MICROBIAL ORIGIN OF THE GUMMY SUBSTANCE OF FUJITA AND GING

Home (Main Menu)

Health - Chronic Disease Differentiation
Pathological Significance
the Gummy Substance in Urine
Possible Identity of the Gummy Substance
Comaprison of the Quat Titer
Oxidation-reduction potentials
Basal Metabolic Rate (BMR).
leukocytes.
the rate of excretion of urine.
Detection of In Vivo Nucleic Acid Hydrolysis
O-R Potential Relationships
RNA, THE SECOND BIO INDICTATOR
The nucleic acid interfering mechanism and malignancy
SUMMARY AND CONCLUSIONS
REFERENCES

  • 9-8-14: Young blood proves good for old brains

    [For more on the impact of infection on human aging, see "Are there spirochetes in the brain in more than 90 percent of Alzheimer's Disease cases?"

    www.avonhistory.org/bug/alz11.htm]

    MICROBIAL ORIGIN OF THE GUMMY SUBSTANCE OF FUJITA AND GING

    By George H. Chapman

    Transactions of the New York Academy of Sciences (11-62)

    DIVISION OF MICROBIOLOGY

    Clinical Reasearch Laboratory, New York, N.Y.

    This paper was presented at a meeting of the division on October 26, 1962. The research reported in this paper was aided by grants from the Conrad Berens Eye Foundation.

    This paper was transcribed to electronic medium by Charles A. Smith.

    The finding of Fujita and Ging (1) that a neurotoxic, brown gummy substance is present in the urine of schizophrenics, and to a less extent of nonschizophrenic neurological patients, prompted me to report studies that I have been making on this substance for the past 15 years; these studies were intimately related to work that I had been doing extensively during the previous 23 years.

    I shall discuss the outstanding findings in those experiments that were related to this substance and shall describe the development of my present concept about its origin and relationships.

    I believe that these findings will demonstrate the profound significance of this substance as a phase of the basic phenomenon which initiates and perpetuates chronic diseases, including cancer and leukemia.

    My primary object throughout this research was to seek the cause of the obscure toxic states that accompany most chronic diseases.

    The existance of a general, obscure form of "intoxication" has been postulated for more than a half century. "Intestinal intoxication," "toxemia," and even the general term "toxic" have been widely used, but never elucidated.

    I was more concerned in exploring different aspects of the subject than I was in making an exhaustive critical study of every possible relationship. However, when a phase appeared to be significant, I made extensive study of thousands of samples.

    I mention this, not to boast, but to point out that although time and space will not permit presentation of all the significant data, the statements which I shall make are based on careful and extensive experimentation, although this still does not preclude the possibility of error. Much of the detailed experimental data and its statistical analysis will be submitted to appropriate journals.

    Top -- Home

    Health-Chronic Disease Differentiation

    Assessment of the possible significance of chronic disease factors is difficult because there are no reliable criteria for differentiating health from chronic disease states.

    Also, "infection" is recognized as a factor in many chronic diseases, but it is difficult to establish that a person has a chronic infection except when there are obvious foci.

    The mere finding of a microorganism or virus in the body does not prove that it has any pathological significance. The organism may have been present in the tissues because some factor reduced local resistance.

    Methods used to determine the possible existence or extent of chronic infection are unreliable, The blood sedimentation rate is within normal limits in 14.7 per cent of persons with chronic streptococcal infection and is of doubtful significance in another 30.5 per cent (TABLE 12). Also, the method does not reflect short term changes in severity of infection. The total leukocyte count is more reliable.

    There is need for complete re-examination and re-evaluation of subjective and objective health - disease - infection relationships, with the aim of establishing more reliable and more significant methods and criteria.

    Differentation of health from disease is difficult because "minor ailments" occur in persons considered healthy, such as medical students and nurses, who are often used as "normal controls."

    Because "minor ailments" occur commonly in "normal" persons, accepted normal biochemical standards are based on findings in persons with a wide range in severity of "minor ailments," but who showed little apparent disturbance in body functions.

    If the "minor ailments" are considered insignificant by the physician, the person is judged to be normal, or a hypochondriac. When they are arbitrarily more severe, he is considered to be diseased.

    Whether or not he is diagnosed as diseased depends largely on clinical opinion because of a lack of precise objective methods.

    This arbitrary difference is untenable because the severity of these "minor ailments" varies considerably, even in the same individual. They may erupt in a "normal" person into acute, subacute or frequently recurring obvious diseases.

    It should be clear, therefore, that persons with "minor ailments," no matter how slight, should not be considered normal. To put it another way, a disease condition is pathological no matter how slight it is. If this view is accepted, it paves the way for better understanding of health-chronic disease relationships.

    In attempting to assess the relative degree of health objectively, it is obvious that accepted normal biochemical standards must be revised on the basis of what has just been discussed.

    When this is done, biochemical values become highly significant, and they point the way to solution of the problems both of etiology and of treatment of the basic biochemical - biophysical disturbance that iniates and perpetuates chronic diseases.

    This biochemical significance increases when it is realized that variations in severity of subjective and objective symptoms, which may occur within short intervals, are the cause of many of the differences in results of repeated biochemical examinations made in the same, or another laboratory, and which are incorrectly attributed to technical error.

    Normal values used by the physicians who have been associated in this research are based on findings in persons who have been restored to robust health by the use of specific bacterial antigens. These values are found in remarkably few diseased persons.

    [Since antigens are things which can induce the body to produce antibodies, what does "restored to robust health by the use of specific bacterial antigens" mean?

    Are "bacterial antigens" substances obtained from bacteria which induce antibodies that will then attack the bacteria? An example may be an outer surface protein from the bacteria.

    Or are "bacterial antigens" the bacteria themselves? How could infecting someone with bacteria "restore" that person to "robust health"?

    Use of one microbe (cow pox) to induce antibodies to fight another microbe (small pox) was Jenner's original vaccination. Is this what Dr. Chapman is talking about? Are most of the antibody IgG (Immunoglobin G) and IgM bands (molecular weights) the same for cow pox and small pox?]

    Top -- Home

    Pathological Significance of Resistant and Reductogenic Variants of Strepococcus salivarius and S. mitis

    In 1947, I presented evidence to this Academy (2) that resistant variants of Steptococcus salivarius and Steptococcus mitis (3) are related to severity of chronic diseases and to differences in the results of a number of biochemical, hematological, bacteriological and other reactions.

    Because of the strong possibility that these resistant streptococci might be significant factors in chronic diseases, I sought simpler and less critical methods for demonstrating them.

    Since many pathogenic bacteria possess strong reducing power, I compared the reducing power of a large number of these streptococci by the following method:

    The reagent, which has and E0 of + .181 volts at pH 7.0, is stable for at least one month when stored in the refrigerator, particularly if air is excluded.

    When freshly prepared it gives an electric blue spot on filter paper but, as the solution ages, the spot gradually becomes grey and the indicator is useless.

    Most resistent cultures reduced the indophenol promptly, whereas nonresistant cultures took more then 30 min., and even then the degree of reduction was slight. Older cultures such as "overnight" or "18 hour," showed relatively nonspecific reduction.

    TABLE 1
    
    REDUCTION OF INDOPHENOL* BY STREPTOCOCCAL CULTURES
    ______________________________________________________
                 |               |              |         
    Organism     | Degree of     | Number of    | % of       
                 | Reduction     | Cultures     | total   
                 |               |              |         
    _____________|_______________|______________|_________
                 |               |              |         
                 |               |              |         
    Streptococcus|    +          |       879    |    89.7
    salivarius   |               |              |         
                 |  + or -       |        57    |     5.8
                 |               |              |         
                 |    0          |        45    |     4.6
                 |               |      _____   |   ______
                 |  Total        |       981    |   100.1
    _____________|_______________|______________|___________
                 |               |              |         
                 |               |              |         
    Streptococcus|    +          |       889    |    67.0
    mitis        |               |              |         
                 |  + or -       |       228    |    17.1
                 |               |              |         
                 |    0          |       214    |    16.1
                 |               |      _____   |   ______
                 |  Total        |      1,331   |   100.2
    _____________|_______________|______________|___________
    
    *Sodium 2,6-dibromobenzenoneindo-3`-methylphenol.
    ["indophenol"]
    
    

    Reducing power in the logarithmic phase has been shown to be a linear function of the number of bacterial cells (4).

    It is possible that reducing variants of S. salivarius and S. mitis are able to grow better, and thus reduce better.

    I made tests of reducing power on growths which appeared to be equally dense, but a few freshly isolated cultures of s. mitis grew poorly, and thus may have contributed to the poorer reducing power in this species (TABLE 1).

    Washed cells of S. salivarius and S. mitis did not reduce the indophenol, showing that the reducing principle is extracellular.

    Extensive study of the reductogenic streptococci showed that they are infrequent in persons with robust health, are present universally in persons with chronic diseases, with obvious exceptions, and fluctuate with changes in severity of the symptoms.

    In disease states there about 10 to 100 times as many reductogenic variants as in normal individuals, and the proportion of reductogenic to total streptococci is from 70 to 100 per cent, compared with about 20 per cent or less in normal persons.(2)

    I had concluded that resistant variants of S. salvarius and S. mitis are the common denominator in chronic diseases, with obvious exceptions, such as those caused by a known specific microorganism, dietary deficiency, etc. This conclusion was based on the following considerations.

    [If "resistant variants of {streptococci} are the common denominator in chronic diseases, with obvious exceptions ...," what about spirochete infections such as syphilis and Lyme disease or viral infections such as herpes and HIV?]

    Hence, the significance of S. salivarius and S. mitis does not lie in their mere presence, but in the possession of properties which are associated with pathogenic effects.

    TABLE 2: COMPARISON OF DECAYED
    MATERIAL AND STREPTOCOCCAL
    CONTENT OF 43 TEETH,
    FRACTURED ON AN ANVIL
    
    ____________________________________________
                     |
    Proportion of    | Average of
    decayed material | resistant streptocci
                     |__________________________
                     |             |
                     |             |
                     | Number      | % of total
                     |             |
    _________________|_____________|____________
                     |             |
    None             |        650  |     25
                     |             |
    Slight           |     19,200  |     34
                     |             |
    Moderate         |  3,465,000  |     85
                     |             |
    Marked           | 19,787,000  |     98
    _____________________________________________
    
    TABLE 3: RELATION OF
    STREPTOCOCCAL COUNTS TO
    VITALITY OF TEETH
    (ALL X-RAY POSITIVE)
    
    ______________________________________
            |         |
            |         |  Average resistant 
            |         |  streptococci
            |         |
    Source  | Vitality|__________________
            |         |            |
            |         |  Number    | % of 
            |         |            | total
    ________|_________|____________|_____
            |         |            |
    Chamber |    +    |   253,000  |    28
            |         |            |
            |    0    | 5,076,000  |    63
            |         |            |
    Root    |    +    |       983  |    18
            |         |            |
            |    0    |    56,000  |    52
    ________|_________|____________|_______
    
    
    TABLE 4: RELATION BETWEEN RESISTANT
    STREPTOCOCCI AND SEVERITY OF GINGIVITIS
    
    _________________________________________________________
               |         |                                  |
    Severity   | Number  |Average number of                 |
     of        | of      | resistant                        |
    gingivitis | persons | streptococci                     |
               |         |__________________________________|
               |         |              |           |       |
               |         |Gingival      |           |       |
               |         |crevices      |Pharynx    |Ratio  |
               |         |(a)           |(b)        |a:b    |
    ___________|_________|______________|___________|_______|
               |         |              |           |       |
    Acute      |  2      |105,400,000   |11,719,000 |8.9:1  |
               |         |              |           |       |
    Severe     | 12      | 39,619,000   |16,503,000 |2.4:1  |
               |         |              |           |       |
    Mild       |  6      |  9,889,000   |16,030,000 |0.6:1  |
               |         |              |           |       |
    None       |  8      |  4,027,000   |10,517,000 |0.4:1  |
    ___________|_________|______________|___________|_______|
    
    ________________________________________________________
               |         |
               |         |Average % of
    Severity   | Number  |resistant streptococci
    of         | of      |________________________________
    gingivitis | persons |              |           |
               |         |Gingival      |           |
               |         |crevices      |Pharynx    |Ratio
               |         |(a)           |(b)        |a:b
    ___________|_________|______________|___________|______
               |         |              |           |       
    Acute      |  2      |59            |48.5       |1.22:1
               |         |              |           |      
    Severe     | 12      |76            |66.5       |1.15:1
               |         |              |           |     
    Mild       |  6      |56            |57         |.99:1
               |         |              |           |    
    None       |  8      |35            |59         |.59:1
    ___________|_________|______________|___________|______
    
    TABLE 5: RELATIONSHIP OF PATHOGENIC TYPES OF
    STREPTOCOCCI AND STAPHYLOCOCCI TO SEVERITY OF SYMPTOMS*
    
    _________________________________________________________
                 |                                           |
    Severity of  |% of cases showing                         |
    symptoms     |resistant streptococci                     |
    _____________|___________________________________________|
                 |          |Oral      |           |Any      |
                 |Nose      |cavity    |  Feces    |focus    |
                 |__________|__________|___________|_________|
                 |          |          |           |         |
    Mild         | 4.0      | 96.6     | 31.6      | 98.9    |
                 |          |          |           |         |
    Moderate     | 9.6      | 97.6     | 28.2      | 98.1    |
                 |          |          |           |         |
    Severe       |13.3      | 98.7     | 37.8      |100.0    |
                 |          |          |           |         |
    Acutely      |10.0      |100.0     | 55.0      |100.0    |
     exacerbated |          |          |           |         |
    _____________|__________|__________|___________|_________|
    
    _________________________________________________________
                 |                                           |
    Severity of  |%of cases showing                          |
    symptoms     |coagulase pos. staphylococci               |
    _____________|___________________________________________|
                 |          |          |           |         |
                 |          |Oral      |           |Any      |
                 |Nose      |cavity    |  Feces    |focus    |
                 |__________|__________|___________|_________|
                 |          |          |           |         |
    Mild         | 64.4     | 48.3     | 5.7       | 71.2    |
                 |          |          |           |         |
    Moderate     | 61.5     | 51.3     | 6.4       | 70.5    |
                 |          |          |           |         |
    Severe       | 60.0     | 42.2     | 6.7       | 64.4    |
                 |          |          |           |         |
    Acutely      | 50.0     | 60.0     | 0.0       | 65.0    |
     exacerbated |          |          |           |         |
    ____________ |__________|__________|___________|_________|
    
    * This Table adapted from Stiles and Chapman (10).
    
    

    Since, in addition to resistance, strong reducing power is characteristic of streptococcal cultures associated with disease, I studied the possible relationship of O/R potential to disease.

