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1. A series of 30 constipated subjects were under observation before, during, and after treatment with B. acidophilus.A comparison of these periods shows that the number of normal defecations has been significantly increased during treatment. The usual daily dose was 1,000 c.c. containing 200,000,000 viable B. acidophilusper c.c.2. The beneficial influence of B. acidophilususually persists for a considerable period of time after treatment has been stopped. Patients have been observed for from one week to about one year after treatment and almost without exception all have had more normal defecations after, than before, treatment.3. The use of lactose during and after ingestion of B. acidophilusdoes much to enhance the beneficial effects.4. A transformation of the intestinal flora from a proteolytic to an aciduric type as shown by microscopic and plate counts may generally be induced. Such transformation is usually accompanied by almost daily defecations regardless of the severity of the constipation.5. Two cases of diarrhea have been successfully treated by the ingestion of B. acidophilus.

Since the beneficial effects of B. acidophilusappear to depend upon a transformation of the intestinal flora, it follows that a mass inoculation is desirable. This means a maximum number of viable organisms per c.c. of pabulum. The usual recommendation on commercial preparations of fermented milk is that such milk be kept in a cool place preferably in the ice-box. Consequently, the influence of low temperature on the number of viable B. acidophilusin milk seemed worth investigating.The number of viable B. acidophilusin milk held in the icebox at about 97° C. was determined daily. The viable organisms were killed as follows: after 1 day—about 50 per cent.; after 2 days—about 75 per cent.; after 3 days about 90 per cent., etc. The obvious importance of having a large number of viable organisms is manifested in the necessity for increasing dosage in severe cases. In fact, it is likely that in cases reported as failures, a sufficient increase in the number of viable organisms administered might have resulted in success. The practice of ice-boxing B. acidophilushas little to recommend it beyond preserving the palatability of the culture and it is therefore more desirable to keep B. acidophilusat room temperature.A study was made of the influence of time and pressure in autoclaving milk, prior to inoculation with B. acidophilur, with respect to the growth of the organism. Milk was sterilized at 15 and 20 pounds pressure in the autoclave for different periods of time and inoculated with B. acidophilus. Subcultures were made from each set of flasks for 3 days on milk identically sterilized, thus obtaining acclimatization. On the basis of these comparative tests it was found that milk to be used for inoculation with B. acidophilus should be sterilized in the autoclave at 20 pounds pressure for 20 minutes, or at 15 pounds pressure for 20 to 30 minutes.

Having obtained relief from chronic constipation and diarrhea by treatment with milk fermented with B. Acidophilus, further studies are in progress to determine whether the essential nature of this phenomenon is physical, chemical or bacteriological.Subject to the limitations of the material under consideration the following points have been established:1. B. Acidophilustherapy is not a physical phenomenon since patients receiving sterile milk were not relieved of constipation.2. B. Acidophilustherapy apparently is not a strictly chemical phenomenon, since patients receiving B. Acidophilusmilk which had been pasteurized to kill all living forms, were not relieved of constipation.3. B. Acidophilustherapy appears to be essentially a bacteriological phenomenon, since patients were relieved of constipation by the ingestion of milk fermented by B. Acidophilus.4. Relief from chronic constipation has persisted for six months after the ingestion of B. Acidophilushas been discontinued.5. Viable B. Acidophilusorganisms in appreciable number have been recovered from the feces of patients six months after the ingestion of B. Acidophilusmilk.

The following method of Gram staining is based upon the satisfactory results obtained by the use of modifications devised by Burke and Atkins:1. Air dry a thinly spread film and fix with least amount of heat necessary to kill the organisms and fix them to the slide.2. Flood slide with dye solution. This is prepared by previously mixing in a beaker about 30 drops of a 1 per cent. aqueoussolution of methyl violet 6B (Coleman and Bell) with 8 drops of a 5 per cent. solution of sodium bicarbonate. Allow the mixture to remain on slide 5 minutes or more.3. Flush off the excess stain with the iodine solution and cover with fresh iodine solution for 2 minutes or longer. The iodine solution consists of 2 gm. iodine dissolved in 10 c.c. normal sodium hydroxide solution, to which is then added 90 c.c. of water.4. Drain off the excess iodine solution, without blotting (no water being used) but the film is not permitted to become dry.5. Add acetone (100 per cent.) drop by drop until no color is seen in the drippings from the slide, which is slightly tilted. This usually requires less than 10 seconds, and should be reduced to a minimum.6. Air dry the slide.7. Counter stain for 10-30 seconds with 0.1 per cent. aqueous solution of basic fuchsin.8. Wash off excess stain by short exposure to tap water and air dry. If slid'e is not dear, bmniersion in xylol is recommended.This method has yielded particularly good results in staining milk slides for Bacillus Acidophilusand in staining Eecal specimens. By this mcthod gonococci and diphtheria bacilli are particularly well differentiated and more easily identified than by the older methods.

A cooperative experiment was undertaken concerned with the direct cultivation of B. tuberculosisby the Loewenstein method from the blood of psychotic male patients at the Psychiatric Institute and Hospital.Seven cc. of blood were introduced aseptically into a tube containing 3 cc. of a 10% solution of sodium citrate, sealed and sent by parcel post to Vienna. Bloods from male physicians and male attendants were sent as controls. Triplicate specimens of blood were taken from each subject at the same time. All tubes were numbered serially in New York. Professor Loewenstein did not know that the specimens were in triplicate. Neither did he know the diagnoses of any patients, nor that control subjects were included.A careful physical examination of allpatients and subjects by Dr. M. M. Harris, Research Associate in Internal Medicine, failed to reveal any clinical signs of active tuberculosis. This was further confirmed by radiographic examination. Dr. L. E. Hinsie, Research Associate in Psychiatry, not only vouched for the psychiatric diagnoses but took all of the blood samples. We are privileged to acknowledge our indebtedness to them.Professor Loewenstein reported all of his findings before being informed of the diagnoses, etc. The results briefly summarized are as follows:Upon visiting Professor Loewenstein in his laboratory this summer after the findings were recorded, we found 13 cultures reported as macroscopically positive which had not yet been discarded. These were brought back to New York and guinea pigs were inoculated. (One culture was dried out.) All of the guinea pigs came down with tuberculosis within 5 weeks after inoculation. All showed strongly positive tuberculin tests, local lesions at the site of injection, gross evidence of tuberculosis in the inguinal glands, spleen and liver, and stained smears revealed typical acid-fast bacilli. Cultures from these organs yielded macroscopic growth in all cases. Histological examinations proved positive.

The treatment of general paralysis by inoculation with malaria has been employed for the past five years at the Psychiatric Institute.1More than 300 patients with general paralysis have been inoculated intravenously with a single strain of malaria which has been therapeutically effective. The present results are concerned with:1. A detailed microscopic study of the life-cycle of this parasite made by blood smears taken hourly from 4 patients inoculated with malaria about 3 years ago.2. The significant finding was the total absence of gametocytes (the sexual forms of the malarial parasite). This agrees with contentions of Gerstmann.2The asexual cycle of this strain of malaria is identical with the asexual cycle of “natural” malaria. From a biological point of view it is interesting to note that our malarial strain has apparently become “sterile”, which means that it has lost its capacity for sexual propagation. The absence of gametocytes would make it impossible to infect mosquitos, thus limiting the life-cycle to man alone. Incidentally, we have failed in our efforts to cultivate this strain of malaria in artificial media.3. A strain of malaria free from gametocytes is of considerable practical value in the treatment of general paralysis in that:(a) It eliminates the possibility of the transmission of malaria to other members of the community.(b) It precludes the occurrence of malarial relapse following adequate quinine administration.

The possible rôle of allergy in the mechanism underlying certain neurological conditions offers an interesting field of investigation. Following the recent demonstration by David and Beatrice Seegal of local organ hypersensitivity in the anterior chamber of the rabbit eye, one of us (L.M.D.) in collaboration with the Seegals produced the same phenomenon in the brain of rabbits. However, the sterile inflammation was observed only after making a second injection directly into the brain, and not, as in the case of the eye, by a simple intravenous injection. It occurred to us that this difference might be accounted for by the fact that in the eye there was more prolonged and more intimate contact of antigen with local tissue in a confined space than in the brain. How to effect this in the brain was our immediate problem.The following procedure was carried out, using dogs as experimental animals:Under local anesthesia a trephine hole was made and somewhat enlarged by rongeurs over the left motor area. The dura was then opened, and into a cavity in the brain tissue there was inserted a 0.5 cm. cube containing normal horse serum to which enough agar was added to make a jelly (0.6%). The wound was then closed. Immediately after the operation each dog was given from 1.5 to 5 cc. normal horse serum intravenously.The brain in each case had been purposely injured on the left side so that immediately after operation and continuing for a few days to a week or 10 days the dogs showed a varying amount of right-sided hemiplegia. In every animal the weakness on the right side had disappeared before the next step in the experiment.

The influence of compounds containing active chlorine on B. tuberculosis(strain H-37 obtained through the courtesy of Dr. S. A. Petroff) suspended in tap water has been determined by guinea pig inoculation. This investigation comprises 3 phases: first, a determination of compounds containing active chlorine germicidal to B. tuberculosis; secondly, the effective concentration of such compounds; and thirdly, the time of exposure required for compounds containing active chlorine in effective concentration to kill B. tuberculosis.The compounds containing active chlorine were tested in amounts ranging from 10 to 100 parts per million, generally at increments of 10 parts per million. The time of exposure usually ranged from 15 seconds, 30 seconds, 1 minute, 3 minutes and 5 minutes.The most practical disinfectant in low concentration appears to be chlorine gas in water in a concentration of 30 to 50 parts per million when applied for 5 minutes. Within the limits of investigation, the results are applicable to disinfection of eating and drinking utensils in restaurants, soda fountains, drinking places, etc., where contamination with B. tuberculosisis encountered.Before reaching a final conclusion it will be necessary to investigate the influence compounds containing active chlorine in B. tuberculosispresent in colloidal suspensions as well as in tap water.

L. acidophilushas been found to produce dextro-rotary lactic acid from lactose when grown in pure culture in casein-digest broth. After an acidity equivalent to 0.6 cc. of tenth normal sodium hydroxide per cc. had developed, it was neutralized by calcium oxide, evaporated under reduced pressure (not heating over 60° C.) to a syrupy consistency, strongly acidulated with concentrated sulfuric acid, filtered and extracted with ether in a continuous extractor. The ether was evaporated under reduced pressure and the syrup neutralized with zinc carbonate to form zinc lactate. This was precipitated with alcohol, filtered, dissolved in water and subjected to fractional crystallization.Using a 5 per cent solution, a polariscope examination of three different fermentations, independently determined by each of us, gave a specific rotation of −2.5° for the second crop of zinc lactate. Therefore the lactic acid, which bears the opposite sign from its zinc salt, is dextro-rotary.The work is still in progress.

Over 150 urinanalyses were made on 40 psychotic patients. The number of daily bowel movements were recorded, and showed a range varying from normal to severe chronic constipation. Various experimental procedures with B. acidophiluswere employed. Such an investigation must be continued for a considerable period of time before any conclusions can be drawn. However, in reporting progress, it is interesting to note that in only five instances did the indican present exceed 20 mg. which is generally regarded as the upper limit of normal excretion. Some of the patients with severest constipation had very small amounts of indican present in the urine. In general patients who received B. acidophilusmilk had a smaller amount of urinary indican than patients receiving B. acidophilusmilk in which the viable organism had been killed.

1. In order to study the influence of physical and chemical factors, B. acidophilusmilk was centrifuged and run through a Mandler diatomaceous filter. Thus the chemical constituents were little altered. When fed to constipated patients it was practically without effect. Regular B. acidophilusmilk ingested subsequently resulted in an increase in the number of normal defecations.2. B. acidophilusmilk was sterilized and lactic acid added, thus again approximating the original chemical composition. When fed to constipated patients, little change was noted. Regular B. acidophilusmilk ingested subsequently resulted in an increase in the number of normal defecations.3. These data indicate that B. acidophilustherapy is essentially bacteriological rather than physical or chemical in nature.

Following the suggestive report of Albus and Holm1on the effect of surface tension on lactobacilli, studies were undertaken with some of the same organisms used by them, and we employed their technic, as described in an unpublished paper generously placed at our disposal. Sodium ricinoleate was used as a depressant, for which we are indebted to Dr. W. P. Larson of the University of Minnesota, who very kindly supplied us with a pure product.Six strains each of L. bulgaricusand L. acidophiluswere tested. While no growth of the former took place in media depressed below 42 dynes, the growth of L. acidophiluswas abundant at this point and considerably below. L. acidophiluswas inhibited at 35 dynes as measured by the drop-weight method.The results, therefore, are in close agreement with those of Albus and Holm, and it appears that surface tension may well be considered an effective criterion for differentiating the very closely allied L. acidophilusand L. bulgaricus.

