Showing papers in "Journal of Experimental Medicine in 1917"

Experimental studies upon lymphocytes : i. the reactions of lymphocytes under various experimental conditions.

Abstract: A simple method is presented by which, with the diffusion of trypan blue into the nucleus as a criterion of cell injury, it is possible to study quantitatively the effect of various agencies upon the small thymus cells and upon the tissue lymphocytes. Preliminary studies with this method have led us to the following conclusions, which, however, unless otherwise stated, may be taken as applying only to the lymphocytes of the rat thymus. 1. The small thymus cells, when suspended in balanced phosphate solutions, show no distinct reaction to variations in hydrogen ion concentrations ranging between P(h) 7.0 and P(H) 7.8. Beyond P(H) 7.0 there is a sudden increase in the permeability of the cells to the dye; plasmolysis of the cells occurs when the alkalinity exceeds P(H) 8.0. 2. Heating to 49 degrees or 50 degrees C. is accompanied by a critical increase in the permeability of the cells to the dye. 3. The injury caused by lack of oxygen can be demonstrated by the increase in the number of stained cells. 4. The addition of serum to suspensions of thymus cells or tonsil lymphocytes greatly inhibits the diffusion of the trypan into the cells. The protection afforded is roughly proportionate to the amount of serum added. Gelatin also exerts a marked protective influence; egg albumin affords a partial protection; starch and gum arabic are inert. Hemoglobin and cholesterol do not modify the stainability of the cells. Arsenious sulfide in weak concentrations partially inhibits the diffusion of the dye. Colloidal iron is without effect, and is precipitated about the cells. 5. The toxicity of the photodynamic substance, hematoporphyrin, and of an impure chlorophyll solution in the presence of sunlight could be strikingly demonstrated by the greatly increased permeability of the cells to the stain. 6. Acute and chronic inanition produces an increased fragility of the cells. The protective power of the serum in acute starvation appears to be increased. 7. The small thymus cells of old animals are more readily injured than are those of young ones, as indicated by the increased proportion of stained cells. 8. The method has been applied to the demonstration of the action of cytotoxic immune sera for rat thymus cells and for human tonsil lymphocytes in vitro. Further experiments dealing with the question of specificity are in progress. The cytotoxins are inactivated by the addition of complement. Thermostabile cytagglutinins have also been produced.

The elaboration of specific soluble substance by pneumococcus during growth

Abstract: 1. A specifically reacting substance of bacterial origin is present in the cell-free fluids of young cultures of pneumococcus. This substance is present when the organisms are growing at their maximum rate and undergoing little or no cell death, and consequently its. presence is not dependent upon cell disintegration but represents the extrusion of bacterial substance by the living organism. 2. The blood and urine of rabbits experimentally infected with pneumococcus contain a similar specific soluble substance during the early hours of the infectious process. 3. Human beings suffering from lobar pneumonia have in their blood and more frequently in their urine a specific soluble substance of pneumococcus origin. The amount of this substance present in the urine varies in different individuals and the presence of a large amount is of unfavorable prognostic import. This specific precipitin reaction in the urine is of diagnostic value. 4. Rabbits injected with soluble pneumococcus material continue to excrete this substance for a considerable period of time. 5. The specifically soluble substance obtained from bacterial cultures and from the urine during infection is not destroyed by boiling, by precipitation with alcohol, acetone, or ether, or by trypsin digestion. 6. Studies are in progress at this time on the degree of toxicity and on the antigenic properties of the substance.

The rat as a carrier of spirochæta icterohæmorrhagiæ, the causative agent of weil's disease (spirochætosis icterohæmorrhagica)

Abstract: 1. On the basis of these findings, we conclude that the extermination of rats and field mice is a highly important prophylactic measure against Weil's disease. 2. The chemical composition of soil and water plays animportant part in the development of Spirochœta icterohœmorrhagiœ , and consequently in the spread of the disease of which it is the causative agent.

Intestinal obstruction : an experimental study of the intoxication in closed intestinal loops.

Abstract: 1. Closed intestinal loops in which bacteria are first removed are compatible with life. 2. Closed intestinal loops in which bacteria are present but in which tissue necrosis is prevented, are compatible with life. 3. Closed aseptic intestinal loops in which the blood supply is completely occluded are compatible with life. 4. The normal secretions and bacterial products of the duodenum and jejunum are not sufficiently toxic to produce any symptoms when allowed to drain into the abdominal cavity. 5. Our results do not support the theory of Draper of a normal toxic secretion of the duodenal mucosa, neutralized by the jejunal mucosa, or the perverted secretion theory of Whipple. 6. Bacterial activity plus necrotic tissue, or the absorption of toxic products resulting from the action of putrefactive bacteria on necrotic tissue is the important factor in the rapid death in simple closed intestinal loops.

The normal fate of erythrocytes : i. the findings in healthy animals.

Abstract: The phagocytosis of red corpuscles, while frequent in the normal dog, rat, and guinea pig, is slight in man, the rhesus monkey, and many rabbits. In cats it is always negligible in amount and frequently absent. Phagocytosis will not suffice as a general explanation of normal blood destruction. When the liver, spleen, and bone marrow of the cat, dog, rabbit, or monkey are slowly perfused with defibrinated blood or Locke"s solution, bodies are given off into the fluid which have the appearance of red corpuscles that have lost their hemoglobin but retained the rest of their cell substance. These bodies possess many of the properties supposedly distinctive of red corpuscles. They are the product of disordered parenchymal cells. By a special method, it has proved possible to search the body, organ by organ, and the circulating blood also, for disintegrating red corpuscles. Shadows of red cells are not present anywhere, nor are hemolyzing red cells found. A hemolytic process, in the ordinary sense of the term, can scarcely play an. important part in normal blood destruction. Instead, it is certain that some red corpuscles, at least, are destroyed in another way; namely, by fragmentation. Normal blood regularly contains small numbers of fragmentation forms—microcytes and poikilocytes—and accumulations of them are regularly present in the spleen, but are found only inconstantly in the other organs. The fragments are in evident process of further subdivision. They occur not only in species in which phagocytosis as a means of cell destruction is negligible (cats), but also in animals in which it is an important process (dogs, some rabbits). The method of study that we have employed is well suited to disclose how the blood is destroyed. The importance of cell fragmentation in this connection is indicated by our failure to find any other means of destruction, save only the phagocytosis already known. Further facts indicating the importance of fragmentation are presented in our second paper, where a general discussion will also be found.