    I had observed that "reductones" in culture media ingredients produced toxic effects on bacteria. When a large number of peptones were dialyzed, they showed a light-colored nonreducing dialyzate and a dark reduced residue, which was highly toxic for bacteria and had a pungent odor. Those peptones with a large amount of the strongly reduced nondialyzable fraction usually gave a long lag phase and poor maximum growth.

    I attributed the toxic fraction to overheating during the drying process, similar to the recent discovery of the tumor producing effect on rainbow trout of feeding high-temperature, flame-dried fish meal (11).

    Although Traxler and Lankford(12) found no significant difference in the Eh of 10 nontoxic and 11 toxic peptones, I found considerable difference among culture media ingredients in power to reduce the indophenol (TABLE 6).

    TABLE 6: REDUCTION OF INDOPHENOL BY INGREDIENTS
    USED IN BACTERIOLOGICAL CULTURE MEDIA.
    ____________________________________________________
                                  |
                                  |        Reduction of
    Ingrediant                    |        indophenol*
    ______________________________|_____________________
                                  |
    Proteose peptone no.3         |         ++
                                  |
    Proteose peptone              |         ++
                                  |
    Proteose peptone              |          0
                                  |
    Tryptose                      |        +++
                                  |
    Yeast extract                 |       ++++
                                  |
    Yeast extract                 |       ++++
                                  |
    Agar                          |          +
                                  |
    Noble agar                    |          0
                                  |
    ______________________________|______________________
     
    

    Also, my homemade brain heart infusion acquired considerable reducing power for the indophenol and increased toxicity for streptococci when it was autoclaved excessively.

    Since I used loss of color of the indophenol as an indicator of reduction, the problem will have to be resolved by electrometric measurements.

    However, it will be shown later that it is not the presence or absence of reducing power per se that is of primary significance as a toxic factor, but the fact that it produces an unfavorable physicochemical disturbance of nucleic acids, causing them to become toxic.

    If reducing ingredients of bacteriological culture media cause them to be toxic for streptococci, I reasoned that the reducing principle produced by variants of S. salivarius and S. mitis might be a major contributing factor in their pathogenicity for man.

    I found (5) that subcultures of arthritis-producing streptococci gradually lost their arthritis-producing power when they were grown with air, but retained it indefinitely when the slants were covered in mineral oil.

    Goebel (13) found that the toxicity of Shigella paradysenteriae can be reduced materially by treating the organisms with an oxidizing agent, such as periodic acid.

    Streptococci gradally lose resisting and reducing power in the logarithmic phase unless steps are taken to exclude air. If a single cell reductogenic variant is planted so as to provide discrete colonies, some of the resulting variants will have poor reducing power because of the influence of atmospheric oxygen.

    When a brain heart infusion, logarithmic culture of reductogenic streptococci is treated with methylene blue, the entire culture becomes colorless, except for the surface, which slowly turns blue, indicating slight oxygen exchange. However, when the culture is treated instead with the indophenol indicator, the color change remains uniform, suggesting that the major reduction is caused by accumulation of electrons.

    Obviously, a far simpler method of demonstrating these reductogenic streptococci would be to use optimally oxidized ingredients plus a suitable O/R indicator, but I have been unsuccessful in inducing a manfacturer to supply satisfactory ingredients for such an experimental medium.

    Top -- Home

    Relationships of the Gummy Substance in Urine

    In 1948, I observed that certain quaternaries produced a precipitate with urine from persons with streptococci infection (14), and that the precipitate is soluble in an excess of the reagent.

    These quaternaries have

    TABLE 7:  CHEMICAL STRUCTURE OF SURFACTANTS
    THAT REACTED WITH THE GUMMY SUBSTANCE
    
    _________________________________________________________
    Sample |      | C in alkyl| Methyl | Benzyl|       |
    no.    | Titer| chain     | groups | group |Halogen|Other
    _______|______|___________|________|_______|_______|_____
     1     | 11   | 8 to 18   |   2    |    1  |  C1   |
     2     | 12   | 8 to 18   |   2    |    1  |  C1   |
     3     | 11   | 8 to 18   |   2    |    1  |  C1   |
     4*    | 11   | 8 to 18   |   2    |    1  |  C1   |
     5     | 13   |   12      |   2    |    1  |  Br   |
     6     | 21   |   16      |   2    |    1  |  C1   |
     7     | 19   |   16      |   2    |    1  |  C1   |
     8     | 14   |   16      |   2    |    1  |  C1   |
     9     | 20   |    2      |   2    |    1  |  C1   |
    10     | 25   |   18      |   2    |    1  |  C1   |
    11     | 38   |    8      |   2    |    1  |  C1   |
    12     | 49   | "poly"    |   1    |naph-  |  C1   |
           |      |           |        |thalene|       |          
    13     | 90   | 8 to18    |   2    |    1**|  C1   |Pyrid-    
           |      |           |        |       |       |inium     
           |      |           |        |       |       |          
           |      |           |        |       |       |          
    _______|______|___________|________|_______|_______|_____
    
    * The standard reagent.
    ** 3,4-dichlorobenzyl.
    
    TABLE 8: CHEMICAL STRUCTURE OF SURFACTANTS
    THAT DID NOT REACT WITH THE GUMMY SUBSTANCE
    
    _____________________________________________________
           |     |          |      |      |       |
    Sample |Titer|C in alkyl|Methyl|Benzyl|Halogen|Other
    no.    |     |chain     |groups|groups|       |
    _______|_____|__________|______|______|_______|______
    14     |     |          |   3  |   1  |   C1  |
    15     |     |    *     |   2  |   1  |   C1  |
    16     |     |   16     |   3  |      |   Br  |
    17     |     |   16     |   3  |      |   Br  |
    18     |     |   16     |   3  |      |   Br  |
    19     |     |   18     |   3  |      |   C1  |
    20     |     |   12     |   3  |      |   C1  |
    21     |     | 8 to 18  |   2  |      |   Br  |Ethyl
    22     |     | 8 to 18  |   2  |      |   Br  |Ethyl
    23     |     |   18     |   2  |      |   Br  |Ethyl
    24     |     |   18     |   2  |      |   Br  |Ethyl
    25     |     |   16     |   2  |      |   Br  |Ethyl
    26     |     | alkenyl  |   2  |      |   Br  |Ethyl
    27     |     |   16     |      |      |   C1  |Pyridinium
    28     |     |   16     |      |      |   C1  |Pyridinium
    29     |     |   16     |      |      |   C1  |Pyridinium
    30     |     |   16     |      |      |   Br  |Pyridinium
    31     |     |   **     |      |      |   C1  |Pyridinium
    32     |     |          |      |      |       |Amine
    33     |     |          |      |      |       |Amine
    34     |     |          |      |      |       |***
    _______|_____|__________|______|______|_______|___________
    
    * Myristamido propyl.
    ** "Complex ester."
    *** "Tertiary amine salt."
    

    The titer with the quarternary chosen (benzalkonium chloride) is closely parallel with the number and proportion of reductogenic streptococci in different parts of the body, and with the results of many biochemical reactions (2).

    The titer drops close to normal after optimum immunization with the specific antigen, but rises when the dose is too high or the interval between doses is too short. The increased titers remain high as long as the effects of overdosage persist (14).

    [How does "optimum immunization with the specific antigen" relate to the the "bacterial antigen" question?]

    I worked out three methods for isolating the active substance from urine:

    In one experiment, 1,280 ml. of urine, which had a titer [with the "chosen quarternary (benzalkonium chloride)"] of 56 units (total 71,680 units), was concentrated as just described. It yielded 15.3 ml. of syrup, with a titer 4,600 units, making a total of 70,380 units, which is equivalent to 98 per cent recovery. This crude syrup is identical to that described by Fujita and Ging (1).

    The syrup contained 0.03 per cent P [Phosphorous?]. Tests for purines and pyrimidines were negative. There was no characteristic U.V. absorption.

    The syrup was separated by dialysis into a dialyzable and nondialyzable fraction. Both fractions were concentrated in vacuo and treated with absolute methanol, providing soluble and relatively insoluble fractions. The methanol was removed, and the residues were diluted with water to make 1.0 per cent solutions and studied.

    The results are shown in TABLE 9. Fractions 3 and 4 are dark brown and nondialyzable. A reduced dark nondialyzable fraction with a pungent ordor was isolated from peptones, as mentioned previously. Urine containing the gummy substance has as unpleasant odor, whereas normal urines are aromatic.

    The different reactions to different reagents suggest the possibility of using them for quantitative determination of the separate fractions.

    For example, flourescence is characteristic of fractions 1 and 2. The aged indolphenol reagent is specific for fraction 1. The difference, representing fraction 2, should coincide with the amount determined by the leuko crystal violet reagent, which is relatively specific for fraction 2.

    Neotetrazoliums should provide quantitative data on fraction 3, which can be subtracted from the quat titer [titer by the "chosen quarternary (benzalkonium chloride)" of] (fractions 3 and 4) to give fraction 4.

    No reagent has been found which is specific for all the fractions of the gummy substance, although several appear to be specific for individual fractions.

    Standardization of such reactions must be precedeed by isolation and purification on the individual fractions, including the "button," in an inert atmosphere.

    Although solution of the quat precipitate is used as an end point in the titration, it is possible that one of the newer quat indicators may give more reliable results. Also, it is possible that another quat, such as one containing substitution of the benzyl group, may give more reliable data, as indicated by the higher titers with numbers 12 and 13 (TABLE 7).

    TABLE 9 
    
    CHEMICAL REACTIONS OF FRACTIONS OF THE GUMMY SUBSTANCE
    
    __________________________________________________________
    Property             |Fraction 1           |Fraction 2    |
    _____________________|_____________________|______________|
    Dialyzability        | +                   | +            |
                         |                     |              |
    Solubility in        |                     |              |
    absolute methanol    | ++++                | +            |
                         |                     |              |
    Physical appearance  | Thick syrup         | Thick syrup  |
                         |                     |              |
    Color of 1% solution | Light brown         | Light brown  |
                         |                     |              |
    Solubility in water  | ++++                | ++++         |
                         |                     |              |
    Fluorescence         | ++++                | ++++         |
                         |                     |              |
    Reduction of fresh   |                     |              |
    indophenol*          | ++++                | +            |
                         |                     |              |
    Reduction of         |                     |              |
    methylene blue       | +                   | ++           |
                         |                     |              |
    Precipitate with     |                     |              |
    aged indophenol**    | ++++                | 0            |
                         |                     |              |
    Precipitate with     |                     |              |
    quat reagent***      | +, redissolves      | +, re-       |
                         |                     |  dissolves   |
    Precipitate with     |                     |              |
    neotetrazoliums****  | ++                  | + or -       |
                         |                     |              |
    Precipitate with     |                     |              |
    leuko crystal        | +, redissolves      | ++++         |
    violet*****          | with blue solution  |              |
    _____________________|_____________________|______________|
    ___________________________________________________________
    Property             |Fraction 3           |Fraction 4
    _____________________|_____________________|_______________
    Dialyzability        | 0                   | 0
                         |                     |
    Solubility in        |                     |
    absolute methanol    | ++++                | +
                         |                     |
    Physical appearance  | Scales              | Amorphous
                         |                     |
    Color of 1% solution | Dark brown          | Dark brown
                         |                     |
    Solubility in water  | ++                  | ++
                         |                     |
    Fluorescence         | 0                   | 0
                         |                     |
    Reduction of fresh   |                     |
    indophenol*          | ++                  | ++++
                         |                     |
    Reduction of         |                     |
    methylene blue       | ++++                | ++++
                         |                     |
    Precipitate with     |                     |
    aged indophenol**    | 0                   | 0
                         |                     |
    Precipitate with     |                     |
    quat reagent***      | ++++                | ++++
                         |                     |
    Precipitate with     |                     |
    neotetrazoliums****  | +++++               | +
                         |                     |
    Precipitate with     |                     |
    leuko crystal        | +, flocculent       | ++
    violet*****          |                     |
    _____________________|_____________________|_______________
    
    * 0.1% Sod, 2,6-dibromobenzenoneindo-3`-methylphenol.
      ["indophenol"]
    
    ** Same reagent, aged until it gives a gray spot on
       filter paper.
    
    *** 1.0% alkyl dimethyl benzylammonium chloride
        (benzalkonium chloride).  [The "quarternary chosen"]
    
    **** Neotetrazolium phoaphate 2B, neotetrazolium
         chloride, neotetrazolium blue.
    
    ***** 4,4`,4``-hexamethyltriaminotriphenylmethane,
          dissolved in a minimum amount of N HCI.
    
    

    Before these separate fractions were discovered, I chose an arbitrary unit in the quaternary titration. When 1.0 ml. of fresh clear urine is titrated with 1.0 per cent benzalkonium chloride, each 0.01 ml. of the quat reagent required is considered as one unit (14).

    The titration method employs an overshot end point to compensate for the insoluble residue of precipitated nucleoproteins; in many high titer specimens, this makes recognition of the end point difficult. Chinard (15) used a quaternary for quantitative determination of protein.

    For the titration, put 1.0 ml. of urine into each of two small tubes. Against a dark background, add reagent to the first tube until the turbidity, which reaches maximum at first then gradually diminishes, does not decrease any further. Add one or two extra drops of reagent to provide an overshot end point.

    Titrate the second tube similary until, after the initial rise in turbidity, it drops to an opalescence equal to that of the first tube. In case of doubt, reverse the order of the tubes. "Q sharp" paper was unsatisfactory for determining the end point.

    The following relationships have been established:

    Top -- Home

    Possible Identity of the Gummy Substance with the Neurotoxic Fraction Isolated from Schizophrenic Blood

    Several investigators, e.g. (16) - (19), have isolated a neurotoxic fraction from the blood of schizphrenic patients that may be similar to the gummy substance in urine.

    In fact, the gummy substance had been isolated by Fujita and Ging (1) from the urine of schizophrenic patients, and to a less extent from nonschizophrenic neurological patients. The blood substance was reported to be nondialyzable, (19) but can be transferred across a cellophane membrane (20).

    These apparently conflicting statements agree with my finding that the gummy substance in urine contains both dialyzable and nondialyzable fractions (SEE TABLE 9).

    The blood substance was reported as proteinaceous (19) "or some substance so closely bound to protein as to survive the methods used for isolation of proteinaceous material" (19).

    This could be a nucleoprotein and agrees with my finding that, when the gummy substance is produced artificially, it may be excreted in the urine for several days, suggesting that the substance becomes attached to a blood fraction and is released slowly.

    The substance isolated from blood is associated with A2 globulin, which disappears on storage of the serum, with increase in B1 globulin (18).

    The substance is extremely sensitive to oxidation (16-18). It is colored (19). It is not a single compound (19).

    No difference was demonstrated between the active fraction isolated from the blood of schizophrenics and that from nonschizophrenics donors (19). In fact, Sanders et al (19) found neurotoxic values of similar magnitude among both groups (TABLE 10). Hence, the substance isolated from blood is similar to that from urine (TABLE 11).

    
    TABLE 10
    
    C.T.D. MIN. SEC. PER MG. PROTEIN*
    ________________________________
                  |            
    From normal   | From
    donors        | Schizophrenics
    ______________|_________________
                  |            
            0.4   |         3.9
            2.6   |         9.2
            3.3   |         9.5
            4.0   |         9.5
            4.3   |        11.9
            6.8   |        19.2
            8.1   |        26.6
            ___   |        ____
    Average 4.2   |        12.8
    ______________|_________________
    
    *This Table adapted from Sanders et al. (19)
    
    TABLE 11
    
    PROPERTIES OF SUBSTANCES ISOLATED FROM BLOOD AND URINE
    
    ________________________________________________________
                                 |           |      
                                 | Found in  |
                                 | blood     | Isolated
    Property                     | fraction  | from urine
    _____________________________|___________|______________
                                 |           |      
    Color                        | "colored" | Brown
                                 |           |      
    Mixture                      | Yes       | Yes
                                 |           |      
    Low O/R potential            | Yes       | Yes
                                 |           |      
    Dialyzability                | Yes and no| Yes and no
                                 |           |      
    Toxic                        | Yes       | Yes
                                 |           |      
    Attached to protein          | Yes       | Probably
                                 |           |      
    Found in both schizophrenic  |           |
    and nonschizophrenic patients| Yes       | Yes
    _____________________________|___________|______________
    
    

    Top -- Home

    Comaprison of the Quat Titer of Gummy Substance in Urine with the Results Obtained by Other Biochemical Methods

    Because a number of factors affect different biological processes, a high degree of correlation cannot be expected among biochemical methods. Also, the methods vary in sensitivity.

    On the basis of the values reported previously (2), moderate, marked, and hypernormal (i.e., abnormal) values were obtained in various proportions according to the method used (TABLE 12).

    TABLE 12  
    
    PERCENTAGE OF POSITIVE FINDINGS IN 1,800 CHRONIC INVALIDS
    
    ___________________________________________________________
                           |          |            |           |
                           | Percentage of patients showing    |
    _______________________|___________________________________|
                           |          |            |           |
    column 1               |    2     |    3       |    4      |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Method                 | Negative |      +     | ++  &     |
                           |          |            |   +++     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Resistant S. salivarius|          |            |           |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Total filamented       | 3,510 to | 2,810 to   | 1,510 to  |
    leukocytes             | 3,700    | 3,500      | 2,800     |
                           |  2.3%    |  9.3%      | 46.3%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Escherichia and        | 75,000 to| 25,000 to  |  1,000 to |
    Aerobacter in feces*   | 990,000  | 75,000     | 25,000    |
                           | 16.1%    | 10.1%      | 34.3%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Total leukocytes       | 7,900 to | 7,000 to   | 5,010 to  |
                           | 8,950    | 7,890      | 7,000     |
                           | 9.6%     | 17.4%      | 37.2%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Nonfil-filament ratio  | 0 to 50  | 51 to 75   | 76 to 135 |
            %              |          |            |           |
                           | 7.7%     | 23.0%      | 39.4%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Quat titer of urine    | 0 to 10  | 11 to 15   | 16 to 40  |
                           |          |            |           |
                           | 9.4%     | 22.7%      | 44.2%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Resistant S. mitis     |          |            |           |
    _______________________|__________|____________|___________|
                           |          |            |           |
    % Polymorphonuclears   |61.5 to 65| 55 to 61   | 40 to 54.5|
                           |          |            |           |
                           | 21.9%    | 19.0%      | 33.3%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Sedimentation rate     | 0 to 4   | 5 to 12    | 40 to 54.5|
    mm./hr.                |          |            |           |
                           | 14.7%    | 30.5%      | 38.6%     |
    _______________________|__________|____________|___________|
    
    
    ___________________________________________________________
                           |          |            |           |
                           | Percentage of patients showing    |
    _______________________|___________________________________|
                           |          |            |           |
    column 1               |    5     |    6       |    7      |
    _______________________|__________|____________|___________|
                           |          |            |           |
                           |          |            |Total cols.|
    Method                 |   ++++   | Hypernormal|  4, 5 & 6 |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Resistant S. salivarius|          |            | 99.7%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Total filamented       | Under    | Over       |           |
    leukocytes             | 1,500    | 3,700      |           |
                           |  32.7%   |  9.4%      | 88.4%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Escherichia and        | Under    | Over       |           |
    Aerobacter in feces*   | 1,000    | 1,000,000  |           |
                           | 34.7%    | 4.8%       | 73.8%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Total leukocytes       | Under    | Over       |           |
                           | 5,000    | 9,000      |           |
                           | 16.8%    | 19.0%      | 73.0%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Nonfil-filament ratio  | Over 135 |            |           |
            %              |          |            |           |
                           | 29.9%    |            | 69.3%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Quat titer of urine    | Over 40  |            |           |
                           |          |            |           |
                           | 23.7%    |            | 67.9%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Resistant S. mitis     |          |            | 67.0%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    % Polymorphonuclears   |Under 40  | Over 65    |           |
                           |          |            |           |
                           | 3.8%     | 22.0%      | 59.1%     |
    _______________________|__________|____________|___________|
                           |          |            |           |
    Sedimentation rate     | Over 35  |            |           |
    mm./hr.                |          |            |           |
                           | 16.2%    |            | 54.8%     |
    _______________________|__________|____________|___________|
    
    

    The high proportion of positive results in severely ill groups is illustrated in TABLE 13, which shows the percentage of positive findings in 24 patients with primary simple glaucoma.

    I have added other data to the Table to illustrate the high proportion of abnormal findings in these severely ill persons. For example, three patients showed Erysipelothrix rhusiopathiae, which is uncommon organism in urban areas.

    TABLE 13 
    
    PERCENTAGE OF POSITIVE REACTIONS IN 24 PERSONS WITH
    PRIMARY SIMPLE GLAUCOMA
    
    ______________________________________________________________
                                            |                     |
    Method                                  |   % Positive        |
    ________________________________________|_____________________|
                                            |                     |
    Blood sedimentation rate above 5mm./hour|   95                |
                                            | Average is 19 mm ** |
                                            |                     |
                                            |                     |
    Hematology:                             |                     |
     Total filamented leukocyte below 3,000 |  100                |
     Low erythrocytes                       |   72                |
     Color index above 1.0                  |   96                |
                                            | Average is 1750     |
                                            |                     |
                                            |                     |
    Thymol turbidity above 4 units          |   77                |
         precipitate                        |   41                |
                                            |                     |
    Cephalin flocculation ++, +++, and ++++ |   55                |
                                            |                     |
    Urine:                                  |                     |
      quat titer above 15                   |   91                |
      Indican                               |   63                |
      Acetone                               |   12                |
      Glucose                               |    8                |
      Mucus                                 |   68                |
      Calcium oxalate                       |   28                |
                                            | Average is 31 units |
                                            |                     |
                                            |                     |
    Pharynx:                                |                     |
             S. salivarius - high number    |   78                |
                      over 70% resistant    |  100                |
             S. mitis - high number         |   87                |
                      over 70% resistant    |   62                |
                                            |                     |
    Gastrointestinal tract - prececal:      |                     |
     S. salivarious - high number           |   73                |
               over 80% resistant           |  100                |
     S. mitis - high number                 |   82                |
               over 80% resistant           |   82                |
     Low Eachorichin                        |   82                |
     Paracoll present                       |   68                |
     Paracoll: Eacharichin ratio            |                     |
                     above 1:100            |   58                |
     S. salivarius: Eacharichin ratio       |                     |
                     above 1:100            |   77                |
     S. mitis: Eacharichin ratio            |                     |
                     above 1:100            |   81                |
                                            |                     |
    Erysipolothrix rhualopathia  *          |   12                |
    ________________________________________|_____________________|
    
    *  ++, +++, and ++++
    ** Stiles (22) found the average in severely ill
      persons 13.9 mm.
    
    

    While investigating relationships of different reactions of urine, I was impressed by the fact that there appeared to be an association among many of the findings. For example, when the quat titer was high,

  • the reducing power was usually strong;

  • the spacific gravity was high;

  • the reaction was strongly acid;

  • and the rate of excretion was low.

    The studies were made after normal intake of ascorbic acid. For preliminary statistical analysis I have assigned different values to varying degrees of severity, as shown in TABLE 14.

    Comparison of the results obtained by different methods is difficult because of widely different methods of expression and widely different ranges between normal and severe cases.

    TABLE 14 
    
    TENTATIVE ASSIGNMENT OF VALUES FOUND BY URINE METHODS
    _____________________________________________________________
                 |
    Method       |        Relative degrees of severity
                 |_______________________________________________
                 |         |         |          |        |
                 |    0    |    +    |    ++    |    +++ | ++++
    _____________|_________|_________|__________|________|_______
                 |         |         |          |        |
    Rate of      | Over    | 0.8     | 0.60     | 0.50   | Less   
    excretion    |   1.0   |  - 0.99 |  - 0.79  |  -0.59 | than 
    (ml/min.)    |         |         |          |        | 0.50
                 |         |         |          |        |
    Specific     | Under   | 1.011-  | 1.016-   | 1.021- | Over
    gravity      | 1.010   | 1.015   | 1.020    | 1.027  | 1.027
                 |         |         |          |        |
    Quat titer   | Under   | 11-15   | 16-25    | 26-40  | Over
    (units)      |   10    |         |          |        | 40
                 |         |         |          |        |
    ml           | 0.00    | 0.025   | 0.050    | 0.075  | .100
    indophenol * |         |         |          |        |  & 
    reduced      |         |         |          |        | more
                 |         |         |          |        |
    Urates on    |         |         |          |        |
    refrigeration|  0      |         |  ++      |        | ++++
    _____________|_________|_________|__________|________|______
    
    *Ml. of 0.10% sod. 2,6-dibromobenzenoneindo-3`-methylphenol
    reduced by 5.0 ml. of urine in 10 min.
    
    

    Becaause the results obtained by each method can now be assigned to different degrees of abnormality, I am converting the observed findings into per cent of the difference between the highest normal value and the lowest value found in severe cases.

    Both hypernormal and subnormal deviations are treated alike. This makes it possible to compare the results obtained by different methods and to determine the statistical significanceof the percentages.

    Schrek (21) devised a nomogram which simplifies the statistical calculations. As an example of the proposed system, the normal range of quat unit is 0 to 10. Therefore, 10 is considered as 0 per cent. Severe cases show more then 40 units. Since all values above 40 units represent ++++ severity, they are considered as 100 per cent. The use of higher percentages to express higher titers leads to confusion.

    By constructing a graph on which the ordinates of 10 and 40 units are connected,with abscissae of 0 and 100 per cent, respectively, it can be determined, e.g., that 30 units corresponds to 65 per cent severity by this method.

    Relationships of the results obtained by different urine methods, but not calculated on the percentage basis just proposed, are shown in TABLE 15.

    TABLE 15
    
    RELATIONSHIPS AMONG URINE METHODS
    ______________________________________________________
    Method           |No. of    |Mean ml.    | Mean      |
                     |Specimens |reduced*    | titer     |
                     |          |            |           |
    _________________|__________|____________|___________|
    Quat titer:      |          |            |           |
    0 to 20          |45        |.110+/-.022 |           |
    21 to 30         |59        |.156+/-.023 |           |
    Over 30          |19        |.175+/-.028 |           |
                     |          |            |           |
    Specific Gravity |          |            |           |
    Less then 1.019  |11        |            |14.2+/-2.63|
    1.020-1.023      |24        |            |19.4+/-5.33|
    1.024-1.028      |58        |            |23.8+/-6.15|
    Over 1.028       |30        |            |28.2+/-5.57|
                     |          |            |           |
    Urates on        |          |            |           |
    refrigeration    |          |            |           |
    0                |56        |            |10.1+/-5.8 |
    +                |67        |            |34.1+/-5.7 |
    H-ion            |          |            |           |
    concentration    |          |            |           |
    Below Ph 6.1     |113       |.145+/-.078 |25.3+/-6.8 |
    Above Ph 6.1     | 10       |.104+/-.073 |20.3+/-5.3 |
    _________________|__________|____________|___________|
    
    * Ml. of 0.10% sod. 2,6-dibromobenzenoneindo-3'
    -methylphenol reduced by 5.0 ml. urine in 10 min.
    

    Persons with low grade chronic illness frequently experience wide fluctuations in health within surprisingly short intervals. The blood sedimentation rate does not reflect such changes, but most other reactions show almost immediate changes with changes in severity of the symptoms.

    If streptococcal intoxication is the common denominator in chronic diseases, as appears likely from what has been discussed, and if the quat precipitable substance is a reaction product of streptococcal intoxication, then the quat titer should rise and fall with fluctuations in severity of symptoms. This was shown to be true in studies of thousands of persons. [Is it possible that "the quat precipitable substance" is also a reaction product of other microbial infections?]

    Since several other biochemical reactions give results parallel with the urine titer, the differences in results obtained by these methods when they are repeated within a short interval in the same, or another laboratory, may not be the result of technical error as is generally supposed, but of fluctuations in severity of the disease processes.

    Top -- Home

    Oxidation-reduction potentials in the body.

    It would be expected that liberation of the extracellular reducing principle by the reductogenic streptococci would lower the O/R potentials in the body.

    Since different tissues and fluids show variations in H-ion concentration, it would be expected that they may show differences in O/R potential also.

    To a certain extent the pH of the urine is related to acid-base relationships elsewhere in the body. Therefore, I studied the reducing power of the urine, believing that it might reflect O/R relationships in other parts of the body.

    In using the term "O/R potential of the body," I intend it to connote general Eh relationships in the sense that, e.g., acidosis which is a general condition has little to do with the pH of the gastric contents or vaginal secretion.

    I found that determination of the reducing power of the urine is significant when the person refrains from smoking and from an excess of ascorbic acid prior to collecting the urine specimen.

    Ascorbic acid in amounts above 75 mg. per day may be excreted for several days after the dose is taken, and this affects the reducing power of the urine for several days.

    Because of the large number of experiments, I measured the reducing power of the urine by the volume of the indophenol solution which was reduced by 5.0 ml. of urine, pH 7.0, in 10 min.

    Strict euthyroids showed no reduction of the indicator, but other persons sometimes reduced more than 1.0 ml. of the indophenol. Those specimens which reduced strongly were retested, using 1.0 ml. of urine diluted with 4.0 ml. of water.

    The reducing power of the urine usually is parallel with the quat titer, etc. (FIGURE 1). This indicates that the gummy substance produces a subnormal state of oxidation of the body.

                                 FIGURE 1
    
           Parallel between Quat Titer and Reducing Power
       ___________________________________________________________
      |                                                           |
    ##|  ## Quat Titer              && reducing power         &&  |
      |     (********)              of the urine as               |
      |                             ml indophenol                 |
      |     rate of                 (XXXXXXXXX)                   |
      |.1   excretion (OOOOOOO)                                   |
      |       O                                                   | 
    50|.2      O   X                        OOOOOOO          .250 -
      |ml/min   O XX                       O       O          ml  |
      |.3        X  X                     O          O            |
      |         X O  X                   O             O          |
    40|.4      X   O  X                 O                    .200 -
      |      X      O  X               O                          |
      |.5  X         O  X             O                   O       |
      |  X            O  X           O                      O     |
    30|X               O  X         OXXXXXXXXX               .150 -
      |        *        O X        OX         X               O   |
      |.7     *  *      O X       O* * * * * * *X              O  |
      |      *    *      OX     *OX             *X             O  |
    20|.8   *      *     O X   *OX               *X          .100 -
      |    *        *     O X *OX                   X * ** * * O  |
      |.9 *          *     O*XOX                      X       *O  |
      |  *            *   * O X                         X      *  |
    10|1.0             * *                                    .50 -
      |                 *                                 X     O*|
      |1.1                                                  X   O*|
      |                                                       X  O|
     0|1.2_______4____GR. THYROID_____4______4______4___________XX|
    
      9      10     11     7      8      9      10     11     12     
                  PM                      AM
    
    

    Stiles (22) showed that about 85 percent of chronic invalids have a subnormal [below 0?] basal metabolic rate. Therefore, I studied the effect of desiccated thyroid on the reducing power of the urine.

    I took 4 grain doses at frequent intervals and found that they improved, not only the oxidation of the urine, but the quat titer, rate of excretion, urates on refrigeration, and H-ion concentration. Changes in the first three are shown in FIGURE 1.

    Simultaneously with improvement in these findings there was dramatic improvement in general health. This indicated that the toxic reduced principles are rendered nontoxic by oxidation, and this may account for the normal quat titers in euthyroid chronic invalids.

    The low O/R potential, particularly in severe cases, may provide an explanation for some unsolved phenomena. For example, "Bowery bums" are able to drink "smoke," which is a water extract of shellac and consists mostly of crude methanol, and yet show no defect in vision.

    When I was highly toxic, I inhaled enormous quantities of methanol vapor with no disturbance in vision. Normally, methanol is oxidized to formic acid, but apparently when the O/R potential of the body is too low, the person is unable to oxidize it and no formic acid is produced to damage the eyes.

    Again, toxic individuals have a high color index of erythrocytes. It is possible that the low O/R potential in the gastrointestional tract converts ferric into ferrous compunds which are more assimilable.

    The reduction of ferric compounds by some fecal specimens was noted by Bergeim (23). A third example is the aggravation of colds following sexual indulgence. Loss of highly oxidized fluids causes a drop in the O/R potential of the body.

    Top -- Home

    Relationships of the basal metabolic rate (BMR).

    Stiles (22) found that the blood sedimentation rate and basal metabolic rate (BMR) are related to the severity of symptoms, regardless of the disease. The BMR was below 0 in 67, 91, and 95 percent, and the mean sedimentation rate was 3.4, 11.4, and 13.9 mm., in mild, moderately severe, and severe cases, respectively.

    Since the BMR is an oxidation phenomenon, it would be expected to show a relationship to the reducing power of the urine. The two were parallel in most cases, when the volume of the indophenol reduced did not exceed 0.100 ml.; but when larger amounts were reduced, the BMR, instead of being progressively lower with progressively greater reduction, actually was closer to normal in some cases (TABLE 16).

    TABLE 16
    
    RELATIONSHIP OF REDUCING POWER OF URINE TO BMR
    __________________________________________________________
    Ml. indophenol reduced|Number of         |Average         |  
    by 5.0 ml. urine      |persons           |BMR             |
    ______________________|__________________|________________|
    0.000                 |16                |-3              |
    0.025 to 0.075        |14                |-8.5            |
    0.100                 | 3                |-17             |
    Over 0.100            |10                |-8 (see text)   |
    ______________________|__________________|________________|
    

    This tendency of the body to respond to severe, but not to mild, foreign stimuli is illustrated by the rate of excretion of urine. The average rate varies inversely with the reducing power up to 0.075 ml. of indophenol, but it is normal when larger amounts of indophenol are reduced (TABLE 17).

    TABLE 17
    
    RELATIONSHIP OF REDUCING POWER OF URINE TO THE 
    RATE OF EXCRETION
    __________________________________________________________
    Ml. of indophenol reduced|Number of|Average rate of       |
    by 5.0 ml. urine         |persons  |excretion (ml./min.)  |
    _________________________|_________|______________________|
    0.000                    |8        |1.03                  |
    0.025 to 0.075           |5        | .46                  |
    0.100                    |3        | .98                  |
    _________________________|_________|______________________|
    

    In eight severe cases the average BMR was only minus 7.5 whereas the reducing power and quat titer showed considerable abnormalities (TABLE 18).

    There was considerable reducing power, but a normal quat titer, in a hyperthyroid case (TABLE 18). Several other hyperthroid patients showed considerably reduced urine.

    This suggests that, at least in some cases, hyperthyroidism may not be an expression of pathological activity of the gland, but a physiological attempt by the thryoid to oxidize the excess reducing substances.

    TABLE 18
    
    COMPARISON OF REDUCING POWER, QUAT TITER AND
    BMR IN SEVERE CASES.
    ________________________________________________
    ml. indophenol reduced|Quat       |             
    by 5.0 ml. urine      |titer      |BMR          
    ______________________|___________|_____________
             .150         | 20        |-6           
             .200         | 25        |-1           
             .280         | 62        |-2           
             .475         | 67        |-8           
             .350         | 33        |-13          
             .375         | 31        |-9      
             .425         | 32        |-5           
             .350         | 33        |-16
             ____         |____       |____
    Average  .388         | 38        |-7.5
             .100         | 10        |+22      
                          |           |(hyper-                
                          |           |thyroid)     
                          |           |                                 
    ______________________|___________|_____________
    
    
    
    TABLE 19 
    
    BIOCHEMICAL RELATIONSHIPS IN PERSONS WITH
    STRONGLY REDUCED URINE
    
    ____________________________________________________________
               |          |          |     |          |         |
     ml.       |          |          |     |          |Sedimen- |
     indophenol|Filamented|Total     |     |Rate of   |tation   |
     reduced   |leukocytes|leukocytes|Quat |excretion |rate     |
     by 5 ml   |per mm.3  |per mm.3  |titer|(ml./min.)|(mm. in  |
     urine     |          |          |     |          | 1 hr,)  |
    ___________|__________|__________|_____|__________|_________|
               |          |          |     |          |         |
         1.200 |  1,240   |  5,650   |  31 |  0.30    |    1    |
               |          |          |     |          |         |
          .725 |    830   |  4,600   |  34 |  0.43    |   13    | 
               |          |          |     |          |         |
          .425 |  1,100   |  4,950   |  42 |  0.35    |   24    |
               |          |          |     |          |         |
          .400 |    900   |  4,200   |  18 |  0.22    |    2    |
               |          |          |     |          |         |
          .225 |  1,770   |  6,000   |  38 |  0.22    |   18    |
               |          |          |     |          |         |
          .175 |  1,240   |  5,650   |  31 |  0.30    |    1    |
         _____ |  _____   |  _____   |  __ |  ____    |   __    |
               |          |          |     |          |         |
     (Average) |          |          |     |          |         |
          .525 |  1,180   |  5,175   |  32 |  0.30    |   10    |
    ___________|__________|__________|_____|__________|_________|
    

    Top -- Home

    Relationships of leukocytes.

