Arthur I. Kendall

OBSERVATIONS UPON THE FILTERABILITY OF BACTERIA, INCLUDING A FILTERABLE ORGANISM OBTAINED FROM CASES OF INFLUENZA

1. The isolation of a filter-passing diplococcus from the blood of certain cases of influenza by means of a special cultural medium is described. The experimental effects of this organism, while in the filterable state, upon rabbits, is discussed. 2. A procedure is formulated for inducing at will both a filterable and a non-filterable state in bacteria. Mention is made of a series of experiments in which both the filterable and the non-filterable state has thus been induced in a series of well-known bacteria comprising a variety of types. 3. It is postulated that a majority, if not all, known bacteria can and do exist in a filterable and in a non-filterable state. 4. A preliminary report of the isolation of microbes in the blood, not only of cases of influenza, but also from common cold, rheumatic fever, arthritis, from Staphylococcus bacteriophage and Besredkas Staphylococcus Antivirus is presented in evidence of the ubiquity of the procedure. 5. An explanation of the chemical basis for the existence of bacteria, both in the filterable and non-filterable states, in the animal and human body, and in culture, is proferred. 6. The relation of this chemical concept to microbic infection, and the state of microbes in the body during infection is discussed. In conclusion, it is a privilege as well as a pleasure to inscribe here my appreciation for the courteous cooperation of Dean Irving S. Cutter, Doctors Charles A. Elliott, Paul Starr, James G. Carr, Walter Nadler, Howard Alt and Herbert Barker, of the department of medicine, and to Northwestern University for the generous facilities and unrestricted opportunity for research which have contributed immeasurably to this investigation.

A Method for Isolation of Ascorbic Acid Fermenting Bacteria

The objectives of this preliminary communication are two: to describe a procedure for the isolation of ascorbic acid fermenting bacteria, from suitable material, and to describe briefly the results obtained with this procedure. This preliminary communication does not presume completeness either with respect to the types or strains of bacteria which may actually utilize ascorbic acid for energy, or with regard to the patterns of fermentation of this substance by specific strains of bacteria. Plain, nutrient broth is reinforced with ascorbic acid.∗ Fifteen to 20 mg. per 10 cc. of broth is a preferable amount, both because this is sufficient to indicate unequivocally that decomposition has taken place, and also because within these limits the initial reaction of the broth is not brought below pH 6.0, a point at which the bacteria thus far encountered grow readily. Also, a reaction slightly on the acid side is conducive to the stability of ascorbic acid. The broth, thus prepared is inoculated with stomach contents, feces, or other suitable material and incubated at 37° for 18-30 hours. Then, if ascorbic fermenting bacteria are present, a considerable growth will be noticed. From this first, or preliminary culture, a second broth culture is made, using 2 to 5 loops for the inoculum. After incubation and growth, plates are made either upon plain, or, preferably, glucose agar. (Ascorbic acid agar is not so well suited for this purpose.) Colonies are picked off, tested for purity and eventually for ascorbic acid utilization.

Action of Formaldehyde Upon Physiologically Active, Histamine-like Substance Produced by Gas Bacillus.

The phenomenon of contracture, induced in surviving sections of guinea pig intestine, suspended in Tyrode solution, by the addition of very small amounts of gas bacillus culture 1 suggests that the active substance, possibly substances, contains one or more free ethyl-amine groups, attached to an aromatic nucleus of the general type R :aromatic nucleus :ethylamine. In such a compound, current theory predicates that its physiological activity is intimately associated with the presence of the free aromatic ethylamine group. It follows theoretically that any chemical change involving the elimination of this free NH2 group from such a compound should reduce, or even destroy the physiological action of the molecule as a whole. The addition of formaldehyde to primary amines changes them according to the following equation, in accordance with the well known “formol titration” of Henriques and Sörenson: 2 It has been found by actual experiment that the addition of 0.1 cc. of neutralized formalin solution releases the contracture in a piece of guinea pig intestine which has been induced by either the soluble, physiologically active substance found in cultures of the gas bacillus, or by histamine. The following graph is the record of such a contracture induced by 0.2 cc. of gas bacillus filtrate, and released by 0.1 cc. of formaldehyde solution. Precisely similar ones have been obtained with histamine and formaldehyde. The formaldehyde-ethylamine compound may be washed out of the preparation with fresh Tyrode solution, and contracture again induced, indicating that the intestinal muscle is not materially injured by the several reagents. The experiments seem to indicate that contracture of smooth muscle may be induced by an aromatic ethylamine group, and that chemical change of this ethylamine group may thereby remove the contracture-inducing capacity of the entire molecule.

Über Wunden, Gasbrand und Sekundären Shock

ZusammenfassungLebende Kaninchen wurden mit Stämmen von Histamin bildenden Gasbacillen infiziert. Aus den Geweben dieser Tiere konnte Histamin isoliert werden.Wurden hingegen Kaninchen mit Stämmen von Histamin nicht produzierenden B. phlegmonis emphysematosae oder mit B. pyogenes foetidus infiziert, so konnte aus den Geweben kein Histamin isoliert werden.Der Zusammenhang zwischen diesen Beobachtungen und dem Auftreten von sekundärem Shock in Fällen von Gasbrand wurde besprochen.

Action of Aldehydes on Histamine and on Active, Histamine-like Substances Produced by Gas-Bacillus.

It was shown in a preceding paper 1 that the contraction induced by histamine, or the histamine-like substance produced by the gas bacillus, in a section of guinea pig intestine, or uterus suspended in Tyrode solution, could be relaxed immediately by the addition of 0.1 cc. neutral formalin solution. The most plausible explanation for this phenomenon is a definite chemical reaction between the amino group of the histamine molecule and the aldehyde according to the equation R.CH2 CH2 NH2 + H.CHO = R.CH2 CH2 N : CH2 + H2O whereby the ethyl amine group is removed. It follows that other aldehydes might react in a similar manner, if this explanation be correct. A series of aldehydes have consequently been tested, using the technique described in the preceding contribution. 1 These comprised, besides formaldehyde, already mentioned, acetaldehyde and its polymer paraldehyde, aldol, glycol aldehyde, glyceric aldehyde and methyl glyoxal, the ketones, acetone and dihydroxyacetone, and acetophenone. The ketones, and those aldehydes having an oxygen atom attached to the carbon atom next the aldehyde group were without obvious effect. This group included acetone and dihydroxyacetone and the following aldehydes : glycol aldehyde, glyceric aldehyde and methyl glyoxal. Acetaldehyde, paraldehyde (which probbly depodymerizes in Tyrode solution) and aldol, which has the oxygen atom attached to the carbon atom twice removed from the aldehyde group, reduced the contractions promptly. FOP reasons that are not quite apparent, acetophenone was also very effective in this regard. These observations are in accord with the hypothesis that certain aldehydes in addition to formaldehyde unite chemically with aromatic amines. They also suggest a pharmacological relationship between the proximity of oxygen upon adjacent carbon atoms and the physiological reactivity of certain aldehydes. This is a preliminary report.

A modification of Teichmann's method for obtaining hemin crystals, with a demonstration of specimens

This modification consists in heating suspected blood in a tightly plugged or sealed tube. Heating is best carried out by immersing the tube in boiling water. The reagents are those usually employed, i. e., sodium chloride and glacial acetic acid. Heating is continued for fifteen minutes, at the end of which time the tubes are removed from the water bath and allowed to cool slowly at room temperature. After cooling, the tubes are broken open and the liquid poured into a watch glass or small evaporation dish and concentrated over the water bath. When the volume of the liquid has thus been reduced to a few drops, it is poured on a glass slide and covered with a cover glass. If sodium chloride crystals appear under the microscope a drop of water will dissolve them and leave the observation of the hemin crystals unobstructed. The treatment with the hot glacial acetic acid in the closed tube completely decolorizes the material on which the stain occurs. Heating in the closed tube keeps the condition of temperature and strength of reagents constant. This method appears to yield larger crystals than those obtained by the old method.