Moyer S. Fleisher

The relative importance of stroma and parenchyma in the growth of certain organs in culture media

In experiments carried out about fifteen years ago, one of us observed that during the regeneration of skin, the epithelial cells are able to penetrate into and to grow in coagula of blood and of blood plasma. 3 This suggested to him that it might be possible to make various tissues grow in culture media outside of the body, in the thermostat, as well as inside the body, in the latter case the body acting as a thermostat. Inasmuch as he noted that the epithelial and also connective tissue cells grew preferably in contact with solid structures as fibres of fibrin, and into solid gelatinous material rather than into fluids he attributed stereotropic sensitiveness to various tissue cells, and he consequently employed more or less solid culture media as agar and coagulated blood serum for his various experiments. At first he carried out experiments in vitro as well as experiments in which the animal body acted as an incubator. Lack of the necessary facilities made it very soon necessary for him to limit himself to the latter kinds of experiments. 1 To our knowledge in these our earlier experiments for the first time the attempt was recorded in the literature to grow tissues of higher animals under artificial conditions in environments that differ from those found in the body under natural conditions, to separate experimentally growing epithelial from connective tissue cells, and furthermore to study the influence of the addition of certain chemicals upon the growth of tissues. 2

Effects of inoculating monilia isolated from psoriatic patients into human beings.

It has been shown by Fleisher and Wachowiak 1 that monilia or monilia-like organisms were found in 85 per cent of the stools of psoriatics, that similar or identical organisms could be cultivated or demonstrated in the skin scrapings in 35 per cent of cases, and that these organisms were present in the blood in 14 per cent of cases. However, in normal individuals such organisms were found in only 6 per cent of the stools, and in our series never on the skin or in the blood. Furthermore, the injection of an emulsion of the killed organisms led, in a certain number of intractable cases, to a startling and rapid clearing of the lesions. Efforts had also been made to reproduce the disease (skin lesions) in rabbits, guinea pigs and dogs, but without success. We believed that this evidence suggested very strongly that there exists some etiological relationship between the monilia and the occurrence of psoriatic lesions. Recently I have studied the effects of applying the monilia isolated from cases of psoriasis to abraded surfaces of the skin of human beings. In normal individuals, two small areas, 2 to 3 mm. square on the upper arm were denuded of the superficial layers of epidermis. One area served as control and this healed rapidly, king practically indistinguishable from the surrounding uninjured skin, after about eighteen to twenty days; at no time was ldesquamation or scaling noted on or about these control areas. To the other areas there was applied a drop of a mixed emulsion of live monilia. Here a scab formed which persisted for about two weeks, and usually during this period there was a varying degree of itching.

Mononuclear Leucocytes in Blood of Guinea Pigs Experimentally Infected with Pneumococcus

In view of the importance attached by many observers to the rôle of mononuclear cells in recovery from infection and especially the concepts advanced by Robertson and his coworkers 1 2 3 4 in pneumococcal infection in dogs, the changes in the mononuclear cells in the circulating blood have been studied in guinea pigs that died and that recovered spontaneously from experimental pneumococcal infection. The animals were infected by intraäbdominal injection of 0.25 cc of an 18-hour dextrose-serum-broth culture of Type I pneumococcus. Of 100 animals so infected 47 survived the infection. In this study of the mononuclear leucocytes 14 animals that died from 60 hours to 30 days after infection (Group C), 18 that recovered at 5 to 9 days after infection (Group D), and 14 that recovered on the fourth day (Group E) are considered. Recovery was assumed to have occurred on the day a negative culture was obtained from the peritoneal exudate. At 6 hours after infection there was a decrease in the number of circulating mononuclears and this decrease was usually still evident or may have been more marked at 24 hours. The changes were observed in all groups regardless of whether death or survival follows (Table I). In Group C, in those animals that died before the third but after the second day, the mononuclears continued to decrease after 24 hours; this change was concomitant with a decrease of total cells and of neutrophils. 5 Those animals that died on the fourth day showed a further decrease on the second day; however, on the third day these latter animals (which died within the next 24 hours) showed a distinct increase in the mononuclear cells.

Serum Sickness in Rabbits.

It is well known that serum sickness appears in a very large proportion of individuals injected for the first time with normal horse serum or with various antisera. For the experimental study of this reaction it is of value to be able to reproduce in laboratory animals a condition analogous to serum sickness in man. We believe that the results reported below are the first ones concerning the occurrence of serum sickness in rabbits. When a single injection of horse serum is given to rabbits in sufficient quantities either intravenously, intramuscularly, subcutan-eously or into the subscapular tissues, there appears, from 3 to 7 days after the injection, a reaction characterized by erythema and edema. This reaction is evident on the rabbits ears; there is noted a generalized flush which may involve the entire lower three-quarters of the ear, and which may be diffuse, confluent and of even intensity, or diffusely distributed, mottled and with discrete darker patches; the edema appears characteristically in the basal third of the ear but may extend upward to involve the entire lower two-thirds of the ear. The edema is frequently so marked that the pitting subsequent to pressure can be noted visually. The erythema and edema may each occur alone but both are usually present. These reactions can easily be differentiated from those occurring in the normal rabbits ears; here hemorrhages, petechial or more extensive. and small circumscribed patches of erythema may be noted. Such appearances are very common. At times edematous areas often associated with trauma or hemorrhage occur; in the normal animal these latter reactions are usually noted in the upper half or twothirds of the ear, and such areas were found in only 35 of 147 normal animals which were observed for periods of 2 to 8 weeks.