The viscous metamorphosis of the blood platelets

Abstract: From the experiments above described it is evident that the viscous metamorphosis of the blood platelets in shed blood is intimately associated with the early stages of coagulation and that the presence of calcium is a very important element, though perhaps not absolutely necessary. No constant differences in this phenomenon could be detected with platelets or coagulating elements from the blood of normal or diseased individuals or from man or rabbits. All the substances, with the exception of egg white, causing this phenomenon were derived from the blood either during the alteration of fibrinogen or after it had been acted on by thrombin. It seems that the substances or mixtures capable of producing the metamorphosis are especially those associated with the early stages of coagulation or capable in the test-tube of forming or liberating that active coagulating element known as thrombin. The substance in serum that is capable of metamorphosing platelets seems to be attached to the globulin fraction rather than the albumin fraction and is destroyed by heat at temperatures which destroy prothrombin, thrombin, and serozyme, and precipitate fibrinogen. The reaction is not caused by pure thrombin or a mixture of pure thrombin and calcium, though substances causing the metamorphosis are intimately related to thrombin. The metamorphosis seems to be caused by serozyme-like substances as shown both by the fact that barium sulfate absorbs the power of serum to cause the reaction and that a serozyme-like substance is probably to be recognized in all the substances or mixtures, including egg white, causing this change in the platelets, except perhaps when thrombin and fibrinogen are reacting. It is to be noted that in serozyme are contained antithrombin, calcium, and potential thrombin, and that a combination of these isolated factors mixed together occasionally allowed the viscous metamorphosis to occur and not infrequently an abortive metamorphosis. The pseudo- or abortive metamorphosis caused by the mixture of pure thrombin, antithrombin, and calcium may be interpreted on the supposition of a close approximation but not a real reproduction of the colloidal state known as serozyme. That such a reaction is related to real viscous metamorphosis is suggested, because it sometimes occurred with the above mixture and when thrombin and heavily oxalated plasmas were reacting, rather than a real metamorphosis. Simple agglutination of the platelets may occur independently of a viscous metamorphosis, though an agglutination of platelets is to be considered an integral part of the viscous metamorphosis phenomenon. The inconstant results seen with the cephalin-treated sera are probably due to the fact that exactly the same mixtures were not obtained. The results with cephalin-soaked and platelet-soaked sera and with dilutions of sera suggest that the reaction of viscous metamorphosis of the platelets is quantitative.

Spirochæta icterohæmorrhagiæ in american wild rats and its relation to the japanese and european strains : first paper.

Abstract: The principal points brought out in the present article are the following 1 Wild rats captured in this country carry in their kidneys a spirochete which possesses the morphological and pathogenic properties characteristic of Spirochata icterohamorrhagia discovered by Inada in the Japanese form of infectious jaundice 2 Cultures of the American, Belgian, and Japanese strains of the spirochete were obtained by a special technique described, the first two strains having been cultivated artificially for the first time 3 Animals actively immunized against the Japanese strain resist inoculation, not only of the same strain, but also of the Belgian and American strains The Belgian strain produces immunity equally effective against all three strains Experiments to ascertain whether the immunity afforded by the American strain also protects against the Japanese and Belgian strains are in progress 4 These findings warrant the conclusion that the spirochetes designated here as the Japanese, Belgian, and American strains are probably identical 5 On account of its distinctive features, a new genus, Leptospira, has been suggested as the designation of this organism

The effects of experimental plethora on blood production.

Abstract: With the purpose of determining whether a diminished activity of the bone marrow could be brought about experimentally, plethora was produced in rabbits by means of repeated small transfusions of blood. Counts of the number of reticulated red cells in the circulating blood were made during the course of the experiments as an index to changes in the activity of the bone marrow. With the development of plethora, the number of reticulated cells in the blood decreased. In the majority of the plethoric animals, this diminution was extreme, and in some instances, reticulated cells practically disappeared from the blood. A comparison of the red bone marrow of these animals with that of normal controls revealed a marked reduction in the content of reticulated cells. After a number of transfusions, there occurred in some of the plethoric rabbits a sudden and marked drop in hemoglobin. The hemoglobin continued to fall until a severe grade of anemia was reached. This was followed by an extremely rapid regeneration accompanied by a striking rise in color index. During regeneration, the reticulated cells were enormously increased in number. Taken together, these facts show that the bone marrow is markedly influenced by plethora. The diminished number of reticulated cells observed, both in the circulating blood and in the marrow, would make it appear that a decided decrease in blood production occurs. The reduction in the number of these cells cannot be due to changes in the constitution of the red cells put out by the bone marrow, as a result of an increased quantity of hemoglobin in the body, because during regeneration from the above mentioned anemia, when the color index was very high, reticulated cells were still present in large numbers. That the activity of the bone marrow does actually diminish during plethora is further evidenced by the occurrence of the anemia. The most reasonable explanation of this phenomenon is that the recipient develops an immunity against the blood of the donors, which results in the destruction of the strange cells that are in circulation. In keeping with this conception is the appearance of isoagglutinins for the donors' red cells in the blood of the recipient, at about the time of the beginning fall in hemoglobin. The occurrence of anemia as a result of the destruction of the alien blood only would seem to be due to the circumstance that, during the period of plethora, blood production is greatly diminished; as a consequence, the blood cells proper to the recipient are gradually reduced in number and replaced by alien cells until the latter come to constitute the bulk of the animal's blood. In those rabbits developing anemia, the initial drop of hemoglobin from the plethoric level to the normal was constantly accompanied by a marked rise in the number of reticulated cells. This brought up a subsidiary problem for study. With the idea that the stimulation of the bone marrow might be due to the presence of an increased quantity of broken down blood, rabbits, were injected intravenously with large amounts of laked blood cells. The procedure had no evident effect on the blood picture. It was then found that simple blood removal from a plethoric animal which brought back the hemoglobin to the normal level, or even to a point somewhat above, sufficed to cause a marked increase in the number of reticulated cells. Although these findings are not conclusive, they suggest an explanation for the increased bone marrow activity accompanying the initial drop of hemoglobin in the plethoric rabbits; namely, that the organism had in some way adapted itself during the period of plethora to the presence of a greater amount of blood and that the result of blood loss in such an organism was a relative but not absolute anemia. The finding that the activity of the bone marrow can be depressed by the introduction of a large quantity of blood into the circulation accounts for the diminished bone marrow activity which sometimes occurs after transfusion in pernicious anemia. In such cases there is a marked drop in the number of reticulated cells and other evidence of bone marrow depression; the patient shows no benefit from transfusion or may grow rapidly worse. The cause of this depression is best explained on the basis that in severe instances of the disease where exhaustion of the bone marrow is imminent, the stimulus of the anemia is only just sufficient to keep the marrow functioning. A sudden lowering of this stimulus is brought about by the introduction of a large quantity of blood into the circulation, and the result is a fall in the activity of the bone marrow. It follows from this that in pernicious anemia with a feebly reacting bone marrow as indicated by the number of reticulated red cells, small transfusions are preferable to large ones.

