Experimental Biology and Medicine

Two principles are realized in the accommodation of an eye that is constructed as a “camera obscura”: (1) Change of curvature of refracting surfaces, principally the lens; (2) change of distance between refracting mediums and image screen, principally distance between lens and retina.

1. There is only increase of curvature, principally of the anterior surface of the lens, during active accommodation. We observe it in mammals, birds, and reptiles (lizards, crocodiles, turtles, a few snakes). Experiments were made before the society to show the increase of curvature of the lens in the eye of the water-turtle—proof of Helmholtz's theory of accommodation.

2. Accommodation by change of the distance between lens and retina is possible in two directions: (a) In cephalopods and fishes, which are normally shortsighted, accommodation for objects at a distance is effected by a movement of the lens toward the retina. In the eye of the fish there is a muscle Musculus retractor lentis (Beer) which draws the lens toward the retina; (b) in amphibia and most of the snakes, the lens is moved toward the cornea, away from the retina, by changes of the intraocular pressure.

About two years ago the authors observed that it was possible to increase the resistance of rabbit blood against trypsin by treating the animal to subcutaneous injections of pancreatic extract. It seemed probable that the resistance of organs against other proteolytic enzymes than trypsin could be increased in an analogous manner.

Rabbits were treated with saline liver extracts. The autolytic powers of the livers of such animals were compared with the autolytic powers of the livers of normal animals. It was found that the autolytic power of the organ was not diminished by the treatment referred to. It was also noted that the organs of the treated animals contained smaller proportions of nitrogen than the livers of the control animals, and further, that the proportions of noncoagulable proteins and of nonbasic nitrogen were higher than in the controls.

In order to accurately interpret the findings, the figures for composition of the livers were compared. It was noted that the amounts of water and carbohydrates were not affected by the treatment, while the proportion of ether-extract was higher in the organs of the treated animals than in those of the control animals. The appended table gives the data of nine experiments, on as many animals.

After it had been proved that the heliotropism of animals and plants is identical, it seemed desirable to find means by which positively heliotropic animals could be transformed into negatively heliotropic ones, and vice versa. Groom and Loeb found that such a transformation was possible in the nauplii of Balanus perforatus at Naples by the influence of light, inasmuch as these animals were positively heliotropic in very weak light and negatively heliotropic in strong light. 1 Later Loeb found that the marine Copepods and young larvas of Polygordius became positively heliotropic on lowering the temperature as well as on increasing the concentration of the seawater, while they became negatively heliotropic under the opposite influences. 2 Moreover Loeb observed that negatively heliotropic Copepods can be made positively heliotropic by mechanical agitation, 2 and Miss Towle showed that the sign of heliotropism in Cypridopsis can be reversed by contact with solid bodies. 3 Holmes made the discovery that the positively heliotropic terrestrial Amphipods, e. g., Orchestia agilis, become negatively heliotropic when thrown into water. 4

The author had tried in vain to change the sense of heliotropism in animals by chemical means, and this gap was felt the more keenly, inasmuch as he was led to believe that chemical changes might ultimately determine changes in the sense of heliotropism. Recently, however, the author succeeded in finding instances in which specific chemical substances were capable of transforming the sense of heliotropism in animals.

The experiments were made with a fresh water shrimp (Gammarus pulex), which can be obtained at any time in large quantities at Berkeley. If one puts a large number of these animals suddenly into distilled water or into common tap water, they all become at first very negatively heliotropic. It is possible that this is caused by the mechanical agitation, connected with the transferral of the animals from one vessel to another, but this has not yet been ascertained with certainty. Half an hour or an hour later, the negative gathering of the animals becomes less dense, and the animals are scattered in the vessel.

The studies made heretofore upon the malarial parasites of birds have shown the existence of four species or types. These are:

Proteosoma.

Halteridium.

Hæmamœba majoris, Lav.

Hsemamœba Ziemanni, Lav.

In the course of an extended study of the parasites of birds, the authors encountered several new species, and, since the number is likely to be still further increased, it seemed desirable to attempt a classification. The authors based their classification largely upon the type of multiplication and the habitat of the parasite. Two genera were given; one, Plasmodium, characterized by formation of segmenting forms in the peripheral blood and invasion of fully developed red blood cells. The injection of blood having these parasites results in an infection. For the other genus the authors used the priority name of Kruse's, Hœmoproteus. This genus is characterized by an entire absence of segmentation-forms in the peripheral blood, and, with the exception of two species which form a transition as it were between the two genera, invasion of young erythroblasts is the rule. Injection of blood having these parasites does not lead to infection.

At the last meeting of the Society of American Pathologists and Bacteriologists an informal statement of this fact was made by Dr. Welch for Drs. Marshall and Knox. The experiments of Dr. Collins and the author are reported here because they were undertaken in a slightly different way and also because a certain number of confirmatory observations are of value.

The maltose-fermenting paradysentery bacillus of Flexner was grown for twenty-four hours on each of eleven consecutive days in fresh bouillon solutions of the serum from a horse immunized through oft-repeated injections of the bacillus. The serum strength in the solutions used was 1.5%, 4% and 15%. The serum agglutinated the culture before its growth in the solutions in dilutions up to 1 in 800, and was strongly bactericidal in animals. After eleven transfers the culture grown in the 15% solution ceased to be distinctly agglutinated by the serum in any dilution and ceased to absorb from the serum any appreciable amount of the agglutinins acting upon the original culture. The cultures grown in the 1.5% and 4% solutions were changed to a less degree and agglutinated in dilutions up to 1 in 100 and 1 in 60 respectively, and continued to absorb agglutinins. The recovery of the capacity to be agglutinated was very slow in the culture grown in the strongest serum solution, when it was from time to time transplanted in fresh nutrient agar. The other cultures recovered this characteristic more rapidly.

The first culture, after growth for sixteen weeks, during which it was transplanted forty-three times, agglutinated in dilutions up to I in ZOO, and after twenty weeks in dilutions up to I in 400. The culture grown in 4% solution of serum agglutinated after sixteen weeks in dilutions up to I in 500, and one in 1.5% agglutinated in dilutions up to I in 800. This diminution and final almost complete lack of development of agglutinable substance in bacteria grown in a serum rich in agglutinin and immune bodies is interesting. It showed not only a rapid variation in bacteria of essential characteristics, but also indicated a possible means of adapting themselves to resist destruction in the living body, since the bacteria which ceased to produce agglutinable substance and probably, also, less substance with affinity for other antibodies, might be considered less vulnerable to these substances.

The essence of Mendelism in inheritance is its alternative character. In this it is opposed to blending inheritance (as in human skin color) which had been regarded as the typical sort of inheritance. At the Carnegie Institution's Station for Experimental Evolution certain new cases of nonblending inheritance have already been found. Among sheep it appears from Dr. Alexander Graham Bell's records that the offspring of two black sheep are (probably always) black, although one or more of the grandparents were white. It looks as if black color (like albinism) might be recessive. Among canary birds it is found that of the offspring of crested and of plain headed birds, some are crested and some are not. Poultry have been studied because of the numerous characters they exhibit. When a Japanese long-tailed, clean-legged cock was crossed on a white bantam hen, the two surviving offspring were highly colored like the father and had abundant feathers on the legs like the mother.

The empirical formula, C₁₀H₁₃NO_3.½H₂O, adopted by Abel for that member of the epinephrin series which he has called epinephrin hydrate (the adrenalin of Takamine) is, at present, the subject of an acute controversy. The authors have been engaged in a repetition of the analytic work on which Abel based the above formula for epinephrin hydrate. In view of the suggestion of Abderhalden and Bergell that this substance should be prepared in a way that avoids oxidation by the air, the authors have undertaken the laborious task of preparing and purifying it in an atmosphere of hydrogen. The results of their work in this direction will soon be published.

The authors emphasized the fact that the ½H₂O of their empirical formula has always been regarded by them to be water of constitution, and not water of crystallization as their opponents have taken for granted. The assumption that this ½H₂O, so easily removable by high heat and by various acids, is water of constitution necessitates doubling the present empirical formula, a procedure which is at variance with the molecular weight determinations of v. Fürth and Jowett. These determinations are, however, open to serious objections, as will be shown at length elsewhere.

Work on the decomposition products of both alkaloidal epinephrin and epinephrin hydrate has been continued. The basic substance, C₃H₄N₂O, which is obtainable equally from both forms of epinephrin, has been decomposed by treatment with caustic potash into ammonia (NH₃), methylamin (CH₃.NH₂), and methylhydrazin (CH₃.NH.NH₂). Of these decomposition products, methylamin is also obtained from both modifications of epinephrin. Methylhydrazin has thus far not been obtained either from epinephrin or its hydrate. This degradation product is of great importance in throwing light on the chemical constitution of the new base, C₃H₄N₂O, for its appearance, under the circumstances referred to, proves that the two nitrogen atoms of this base are directly combined one with the other, and suggests, among other things, for this base a ring structure, such as is found in bodies of the pyrazolon series.

Mr. Lieber gave an account of the discovery of radium by Mme. and Professor Curie, and demonstrated many radioactive phenomena. Special attention was drawn to recently discovered facts bearing on radium emanation. For a time it was thought that radium discharged directly (a) the so-called “emanations,” which had practically no penetrating power and which, like a gas, were readily carried from one point to another by an air current; and (b) the so-called “rays”—alpha rays of very low penetrating power, beta rays of considerably greater penetrating power, and gamma rays of enormous penetrating power. Later investigations have shown, however, that radium discharges primarily emanations and alpha rays only. However, the emanations soon disintegrate, and the disintegration products yield the beta rays and the gamma rays. Consequently the powerful beta and gamma rays are the products of a decomposition product of radium. The proportions of the radiations given off by a certain quantity of radium and its disintegrated emanations are about 95% alpha rays and about 5% combined beta and gamma rays. Because of their nearly negative penetrative power, the alpha rays, as well as the emanations, are practically unavailable for therapeutic purposes when the radium is used in glass tubes or similar containers. Even the superficial layers of a given radium preparation are relatively impervious to both the emanations and the alpha rays proceeding from the underlying portions of the preparation. Therefore, it is essential, in order to obtain full radioactive effects, that (1) the given quantity of radium should be spread so thin that, from the practical standpoint, an upper layer would not exist and (2) should be held in a container with walls that would be permeable both by the alpha rays and by the emanations.

The investigation of the subject has been continued by the employment of a method by which the isotonic curves of all the contractions of an excised non-curarized muscle stimulated at regular intervals, are superimposed upon a recording surface. The differences which were previously pointed out in the mode of fatigue of the muscles of the frog, the turtle and the mammal, have been confirmed. Lohmann's work, in which a frog's gastrocnemius on being heated to a mammalian temperature, shows a course of fatigue similar to that of mammalian muscle, has been repeated and found incorrect. Both that muscle and the turtle's coracoradialis profundus, similarly heated, continue to give their characteristic curves of fatigue. [See page 60 (124).]

Kaiser's method for determining the point of the isotonic curve where the contractile stress terminates, has been employed for the frog's gastrocnemius, and it has been found that as the height of the curve diminishes in the course of fatigue, the contractile stress terminates at progressively lower and lower points. The lowering of the latter does not, however, seem to keep pace with the lowering of the summit of the curve. Hence the two points seem to approach one another.

In a series of experiments on dogs the starvation nitrogen was first determined during fasting periods. Varying amounts of gelatin, containing from one fourth to two thirds of this amount of nitrogen were then fed, the remaining three fourths to one third of the starvation quantity being supplied in meat or other proteins. The calorific requirement of the animal, estimated from Rubner's tables, was made up in each experiment with fats and carbohydrates. Results show an equal sparing of the body-protein, whether one fourth, one third or one half of the starvation nitrogen was fed in the form of gelatin, the coincident sparing of protein by fats and carbohydrates being the same. When the coincident sparing of protein by non-nitrogenous food was increased by feeding a larger percentage of carbohydrates and less fat, the fraction of the starvation nitrogen fed in the form of gelatin could be raised to two thirds, the other one third being fed in meat. Nitrogenous equilibrium was maintained on this diet for several days.

The same result was obtained on man. The starvation nitrogen was obtained by analysis of the urine and feces during a fasting period of three days, and equilibrium was then established at this level on a mixed diet containing two thirds of the nitrogen in meat, the other one third in cereals. Then for two days the meat nitrogen was replaced entirely by gelatin nitrogen, the other one third remaining the same, and the potential energy supplied was increased from 40 to 48 cal. per kilo of body-weight by giving more cane-sugar, which served at the same time to make the gelatin more palatable. The nitrogen equilibrium was not disturbed during these two days nor on the two following days, when the diet was exactly the same as before the gelatin period.

The float consists of an aluminium cylinder with very thin wall, supporting a writing arm of fine aluminium wire. For manometer tubing of in inside diameter, in. or ¼ in. aluminium tubing 2½ in. long is used. This is bored out until the walls are sufficiently light. In the upper end is forced a solid cap of aluminium with a small hole in the center into which the wire for the writing lever is driven. The lower end is plugged with cork. Both ends are painted over with hot paraffin to prevent leaking. For use in alcohol a somewhat larger tube is necessary. Three crossed hairs held in place across the open arm of the manometer tube by a strip of adhesive plaster keep the writing arm centered with sufficient accuracy.

The value of the float consists in its cheapness, the ease with which it can be made, its very slight inertia, and its convenience in estimating delicate changes in pressure for which a water or alcohol manometer is needed.

An apparatus was demonstrated which had been devised for the purpose of measuring the reducing processes of the different kinds of cells in vitro. Definite quantities of organ pulp were placed in specially constructed tubes and anærobic conditions were established by the passage of nitrous oxid gas. Definite quantities of methylene blue of known strength were then added. The rate of reduction was indicated by the disappearance of the blue color owing to the reduction of the animal cells. It was shown that in vitro the influence of temperature is the same as that observed in the living organism. The influence of alkali in accelerating reduction was also shown. The action of salts and various poisons is at present the subject of investigation.

