser removed. 
 
 Fig. 10. Portion of a fresh villus of a guinea-pig killed five 
 hours after being fed with stale yolk. The epithelium has been re- 
 moved. Ammonium hydrogen sulphide, glycerine, x 920. 
 
 Fig. 11. Intestinal epithelial cell of a villus from the same 
 animal. Alcohol, ammonium hydrogen sulphide, glycerine, x 820. 
 (Zeiss oc. 4, apoehr. imm. 1.5 mm.) 
 
 Fig. IS. Epithelium and underlying leucocytes of a villus from 
 the same animal, to show the absorption of the yolk chromatin. 
 Alcohol, ammonium hydrogen sulphide, glycerine, x 620. 
 
 Fig. 13. A liver cell of an Amblystoma fed artificially for four 
 days with egg-yolk. Alcohol, sulphuric acid alcohol, acid ferro- 
 cyanide mixture, balsam, x 820. Drawn with the diaphragm of 
 
 46 
 
the condenser removed. 
 
 Fig. 14. Yolk spherules from a hard-boiled egg, to show the 
 distribution of the iron (of the hasmatogen). In the spherule on the 
 left the elements are fewer and coarser. Sulphuric acid alcohol 
 (for 48 hours), ammonium hydrogen sulphide, glycerine, x 820. 
 (Comp. oc. 4, imm. apochr. 1.5 mm.) 
 
 Fig. IS. Free border of an intestinal epithelial cell of an 
 Amblystoma fed with yolk. "P," cell protoplasm, "h," hyaline 
 border, "y," elements of a yolk. Alcohol, ammonium hydrogen 
 sulphide and glycerine (at 60 c. for eight hours). (Comp. oc. 4, 
 apochr. 1.5 mm.) 
 
 47 
 
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PART III. 
 
 The Haemoglobins and Related Pro= 
 ducts of the Market. 
 
 BY CHARLES H. WILLIAMS, Ph. B., Ph. C. 
 
 School of Pharmacy, University of Michigan. 
 
THE HEMOGLOBINS AND RELATED PRODUCTS OF THE 
 
 MARKET. 1 
 
 Br CHAELES H. WILLIAMS, Ph. C, 
 School of Pharmacy, University of Michigan. 
 
 The Haemoglobins of the market are comparative innovations. 
 Side by side with ferric tablet triturates, troches, etc. , the Haemo- 
 globins have come, adding their quota to the elegance of medicinal 
 ingestion. 
 
 It is well known that Haemoglobin contains iron, naturally 
 combined as it exists in the normal blood. Physiologists disagree 
 as to the absorption and disposition of iron when taken into the 
 system. "The iron salts,'' says Bunge, "seem not to be absorbed 
 at all in the intestinal canal, or only to such a small extent that 
 it is questionable whether their absorption has any mentionable im. 
 portance." "It seems more probable that the absorption of iron 
 from the food takes place in the form of proteid bodies containing 
 iron." The only importance that Bunge attributes to the ingestion 
 of these bodies is that they unite with the hydrogen sulphide of the 
 intestine to form iron sulphide, thus conserving for absorption the 
 iron which the food contains. 2 
 
 If nature thus provides sufficient iron in the food we eat, why 
 give iron at all? Why not administer the salts of some other metal, 
 such as manganese or bismuth, which would with equal readiness 
 unite with the intestinal gases? In chlorosis, which is pre-em- 
 inently the result of a lack of iron, the salts of manganese and bis- 
 muth have been substituted, but the disease will not yield to them 
 This fact certainly offers some evidence in support of the theory 
 that iron plays some other part aside from mere sulphide combina- 
 tion and ejection. Physiologists have told ub that the soluble forms 
 of iron are readily absorbed and unite with' the albumen of the blood 
 to form albuminates, and that the insoluble preparations do like- 
 wise after having been changed into chlorides by the gastric juice. 
 
 i From the Pharmaceutische Rttndschatt, September, 1894. 
 2 See also Pharm. Rt/ndschau, vol. 9, p. 114, and vol. 10, p. 267. 
 
