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5. Albumen Preparations (Prep. 6)

Based on the tension-charge formula, the substances necessary to complete the [development of] function in the structures had to possess the property of swelling. Since, according to other studies, lecithin behaves like potassium and cholesterin like calcium (the former causes tumescence, the latter detumescence), I proceeded to add lecithin and cholesterin to the earth preparations. The result was as follows: In addition to the already described phenomena, there were a few others which had not been detected previously. First of all, it was noticeable that the preparation was full of tubular, regularly formed structures, which slowly changed their shape (Figures 38, 39, 40).

 
Figure 38.   (left) Lecithin, spread dry on cover slip, 400x.
Figure 39.   Cholesterin crystals, 400x.

 
Figure 40.   Tabular structures from lecithin. Lecithin in 0.1 N potassium chloride, 1200x.

Control tests involving identical treatment of each substance individually soon revealed that these structures had to be attributed to lecithin. They had the following characteristics: The tubular, often regularly notched structures elongated and thickened, bent, changed their position, and in particular showed budding; i.e., in some places, another tubular structure grew out of the original one, which again branched, etc. When I added only KCI to lecithin, I obtained the identical structures. They did not display any organic movement. The "growth" and budding observed I believed to be attributable to the intake of fluid, as into a sac. It ceased after approximately forty-eight hours. However, lecithin, without doubt, was capable of forming structures. The movement connected with this growing and budding was fundamentally different from that of the pseudoamebae.

 

x   Elongating tubular structure.
xx   Luminating green expanding and contracting; when current is applied, elongation is towards the anode.

I now added chicken albumen to the earth-lecithin preparation. The result was beyond all expectations. In unheated preparations, round cells formed after a few minutes; they contained dark nuclei which divided frequently and in rapid succession. The cells divided either through a central cleavage furrow, through which the nucleus also divided, or by budding they produced smaller cells, which then detached from the mother cell. In some areas, one could see distinct movements of breaking away. What I am attempting to describe here briefly was unfolded in reality only after long, laborious observations.

Cell-like structures obtained upon addition of chicken albumen.

Nucleus-like formation inside tubular structure, and cell-like structures.

To the extent that I learned to observe and follow the phenomena, they became more consistent, for I gained confidence in identifying them. Thus, actual cells were formed upon the addition of albumen. Cell formation did not occur without the addition of lecithin and cholesterin; otherwise, only heaps of flaky albumen were observed in the preparation, as can also be seen when unboiled albumen is examined under the microscope. Neither did lecithin with water alone give rise to motile cells.

We also observed that in the unheated albumen preparations the formation of motile substances occurred much more slowly and sparsely than in the heated preparations. The latter could also be distinguished outwardly from the unheated ones by the colloidal turbidity of the solution.

The unheated preparations were far less cloudy and showed a greater amount of sediment. The colloidal particles formed after heating were therefore in suspension, which, according to all previous knowledge, must be attributable to their electrical charge. I let the preparations stand for weeks and was unable to observe that there was a tremendous increase in the number of individual formations. However, after a certain time, about six to eight weeks, the solution started to clear again and the particles sank to the bottom where they formed a thick, whitish layer. Microscopic examination showed that, when the colloids were no longer in suspension, they also lost their motility and their reaction to electricity. The structures were "dead." Let me recapitulate briefly:

Lecithin with KCI alone does not produce cells, but only various kinds of regularly formed tubular structures. Moreover, there is no organic motility, but only growing and budding which is obviously due to fluid intake. Adding only KCI to albumen does not result in cell formation. But albumen plus lecithin plus KCI plus cholesterin results in cell formation.

Figure 41.   Bions immediately after their formation
+ = structures formed upon addition of coal bion prep. 6b, 1000x.

Figure 42.   Fresh sterile bions, bion prep. 6c.

I now complicated the experiment by adding gelatin also. Therefore, I was now mixing together albumen, earth, lecithin, cholesterin, gelatin, and KCI. Addition of gelatin caused plasmoid vesicular structures to be visible immediately after heating; they moved vigorously and displayed a much finer structure and far more life-like vegetative motility than the pseudoamebae in the earth preparation described earlier. It was no longer jerky, no longer mechanical, but organically flowing. (Figure 41). The longer the preparation was heated, the more complex were the ameboid creatures obtained in this way; they possessed many more [life] functions than the heated earth preparations. While the pseudoamebae in the earth preparations simply moved from place to place, displayed minimal or no movement of the particles within, and showed only very weak and occasional streaming, contraction, and expansion; the ameboids in the albumen preparations showed streaming, internal movement, contraction, expansion, locomotion. The function of division was present in both, as was the function of "ingestion" (Figures 42, 43).

The ameboid structures in the albumen preparations showed a much finer vesicular structure. Also, after a few days, one could see structures with a homogenous plasma that showed flowing, pseudopodal movements. Also present were contraction, expansion, division, budding, "ingesting," locomotion. Still unanswered were questions of metabolism, culturing, staining, and the bacterial nature of the structures we had produced.

Figure 42.   Vibrating amebae moving in place. Bion prep. 6. 2300x.