The author called attention to the influence of temperature on the activity of reduction in the living organism as indicated by intravital infusion of methylene blue. Elevation of the body temperature greatly accelerates the rate of reduction in the tissues. This was demonstrated by means of an intravital infusion of methylene blue in a rabbit, whose body temperature had been elevated to 42° C. through the external application of heat. Simultaneously with this infusion, another infusion was made in a rabbit of approximately equal weight, in which the temperature was maintained at about 39° C. In other respects, the conditions of the infusion were as nearly alike as possible in the two animals. A definite contrast was noted at the close of the infusion between the organs of the two animals as respects their color, the normal rabbit showing more color than the one in which the temperature had been elevated. The differences in the nervous system and the muscles were particularly striking. Even during life, an inspection of the muscles indicated that the reduction was carried on with greater rapidity in the heated rabbit than in the normal one. Previous observations on the reducing action of the animal body under the influence of cold were referred to.

The author demonstrated a substance of singularly great powers of condensation with other organic substances, this condensation resulting in the formation of colored bodies. He demonstrated especially the reactions of naphthoquinon sodium mono-sulfonate with anilin and various amins, with nicotin, conin, piperidin, and finally with indol, skatol and pyrrol. The reactions with indol, skatol and pyrrol possess unusual physiological and chemical interest and will form the subjects of future publications.

The reaction with pyrrol occurs in the cold and is evidenced by the deepening red which on the addition of alkali changes to purple, violet, blue and finally reddish-brown. The addition of acid to the red solution obtained without alkali is followed by the development of a green and finally brown color. These color reactions (and particularly the one dependent on acids) occur with such rapidity if one uses concentrated heated solutions of pyrrol, that the characteristic color stages may be of extremely short duration. This reaction with pyrrol is a highly characteristic one, and should prove of service to chemists.

Among the biological and medical applications of the naphthoquinon sodium mono-sulfonate reactions, the author mentioned the study of various aromatic compounds in the organism, the occurrence of certain intravital syntheses, the detection in the urine of organic compounds, such as para-amidophenol, and the development of a method of staining the bile capillaries DY means of intravenous infusion of the derivatives of the naohthoquinon compound. The author also stated that these substances facilitate the study of the relation between the chemical constitution and distribution of poisons in the body.

In physiology no distinction is made between absorption from the subcutaneous tissue and absorption from muscles. In experimental infection and immunity, injections of virulent toxic and antitoxic materials are being extensively employed, but intramuscular injection has not yet even been thought of. In therapeutics it is practised promiscuously, and for the reason, as pharmacologists and clinicians expressly state, that it gives less pain and causes less frequently the formation of abscesses.

The authors came upon the observation that absorption from the muscles is incomparably more rapid and efficient than from the subcutaneous tissue and tested the matter with several substances. With suprarenal extract, it was tested in three ways.

1. By the effect upon blood-pressure.—A subcutaneous dose of 0.6 c.c. adrenalin or less per kilo (rabbit) exerts no effect, and the variable effects of larger doses consist in a rise of pressure of from about 10 mm. to 20 mm. of mercury, which sets in late and develops slowly. An intramuscular injection of 0.5 c.c or 0.4 c.c. per kilo, or even less, invariably causes, on the other hand, a considerable rise of pressure, which sets in after a very short latent period and reaches its maximum in a few seconds. The curve obtained after intramuscular injection is very similar to that after an intravenous injection. The increase has been as high as 50 mm. or 60 mm. of mercury and may go even higher. The course of the curve is frequently interrupted by “vagus pulses.”

2. By the effect upon the pupil on the side from which the superior cervical ganglion had been previously removed.— An intravuscular dose of 0.5 c.c. or 0.4 c.c. of adrenalin per kilo causes dilation of the pupil in less than a minute, while such a dose given subcutaneously rarely produces any effect. The effect of a larger subcutarjeous dose sets in only after 10 or 15 minutes.

The author called attention to the usefulness of the antitoxin unit furnished by the Institute for Experimental Therapy under the direction of Professor Ehrlich in the routine testing of the strength of diphtheria antitoxin. The one uncertain element is the relative resistance of the guinea-pigs to diphtheria toxin.

In the course of the past nine years the author has given considerable personal attention to this subject and found that different dealers furnished guinea-pigs of slightly different susceptibility. This difference was attributed to environment and care. The animals bred under the author's supervision generally showed maximum resistance. Irregularities in the routine tests during the past year led the author to look up the genealogy of the pigs used and he found that the different degrees of resistance belonged to certain families or litters and were constant for those families. Thus, one mother gave birth to young which did not react to what was the usual fatal dose. Four successive litters possessed the, same resistance. As each pig could be tested but once the, precise degree of resistance could not be measured, but it appeared probable that this family could stand 40% more toxin when mixed with the antitoxic unit than those of average susceptibility. Other mothers were traced whose offspring possessed less resistance than the ones described, but could still neutralize 20% more toxin when mixed with the antitoxic unit than the average.

It would seem from these observations that different degrees of susceptibility to toxin are to be found among guinea-pigs and that the special degree possessed by any one is not to be attributed to individual variation, but to a family trait or character. The resistance in the cases cited could not beattributed to any preliminary treatment with toxins and antitoxins. Experiments are now under way to determine the part played by the male in the transmission of toxin resistance. In the case of the most resistant family, the four litters were the offspring of two males.

That concentrated solutions of venom fail to destroy and tend to preserve blood-corpuscles was noted by Mitchell and Stewart. Among the recent writers who have paid especial attention to the interpretation of this phenomenon are Kyes and Sachs. They ascribe it to deviation of the hemolytic complement through the excess of venom amboceptors. The study which forms the basis of this brief communication shows the hypothesis of Kyes and Sachs to be untenable, since it could be demonstrated that (1) the protective action fails to occur with venom in which, through heating to from 95° to 100° C., the hemolytic principle has been preserved, but certain other constituents have been coagulated, and (2) the action extends to protection of the corpuscles from laking by water, ether, saponin, etc. The conclusion which has been reached by the author is that venom unites with the globulins and especially with the hemoglobin of the red corpuscles, yielding a water-insoluble compound to which the protection is due. Various substances, such as salts, acids and alkalis, restore the hemolyza-bility of the corpuscles by dissolving the venom-hemoglobin compound. The permeability of the corpuscles is not markedly altered.

During the year 1904 an effort was made in the pathological laboratory of the University of Buffalo to make studies on hematozoa in the lower animals. In a considerable number of normal cats, dogs, rabbits and guinea-pigs no hematozoa were found. The results of other examinations were as follows: 51 English sparrows {Passer domesticus), half in the winter, half in the spring, all negative; 27 mud-puppies (Necturus maculatus) in March, all negative; 40 toads in the summer, all negative. In 140 frogs from the Niagara river there occurred the following infections: 14 with Trypanosoma, 5 with Drepanidium, 1 with Filaria. Drepanidium was found both in the summer and fall. The infections with Trypanosoma were distributed as follows:

In July, of 15 frogs, 2 showed trypanosomes.

In August, of 26 frogs, 10 showed trypanosomes.

In September, of 14 frogs, 2 showed trypanosomes.

From October to December, of 85 frogs, none showed trypanosomes.

In one case Trypanosoma and Drapanidium occurred in the same blood. The trypanosomes had the usual characters of Trypanosoma rotatarium (ranarum). They were in no case numerous; two were rarely seen in one low-power field. The frogs appeared healthy. Eight attempts to inoculate normal frogs by way of the peritoneum with the blood of infected frogs gave negative results.