 51 
 
The administration of iron as it exists in Haemoglobin, seems to 
 be based upon a combination of the above rival ideas ; for according 
 to Bunge, absorption only occurs in the case of proteid bodies 
 containing iron. Haemoglobin is pre-eminently a proteid body, 
 and if iron must eventually become an albuminate and exist 
 as such, then its ingestion as an albuminate removes the bur- 
 den of an intermediate formation of a chloride, for gastric diges- 
 tion does not form a chloride of the natural proteid iron, 
 but rather converts it into acid-hsematin. Hammerston, in his 
 physiological chemistry, says, "Hrematin is formed by the action of 
 the gastric juice upon oxyhemoglobin." The writer's own experi- 
 ments confirm this statement, for, of the six proteid irons of the 
 market which were subjected to gastric digestion, all but one were 
 so converted. That one was not a natural proteid compound, but a 
 synthetical product. The haematin so formed is said by Hammer- 
 ston to be changed into bilirubin by the liver ; this change necessi- 
 tates the setting free of the combined iron. The question naturally 
 arises, "is the iron entirely eliminated by the bile?" Kunkel says 
 not; for "in 100 parts of bilirubin, eliminated by the bile, there are 
 only 1.4 to 1.5 parts of iron; while 100 parts of haematin contain 
 about 9 parts of iron." 
 
 The terms Haemoglobin and Oxyhemoglobin are often used in- 
 terchangeably ; whereas oxyhemoglobin is a molecular combination 
 of the former with oxygen; the oxygen is loosely combined and 
 easily disassociated. Arterial blood is nearly all Oxyhemoglobin, 
 with some Haemoglobin, while venous blood contains large quanti- 
 ties of the latter. Methaemoglobin is a transformation product of 
 Oxyhaemoglobin, while Hematin is a decomposition product. 
 Characteristic absorption bands are given by each of the above sub- 
 stances with the spectroscope and are of great importance in their 
 identification. 1 Normal, freshly, drawn blood, which is mostly 
 Oxyhaemoglobin, shows two absorption bands between the lines D 
 and E. Band a is narrower but darker and more clearly outlined, 
 and lies on the line D. Band b is broader, less dark and less denned 
 and lies at E. Haemoglobin readily absorbs oxygen from the atmos- 
 phere and is converted into Oxyhaemoglobin ; its spectrum, however, 
 shows a broad not clearly defined band between D and E. Methaem- 
 oglobin has the same composition as Oxyhaemoglobin. If arterial 
 blood be sealed in a tube, it gradually consumes its oxygen and be- 
 comes venous, and by this absorption of oxygen a little methsemo- 
 globin is formed. Methaemoglobin also presents two absorption 
 bands; but band b is stronger and more definitely outlined than band 
 
 i See Pharm. Bundschau, vol. 10, pp. 56—59. 
 
 52 
 
a ; besides this difference from Oxyhaemoglobin, the bands are more 
 toward the red part of the spectrum. Haematin, in acid solution, 
 absorbs more of the violet than of the red. It shows a sharply de- 
 fined band between c and d, and a broader but less clearly distinct 
 band between d and /. On diluting the acid-hsematin solution, four 
 bands manifest themselves; the broad one between d and/ dividing 
 into two, while a very indistinct band appears on the e side of d. 
 The Haemoglobins examined by the writer were subjected to spectro- 
 scopic analysis and the above absorption bands determined. 
 
 What chlorophyl is to the leaf, Haemoglobin is to the blood, 
 namely : its coloring substance. Iron is an integral constituent of 
 this coloring substance and its function is to take up oxygen. Its 
 presence, therefore, is of vital importance. It necessarily follows 
 that an insufficient supply of iron causes an inadequate formation 
 of Haemoglobin. As Haemoglobin, after having been oxidized into 
 Oxyhaemoglobin in the lungs, is the oxygen carrier of the blood, 
 the sequence of this blood poverty is lessened tissue oxidation. 
 
 Before beginning experimental work upon the Haemoglobins 
 and their related products of the market, Haemoglobin was pre- 
 pared. The method employed was that recommended by Hoppe- 
 Seyler. Sheep's blood was used. This was defibrinated immediate- 
 ly upon withdrawal from the body. In the absence of a centrifugal 
 machine the defibrinated blood was allowed to settle and the serum 
 separated from the corpuscles. These were put into three volumes 
 of water and heated to 35° C. The temperature was then reduced 
 to 0° C. and the mixture washed with ether and this was afterward 
 replaced with one-fourth its volume of alcohol. From this alcohol 
 solution, Haemoglobin separated, after having been placed in a 
 freezing mixture during 24 hours. The Haemoglobin thus obtained 
 was dried in vacuo upon an absorbing plate over sulphuric acid. It 
 occurred as an amorphous powder, reddish-brown in color, bloody 
 odor and. albuminous taste. The spectroscope revealed it to be. a 
 mixture of Oxyhaemoglobin and Methaemoglobin. 
 
 Similar experiments were made upon the blood of the ox ; also 
 upon the blood of the cat. Eepeated attempts upon the ox-blood 
 failed of any satisfactory results in the formation of crystals. From 
 the caf s blood, crystals of Oxyhaemoglobin were obtained. 
 
 The Haemoglobins of the market have been examined with re- 
 spect to external form, color, o'dor, taste, solubility, reaction, per- 
 centage of iron, artificial gastric digestion and composition. The 
 percentage of iron obtained in each case is not the result of merely one 
 estimation, but of several, and in some cases, estimations from dif- 
 ferent samples. 
 
 53 
 
The more recent proteid irons of the market which have come 
 to notice are the following : "a" Haemoglobin (Merck) ; "b" Hcemofer- 
 rum; "c" Ferroglobin; "d" Ferrum Sanguinis (Chapoteaut) ; "e" Fer- 
 ratin; "f" Hcemogallol; "g"' Desiccated Deflbrinated Blood; "h" 
 Haemoglobin Compound. These will be considered in the order 
 named. 
 
 a) Haemoglobin from Merck, occurs in fine, chocolate brown, 
 amorphous powder. The odor is somewhat putrid, the taste album- 
 inous. It is very slowly soluble in water and leaves considerable 
 residue. Its reaction is neutral. Several estimations were made for 
 iron on two different samples, one purchased in Detroit and the 
 other in New York. The result shows . 504 per cent, of metallic iron. 
 Upon subjection to artificial digestion, it was converted into acid 
 Hsematin. The spectroscope reveals it to be a mixture of about 
 equal parts of Oxyhaemoglobin and Methaemoglobin. It possesses 
 the common property with other Haemoglobins of burning with an 
 empyreumatic odor. Artificial digestion with this sample, as with 
 the others, was conducted in a pepsin tester, kept between 37° to 40° 
 C and subjected to constant agitation. "With this Haemoglobin, 
 as with the remaining proteids, the iron estimation was conducted 
 volumetrically. The Haemoglobin was first incinerated in a plat- 
 inum crucible. The ash was then dissolved in hydrochloric acid, 
 afterward replaced by sulphuric acid. The resulting iron salt was 
 then completely reduced by nascent hydrogen to a ferrous condi- 
 tion; this was then titrated with N / 50 potassium permanganate. 
 
 b) Haemoferrum comes upon the market in three-grain gelatin 
 pilloids, inclosing a "pseudo-crystalline'' powder. The color is red- 
 dish-brown, while the odor is like that of blood. The taste is sim- 
 ply albuminous. It is almost entirely soluble in water and has a 
 neutral reaction. Iron estimation shows .411 per cent. Artificial 
 gastric digestion converts it into acid-haematin. It is mostly Oxy- 
 haemoglobin, but shows traces of Methaemoglobin. 
 