Further studies on the epidemiology of lobar pneumonia.

Abstract: 1. Pneumococci of Type I and Type II are responsible for the majority of the cases of lobar pneumonia. 2. Among the pneumococci found in the mouths of healthy individuals Type IV predominates, Type III is frequent, and atypical organisms of Type II are occasionally found. 3. Healthy persons intimately associated with cases of lobar pneumonia may harbor in their mouth secretions the highly parasitic pneumococcus of Types I and II. 4. Occasionally a carrier of Type I or Type II pneumococcus is encountered in whom it is impossible to trace any contact with an infected patient. 5. From the dust of homes where cases of pneumonia due to Types I and II have occurred, pneumococci of the same type may be recovered.

The relation of the meninges and choroid plexus to poliomyelitic infection

Abstract: Among the mechanisms which defend the body from infection with the virus of poliomyelitis is the meningeal-choroid plexus complex, which normally is capable of excluding the circulating virus from the central nervous organs. The complex plays a part also in preventing infection from virus present upon the nasal mucosa. Aseptic fluids which irritate, inflame, or even slightly alter the integrity of the meninges and choroid plexus diminish or remove their protective function. Normal monkey or horse serum, isotonic salt solution, and Ringer's and Locke's solutions, when injected into the meninges, promote infection with the virus of poliomyelitis introduced into the blood, the nose, or the subcutaneous tissues. Simple lumbar puncture and the withdrawal and return of the cerebrospinal fluid in normal monkeys, hemorrhage having been absolutely avoided, do not promote infection with virus injected into the blood; while the replacement of the cerebrospinal fluid of one monkey with that of another does in some instances lead to infection. Simple lumbar puncture attended with even very slight hemorrhage opens the way for the passage of the virus from the blood into the central nervous tissues, and thus promotes infection. Hence, changes in the structure or function of the meningealchoroid plexus complex, too slight to be detected by chemical and cellular changes in the cerebrospinal fluid or by morphological alterations, suffice to diminish in an essential manner its protective powers. Of all the irritant fluids tested, immune serum alone injected into the meninges is not succeeded by infection from the virus introduced into the blood. The protective property of the immune serum is capable of overcoming the promoting action of normal monkey and horse serum and the other irritants mentioned. The importance first of the meningeal-choroid plexus complex in preventing infection with the virus of poliomyelitis, and next of immune serum in offsetting the disadvantages and dangers arising from defects in the mechanism is apparent, as is the bearing of the experiments reported on the serum therapy of epidemic poliomyelitis.

Toxin and antitoxin of and protective inoculation against bacillus welchii.

Abstract: Five cultures of Bacillus welchii have been studied and compared Four came from infected wounds in the western theatre of war, and one was obtained from a personal article of clothing. Each culture possesses the essential characteristics ascribed to that group of bacteria. The infectious processes caused by the five cultures in rabbits, guinea pigs, and pigeons, are local in character; and very few or no bacilli enter or are found in the general blood stream during life or immediately after death. Glucose broth cultures, injected intravenously, are fatal to rabbits. Death occurs, almost immediately or after a few hours. Agglutinative bacterial emboli have been ruled out as the cause of death, as has been an acid intoxication. The fluid part of the culture acts in the same manner as the full culture and irrespective of neutralization with sodium hydroxide. The full cultures and supernatant fluid are hemolytic when injected directly into the circulation of rabbits and pigeons, and the acute death produced may be ascribed to a massive destruction of red corpuscles. The passage of the fluid portion of glucose broth cultures through Berkefeld filters reduces materially the hemolytic and poisonous effects. Cultures of the Welch bacilli in plain broth to which sterile pigeon or rabbit muscle is added are highly toxic, and the toxicity is not noticeably diminished by Berkefeld filtration. The filtrates are hemolytic when injected intravenously and inflaming and necrotizing when injected subcutaneously and intramuscularly. The local lesions produced in the breast muscles of the pigeon closely resemble those caused by infection with the bacilli. The toxicity of these filtrates is not affected by neutralization with sodium hydroxide, but is materially reduced by heating to 62°C. and entirely removed by heating to 70°C. for 30 minutes. Successive injections of carefully graded doses of this toxic filtrate in pigeons and rabbits give rise to active immunity. The blood taken from the immunized rabbits is capable of neutralizing the toxic filtrate in vivo and in vitro. The filtrate has therefore been designated as toxin and the immune serum as antitoxin. The antitoxin neutralizes the toxin in multiple proportions. Hence the latter would seem to possess the properties of an exotoxin. Moreover, it neutralizes the hemolytic as well as the locally .injurious toxic constituent. Antitoxic serum prepared from a given culture of Bacillus welchii is neutralizing for the toxins yielded by the other four cultures of that microorganism. The antitoxin is protective and curative against infection with the spore and the vegetative stages of Bacillus welchii in pigeons. The limits of the protective and curative action are now under investigation.