Attemps ad cultivation.— The blood of frogs and toads was taken to make blood-agar (used by Novy and MacNeal for the cultivation of trypanosomes). The blood was first examined carefully to see that it was free from parasites. The animal was etherized and placed in HgCl₂ solution I to 1,000 for 15 minutes, rinsed with distilled water, opened with a11 precautions, the blood from the heart taken wit11 a sterile pipette, and mixed rapidly with the water of condensation on slanted agar tubes (made with meat extract and peptone, and slightly alkaline to litmus).

The search for the cause of an infectious disease like measles becomes greatly simplified when we learn how to secure the unknown “virus” in relatively pure form unmixed with common microbes. Various methods may now be applied to the investigation of the virus. The transmission of measles from mother to fetus would seem to point to the presence of the cause of the disease in the blood. In the twenty cases of fetal measles collected by Ballantyne, it seemed that the infection of mother and fetus must have been simultaneous, because the eruption in both corresponded in character. In order to learn something further as to the presence in the blood of the cause of measles, inoculations of human beings would seem to be necessary; because, so far as we now know, this disease is probably not communicable to animals. Grünbaum's experiments with measles in the chimpanzee appear to have given negative results.

Critical review of the literature shows that almost without exception the recorded experiments in the inoculation of measles, for which positive results have been claimed, are without real significance. The claims that the experiments of Home, of Wachsel, of Speranza, of Katona, of McGirr, of Bufalini gave definitely positive results do not stand close scrutiny in the light of the evidence at hand : In many instances the rubeolous nature of the sickness, sometimes very mild, following the inoculation and regarded by the experimenters as measles, is not at all securely established, and in practically all cases the possibility of natural infection was not excluded. These experiments, practically all of which were undertaken with the idea of producing a modified form of the disease, consequently permit no conclusion as to the infectiousness of the blood or other substances in measles. If we accept Mayr's results as they are given by him it may be concluded that in measles, nasal mucus and cutaneous scrapings (containing blood, epithelial dhbris, and tissue juices) may contain the cause of measles at or near the height of the eruption.

To test whether the centrosome is a permanent cell organ or not, Professor E. B. Wilson (1901) made an experiment on the sea urchin egg by treating, with a salt solution, enucleated egg fragments obtained by shaking. He observed that asters containing centriole and capable of division were produced in the enucleated fragments. He, therefore, came to the conclusion that at least some of the centrioles in the asters thus formed must have arisen de novo. Some writers criticized his results, saying that the formation of the centrioles in the enucleated fragments observed by him might have been due to the shaking-out of the nuclear fluid into the cytoplasm. Wilson, therefore, suggested that his experiment be carried out by the author in a somewhat different manner—instead of shaking, to cut eggs singly and to treat the nucleated and enucleated pieces separately. The author tried this experiment on the egg of Cerebratulus in the summers of 1903 and 1904. Strict precautions were taken to prevent accidental fertilization, everything used for the experiment being sterilized. Individual eggs were cut into nucleated fragments (i. e., fragments containing the first maturation mitotic figure) and also enucleated fragments. The latter were kept for an hour in a solution of calcium chlorid. Then they were transferred to sterilized sea water. Asters were produced in almost all enucleated fragments thus treated. What is more striking, all the asters had centrioles which were identical with those found in the whole eggs subjected to the same treatment. The nucleated half was stained and was shown to have had the mitotic figure intact. From these experiments no other conclusion can be drawn than that the centrosomes, with centrales of the enucleated fragments, were formed de novo.

One of the few points on which all observers of vaccine bodies are agreed is that these structures are extremely susceptible to artificial changes. The author has for some years endeavored to find a method of examination of these bodies by which artificial changes could be avoided; and this object seems to have been accomplished by the very simple procedure of making Klatsch preparations of corneal vaccine ulcers.

A glass slide is cleaned with soap and water, and thoroughly heated in a Bunsen flame. It is then found to be unusually adhesive. The cornea of an anesthetized rat or rabbit, presenting a vaccine ulcer at any stage, is exposed by holding back the eyelids and protruding the eyeball. The cooled slide is then lightly applied to the ulcer and quickly withdrawn without lateral motion. The slide carries away with it an impression of the ulcer in the form of isolated cells or groups of cells loosened by edema. In this way ten to twenty impressions may be taken in serial order and the minute ulcer may be completely excavated without sacrificing the animal. The isolated cells dry instantly and may be fixed by gentle heat, and afterward by methyl alcohol, and then stained by various methods, preferably by Nocht-Romanowsky for ten minutes. The vaccine bodies are then presented with a clearness equal to that of the malarial parasite in blood spreads.

In the Klatsch preparations stained by Nocht's method the following features of the vaccine bodies appear to be demonstrated. The vaccine body is a portion of the cytoreticulum, its reticular structure being continuous on the one hand with the cytoreticulum and on the other usually with the nuclear reticulum. The clear zone surrounding the vaccine body in sections of tissue is an artifact. The reticulum of the vaccine body takes the chromatin stain, indicating that it contains chromatin, and many of the bodies are so intimately connected with the nucleus, the meshes of one passing insensibly into the other, as to force the conclusion that these particular bodies have arisen by recent extrusion of nuclear chromatin into the cytoreticulum.

The toxic effects of bile are manifold, and have been the subject of numerous investigations. The authors referred only to the general effects: coma and convulsions. Of the early investigators of the effects of injection of bile into animals, some observed only coma, others convulsions, and still others stated that they observed both. The last work on this subject, the work which is now frequently quoted, was done by Rywosch about fourteen years ago. Rywosch claims that coma is the only effect of the two which the injection of bile or bile salts produces.

In their extensive series of experiments on frogs the authors established the fact that the injection of bile can produce coma as well as tetanus. Coma is the frequent and the more reliable result. By a certain device, however, they were able to demonstrate the presence of the tetanic element even in bile which infallibly produced coma; it was by the addition of a subminimum dose of strychnin. A frog of medium size will not respond, even with the slightest hyperesthesia, to an injection of a hundredth of a milligram of strychnin. When such a small dose, however, is injected into a frog which has received a certain quantity of bile, the animal reacts, sooner or later, with a distinct tetanus. The effective dose of bile varies with the animal from which it is obtained. For instance, of ox bile hardly more than 0.3 c.c. need be used, otherwise the coma will completely mask the tetanic element. Rabbit's bile, on the other hand, may be given sometimes even in doses of 2 c.c. or 3 c.c., without supressing any of the tetanic features. The setting in of complete coma usually masks the tetanic element, as already stated. A close observation, however, will reveal in many cases some distinct differences between the coma of animals which received a subminimum dose of strychnin and that of animals which had not received any strychnin.

Thorium nitrate precipitates proteins and is intensely astringent. Its intravenous injection is promptly fatal by embolism. Applied subcutaneously, it causes local necrosis. Administered by the stomach, even large doses have no appreciable effect.

Solutions in sodium citrate were found to be nonprecipitant and nonastringent. As much as 1 gm. of thorium nitrate, per kilogram of dog, injected subcutaneously and intravenously in citrate solution, had little acute action; however, the animals appeared depressed and became emaciated. The postmortem examination, made after several weeks, showed extensive and widespread calcification of tissues. Thorium could not be demonstrated in the calcified areas.

A method for the quantitative estimation of thorium was elaborated; this gave satisfactory results with urine, to which known quantities where added. But in actual experiments on animals it was found inaccurate, a large proportion of the injected thorium escaping detection. However, it was found that on intravenous or subcutaneous injection, the thorium appeared in the urine, and not in the feces. When administered by mouth, it appeared in the feces, but not in the urine. The conclusion appears justified that absorbed thorium is excreted by the kidneys, but that the metal is neither absorbed nor excreted through the intestine.

Our experiments comprised the third series in a study, still in progress, of the toxicology of rare elements. 1 They were twenty-seven in number, and were performed on as many animals (frogs, mice, dogs). They were carried out before Baskerville's announcement of his discovery that thorium contains two new elements, named by him berzelium and carolinium. Publication of our results was deferred because of our desire and intention to complete the work with a study of the toxicological effects of these two new elements, which Professor Baskerville has generously agreed to furnish at a later stage in his investigations. The foregoing communication by Professor Sollmann has induced us, however, to present our results as they stand.

In some of the early experiments (1900) it was found that thorium (nitrate) had a uniform precipitative effect on various connective tissue mucoids. In a study with Professor Loeb (1902), on the antitoxic influence of ions, thorium (nitrate) was used as a tetravalent element, and was found to exert only very slight, almost inappreciable antitoxic effects in mNaCl, with fertilized Fundulus eggs as the indicators. At that time we observed a strong precipitative effect of thorium on protoplasm, and a marked toxicity on various fishes, and on both fertilized and unfertilized Fundulus eggs in sea water, although these facts were not recorded in our paper.

In the experiments on frogs and warm-blooded animals the tetrachlorid was used exclusively. Of our results the following were in harmony with those reported by Brown and Sollmann : Thorium exerts marked astringent action. The chlorid is acid in reaction (in water). The aqueous solution of the chlorid blanched and hardened tissues, proteins were precipitated by it, and blood not only precipitated but blackened. Injected directly into the circulation even very small doses caused intravenous precipitation, and resulted fatally. Subcutaneous injection resulted in local necrosis. We have had no experiments with thorium in citrate solution nor on the excretion of thorium.

The method of development of the frog's egg may be changed by a number of external agents. If the eggs are revolved at the rate of 180 revolutions per minute; if they are put into salt solutions of definite strengths; if they are subjected to a low or to a high temperature; if they are deprived of sufficient oxygen or surrounded by carbon dioxid in solution; if they are placed on wet filter paper instead of developing under water—in any of these ways abnormal embryos result.

An examination of the effects of these external agents brings out two points of especial interest. First, that the effects are not gradual, i. e., corresponding in degree to the increasing strength of the agent employed, but that no effects appear up to a certain point and then suddenly the agent begins to act. Increasing the strength of the agent above this point may for a small range increase the effect, but this occurs within extraordinarily narrow limits compared with the lower range of non-action. The most plausible explanation of this mode of behavior in most of the cases is as follows: The agents act by coagulating certain parts of the egg, thereby preventing their further development. Other parts of the eggs that are made up of different colloids or of different concentrations of colloids remain unaffected, and proceed to carry out their development as far as the presence of the injured region allows.

The second point was the one that the author spoke of especially. Despite the great diversity in the form of the abnormal embryos, most of them may be reduced to modifications of the same type. For example, in many cases the dark cells of the upper hemisphere do not grow down over the lower hemisphere to produce there the embryo, but, remaining at the top of the egg, partially constrict off from the yolk cells at, or even above, the equator of the egg. Out of these dark cells the abnormal embryo develops usually in the form of a ring. Sometimes one side only of the ring develops and a half embryo appears; sometimes only the anterior end of the ring develops and an anterior embryo appears (often more or less “ open ”), etc.

It has been pointed out previously by the author and others that the contraction process of the muscles of cold-blooded animals in the course of fatigue becomes greatly slowed, while those of warm-blooded animals show no such phenomenon. Lohmann has recently claimed that a cold-blooded muscle on being heated to a mammalian temperature shows a course of fatigue similar to that of mammalian muscle, and on the other hand, that a warmblooded muscle on being cooled, fatigues like the muscles of coldblooded animals at a similar temperature. From the supposed effects he concludes that in the matter of fatigue there is no real physiological difference between the two groups of muscle.

The author has investigated the question by very careful methods in a considerable variety of animals, and has not been able to confirm Lohmann's conclusions. The muscles of the frog and the turtle show their characteristic method of fatigue whatever the temperature. The muscles of warm-blooded animals on being cooled and then fatigued, show either no slowing of the contraction process or only a slight slowing. The latter seems to be most pronounced in the rodents, namely, the rabbit, the mouse and the rat. [See page 37 (101).]

The process of conjugation in protozoa involves either temporary or permanent union of two individuals. During this union there is a fusion of nuclear material from both organisms resulting in the formation of new cleavage nuclei in each exconjugant. The process is directly comparable with fertilization of an egg by a spermatozoön, and the biological significance of the phenomena involved is probably identical in all living things.

Since 1876 it has been generally assumed that one effect of conjugation is rejuvenescence or renewal of vitality in both of the exconjugants. This assumption has never been submitted to experimental proof. In his Paramecium work, begun in 1901, the author almost had the proof, but allowed the opportunity for obtaining it to slip through his fingers without realizing its importance at the time. The author's object in bringing this up at the present time is to announce that on the last day of February (1905) he started a new series of experiments with Paramecium, consisting of three different lines at present in about the fortieth generation after conjugation, mainly for the purpose of completing his earlier work.

Another point of general biological importance will also be investigated. In his original experiments the author found strong evidence that the old view that both exconjugants are rejuvenated is erroneous. In twenty pairs which were cultivated after separating from conjugation, one individual of each pair invariably out-lived the other, thus indicating an incipient fertilization like that in metazoa. This phenomenon will be given careful study in the experiments now under way.

By means of a cannula placed in the ureter and retained without ligatures, and which did not materially interfere with the peristalsis of the ureter, the intraureteral pressure and its relation to the peristaltic movements of the ureter were ascertained.

In nine experiments on dogs narcotized with morphin and atropin, the pressure in the ureter arose only to a minute degree, the average being a negative pressure, more pronounced under the influence of diuretics. In five, in which chloroform was used, the pressure was always positive; the irritability and contractility of the ureter were noticeably diminished. In six, under ether, the ureter was noted to be irritable and contractile three hours after the anesthesia was commenced; the pressure was low. In four, in which ether followed the administration of chloroform, ether showed a stimulating effect on the peristalsis, running the pressure rapidly down. In three, in which morphin and atropin, chloroform, and ether were successively tried during the same experiment, the specific effect of each as above noted was again observed. In an animal in which anesthesia was produced by decerebration, irritability and contractility of the ureter muscle were noted; the pressure was low, tending to negative on stimulating the ureter distal to the cannula. In one animal, anesthetized with cocain by the lumbar puncture method, the same results as with morphin were obtained. In two artificially constructed systems, which were demonstrated, phenomena analogous to those observed in the animals were produced and the causes indicated.

The experiments, which also were demonstrated in part, seem to justify the following deductions:

1. A suction normally follows the peristaltic wave of the ureter; at the same time a force is exerted on the fluid in front of the wave.

2. When the ureter is normally acting, the pressure in the pelvis of the kidney remains very low, fluctuating about a neutral point, this condition obtaining through the anatomical arrangement of the pelvis, which prevents it from collapsing under negative pressure. The rhythmic movements of the pelvis of the ureter effect a milking of the portion of the pyramid which projects into it.