Since the gastro-intestinal tract has long been under suspicion in epilepsy, it appeared worth while to study the transformation of the intestinal flora effected by L. acidophilus, and any influence this effect might have upon the number of convulsions. A series of 15 epileptics were carefully observed for one month. Twelve of these then received one litre of sterile milk daily for 42 days. All 15 patients were then given one litre of L. acidophilusmilk daily for 2 to 3 months. The methods employed have been described elsewhere.1Three patients had only one convulsion per month, or less, and are therefore disregarded. Of 12 other patients studied, 4 showed a rather striking reduction in the number of convulsions. One of these, who had 122 convulsions in 23 days during the 28-day preliminary period, had an average of 5 convulsions per month for the three months during which he received L. acidophilusmilk.Epileptics are notoriously prone to show improvement under almost any form of treatment, but in view of the sterile milk control and another control group of epileptics who showed little benefit from kaolin administration at the same time, these results seem encouraging enough to enlarge the series. Further work is now in progress.

Having demonstrated that constipation and diarrhea are alleviated during the ingestion of L. acidophilusmilk, it is of interest to establish whether such benefits are permanent or transitory. In other words, is it necessary to continue the ingestion of L. acidophilusmilk in order to maintain the benefits of treatment?Of 34 cases where daily observations have been carefully recorded after discontinuing treatment, 27 cases, or 80 per cent continued to have more normal defecations; 6 cases, or 18 per cent, had approximately the same number, and 1 case, or 3 per cent, had fewer normal defecations than before treatment. One case which prior to treatment averaged one normal defecation per week, had daily normal defecations without exceptionfor 924 days after treatment was discontinued. Three other cases showed benefits over a year after treatment was discontinued, and 10 cases showed benefits from 3 to 6 months afterwards. There were 14 cases where the treatment was of relatively short duration, i. e., about 2 months or less, and the benefits persisted for considerably longer than that time.While the results of L. acidophilustherapy obviously vary from individual to individual, there is here good evidence for believing that in most instances the beneficial effects of L. acidophiluspersist after treatment has been discontinued.1

Experiments of this kind have been carried out on groups of four human subjects by Rahe,1and by Rettger and Cheplin.2Because of its importance, 12 psychotic subjects showing on repeated fecal examination no acidophilus-bulgaricustype of colony were fed 3 strains of L. bulgaricusmilk for 14 days, after a preliminary observation period of 14 days. L. bulgaricuswas not recovered from the feces, except in one instance.After an interval of 14 days, L. acidophilusmilk was fed for 14 days. The constipated cases were greatly benefited, and the non-constipated cases also had a greater number of normal defecations. L. acidophiluswas recovered from the feces of all subjects. In three cases the per cent was 99-100, and in all cases but one, it was over 67.Subsequently, some cases continued to show marked improvement over their original condition, while others showed a tendency to revert to it. Seven days after discontinuing treatment, there were no L. acidophiluspresent in the feces.It is thus clearly established that L. acidophilusdiffers from L. bulgaricusin its ability to live in the gastro-intestinal tract. In a further inquiry into the nature of the activity of L. acidophilus,3we are studying the enzymes of this organism, and have good reason to believe that they differ from those of L. bulgaricus.Another interesting chemical problem being considered is the stereoisomerism of the lactic acid produced by these Lactobacilli.

TO THE EDITOR:— Dr. M. C. Petersen in a recent article (The Journal, March 7, p. 775) refers to the "recurrence of inoculation malaria." This emphasizes the importance of using a malarial strain in which no sexual forms of the parasite are found. In a series of articles published in the American Journal of the Medical Sciences176:664 (Nov.) 1928, 179:800 (June) 1930 and 184:262 (Aug.) 1932, my associates and I have shown that approximately two and a half years is required to obtain a true biologic adaptation of the malarial parasite from a strain in which gametocytes occur to one in which they are absent. This has been done under the auspices of the New York State Psychiatric Institute and Hospital by continuous human passage of a sexual strain of malaria.By using a strain of malaria in which no sexual forms are present, the possibility

The article presents a list of books on art, architecture, biography, memoirs and letters. These include "Western Art and the New Era," by Katherine S. Dreier; "Early American Prints," by Carl W. Drepperd; "Mahatama Gandhi's ideas" by C.F. Andrews; "Bach" by Rutland Bougghton; "Memoirs of a Victory," by Georges Clemenceau; "Freud and Carlyle," by Waldo H. Dunn; "Life of John Keats" by Albert Erlande; "Seven Horizons" by Charles J.Finger; "Youth in Revolt" by Levin Shmarya; "Lincoln" by Emil Ludwig.

The article presents a list of contributors to the April 16, 1930 issue of "The Nation." The list includes James Dunbar, Irwin Edman, Joshua Kunitz, Granville Hicks, Smith Ely Jelliffe, Matthew Josephson, Sidney B. Fay and Mark Van Doren.