 c) Ferroglobin occurs in two-grain, compressed, sugar-coated 
 tablets. The color is reddish-brown, the odor that of blood. The 
 taste resembles that of burnt albumen. It is composed of a soluble 
 portion and an insoluble portion. The soluble portion is all Oxy- 
 haemoglobin, and constitutes about one-third of the powder, while 
 the insoluble portion is Haematin. Titration revealed .299 per cent, 
 of iron. * 
 
 d) Ferrum Sanguinis is in gelatine globules inclosing minute, 
 brownish-black scales. The odor is putrid, the taste albuminous, 
 and upon dissolving in water leaves a large albuminous deposit. 
 Its reaction is neutral. Iron is present to the amount of .418 per 
 
 54 
 
cent. Is converted into acid-haematin by digestion. The spectrum 
 shows it to be mostly Methaemoglobin with a trace of Oxyhemo- 
 globin, 
 
 e) Ferratin is not a natural proteid form of iron, but a synthetic 
 product. It "is artificially prepared from egg albumen and chemi- 
 cally pure iron salts in alkaline solution." It occurs as a very fine, 
 gritty, cinnamon-brown powder ; the odor is somewhat alcoholic at 
 times ; when confined, it resembles that of brown sugar. The taste 
 is somewhat ferruginous. It is insoluble in water but soluble in a 
 faintly alkaline mixture to a reddish-brown solution. Several titra- 
 tions on different samples show the amount of iron to be 5.707 per 
 cent. It shows no absorption bands, while artificial digestion con- 
 verts its iron into a chloride. 
 
 f ) Haemogallol is made by the action of pyrogallol upon Haemo- 
 globin. It is a fine, slightly gritty, snuff-brown powder, with an 
 ethereal-alcoholic odor and a slight ferruginous taste. It is insolu- 
 ble in water, but soluble in alkaline mixture to a greenish-yellow 
 solution. It shows no absorption bands and digestion converts it 
 into acid-haematin. Its percentage of iron is .444$. Haemol, a pro- 
 teid iron, is closely allied to Haemogallol. . It is obtained similarly 
 to Haemogallol; the only difference in its manufacture being the 
 use of zinc dust instead of pyrogallol. 
 
 'g) Desiccated Defibrinated Blood occurs in long brownish black 
 scales. Its odor is putrid, its taste is albuminous, and it is very slowly 
 soluble in water. If the scales be placed into hot water, they swell 
 and become plastic and gelatinous. The reaction is neutral; the 
 amount of iron .149 per cent. Its spectrum shows Methaemoglobin 
 with a trace of Oxyhaemoglobin. 
 
 h) Haemoglobin Compound is a brownish black liquid. The 
 odor is that of burnt malt, while the taste is sweet, alcoholic and 
 malty. It is readily miscible in water and has an acid reaction. It 
 contains .015 per cent, of iron. Its spectrum shows it to contain a 
 mixture of Oxyhaemoglobin and Methaemoglobin. 
 
 Considerable evidence is at hand of the efficacy of the Haemo- 
 globin, treatment. * Especially so from hospital service. Dr. LeBon 
 in speaking of the use of Haemoglobins, says, — "desiccated ox blood 
 and Haemoglobin has been thoroughly tried in the hospitals of Paris 
 and found very efficacious in treating debilitated patients." Also 
 Benczur (Disch. Med. Ztg.) reports upon the Haemoglobin treatment 
 as carried on in von Liemssen's clinic. "Troches were prepared 
 from ox blood. The daily amount of Haemoglobin given was about 
 25 grains. Not only was there marked improvement in the cases of 
 anaemia thus treated but no gastric disturbance was observed." 
 
 55 
 
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