Last winter, at a meeting of this society (Proceedings, Vol. I, page 39), the author reported observations showing that extracts from dried kidneys, obtained with the Rosenfeld alcohol-chloroform method, contained from a third to two-thirds of their weights of lecithins. Rubow, 2 of Copenhagen, reported similar results of more extended studies, printed in Danish some months earlier.

During the last few months the author has learned that it is not necessary to boil with absolute alcohol and extract with chloroform in order to obtain the large quantities of extract yielded by the Rosenfeld method. Repeated extraction of the fresh, undried organ, with 85% alcohol at 45∗deg; C., will accomplish practically the same result. When making these extracts, it was found that upon cooling, the alcoholic solutions yielded a precipitate from which a substance resembling the protagon of Liebreich could be obtained. It is to this substance that the author directs attention in this report. The yield is from about o. 14% to 0.20% of the fresh organ, or from about 0.6% to 1.0% of the dried kidney.

“In order to obtain sufficient material for analysis, the author employed the method used by Cramer” in preparing protagon from the brain. The method employed was, in brief, as follows : The minced kidney, freed from obvious fat, was treated twice with 5% sodium sulfate solution at 85 ° C. to 90 ° C.; the filtrates were discarded and the coagulum extracted, first with 95% alcohol and then repeatedly with 85% alcohol, at the boiling points, and the extracts filtered from the coagulum on a hot-water funnel. The filtrates were cooled to from 0 ° C. to − 5 ° C., the precipitate was filtered out and purified by boiling with absolute alcohol, diluting the filtrate with water to make 85 % alcohol, chilling, filtering, treating the precipitate repeatedly with cold ether to remove cholesterin, dissolving in hot chloroform, reprecipitating by chilling, filtering and expressing all possible traces of chloroform.

The experiments were undertaken to investigate the comparative physiological action of three elements, which are of equal valency and of approximately the same molecular weight, and whose chemical properties are closely related. The experiments were carried out in vitro and on unicellular organisms, bacteria and infusoria, frogs, pigeons, rats and guinea-pigs. The solutions used were the chlorids, isotonic with 0.6% sodium chlorid. In one case the propionate was used without any marked difference in the result being observed.

The chlorids coagulate egg and serum albumins, but neither the purified albumoses from Witte's peptone nor peptone are precipitated.

Dilute solutions delay the growth of bacteria and eventually kill. The solutions are not very toxic to spores. Opalina, para-mecia, and vorticellae are killed quickly, equivalent solutions of the chlorids acting in the following order of strength: Neodymium, prseseodymium and lanthanum.

In frogs voluntary and involuntary muscle are quickly put out of action. This is particularly the case with perfused muscle. The solutions act in the same order as with unicellular organisms. Intravenous injection causes almost instant death, due to multiple embolism.

Attempted chronic poisoning was unsatisfactory. The solutions were introduced both subcutaneously and intraperitoneally. Some of the animals died with ill-defined symptoms. Others remained well, except for areas of induration at the seat of injection. Experiments with oral administration and on elimination will be conducted.

As all the solutions, owing to hydrolysis, are acid in reaction, the authors are inclined to attribute a large share of the acute effects to the acid present. The salts range themselves in their toxicity according to their molecular weights.

There have not been very many studies regarding the influence of bile upon blood-pressure, and among these the statements are conflicting. Thus, Traube, who was the first to study it upon the kymograph, states that the intravenous injection of bile salts causes a considerable fall of the blood-pressure, while Edmunds states, in a recent report from Halliburton's laboratory, that the effect is an insignificant one.

Nearly all of the investigators of this question within the last fifty years have employed bile salts in their experiments. The results of the authors' experiments were derived from intravenous injections of filtered ox bile into rabbits. Of the several reasons for employing bile and not its salts, one should be mentioned: It is the belief of the authors that for biological phenomena we have as yet no right to assume that the sum of the known parts is equal to the whole.

In these experiments all degrees of effects have been observed, from an insignificant one to a considerable and even a fatal fall of blood-pressure. But these different degrees could be produced at will. Besides the quantity and the concentration of the bile, it was found that the rate at which it is introduced into the circulation is the most effective factor in the result. A quantity of bile of a given concentration, which, when injected slowly, would cause only an insignificant depression, brought about a tremendous fall of the blood-pressure when injected rapidly. By injecting normal salt solution speedily the fact has been established that neither the mechanical influence of the rate of injection nor the temperature of the injected fluid can have anything to do with the pronounced effect which is invariably produced by the rapid injection of bile.

The fistulous opening between the portal vein and the inferior vena cava was made in six dogs by Dr. J. E. Sweet. Observations were made as to the behavior of the animals when fed on a diet of proteid food. One typical experiment may be summarized as follows: During eleven days on a mixed diet there were no abnormal symptoms. On the four succeeding days beef meal and milk were fed, with the result that on the fourth day pronounced ataxia, loss of sight and hearing, complete anesthesia, and catalepsy were observed, recovery occurring on the next day. After fasting for 24 hours the animal was placed on a diet of fresh lean beef. In five days a recurrence of the above symptoms was noted. The death of the animal occurred on the fifty-ninth day of the experiment, after the dog had undergone a loss of 42% of his weight. Autopsy showed a fistulous opening 2 cm. in length and no collateral circulation. In other cases the symptoms described occurred only after the addition of Liebig's extract to the meat diet.

The administration to normal dogs of sodium carbamate either by mouth or by intravenous injection, gave rise to none of the symptoms observed by Pawlow and associates.

1. In two previous publications the author mentioned the fact that by applying two different methods of treatment to the unfertilized egg of the sea urchin, this egg could be caused to develop in a way which resembled in all its essential features the development of the eggs fertilized with sperm. These two methods consisted, first, in putting the eggs for about two hours in hypertonic sea water (the method used in the early experiments) and, second, in exposing the eggs for from one to two minutes to sea water, to which a certain amount of acetic acid or formic acid had been added. When the old method alone was used the eggs did not form a membrane, nor did the larvas rise to the surface. When the acid treatment alone was used, the eggs formed a membrane and after about six hours divided into from two to six cells, but then died. When the eggs were exposed to the acid for only a short time, e. g., for three-fourths of a minute, not all the eggs formed a membrane when put back into normal sea water; and in this case only those divided into two or four cells and subsequently died within 20 hours, which had formed a membrane, while those eggs which had not been exposed long enough to the acid to form a membrane neither segmented nor died. If both methods of treatment were combined, however, those eggs which had formed a membrane developed at about the same rate as the eggs fertilized with sperm. A certain percentage of these eggs rose to the surface of the water in the usual way, while the eggs which had not formed a membrane either did not develop at all, or developed in the somewhat abnormal and slow way characteristic of the treatment by hypertonic sea water alone.

Baumann and his pupils investigated the effect of the administration of the halogen derivatives of the hydrocarbons, and found that one of the results in the disturbance which followed was the elimination of halogen aromatic mercapturic acids, which were regarded as derivatives of cystin. Later, Friedmann investigated these compounds and was able to confirm Baumann's view. The mercapturic acids are derivatives of cystein, the reduction product of cystin.

This study is a part of an investigation of cystinuria, the view, which has been advanced by previous observers, being taken that the process in brombenzol poisoning is an experimental cystinuria. Dogs were used in these experiments, and were fed on a uniform diet. The animals were catheterized once a day. Analyses of the urine, as complete as possible, were made. The feces were examined for nitrogen and fats.

During the period of administration of the brombenzol the nitrogen and urea were somewhat increased. The urea followed the total nitrogen closely. The preformed ammonia remained at a level below that of the fore period. The kreatinin estimations did not give results of any distinctness.

The investigation of the partition of sulfur led to the result that, while the total sulfur excretion was not increased during the experiment, there was almost complete suppression of the alkaline sulfates. The excretion of neutral sulfur, represented for the most part in this case by parabromphenyl-mercapturic acid, was increased 400 %. The curves representing the alkaline and neutral sulfur were antipodes. The ethereal sulfates rose markedly during the feeding period. In one experiment it was shown that during the first two days of the administration the total sulfur excretion remained constant, while the total sulfate-sulfur fell rapidly. This period is being made the subject of closer study.

Inoculations of different tumors (sarcomas of the thyroid, mixed tumor of the submaxillary gland) through several or many generations have shown that, under the influence of experimental conditions, the energy of tumor growth varies in a definite way. The rate of growth is relatively slow in the animal originally affected by the tumor; after the inoculation into the first generation there is a certain latent period, after which the tumor begins to grow. The growth in the first generation is more rapid than in the original animal. After the inoculation into the second generation the latent period is abbreviated, more or less, and the succeeding growth is likewise more rapid than the growth in the original animal, or in the first generation. A further shortening of the latent period, or an increase in the rapidity of tumor growth, does not take place in the succeeding generations. Duration of the latent period and rapidity of growth may remain stationary through many generations, or the energy of the tumor growth may even somewhat decline.

These facts permit the conclusion that transplantation of a tumor has a tendency at first to increase the energy of tumor growth, and that this increase may be cumulative. That this increase does not continue in succeeding generations may perhaps be explained by the existence of counteracting influences, the actual existence of which can be demonstrated, as will be shown later.

The energy of tumor growth can be increased directly, and not only indirectly, merely by removal of the tension of the surrounding capsule or by better conditions of nourishment. Such a direct stimulating effect of the wound upon the cell growth causes probably a phenomenon not infrequently observed by surgeons, namely, the increase of malignancy in recurrent tumors. It is also possible to diminish the energy of tumor growth. In the course of tumor inoculations it not rarely happens that certain tumors remain stationary or apparently even retrogress spontaneously. This is especially found in the course of later inoculations, and it probably indicates that after many inoculations one or several of the factors determining a vigorous tumor growth become gradually weakened. In such cases one can observe that even a long time after the expansive growth of such a tumor piece has ceased, many mitoses are present in the cells of the stationary or retrogressive tumor.

Dr. Davenport exhibited photographs and plumage-charts of four hybrids between different races of poultry, and also of their parents, and remarked on the nature of the inheritance illustrated by each example.

The experimental studies upon which this communication is based were suggested by an investigation of the necrosis produced in the liver of the dog as the result of injecting hemolytic immune sera of high hemagglutinative power. 1 These necrotic lesions, which are due apparently to an obstruction of the circulation by thrombi composed of fused red blood-corpuscles, vary in position and extent, according to the dose of serum administered. Small doses cause more or less isolated lesions which may occupy any portion of the lobule; large doses produce a diffuse necrosis which spares only the tissue about the larger portal spaces. The uniformity of this necrotic lesion suggested the importance of a study of the repair process which would naturally follow in animals recovering from the acute toxic effects of the injected serum. The extent of the destruction precluded complete regeneration of liver parenchyma, and if the defect was repaired by connective-tissue proliferation, the resulting histological picture would be, except for a difference in the relation between the new tissue and the remainder of the lobule, analogous to cirrhosis in man.

Methods.— Dogs were injected either in the smaller branches of the femoral vein, or in the abdominal cavity, with serum obtained from rabbits which had received repeated injections of red blood-corpuscles of the dog. The dose usually employed was I c.c. of serum to from 500 gm. to 800 gm. of body-weight, and the animals were killed at intervals varying from 48 hours to 36 days

Results.— A majority of the animals die after intervals varying from 4 minutes to 48 hours. In those surviving, hemoglobinuria was a constant phenomenon usually appearing within 18 to 24 hours, persisting 3 to 4 days, and followed for several days by the presence of bile pigment in the urine. The first evidence of repair was mitosis of the liver cells lying at a slight distance from the necrotic areas. The earliest period at which this was seen was 38 hours after injection.

Within the past two years several French and German writers (Josué, Erb and others) have described under the various names of calcification, atheroma or arteriosclerosis, a lesion of the aorta of rabbits produced by the intravenous injection of adrenalin. These experiments have been repeated for the purpose of making detailed histological studies and in the hope of throwing some light upon the histogenesis of arteriosclerosis in man.

Methods.—Rabbits have received injections of a 1 to 1,000 solution of adrenalin in the ear vein. An initial dose of 3 m. repeated every other day has been the usual procedure. In other instances, the dose has been gradually raised until a dose of 20 m. to 25 m. was given every day. The animals have been killed after periods varying from a few days to eight weeks.

Results.—The vascular lesions produced are limited to the aorta and exhibit a more or less definite sequence. Rabbits receiving five to six injections show no gross lesions, but histologically important changes in the media are evident. These consist of focal areas of degeneration in which the muscle fibers are destroyed without alteration of the elastica. Later the degeneration is more extensive and involves the greater portion of the middle zone of the media. At this time changes in the elastic tissue appear; the fibers become swollen, stain irregularly and in some places appear to be fused together. Special stains show a small number of minute fat droplets in such areas. After twelve to fifteen injections very definite lesions are evident macroscopically. The aorta is more or less distorted, rigid and nonelastic. Irregular dilations alternate with elevated brittle areas of calcification. Distinct atheroma with ulceration is seldom seen. In the experiments continued for six to eight weeks, the process becomes very diffuse and small dilations of the thinner portions of the aorta assume the appearance of aneurysms.

A chemical study of the castor bean has indicated that this seed contains proteins of the same character as the other oil-seeds which have been examined, namely, (1) a considerable quantity of a globulin which can be obtained in octahedral crystals; (2) a much smaller quantity of an albumin, coagulating at about 60° C. to 70° C., the temperature at which it separates depending to a large extent on the rate of the heating and other conditions; (3) proteoses which appear to belong to several of the now recognized groups of this class of substances. The satisfactory separation of the various types of proteins was accomplished largely by the use of fractional salt precipitation and dialysis.

The toxic constituent of the castor bean has been investigated under Kobert's guidance by Stillmark, who applied the name ricin to protein material which he separated. The product which Stillmark regarded as relatively pure must have been a mixture of proteins and have contained only a small proportion of the toxic compound. Cushny made a more careful study of ricin and obtained a substance of sufficient toxicity to produce death in animals with a dose of 0.04 mg. per kilo of body-weight. He regarded the toxic compound as protein in nature. Among subsequent investigators, Jacoby has denied the protein character of ricin. He digested his toxic preparations with trypsin and obtained solutions which retained their toxicity although apparently no longer giving protein reactions. Brieger, however, failed to prepare toxic preparations free from protein material.

The efforts of the authors have been directed especially to the possibility of isolating the toxic constituent of the castor bean and determining its chemical nature. The toxic action has been found to be associated wholly with the preparations containing the coagu lable protein and never with those free from the albumin already mentioned

The method described by the authors constitutes a rapid and accurate means of determining indol. It is based on the fact that indol, in slightly alkaline solution, readily condenses with naphthoquinon sodium mono-sulfonate, and forms a blue crystalline compound which is only very slightly soluble in water and is readily extracted by chloroform from a watery solution or suspension. The condensation compound results from the union of two molecules of indol with one of the naphthoquinon compound. The union does not occur as in the case of compounds with amins, with the elimination of the sulfonic acid group, but occurs between one of the carbonyl groups of the naphthoquinon compound and the imid group of the indol. The new compound is, therefore, a di-indyl naphtho-ketone mono-sulfonate. The solubility of this substance in chloroform is about one part in 4,000 of the solvent, and is sufficiently great to permit a rapid and thorough extraction of the substance. Chloroform containing the di-indyl compound has a red color, very like that of hemoglobin. Owing to this circumstance, the condensation compound in chloroform can be approximated colorimetrically in a convenient manner by comparing the tint of the solution with that of the orange-red glass scale of the Fleischl hemoglobinometer. When more accurate results are desired, the chloroform is evaporated and the residue of the di-indyl compound weighed.

It was found that the method here indicated serves for the recovery of a very large percentage of indol from peptone solutions or bouillon. From solutions containing a little protein, the indol may be recovered almost quantitatively. The presence of a large proportion of protein may cause the retention of considerable indol. The distillation should be carried on directly, without steam, from the acidified fluid. The presence of indol in a small fraction of distillate is best ascertained by boiling the acid solution with a few drops of a 2 per cent. alcoholic solution of di-methyl amido-benz-aldehyde.

The authors exhibited to the society two guinea-pigs, which were deeply narcotized by injections of magnesium sulfate. One of these animals had been similarly narcotized twice before, and fully recovered each time. In their physiological and toxicological studies of magnesium salts, the authors found that by subcutaneous injections of certain quantities of sulfate or chlorid of magnesium, animals can be brought into a state of deep anesthesia, during which any operation can be performed upon them without the least resistance. If the dose of the salts is not too large, heartbeat, blood-pressure and respiration remain nearly normal. It was tested on dogs, cats, rabbits, guinea-pigs, white rats and frogs. A gram and a half of magnesium sulfate is about the effective dose for most of the animals. The chlorid has to be used in smaller doses in proportion to its smaller molecular weight. Particulars will be reported later. The authors emphasized the fact that these salts are very poisonous when certain maximum doses are exceeded.

Exudates obtained by injecting suspensions of aleuronat into the pleural cavities of dogs and rabbits were subjected to autolysis. The Kjeldahl method was used to determine the nitrogen of coag-ulable proteins converted by digestion into soluble form.

Inflammatory exudates removed one or two days after injection of the irritant undergo very little change, while those removed three or four days after the onset of inflammation exhibit appreciable though slight autolysis. There is no relation between the amount of digestion and the number of cells which are present. If the cells are separated by centrifugalization from the serum and suspended in normal salt solution, well-marked autolysis is demonstrable. By recombining cells and serum it can be shown that the serum inhibits this autolysis. When this inhibitory action is prevented by heating serum to 100° C., leukocytes acting upon the coagulated serum cause very active digestion. In the following experiments nitrogen of uncoagulable substances is represented by cubic centimeters of sulfuric acid:

The anti-enzyrnotic action of the serum is unaffected by a temperature of 65 ° C., but is prevented at 75 ° C. The proteolytic ferments of the leukocytes act both in an acid and in an alkaline medium, but are most efficient in the latter. The anti-enzymotic action of the serum is favored by an alkaline reaction, but is completely prevented in an acid medium, The serum of the exudate contains a proteolytic ferment, which is active only in an acid medium.

Many streets of Brooklyn are without a public water-supply and a sewage system. The residents of these streets are therefore dependent upon wells for their water-supply, and upon privy vaults and cesspools to remove the sewage and waste water of their homes. The soil is uniformly sandy and water may be had by driving a pipe or digging a few feet below the surface. The water obtained is to a certain extent surface water. The underground water is necessarily influenced by the sea water. This influence is very marked in some instances, as is shown by the high chlorin content, accompanied by the low contents of other constituents that could indicate sewage contamination.

The following tables present average analytic data regarding condemned shallow wells, also regarding wells considered to be of a suspicious quality and wells which were passed as being of fair quality. Very few of the latter class were considered to be of good quality, and some might possibly have been classed as suspicious upon their high nitrate contents, considered with the proximity of the sources of contamination.

Tables giving average, and high and low results of analyses of 438 shallows wells in Brooklyn N. Y., which are used for domestic purposes; also sic data obtained for 14 deep wells used for manufacturing purposes. The figures represent parts in 100,000.

The highest and lowest figures which enter into the averages are also given. These figures do not represent any particular analyses, but are selected from the different results from which the averages were made. Table D gives the average data for deep wells that supply water for manufacturing purposes. These data may be used as standards in judging the purity of wells whose waters are used for domestic purposes.

It was proved by a series of determinations that the viscosity of the “living” blood can be greatly influenced by changing the temperature of the surrounding medium. The viscosity was found markedly increased, if the dogs used in the experiments were immersed in water at 25° C. Warm water baths (42° C. to 45° C.) produced a corresponding decrease in the viscosity. The specific gravity of the blood was changed in a corresponding manner.

Determinations of the viscosity of the “living” blood were made during deep and light ether and chloroform narcosis. It was found that the viscosity is increased by deep and lessened during light narcosis. The specific gravity of the blood also shows regular variations. It is increased by deep and lessened by light ether narcosis. Chloroform, on the other hand, produces a slight decrease during deep and an increase during light narcosis. Hence the specific gravity cannot be regarded as an accurate index of the viscosity.

The various studies of the effects of radium that are included in this communication were carried out at the writer's suggestion and under his general direction. All of them are still in progress. They were made possible by the generosity of Mr. Hugo Lieber, who gave the writer an abundant supply of radium bromid for each series of experiments. Professor William Hallock also encouraged the work by permitting the use of some of his valuable samples of radium bromid and radioactive substances. Dr. G. B. Pegram has given advice freely on physical matters connected with radioactivity. The studies included in this plan were the following (I-V): 1

1. “Preliminary notes on the effects of radium rays on plants”: C. STUART GAGER (New York Botanical Garden).

The radium (bromid) has been employed in several forms, and in degrees of activity ranging from 10,000 to 1,500,000. Experiments so far indicate that the effect is the same in kind, whether the plants are stimulated with gamma rays only, or with alpha and beta rays as well. When three kinds of rays are employed the effects, within the same time, seem to be increased. The results already obtained justify the following statements:

The early stages of seed germination are accelerated, if stimulation ranges between the minimum and optimum points, otherwise they are retarded. Seeds are less sensitive to the rays when dry than when soaked. When germinating seeds are exposed to radium at short distance, germination and subsequent growth are retarded, but when the distance between the radium and the seeds is increased and a screen of metal is interposed, growth is accelerated, Radium rays acting through soil in which plants are growing accelerate both germination and subsequent growth of the shoot, and increase the number and length of root hairs.