Basic evidence for the possibility of creating the living from the non-living is the fact that when various substances are used, the addition or omission of one or another substance can change the biological nature, the composition, and the activity of the motile structures.

It is not necessary to mix the substances in exact ratios. It is as if the structures form according to still unknown laws, as if the ration of substances that combine to produce motile life, motile forms, is governed by a self-activating principle. Here everything is in the dark.

Subsequently, I also added meat bullion, milk, and egg yolk as nutrients to the bion mixture. Also, carbon in various forms: finely pulverized coal sterilized dry; coal dust autoclaved in KCI; incandescent coal dust, KCI, dry or in bullion; finally, dry, sterilized or incandescent soot. The best results were achieved upon addition of soot; extremely small, but exceedingly motile amebae of the finest structure were produced. Bion mixture 6 is now prepared in various ways:

6a: Unsterile mixture;
6ab and 6ac: The unsterile mixture is heated (100 degrees, 1/2 hour) or autoclaved (120 degrees, 1/2 to 1 hour);
6c: The substances are previously sterilized;
6cb: The sterilely produced mixture is heated afterwards;
6cc: The sterile mixture is autoclaved again;
6ccc: The sterile mixture is autoclaved twice.

The best culture results are obtained with mixture 6cc. Inoculation is done for the first time after three to four days.

Can one speak here about artificially created life? Vanity would like us to answer in the affirmative. But correct consideration leads to a no! If the living were a distinct metaphysical realm totally separate form the nonliving, then I would have the right to say that I create "artificial life." However, my experiments demonstrated that there are developmental stages ranging from the nonliving, nonmotile, to the living; that, in nature, the creation of life from inorganic matter probably occurs every hour, every minute. In this case, one can hardly talk about artificial life. I merely succeeded in uncovering the developmental process of life experimentally. Perhaps a new kind of organism was thereby also produced artificially.

On 1-8-1937, I sent the following report to Professor du Teil:

 

First, at the present experimental stage, I mix 100 cc of sterile Ringers solution with 0.1 N KCI solution. To this mixture I add dissolved red gelatin until a faint pinkish -red color appears. One tweezer-tip full of coal dust is added to this mixture, and the same amount of cholesterin crystals. The total experiment is based on the principle of adding together in a specific sequence those that are necessary for cell formation.

We then dissolve one teaspoon of fresh, clear chicken albumen in about 50 cc of sterile KCI solution, and add both to the previous mixture. After brief stirring, the albumen is dissolved. Microscopically, no structures, movement, or plasmatic forms of any kind are visible. At high magnification (1000x), we see merely the typically motionless coal and cholesterin crystals.

At this point, we add about 1 to 2 cc of milk and some egg yolk. The latter causes the previously clear mixture to become cloudy.

We now prepare a second solution: lecithin ointment is stirred into KCI solution. Under the microscope, at a magnification of about 5-900x, we can observe unusual structures forming and growing: tubular structures which are swelling, budding, bending. Inside the tubular structures, we see no organization at all. However, heaps of vesicle are found occasionally. Organic movement is lacking. They are the purely physical phenomena of swelling corresponding to changes between internal pressure and surface tension.

If lecithin solution is now added to the first mixture, we immediately see a progressively increasing cloudiness that is yellowish-grey in color. Under the microscope, the surprising picture of motile life unfolds: vibratory movement from place to place, budding, division of cell-like structures, jerky to flowing motion. The organic motility can be observed from 1500x, but is best seen at 2300x to 3000x with a binocular microscope. At a magnification of about 3000x, while focussing continually on one structure, one can see luminous vesicular, nuclear structures which are [constantly] changing form. Basically, one can distinguish four groups of active, motile structures: spherical, nuclear-like vesicles; rods; round, cell-like structures containing nuclei, which move in solution but do not show any movement within; and finally, ameboid structures. The latter are especially striking because they show movements of contraction and expansion within the organisms at a magnification of approximately 3000 to 3500x. Indeed, their locomotion is already visible at a magnification of only 2000x.

At least according to past and current experiments, the structures produced a negative electric charge; they move toward the anode. When the colloid mixture is allowed to stand for 6 to 8 weeks, a thick sediment is formed on the bottom; the solution becomes increasingly clear. Upon examining the sediment at the previously mentioned magnifications, it is shown that the described motility has been lost; the structures are "dead." I will report on the control experiments performed in my detailed discussion. Currently, I am working on tests concerning metabolism and sensitivity to stains. It is certain that the structures can be cultured; for when the mixture is heated until all the liquid has been evaporated and the solid matter is extracted, inoculation of this extracted residue with sterile, colloid mixture produces growth [of structures] which have already propagated to the fifth generation.

In animal tests, these structures, if kept sterile, proved to be harmless when injected subcutaneously into mice and guinea pigs.

The question as to whether we are dealing with complete, living organisms can only be answered in conjunction with other experiments that I will discuss at another time, and only after all tests and controls have been performed. The experiment described was recorded on film.

We called the artificial, lifelike structures "bions."

(To be continued)

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