The transplantation of splenic tissue into the subcutaneous fascia of the abdomen in rabbits

Abstract: We have not found in the literature a report of an instance of permanent homoio or autotransplantation of the spleen, or of the probably closely related spleno and hemolymph glands. Spleen autotransplants with considerable difficulty as compared with thyroid, parathyroid, ovary, or adrenal cortex. This may be due to its complex anatomical structure. An instance of a permanent autotransplant has been observed. None of our attempts to homoiotransplant it were successful beyond the usual taking and persistence for 2 or 3 weeks, common to all homoiografts. The successful permanent subcutaneous autotransplantation had all the morphological characteristics of a fully differentiated and functionally active spleen. This method of transplantation would seem to offer a means of learning more of the normal development, regeneration, and function of this complex tissue.

Experiments on the causation and amelioration of adrenalin pulmonary edema.

Abstract: The intratracheal injection of one moderate dose of adrenalin in rabbits whose vagi are divided produces a marked pulmonary edema in a large percentage of cases. The same dose in normal animals causes only slight effects. Artificial respiration greatly reduces the production of pulmonary edema in vagotomized rabbits after adrenalin. As adrenalin can exert a bronchoconstrictor effect, evidence is submitted to show that the aspirating action of the lung alveoli under this condition apparently plays an important part in the production of adrenalin pulmonary edema. On the basis of this mechanism the protective action of artificial respiration is explained. Stenosis of the trachea facilitates. the production of adrenalin pulmonary edema in rabbits whose vagi are intact. The intratracheal injection of adrenalin in vagotomized rabbits produces a temporary incoordination between the heart ventricles, visible on inspection, so that the left ventricle beats apparently half as fast as the right, causing hyperemia of the lungs and hemorrhages. Atropine injected intratracheally in vagotomized rabbits exerts a protective action against adrenalin pulmonary edema.

The cyclic changes in the mammary gland under normal and pathological conditions : i. the changes in the non-pregnant guinea pig.

Abstract: 1. A definite cycle exists in the mammary gland of the non-pregnant guinea pig which corresponds to the cycle in the ovary and uterus. This cycle can be presented through a curve in which the ordinates represent the degree of activity of the gland in a series of animals, and the abscissae the time since ovulation (period of sexual cycle). The curve passes through a first maximum at the time of heat and ovulation and gradually falls. The minimum is reached on the 6th day and continues until the 15th day after ovulation. Next begins the period when a new ovulation is imminent and the number of the proliferating glands again increases. We see. then that during the normal cycle the presence of well preserved, functioning corpora lutea does not lead to proliferation, neither do mature follicles have such an effect. On the other hand, the absence or degeneration of the corpora lutea is required to insure the proliferation of the mammary gland in the first period of the sexual cycle. If the sexual period is experimentally prolonged, we find in some instances proliferation, while in others it is absent. As far as we can determine at the present time, two factors seem to favor proliferation of the mammary gland under these conditions: (1) the presence of well preserved corpora lutea, particularly if they are associated with well preserved experimentally produced deciduornata, and (2) the imminence of a new period of heat. The connection between good corpora lutea and good deciduornata and the presence of proliferating mammary glands at this stage of the sexual cycle is, however, not absolute. There are cases in which a proliferating gland is associated with some degeneration of the corpus luteum. Or on the other hand a well preserved corpus luteum is associated with a non-proliferating gland. In some of the latter cases the simultaneous presence of a necrotic deciduorna may perhaps explain the lack of proliferation in the mammary gland. However, in the majority of cases we found the presence of good corpora lutea and good deciduomata associated with a proliferating mammary gland. Whether a living corpus luteum as such is able to produce proliferation of the gland is as yet doubtful. 2. Extirpation of the ovaries prevents not only the proliferation of the mammary gland associated with the first stage of the sexual cycle, the condition of heat and ovulation no longer taking place in castrated animals, but in all probability also inhibits the proliferation of the mammary gland which occurs under certain conditions towards the end of the sexual cycle, or in instances of experimentally prolonged sexual cycle in which well preserved corpora lutea and deciduornata are present. 3. In animals in which the ovaries were hypotypical, the mammary glands were in an inactive condition. The presence of hypotypical ovaries has the same influence on the mammary gland as castration. In the majority, but not in all of these cases well preserved corpora lutea were absent. 4. Complete extirpation of the corpora lutea seems directly or indirectly to prevent the secondary proliferation of the mammary gland, which occurs during the latter part of the sexual cycle or during an experimentally prolonged cycle, in cases in which the extirpation is not followed at once by a new ovulation. This conclusion we consider, however, merely as suggested, not yet as definitely established through our results. On the other hand, the primary proliferation of the mammary gland, during the first stage of the sexual cycle, as well as ovulation and the objective signs of heat, is accelerated through complete extirpation of the corpora lutea. Thus the effect of extirpation of the corpora lutea differs from the effect of castration, in that after the latter neither a new heat nor the primary proliferation of the mammary gland occurs. As one of the authors has pointed out previously, the absence of functioning corpora lutea and the presence of either well developed ovarian follicles or of mature follicles are necessary for the occurrence of heat and ovulation. The same conditions are prerequisites for the primary proliferation of the mammary gland. 5. In cases in which the whole or almost the whole uterus had been extirpated, the corpora lutea were well preserved and the mammary gland was proliferating.

Carriage of the virus of poliomyelitis, with subsequent development of the infection

Abstract: A family group containing four children of whom all showed in varying degree symptoms of poliomyelitis is described. The source of infection and periods of incubation have been followed. Two of the children were proven by inoculation tests to carry the virus of poliomyelitis in the nasopharynx. Of these, one was detected to be a carrier after recovering from a non-paralytic attack of the disease, and the other was discovered to be a carrier about 5 days before the initial symptoms, attended later by paralysis, appeared. The original case from which the three others took origin was fatal; the youngest child, after quite a severe onset, was treated with immune serum, and made a prompt and almost perfect recovery. The nasopharyngeal secretions of two of the cases, taken 1 month after the attack, proved incapable of neutralizing an active poliomyelitic virus. The proposition is presented that every case of poliomyelitis develops from a carrier of the microbic cause, or virus, of poliomyelitis.

Experimental studies on the relation of the pituitary body to renal function.

Abstract: 1. The inconstant results of past observations on the relation of pituitary extracts to renal activity have been due chiefly to unsuitable methods. 2. A standard curve of artificially induced polyuria may be plotted for rabbits, giving 200 cc. of water by mouth. 3. Extracts of the pars intermedia and posterior lobe of the hypophysis, given by mouth, subcutaneously, or intravenously, are able definitely to check polyuria thus induced. Extracts of the anterior lobe show a similar effect, but only to a slight degree. 4. This antidiuretic effect is constant, and is independent of (a) changes in blood pressure, (b) intestinal absorption, and (c) the vagi. The effect is apparently prevented or delayed by division of the splanchnics, and is diminished by division of the renal nerves near the hilus. 5. A similar antidiuretic property is possessed: (a) by β-imidazolylethylamine, (b) by p-oxyphenylethylamine, (c) by a preparation from Secale cornutum, (d) by small doses of nicotine, (e) by large doses of caffeine, and (f) by extracts of the adrenal cortex. 6. No effect on the polyuria is produced: (a) by strychnine (b) by morphine, (c) by adrenalin, or by extracts of (d) thyroid, (e) thymus, (f) pineal, (g) pancreas, or (h) corpora lutea. 7. In animals under chloral or paraldehyde anesthesia a short and inconstant initial increase in flow of urine is seen. 8. The antidiuretic effect is absent or only slightly marked in checking the so called salt diuresis.

The pathologic effects of streptococci from cases of poliomyelitis and other sources.

Abstract: Streptococci cultivated from the tonsils of thirty-two cases of poliomyelitis were used to inoculate various laboratory animals. In no case was a condition induced resembling poliomyelitis clinically or pathologically in guinea pigs, dogs, cats, rabbits, or monkeys. On the other hand, a considerable percentage of the rabbits and a smaller percentage of some of the other animals developed lesions due to streptococci. These lesions consisted of meningitis, meningo-encephalitis, abscess of the brain, arthritis, tenosynovitis, myositis, abscess of the kidney, endocarditis, pericarditis, and neuritis. No distinction in the character or frequency of the lesions could be determined between the streptococci derived from poliomyelitic patients and from other sources. Streptococci isolated from the poliomyelitic brain and spinal cord of monkeys which succumbed to inoculation with the filtered virus failed to induce in monkeys any paralysis or the characteristic histological changes of poliomyelitis. These streptococci are regarded as secondary bacterial invaders of the nervous organs. Monkeys which have recovered from infection with streptococci derived from cases of poliomyelitis are not protected from infection with the filtered virus, and their blood does not neutralize the filtered virus in vitro. We have failed to detect any etiologic or pathologic relationship between streptococci and epidemic poliomyelitis in man or true experimental poliomyelitis in the monkey.

The cyclic changes in the mammary gland under normal and pathological conditions : ii. the changes in the pregnant guinea pig, the effect of lutein injections, and the correlation between the cycle of the uterus and ovaries and the cycle of the mammary gland.

Abstract: 1. In the pregnant guinea pig proliferation of the mammary gland becomes regular only at a later stage of pregnancy; namely, during the period following the 24th day of pregnancy. Previous to this period proliferation was absent in the majority of cases. Proliferation of the mammary gland during pregnancy becomes regular only at a period of time which exceeds the duration of the normal sexual cycle unaccompanied by pregnancy. It is probable that pregnancy as well as the presence of living deciduornata and corpora lutea increases the proliferative activity of the mammary gland as compared with the ordinary cycle in non-pregnant animals or in animals lacking corpora lutea and deciduornata. 2. After the completion of pregnancy and in the beginning of secretion some mitotic proliferation may still be present, but it soon ceases, probably as the result of those processes that lead to secretion. While during the period of secretion, notwithstanding the presence of a new pregnancy, mitotic proliferation soon ceases, some proliferative stimulus seems still to be active, which, however, under existing conditions apparently leads only to a mitotic multiplication of nuclei. The latter conclusion is only suggested at the present time and needs confirmation through further studies. 3. In cases in which abortion took place in the first half of pregnancy secretion in the gland was not established; secretion occurred in two animals aborting toward the latter part of pregnancy. In one of these cases, mitotic proliferation of some gland cells was associated with the microscopic appearances of secretion. 4. In guinea pigs castrated during an early period of pregnancy in which pregnancy continued for some time, proliferative changes were absent in the mammary gland. In conjunction with a partial similar effect observed after extirpation of the corpora lutea during pregnancy, we may perhaps attribute the lack of proliferation in some of these cases to the absence of the ovaries. 5. Extirpation of the corpora lutea during pregnancy induces a new ovulation and with it the primary proliferation in the mammary gland; abortion does not necessarily prevent these proliferative changes. Extirpation of the corpora lutea during pregnancy perhaps prevents the secondary proliferative changes in the mammary gland. 6. Five injections of cow's lutein given in relatively large quantities intraperitoneally do not produce proliferation of the mammary gland in the guinea pig.

STUDIES ON THE METABOLISM OF CELLS IN VITRO : I. THE TOXICITY OFalpha-AMINO-ACIDS FOR EMBRYONIC CHICKEN CELLS.

Abstract: Summing up these results, we found that all the ten α-amino-acids used inhibited the growth of the cells and finally killed the cultures. This inhibition is preceded by a short period of activity. The typical effect on the cells is shown in Figs. 1 and 2. The first (Fig. 1) is a control culture showing the usual growth of cells and their typical spindle shape form. The second (Fig. 2) is a culture in plasma plus asparagine showing the cells rounded off and beginning to undergo dissolution. We do not wish to draw too extensive conclusions from these experiments, but we believe that toxicity of α-amino-acids towards growing cells has been shown beyond a reasonable doubt; while we have found that compounds of higher molecular weight, namely, the peptones of egg yolk and proteins, are non-toxic. This toxicity depends upon the concentration and the time that the cells are exposed to their action. As these factors are reduced, the toxicity is decreased. In this respect, these substances are similar to all cell poisons. Applying these results to the work done on the intravenous injection of digestion mixtures, we believe that we have found a reason for the death of the experimental animals when the hydrolyzed proteins were injected too rapidly. Buglia found that large amounts of α-amino-acids could be injected into the circulation without causing deep-seated changes in the renal and intestinal functions, provided they were injected slowly enough; in fact, that enough of these mixtures could be injected in this way to cover the nitrogen consumption of the body. This injection, however, was always accompanied by an α-amino excretion through the urine and an increase of the peristalsis, of the intestine, with resultant liquid stools. As is well known, a sudden great concentration of these substances in the blood of an animal causes death. These results agree with our findings Folin and Denis demonstrated the fact that α-amino-acids probably pass into the circulation through the intestines. Van Slyke and Meyer, by means of Van Slyke's nitrogen method, have practically proven this, and Abel, Rowntree, and Turner, and Abderhalden have lately succeeded in obtaining α-amino-acids in crystalline form from the blood. Van Slyke and Meyer have shown that the tissues take up α-amino-acids to a certain point, but that after this the limit of saturation is reached. This is not so in the liver, which continually desaturates itself by metabolizing the α-amino-acids that it has absorbed, and consequently maintains indefinitely its power of removing them from the circulation, as long as they enter it no faster than the liver can metabolize them. Marshall and Rowntree have shown that there is an increase of the α-amino-acid concentration in the blood after injuries to the liver, which have caused deep-seated anatomical changes. Our experiments prove that tissue cells in general are unable to live in the presence of any great concentration of these acids. At the present time we do not feel able to give an explanation of the significance of this evident toxicity. However, the fact in itself seems to indicate that we should expect stimulation from a certain increase of the α-amino-acid concentration in the body, or the concentration of any one of the acids, while a greater increase would lead to marked disturbances of the metabolism.

Experimental studies upon lymphocytes : ii. the action of immune sera upon lymphocytes and small thymus cells.

Abstract: The work of previous investigators gives the impression that it is easy to produce sera which both in vitro and upon injection are leukotoxic. At the same time the specificity of these leukotoxic sera for the particular type of cell used as antigen, and even for leukocytes in general, has been doubtful. The methods used have made certain possible factors of error unavoidable. Even careful washing of an organ or suspension cannot render it wholly blood-free, so that it is not surprising that the sera should be moderately hemolytic and hemagglutinative. Pearce has shown that the injection of very small amounts of blood is sufficient to evoke the production of immune hemolysins. When such sera are injected the lesions, as Pearce states, may be due in part to the production of hemagglutinative thrombi, although this hardly seems to apply to the changes in lymphoid tissue described by Flexner. On the other hand, the lymphotoxic effect of hemolytic sera may be due to the lymphocytes injected with the red cells. Our own experiments indicate that the lymphotoxic and agglutinative factors are to a considerable degree distinct from the hemolytic and hemagglutinative ones, since they can be separated from one another by absorption. Further evidence is presented that the small thymus cells are biologically related to, if not identical with the lymphocytes derived from lymph glands.

Neutralization of the virus of poliomyelitis by nasal washings.

Abstract: 1. The results of 56 experiments have shown that washings of the nasal and pharyngeal mucosas possess definite power to inactivate or neutralize the active virus of poliomyelitis. 2. This power is not absolutely fixed, but is subject to fluctuation in a given person. Apparently inflammatory conditions of the upper air passages tend to remove or diminish the power of neutralization. But irregularities have been noted, even in the absence of these conditions. 3. Too few tests have been made thus far to ascertain whether adults and children differ with respect to the existence of this neutralizing property in the nasal secretions. While the inactivating property was absent from the secretions of one child during the first days of poliomyelitis, it was present in another to whom immune serum was administered, and in still another on the 15th day of illness when convalescence was established. 4. The neutralizing substance is water-soluble and appears not to be inorganic; it appears to be more or less thermolabile, and its action does not depend upon the presence of mucin as such. 5. It is suggested that the production of healthy carriers through contamination with the virus of poliomyelitis may be determined by the presence or absence of this inactivating or neutralizing property in the secretions. Whether this effect operates to prevent actual invasion of the virus and production of infection can only be conjected. Probably accessory and not the essential element on which defense against infection rests. It is more probable that other factors exist which help to determine the issue of the delicate adjustment between contamination and infection.

Spirochaeta morsus muris, n.sp., the cause of rat-bite fever : second paper.

Abstract: 1. Since our first report on the discovery of the cause of rat-bite fever, we have been able to prove the existence of the same spirochete in five out of six more cases which have come under our observation. 2. The clinical symptoms of rat-bite fever are inflammation of the bitten parts, paroxysms of fever of the relapsing type, swelling of the lymph glands, and eruption of the skin, all occurring after an incubation period usually of from 10 to 22 days, or longer. 3. Our spirochete is present in the swollen local lesion of the skin and the enlarged lymph glands. But as the spirochetes are so few in number it is exceedingly difficult to discover them directly in material taken from patients. It is therefore better to inoculate the material into a mouse. In some cases the organism is found in the blood of the inoculated animal after a lapse of 5 to 14 days, or at the latest 4 weeks. 4. Generally speaking, the spirochetes present thick and short forms of about 2 to 5 µ and have flagella at both ends. Including the flagella, they measure 6 to 10 µ in length. Some forms in the cultures reach 12 to 19 µ excluding the flagella. The curves are regular, and the majority have one curve in 1 µ. Smaller ones are found in the blood and larger ones in the tissues. 5. The spirochetes stain easily. With Giemsa's stain they take a deep violet-red; they also stain with ordinary aniline dyes. The flagella, too, take Giemsa's stain. 6. The movements of our spirochetes are very rapid, resembling those of a vibrio, and distinguish them from all other kinds of spirochetes. When, however, the movements become a little sluggish, they begin to present movements characteristic of ordinary spirochetes. 7. For experimental purposes, mice, house rats, white rats, and monkeys are the most suitable animals. Monkeys have intermittent fever after infection, and spirochetes can be found in their blood, but they are not so numerous as in the blood of mice. Mice are the most suitable animals for these experiments, and they appear, as a rule, to escape fatal consequences. 8. The spirochete is markedly affected by salvarsan. 9. The organism is not present in the blood of all rats, and there is no relation between the species of the rat and the ratio of infection. We have never found the spirochete in healthy guinea pigs or mice. By permitting a rat infected with the spirochete to bite a guinea pig, the latter develops the disease. 10. We have succeeded in cultivating the spirochete in Shimamine's medium. 11. Among the spirochetes described in the literature or discovered in the blood of rats and mice, there may be some resembling our spirochete, but none of the descriptions agree with it fully. Hence we have named our organism Spirochaeta morsus muris and regard it as belonging to the Spironemacea (Gross) of the nature of treponema. 12. The spirochete can be detected in the bodies of patients. In seven cases out of eight, it disappears on recovery, only to reappear during the relapse. 13. The spirochete can be detected in about 3 per cent of house rats. These facts enable us to identify the cause of the disease. 14. There may be other causes than the spirochete for diseases following the bite of a rat. The cause, however, of rat-bite fever in the form most common in Japan is, we believe, the spirochete which we have described.

The drug-fastness of spirochetes to arsenic, mercurial, and iodide compounds in vitro.

Abstract: In the foregoing experiments we attempted to determine whether or not, by subjecting several varieties of spirochetes to increasing doses of certain chemotherapeutic agents, a gradual increase of resistance to the latter could be shown. For this purpose, pure cultures of Treponema pallidum, Treponema microdentium, and Spirochœta refringens were used against the action of salvarsan, neosalvarsan, bichloride of mercury, and iodine-iodide potassium solution in vitro. For culture media, the usual ascites-broth-tissue medium as well as solid ascites-agar-tissue medium was used. After permitting the spirochetes to grow for a fortnight in media containing certain quantities of each drug, transfers were made from tubes showing various degrees of growth to the next series of tubes containing the same drug in still higher concentrations, and similar transfers repeated every 2 weeks. The results of the experiments may be briefly summarized as follows: 1. Treponema pallidum and Treponema microdentium have, within 3 to 4 months, increased their tolerance to salvarsan and neosalvarsan to five and one-half times their original mark. With Spirochata refringens the increase was about three times. 2. Against the action of bichloride of mercury, the amount of increased tolerance of Treponema pallidum was about 35 to 70 times the original, while that of Treponema microdentium was about 10 times as much and was reached within 10 weeks. Spirochata refringens resisted 30 times the original dose. 3. There was an unmistakable increase of resistance of these spirochetes to the action of the iodine-iodide solution (Lugol's solution) when they were grown for several generations in fluid media containing the iodine solution, but the rate of increase between the initial and the acquired tolerance was slight. In general, the addition of Lugol"s solution to fluid media has a weak inhibitory influence upon the growth of the spirochetes, requiring for the total suppression of growth a quantity of over 0.7 cc. to 5 cc. of the culture media. The tolerance reached was for about three times that amount. 4. A similar tolerance phenomenon has not been established when employing a solid instead of a fluid medium containing the drugs. No explanation is offered except a suggestion that the drugs held in the agar do not enter into combination with certain tissue constituents of the medium as they are able to do with tissue elements in fluid media. This may be a factor necessary for inducing drug tolerance in these organisms in vitro. 5. The increased drug-fastness in vitro has a limit beyond which no further advance can be made. This limit varies with different species of spirochetes. 6. The acquired drug-fastness in vitro gradually disappears when the spirochetes are cultivated again in the drug-free media for several generations.

The production of antipneumococcic serum

Abstract: In the production of immune serum for therapeutic purposes strict attention must be paid to the immunological specificity of the bacteria used for immunization. At present the only serum of which the therapeutic value has been proven is that effective against Type I pneumococcus infection. This serum should have agglutinating power for Type I pneumococcus and should have the power of protecting mice against large amounts of virulent culture. Experiments have shown that for producing the primary immunity most rapidly several series of small doses of dead cultures should be given, the injections being made daily for 6 to 7 days, followed by a week in which no injections are made. To produce the highest type of immunity probably living organisms are required. These should be given in moderate doses daily for 3 days, with an interval of a week between each series of injections. By following accurately the methods described, horses may be made to produce rapidly a high grade of specific serum. The observations so far made indicate the importance of employing small doses of culture frequently repeated in this form of immunization.

Pulmonary embolism : an experimental study.

Abstract: Emboli made of paraffin and the animal's own blood were sent into the venous circulation of dogs. Death did not occur until the pulmonary circulation was practically occluded. The results were the same whether the blood pressure of the animal was normal or depressed by ether or disease and whether the procedure was carried out under ether or local anesthesia.