Skin Reacting Substances in Filtrates of Cultures of Eberthellia Typhi

If we inject 0.1 cc. of the filtrate of a broth culture of typhoid bacilli intracutaneously in the arm of a human being, there appears in from 12 to 24 hours a reaction similar to that produced by the injection of Streptococcus scarlatinae toxin. The reaction fades after 24 to 48 hours, but may leave a slight pigmentation. This reaction has been produced with filtrates of 1, 2 and 5-day cultures, being approximately the same with all. The filtrate was obtained by passing the culture through either Berkefeld W, N or V filters, but usually the N filter was used. Several strains of the organism gave similar results. The filtrates varied in activity, and we have obtained reactions with dilutions of even one to two thousand. Usually a dilution of 1 to 500 was used. The reactions produced in normal subjects (who neither had had typhoid fever nor were ever immunized against typhoid fever) are very variable, and while reactions can be produced in all, the area and intensity of the response seem to be individual characteristics. In individuals who had been immunized against typhoid fever, either recently or several years previously, the reactions were similar to and quite as varied as those in the normal subjects. The same statement holds true of individuals who had had typhoid fever some years previous to the injections. No evidence of any ability to inhibit the local reaction was noted in persons who had had typhoid fever, or who had been immunized against typhoid fever. In individuals having typhoid fever, the reaction to the injection of the filtrate was markedly diminished (as compared with weakly reacting normal inclividuals) or was negative.

The Bactericidal Influence of Various Substances Upon Gram-Positive and Gram-Negative Bacteria.

We have found that ether is more rapidly bactericidal in relation to gram-negative organisms (Escherichia coli, Eberthellia typhi, Eberthellia dysenteriae, Eberthellia paradysenteriae (Flexner), Alcaligines fecalis, Salmonella paratyphi, Salmonella Schotmulleri, Pseudomonas aeruginosa, Pseudomonas fluoresoens, Proteus vulgaris, Serratia marcescens, Encapsulatus pneumoniae, Neisseria intracellularis) than in relation to a number of gram-positive organisms (Staphylococcus albus, Staphylococcus aureus, Staphylococcus tetragenus, Diplococcus pneumoniae (Types II and III), Streptococcus pyogenes). It appears at present that acid fast gram-positive organisms (Mycobacteria), diphtheroids (Corynebacteria) and possibly the Lactobacilli, stand midway between the two first mentioned groups in sensitiveness to this reagent. We further tested a number of substances containing hydrocarbon chains and possessing varying degrees of lipoid solvent activity on gram-negative and gram-positive bacteria. Our results are shown in Table I. In general it will be seen that all of these substances act in much the same manner as does ether. From Table 11, it is seen that alkalis act as do the substances containing hydrocarbon chains, but acids act more rapidly on gram-positive than on gram-negative organisms. Finally we tested two inorganic salts which, however, acted with about the same degree of rapidity on both types of organisms. Our results so far do not permit of any definite explanation of these differences in the action of the various agents upon the gram positive and gram negative bacteria.

The effect of the administration of thyroxin upon the surface tension of blood

In a previous communication 1 we have shown that thyroidectomy brings about in the course of nineteen to twenty-two days an increase in the value of the surface tension of the blood of guinea pigs. Since hypothyroidism causes this increase in the surface tension, it was of course of interest to ascertain whether the presence of excessive quantities of thyroid secretion or hormone would have the reverse effect upon the surface tension of the blood. We therefore fed thyroxin to normal guinea pigs. The feeding was continued over a period of three to four days, and from 1 to 1.2 milligrams were fed to each animal. The animals were bled before feeding, and again on the fourth or fifth day after beginning the feedings. Determinations of the surface tension of the blood plasma were made on both of these samples. At the same times normal animals were bled and, in order to have controls of unfed animals, they were kept under the same conditions as the ones used in the experiments. Twenty-seven animals were fed thyroxin; of these, twenty showed a rail in the value of the surface tension of the blood which was greater than 1 dyne. The remaining seven animals should a change of less than 1 dyne, some even showing a slight increase of the surface tension. The fall in the surface tension in the twenty animals varied from 1.9 dynes to 13.9 dynes. The average fall of surface tension between the first and second samples in all twenty-seven animals was 3.7 dynes; if we consider only those which showed a fall greater than 1 dyne, the average for these twenty is 5 dynes. Of the seven control animals kept under the same conditions as the fed animals, six showed variations between the first and second determinations of less than 1 dyne, showing either a rise or a fall; one animal, however, showed a fall of 1 dyne.

THE PATHOGENESIS OF CARDIAC HYPERTROPHY AND MYOCARDITIS

In several former papers 1 we described a simple method which permitted us with great certainty to produce a combination of hypertrophic and myocarditic changes within a relatively short time in the heart of the rabbit. At the same time we suggested an explanation of these phenomena without, however, going into any detailed discussion regarding the basis for our theoretical conceptions. Inasmuch as it appears to us that the principle underlying our explanation of the phenomena observed in the rabbit may, perhaps, be extended to the pathology of the heart in man, we believe that a somewhat more detailed discussion of our theory might be of advantage. The method employed by us in the production of hypertrophic and myocarditic changes in the heart of rabbits was, briefly, as follows: We injected into the earvein of a rabbit a single small dose of epinephrin (0.2 c.c.) preceded by spartein sulphate or

The influence of calcium chloride and of adrenalin upon the secretion of urine and upon absorption from the peritoneal cavity

I. Intraveneous injection of calcium chloride diminishes the secretion of urine. Porges and Pribram ascribed this effect to the lowering of blood pressure which follows the intravenous injection of this substance. Our experiments, we believe, show such an interpretation to be erroneous for the following reason: