The Goose That Laid Golden Eggs

Enzymatic Catalysis of a Nuclear Fusion Reaction Converting Iron to Gold in the Liver of a Goose

Abstract

The tale of a goose that laid golden eggs in which the goose was killed for greed and none of its eggs hatched to give rise to another goose that laid golden eggs is very familiar. That such goose might actually exist is consistent with the hypothesis that the phenotype of laying golden eggs arose in the goose due to an accumulation of multiple independent mutations in its somatic tissues after development, a situation common in cancer. We conducted a study on commercial geese in which multiple mutations in random tissues were introduced by inserting P-elements and the eggs were screened for the presence of gold. We report here the isolation of one Goose that lays golden eggs. Biochemical analyses suggest that the gold in the eggs is being produced by a nuclear fusion reaction involving conversion of iron to gold rather than by ingestion of food containing gold. Tracing the origin of gold by examination of blood from various tissues revealed that the conversion of iron to gold occurs in the liver of the Goose, the gold being subsequently transported to the ovaries for excretion as eggs. This is the first report of a nuclear fusion reaction occurring under physiological conditions.

Introduction

In a survey of common folktales in independently evolved languages, Carpenter et al¹ found the story of the goose that laid golden eggs to be one of the ten most frequently told stories, even in the most secluded of tribes. They attributed the phenomenon to the dual coincidence of presence of geese and the presence of gold in the proximity of human civilization. We wondered if there were a more biological explanation possible and obtained permission from FDA (Permit ID USFDATX1967QF474387) in 1967 to conduct a study in McGregor’s farm in San Antonio, Texas, USA on commercially used geese.

Barbara McClintock recently described the occurrence of randomly hopping fragments of DNA called P elements (brought about by an enzyme called transposase) in rice². We decided to isolate geese expressing P elements or transposases and use them to generate mutant geese which could potentially lay golden eggs. After examining more than 10 million eggs in this fashion, we indeed found a goose that laid eggs, all of which had traces of gold in it. We now report the results of further tests carried out on the goose that laid it.

Materials and Methods

Geese strains

Geese were screened by Southern Blotting using probes for P element (Toba et al, 1964). Geese carrying exactly one P element coding for transposase were mated with each other to generate a flock of geese called T flock. Each progeny was selected for the presence of the transposase by Southern Blot.

Geese containing only P elements but no transposase were also identified by Southern Blotting and interbred to generate a flock of geese called the P flock.

Southern Blotting

Genomic DNA from the eggs were digested and DNA was separated on 1% agarose TBE gel using a CHEF DRII pulse field apparatus (Bio-Rad Labs., Hercules, CA) set for 10 hrs at 200 volts and 2 second pulses. A sizing ladder purchased >from Life Technologies was used for size determination. Nitrocellulose lifts of the DNA were hybridized to ³²[P] labeled transposase cDNA.

Egg Collection

One female goose from the P flock and one male goose from the T flock were paired for 3 days and the female goose was subsequently isolated. Embryos at the 2-16 cell stages were partly dejellied with fine-tipped forceps and dissolved with Ringer's solution containing 111 mM NaCl, 1.9 mM KCl, 1.1 mM Cacl,, 0.8 mM MgS04, 0.08 mM NaH,PO,, and 2.4 mM NaHC03 at pH 7.4. Only the first 10 embryos were collected and used for analysis.

Measurement of 197Au in fly embryos

Unless otherwise specified, all NMR measurements were made at 53 MHz and 21 "C using a Varian XL-200 NMR spectrometer with a 5/10-mm diameter concentric combination of sample tubes. 50 eggs were placed in the inner tube, and 50 (for field-frequency locking) in the annular space between the inner and outer tubes. The intensities of the 197Au resonances directly yielded the concentration of intracellular 197Au.

Data Analysis

NMR free induction decay signals from about 50 eggs contained in a 5-mm sample tube were time-averaged for 2 min to obtain the spectra shown. Comparable spectral signal-to-noise could be obtained in only 10 s with a 10-mm diameter sample tube containing about 500 eggs.

Results

Identification of geese carrying transposons and transposases

Genomic DNA was isolated from geese tissues and subjected to a Southern analysis for presence of a transposase flanked by P elements. We isolated one goose that carried such a transposase. We backcrossed its progeny to generate a flock of geese carrying this transposase (T flock). Since it is impossible to generate a homozygous line of geese carrying a transposase, we performed a Southern analysis on all the geese that were used for subsequent crosses. Fig 1 shows a typical Southern blot performed on genomic DNA isolated from a transposase containing goose. The hybridized bands of digested DNA derived either from a transposase clone or from a test goose show a similar pattern, confirming that the goose contains a transposase.

Since all geese carry multiple P elements naturally, we isolated lines of geese that did not express any transposase and generated a flock of them (P flock). Virgin females from this flock were used for subsequent crosses.

Isolation of the Golden Egg

One male from the T flock and one 1-month old virgin female from the P flock were incubated together in isolation at room temperature for three days. After this, the male was used for further crosses whereas the female was isolated and its eggs tested. Only the first 10 eggs were tested for the presence of gold and if no egg was detected, this goose was transferred to the commercial stock.

More than 10 million eggs were screened before an egg with some gold contents was identified. The wild type eggs and the “golden” eggs were subjected to spectrographic analysis and compared to spectra derived from a pure gold sample and distilled water (Fig 2). The spectra of golden eggs showed a peak at the gold resonance point whereas that of the wild type eggs remained flat, similar to spectra of pure gold and distilled water. This conclusively demonstrated that gold is present in the golden eggs and that the isotope of gold present in them is 197-Au.

Physical and Chemical properties of the eggs

The eggs weighed 825.6 grams on average. They had an outer appearance of white color. When dropped from a height of 2 meters, the eggs did not smash, nor was there any splash of white and yolk. In fact the eggs lay where they had fallen with the bottom of the egg caved in. Upon reexamining these eggs, we saw that the eggshell had shattered where the egg had struck. Pieces of it had flaked away and what shone through was a dull yellow in color. We excluded the possibility of the yellow substance being brass by confirming that it remained inert to concentrated nitric acid. The spectrographic analysis of the material confirmed that it was gold. The eggs contained only a shell of gold because they could be bent with moderate pressure. Besides, if the eggs were all solid gold, then they would weigh over ten pounds. We also reconfirmed that the egg shell was pure calcium carbonate. The average radius of a typical egg was 35 millimeters (major axis = 72 millimeters, and minor axis = 68 millimeters). The gold shell was 2.45 millimeters in thickness. This value is rather high compared to the average thickness of normal eggs (2.1 millimeters). Inside of the shell was egg. It looked like egg and smelled like egg.

Aliquots were analyzed and the organic constituents were found to be reasonably normal. The white was 9.7 percent albumin. The yolk had the normal complement of vitellin, cholesterol, phospholipid and carotenoid. Strangely, the eggs’ behavior on heating was abnormal – it coagulated at once, indicating that the proteins were partially denatured to begin with and were most likely so due to contamination by gold. An examination of yolk for inorganic constituents revealed an abnormally high presence (0.32%) of chloraurate ion, which is a singly-charged ion containing an atom of Gold and four of chlorine, the symbol for which is AuCl₄. Also, the iron content of the yolk was double the normal.

Physiology of the Mother Goose

So far only one goose has been found to lay golden eggs in our experiments and the following discussion refers to data only from that one goose. Blood samples of the goose were obtained without anaesthetizing the goose because we did not want to alter its metabolism. It carried 2 parts per hundred thousand of chloraurate ion. Blood taken from the hepatic vein was richer than the rest, almost 4 parts per hundred thousand. This result prompted us to take an X-ray of the liver.

On the X-ray negative, the liver was a cloudy mass of light gray, lighter than the viscera in its neighborhood, presumably because the liver was less penetrable to the X rays because it contained more gold. The blood vessels showed up lighter than the liver proper and the ovaries were pure white (Supplementary online figure S1).

Origin of gold

Where was the gold coming from? In the beginning there was only negative evidence. There was no perceptible gold in the Goose’s feed, nor were there any gold-bearing pebbles about that it might have swallowed. There was no trace of gold anywhere in the soil of the area and a search of the grounds revealed nothing golden. In fact we found very remarkable that indeed it was possible to have a goose laying golden eggs, in spite of there being no natural source of gold.

We forced gastric tubes into the goose, with the idea of testing the contents of the alimentary canal in search of exogenous gold. Indeed, gold was found, but only in traces and was most likely due to the digestive secretions and therefore, endogenous in origin. However, something else was revealed by the test – the goose was low in bile pigment. Duodenal contents showed virtually no bile pigment. We wondered if the liver was malfunctioning due to a high concentration of gold in it, but further fecal tests confirmed that bile acids were present, only the bile pigment was missing. It appeared as though the porphyrin catabolism responsible for producing the bile pigment was not following the proper course in the liver.

We took a liver biopsy of the goose, isolated hemoglobin from the blood and small quantities of cytochrome (the oxidizing agents, also containing iron), and precipitated some of it as brilliant orange substance. Unlike ferrous or ferric ion, we had isolated a triply charged auric ion in the hemoglobin complex. We called this aureme, short for auric heme. It appeared that the liver was not breaking heme to bile pigment; instead it was converting it to aureme.

Further analysis showed that 29 percent of the gold in the blood stream was carried in the plasma in the form of chloraurate ion. The remaining 71 percent was carried in the red blood corpuscles in the form of auremoglobin. An attempt was made to feed the goose traces of radioactive gold with the idea that we pick up radioactivity in plasma and corpuscles and see how readily the auremoglobin molecules were handled in the ovaries. It seemed to us the auremoglobin should be much more slowly disposed of than the dissolved chloraurate ion in the plasma. The experiment failed, however, since we detected no radioactivity. We initially put it down to our inexperience with nuclear physics, but it turned out to be a very significant result. The goose was, in fact, absorbing all nuclear radiation in the background, presumably converting it to some usable form of energy, as if it were a living nuclear reactor.

Isotopic analysis of Iron and Gold

We isolated fresh samples of heme from the goose, precipitated the iron oxide and sent it to Brookhaven Labs for isotopic analysis. The results showed that there was no Fe⁵⁶, suggesting that it was this isotope of iron that was being converted to gold. Since the reaction was endothermic, we hypothesized that it would be coupled to an exothermic reaction and searched through the tables of nuclear reactions to identify a series of reactions that would yield just enough energy to support conversion of Fe⁵⁴ to Au¹⁹⁷. We found the following reactions in the table.

Xe¹³⁵ + 2Fe⁵⁴ => Pu²³⁹

2 Pu²³⁹ => 2 I¹³⁵ + Au¹⁹⁷ + O¹⁸

I¹³⁵ => Xe¹³⁵

3 O¹⁸ => Fe⁵⁴

12 O¹⁸ => Au¹⁹⁷

Genetic analysis of the parent goose

The grandfather goose was from the T flock and hence contained the transposase. Sequencing the genome around the site of insertion in the mother goose revealed that the transposon had hopped near the promoter of the period gene (Supplementary online figure S2). Period gene is involved in circadian rhythms in the fruit flies and birds and most likely has a similar role in the geese. It is expressed in many tissues throughout the body, including in brain, liver and gonads.

The mother goose contained multiple transposons. Initial sequencing from the mother goose, as expected, has not given a clear indication of the sites of insertion of the transposons as different tissues have a different location of the insert. We have made primary cultures from many tissues of the mother goose and are conducting detailed genetic analysis on them.

Discussion

Since the blood from goose liver contained more gold than blood from other parts, we hypothesized that the chloraurate ion is secreted by the liver into the blood stream. The ovaries acted as a trap for the ion, reducing it to metallic gold and depositing as a shell about the developing egg. Relatively high concentrations of the unreduced chloraurate ion penetrate the contents of the developing egg.

There is little doubt that the goose finds this process useful as a means of getting rid of the gold atoms which, if allowed to accumulate, would undoubtedly poison it. Excretion by eggshell may be novel in the animal kingdom, even unique, but it is in fact this very mechanism that is keeping the goose alive.

Unfortunately, however, the ovary is being locally poisoned to such an extent that few eggs are laid, probably not more than what will suffice to get rid of the accumulating gold, and those few eggs are definitely unhatchable.

Gastric examination of the goose revealed that the bile pigment was missing in the goose. Bile acids are steroids secreted by the liver into the bile and poured into the upper end of the small intestine. These bile acids are detergent-like molecules, which help to emulsify the fat in our diet and distribute them in the form of tiny bubbles through the watery intestinal contents. This distribution makes the food easier to be digested. Bile pigments, on the other hand, are completely different. Liver makes bile pigments out of hemoglobin, the red oxygen-carrying protein of the blood. Worn out hemoglobin is broken up in the liver, the heme part being split away. The heme is made up of a square shaped molecule – called porphyrin – with an iron atom in the center. The liver takes the iron out and stores it for future use, then breaks the square shaped molecule that is left. This broken porphyrin is bile pigment. It is colored brownish or greenish – depending on further chemical change – and is secreted into bile. The bile pigments are of no use to the body and are poured into the bile as waste products. They pass through the intestines and come out with the feces. In fact, the bile pigments are responsible for the color of the feces.

Liver biopsy revealed that the liver of the goose was not breaking the heme to bile pigment; instead, it was converting it to aureme. The aureme, in equilibrium with chloraurate ion, entered the blood stream and was carried to the ovaries, where the gold was separated out and the porphyrin portion of the molecule disposed of by some as yet unidentified mechanism. The auremoglobin was, of course, useless as far as carrying oxygen was concerned, but it only made up about 0.1 percent of the total hemoglobin of the red blood cells, so there was no interference with the respiration of the goose.

The goose was excreting gold at the rate of 38.9 grams of gold a day and had been doing it over a period of months. That gold had to come from somewhere and failing that, it had to be made from something. Due to the mere fact that we were face to face with the goose that laid golden eggs, we had to seriously consider the possibility that while it was hemoglobin that entered the liver, not all of what came out was hemoglobin – a fraction of it was now auremoglobin. The gold shell of the eggs had iron as its only impurity. The egg yolk was high in only gold and iron. It all suggested that somehow iron was being converted into gold in a nuclear reaction.

However, there were a great many serious problems involved in this iron to gold idea. For one thing, the total quantity of iron in the goose can only be of the order of half a gram, yet nearly 40 grams of gold a day were being manufactured. There was a worse problem than that – Iron is about the bottom of the packing fraction curve. Gold is much higher up. To convert a gram of iron to a gram of gold takes just about as much energy as is produced by the fissioning of one gram of Plutonium-239.

Isotopic analysis of heme revealed a lack of Fe⁵⁴, while showing a presence of Fe⁵⁶, Fe⁵⁷ and Fe⁵⁸ in normal proportion. The implication was obvious – a nuclear reaction would use only one specific isotope and not others whereas an ordinary chemical reaction would dispose all isotopes equally. However the fact remained that such a reaction seemed energetically impossible. Even though many reactions requiring an input of energy go on in the body because they are coupled to an energetically favorable reaction, the amount of energy involved is very low – few kilocalories per mole. Nuclear reactions, on the other hand, consume millions. For the fusion to work, we needed an energy producing nuclear reaction.

A search of the nuclear reaction database revealed a set of reactions involving iron, oxygen, xenon, iodine and plutonium so that the energy produced by the exothermic reactions were just infinitesimally greater than the energy absorbed by the endothermic ones.

Genetic analysis of the geese revealed that the transposase had acted in the tissues expressing the period gene. Period gene is one of the core set of genes responsible for generating circadian rhythms. These are expressed in the brain, eyes, gut, liver and ovaries. This expression pattern correlates with the sites involved in generating gold due to altered catabolism.

Interestingly, in the 1940s, it was speculated that the circadian rhythms might be generated by the changing magnetic field, solar winds or other naturally oscillating quantum mechanical phenomena³. In fact, on Tuesday evening, September 25, 1944, during the building of the atomic bomb, Enrico Fermi had created a pile of Plutonium and was creating the first chain reaction when something bizarre happened, resulting in an oscillation of free neutrons with a period of 24 hours⁴. These oscillations involved Xenon, Iodine and Plutonium (See supporting online material). We wonder if those oscillations and circadian rhythms have any unknown connections.

Very recently, a group led by Kondo T has shown that the three proteins KaiA, KaiB and KaiC along with ATP are capable of generating circadian oscillations in the phosphorylation levels of KaiC in vitro. The Kai proteins are responsible for generating circadian rhythms in cyanobacteria, whose whole transcriptome is under circadian control. The evidence of a goose displaying nuclear reactions in the tissues where circadian clocks are expressed, coupled with the fact that these nuclear reactions display an ability to generate a circadian rhythm in neutron levels in vitro suggests that the circadian clock may indeed be constituted by core enzymes regulating nuclear reactions. Traditionally, one would have argued against such a possibility as the circadian rhythms were supposed to have been generated from negative feedback of proteins on the transcription of their coding genes. In the light of work done by Kondo’s group, it would appear that circadian rhythms indeed might be generated by proteins in the cytoplasm and the transcription factors such as CLOCK and PERIOD are involved only as a means of telling the cell the time of the day.

References

1. “Ten most frequently told stories in the world” Carpenter el al, Journal of Literature, Vol 45, Nov 1940, Pages 126-132

2. “Mobile elements in genome” Barbara McClintock, Journal of Evolutionary Genetics, Vol 16, Jun 1959, Pages 67-70

3. “Reconstitution of circadian oscillation of cyanobacterial KaiC phosphorylation in vitro” Nakajima et al, Science. 2005 Apr 15;308(5720):414-5

4. “The making of the atomic bomb” Richard Rhodes, 1976, pages 557-559.

Supplementary Online material

Description of events of September 25, 1944.

Briefly, the largest atomic pile yet assembled on earth was ready, having reached dry criticality – the smallest loading at which it would have gone critical without cooling water if its operators had not restrained it with control rods – the previous Friday. The operators withdrew the control rods in stages and gradually, the pile started working, as indicated by the gauges that showed that the cooling water warmed, flowing in at 50^o F and out at 140^o F. The pile went critical a few minutes past midnight. By 2 A.M. it was operating at a higher level of power than any previous chain reaction. For the space of half an hour all was well. Then the pile reactivity started steadily declining with time, necessitating withdrawal of control rods, until all the rods had been completely removed and even then the power kept declining, finally dying by early Wednesday evening. Early Thursday morning, the pile came back to life. By 7 A.M. it was running well above critical. But twelve hours later, it began another decline.

Princeton theoretician John A. Wheeler had been concerned about fission product poisons. The mechanism would be compound – A non neutron absorbing mother fission product of some hours’ half life would decay into a daughter dangerous to neutrons. This poison itself would decay with a half-life of some hours into a third nuclear species, non-absorbing and possibly even stable. So the pile would chain react, making the mother product; then the mother product would decay into the daughter; as the volume of daughter product increased, absorbing neutrons, the pile would decline; when sufficient daughter product was present, enough neutrons would be absorbed to starve the chain reaction and the pile would shut down. Then the daughter product would decay into a non-absorbing third element; as it decayed the pile would stir; eventually too little daughter product would remain to inhibit the chain reaction and pile would go critical again.

If this explanation made sense, then an inspection of the chart of nuclei showed that the mother had to be 6.68 hr [Iodine]¹³⁵ and the daughter 9.13 hr [Xenon]¹³⁵. Furthermore, the cross section for absorption of thermal neutrons by Xe¹³⁵ was roughly 150 times that of the most absorptive nucleus previously known, [cadmium]¹¹³. Almost every Xe¹³⁵ nucleus formed in a high flux reactor would take a neutron out of circulation. In fact Xe¹³⁵ was the poison. This phenomenon was observed in another nuclear reactor in Chicago three days later, which also showed a shutting down of the pile twelve hours after it had gone critical.