The effect of alcohol on the reproductive tissues

Abstract: Administration of alcohol in the food of male white rats for 2 or more months, in daily quantities of 025 to 225 cc, results almost constantly in the appearance of marked degenerative alterations in the testicles These changes affect the steps of spermatogenesis in inverse order to their occurrence, so that for some time before sterility and complete aspermia result, the animal is producing spermatozoa with all possible degrees of abnormality and deficiency The possible relation of this abnormal spermatogenesis to the production of defective offspring is obvious Individual rats show marked differences in the degree of change produced by equal amounts of alcohol The fibrous, interstitial, and vascular elements of the testicle are not affected, except for intertubular edema compensating for tubular atrophy These experimental observations harmonize with the necropsy findings in human alcoholics No other tissue was found to be noticeably affected by the alcohol; especially to be remarked is the absence of cirrhosis or fatty infiltration in the liver

A report on the serum treatment of twenty-six cases of epidemic poliomyelitis

Abstract: 1. Serum taken from recently recovered cases of poliomyelitis may be employed in its treatment and probably yields the best results. 2. When sterile for ordinary bacteria, free of corpuscles and hemoglobin, and when injected by the gravity method, observing well known rules of caution, it may be employed without danger. 3. The serum should be injected both intraspinally and intravenously, the latter either directly or by way of the subcutaneous tissues. 4. The earlier in the course of the disease the serum is employed in suitable doses, the more promise there is of benefit. 5. The action of the serum appears to be more precise and definite in arresting paralysis than in rapidly bringing about its retrogression. 6. The decision to employ the serum should rest upon a clinical examination supported by the results of the microscopic and chemical study of the cerebrospinal fluid. 7. The question of multiple and repeated injections of the serum has not yet been worked out. In the cases here reported and especially in the group in which no paralysis existed at the time of the first injection, the pathologic process either did not progress at all, or where there was extension, as in Cases 14 and 15, the upper segment of the spinal cord became rapidly involved, and was followed by respiratory paralysis and death. Probably in cases in which some degree of muscular weakness develops soon after the injection of serum, reinjection 12 to 24 hours later may be advantageous. The temperature curve may serve to indicate the time for reinjection. 8. The favorable results thus far achieved in human beings by means of the immune serum support and extend those obtained experimentally in monkeys and indicate, as was foreseen, that the milder or less fatal form of poliomyelitis appearing in man is even more amenable to the serum treatment than is the highly fatal disease produced by inoculation in monkeys.

The focal pulmonary tuberculosis of children and adults.

Abstract: Evidence of tuberculous infection has been found in the lungs of all of fifty adults who have been examined. Approximately one-half of all adults have encapsulated lesions of the lungs or bronchial lymphatic nodes, whereas in one-third pulmonary and lymphatic lesions are firmly calcified and completely healed. Tuberculous pulmonary lesions of adults who have died of diseases other than tuberculosis are of two types: (1) apical tuberculosis similar to the usual type of fatal phthisis and unaccompanied by caseation, of the regional lymphatic nodes; (2) focal tuberculosis not more commonly situated in the apices of the lungs than elsewhere and accompanied by caseation (or calcification) of the adjacent lymphatic nodes. Focal pulmonary tuberculosis of adults is identical with the tuberculosis of childhood. It occurs in at least 92 per cent of all adults. It may be acquired between the ages of 2 and 10 years but in more than half of all individuals (in this city) makes its appearance between the ages of 10 and 18 years. Tuberculosis of children does not select the apices of the lungs, is accompanied by massive tuberculosis of regional lymphatic nodes, and exhibits the characters of tuberculosis in a freshly infected animal, whereas tuberculosis which occurs in the pulmonary apices of adults has the characters of a second infection. Almost all human beings are spontaneously "vaccinated" with tuberculosis before they reach adult life.

A study of the acidosis, blood urea, and plasma chlorides in uranium nephritis in the dog, and of the protective action of sodium bicarbonate

Abstract: 1. The presence of an acidosis in dogs with experimental uranium nephritis is demonstrable by the Van Slyke-Stillman-Cullen method and that of Marriott. It is detected more readily by the former method. 2. This acidosis is associated with increase in the blood urea and plasma chlorides and with the appearance of albumin and casts in the urine. 3. The oral administration of sodium bicarbonate diminishes the acidosis, the increase in plasma chlorides, the amount of albumin and casts in the urine, and, to a lesser degree, the increase in the blood urea following the administration of uranium. It also diminishes the severity of the changes produced by uranium in the kidneys. 4. The oral administration of sodium bicarbonate to normal dogs raises the carbon dioxide content of the plasma as determined by the. Van Slyke-Stillman-Cullen method.

The normal fate of erythrocytes : ii. blood destruction in plethoric animals and in animals with a simple anemia.

Abstract: 1. The increased destruction of red cells in animals rendered plethoric by transfusion takes place predominantly by a fragmentation of the corpuscles without loss of hemoglobin. 2. The microcytes and poikilocytes observed in animals with a severe anemia due to hemorrhage are not put forth as such by the bone marrow, but are portions of cells fragmented while circulating. 3. The cells thus fragmented are for the most part those newformed to meet the exigencies of the situation. Such cells are in large part unable to withstand the wear and tear of function. There results a vicious circle. The anemia renders the bone marrow unable to put forth proper cells, and those it does produce are soon destroyed, thus prolonging the condition. A similar state of affairs probably exists in many human anemias. 4. The occurrence of large accumulations of microcytes and poikilocytes in the spleen of anemic and plethoric animals indicates that the organ exercises some important function in connection with these forms. The same is true of normal animals, for the findings in them are similar, though less striking. 5. The normal fate of the red corpuscles, in those species in which phagocytosis is negligible, is to be fragmented one by one, while still circulating, to a fine, hemoglobin-containing dust. The cell fragments are rapidly removed from the blood, but their ultimate fate remains to be determined. The facts indicate that they are removed from the blood by the spleen, and under exceptional conditions, by the bone marrow.