    Usual differentation of polymorphonuclear neutrophilic leukocytes is based on the morphology of the nucleus.The cell is considered to be mature when the lobes of the nucleus are seperated by no more than a narrow band.

    My experiments support the claim of Crocker and Valentine (24) that leukocytes are truly mature when the lobes of the nucleus are separated by a thin filament or thread.

    The proportion of filamented leukocytes is low in most chronic invalids but the significance is increased considerably when they are considered on the basis of the total number per mm^3. Strictly normal persons have 3,600 filamented leukocytes per mm^3, but the average chronic invalid has about one-half of this number (FIGURE 2).

    Figure 2
    Total and Filamented Leukocytes

    It was shown in TABLE 12 that the total number of filamented leukocytes gave the second highest proportion of positive results of any of the methods reported. Therefore, it is a highly significant method.

    A person may have a leukocytosis of 16,000 per mm^3 and yet have only 2,400 filamented leukocytes (FIGURE 2). I refer to such deficient leukocytosis as "inefficient stimulation." The majority of persons with a strictly normal number of leukocytes (7,500 to 8,900) have less than 3,600 filamented leukocytes.

    Top -- Home

    Relationships to the rate of excretion of urine.

    Several disease conditions (e.g., myxedema, adrenal insufficiency, diabetes, hyperaldosteronism, etc.) are known to affect the rate of excretion of urine.

    With these exceptions, the rate of excretion is inversely proportional to the severity of the intoxication. It is above 1.0 ml. per minute in normal persons and in euthyroid chronically diseased persons, but it is diminished in the toxic states under discussion in proportion to the severity of the intoxication.

    Fujita and Ging (1) found that schizophrenics excrete about 20 to 25 percent less urine than do nonschizophrenic patients. [Does this mean that some cases of schizophrenia may be the result of microbial infections?]

    Fluid intake does not increase the rate of excretion of urine when it is less than 1.0 ml. per minute, but it does increase the rate severalfold in most cases after fluid intake, when the rate of excretion is more than 1.0 ml. per minute.

    It was shown in TABLE 17 that the average rate of excretion was almost normal in three persons who showed markedly reduced urine. However, other persons with markedly reduced urine showed a low rate of excretion (TABLE 19), possibly because the toxic stimulus was too severe to permit response.

    A similar relationship in thyroid activity was just mentioned. It is similar to the stimulation of antibody production by highly pathogenic microorganisms but not by relatively innocuous ones. Hence, the parallel between severity of the toxic stimulus and the biochemical findings may be poor in some severe cases.

    [Does this mean that a severe infection would stimulate the production of fewer antibodies if the infecting agent were not "highly pathogenic," that is, if the severe infection were not capable of killing or disabling the patient in a relatively short period of time?

    Does "the toxic stimulus was too severe to permit response" mean that, whether or not the microbial infection were "highly pathogenic," antibodies would not be produced in significant numbers because the immune system is overwhelmed?]

    In the reduced state, the gummy substance has strong anti bacterial power. For example, when urine is allowed to stand in an open vessel, it will decompose rapidly if the quat titer is below 15 units. The higher the titer is above 15 units, the longer it will remain clear. High titer specimens show no bacterial decomposition in several days.

    This relationship is reflected in the genitourinary tract. The titers of clear supernates of urine from persons with genitourinary tract infections were 9, 15, 12, 10, 17, 11, 13, 14, 7, 15, and 13 units. It is probable, then, that the presence of the gummy substance in urine in more then about 15 units prevents multiplication of bacteria in the genitourinary tract.

    A similar relationship was found in the gastrointestinal tract. In a freshly passed, mushy (not merely "soft") stool obtained after a laxative, such as a compound cathartic, which I refer to as a "prececal" specimen, the higher the number and proportion of reductogenic strepococci, the smaller the number of Escherichia and Aerobacter, but the larger the number of paracoli.

    Chapman et al(25) found that the average number of Escherichia in millions per 100 gm. of dry feces was 57,000, 162,000, and 668,000 when the resistance of the fecal strepococci was high, moderate, and nil, respectively. This is an illustration of the toxic inhibiting effect of strepococci on these organisms.

    After the feces passes the ileocecal valve, liquids are absorbed and the specimen becomes formed. This is accompanied by marked changes in the flora, which indicates that the gummy substance is absorbed in the colon.

    The number of califorms rises, streptococci disappear, and enterococci, which are rare in prececal stools, become abundant. The presence of enterococci raises suspicion that the specimen was postcecal.

    Rawls and I (26) found that the inhibition of coliforms does not occur in the small intestine when the person has achlorydria because the relatively low gastric acidity permits such rapid multiplication of coliforms in the upper gastrointestinal tract that the reductogenic streptococci are unable to inhibit them.

    In addition to the number of escherichia and Aerobacter in the small intestine, there is an increase in the number of paracoli. Previously they were considered to be variants of Escherichia and Aerobacter, but recently there had been a tendency to form a new genus - Paracolobactrum.

    However, typical Escherichia and Aerobacter can be converted into paracoli by growing them in the presence of the gummy substance. Morphologically, the colonies of paracoli resemble those of the present species. Not only do the paracoli so derived lose power to ferment lactose but they acquire the strong reducing power of the reductogenic streptococci.

    This genetic mutation is readily demonstrated on Tergitol-7 agar with TTC [neotetrazolium chloride?] (27) which is reduced by paracoli and a few other lactose nonfermenters.

    The gummy substance can be extracted from feces by adding water, removing solid particles, and treating the clear, highly colored extract as described for urine.

    Top -- Home

    Detection of In Vivo Nucleic Acid Hydrolysis

    If ingested nucleic acid or nucleoprotein is not assimilated, it would be expected that it would be hydrolyzed and the hydrolytic products (purines, pyrimidines, ribose or deoxyribose, and phosphoric acid) would expect to find in the urine:

    • an increase in uric acid or urates, from purines;

    • oxalates, from pyrimidines (28);

    • and excess phosphates.
    Before I realized that I was studying products of nucleic acid hydrolysis, I had found the following simple urine methods highly useful in following the progress of toxic effects on the body:
    • rate of excretion;

    • specific gravity;

    • quat titer;

    • reducing power;

    • H-ion concentration;

    • urates on refrigeration;

    • microscopic examination for mucus, RBC and WBC and crystals;

    • and examination for crystals after storage in the refrigerator.
    Quantitative methods can now be substituted for the semiquantitative ones, and additional methods can be developed.

    With these refinements, it should be possible to determine the extent of nucleic acid decomposition in the body with considerable precision by simultaneous use of several of these quantitative methods.

    The concept of hydrolysis of unassimilable nucleic acids causes these methods and interpretation to assume a new and highly significant role in health-disease relationships.

    Top -- Home

    Oxidation-Reduction Potential Relationships of Nucleic Acids

    Commercially available nucleic acids and derivatives are relatively inneffective in improving the number and maturity of leukocytes. I considered the possibility that the low O/R potential produced by the reductogenic streptococci may interfere with nucleic acid assimilation.

    Early experiments in oxidizing yeast RNA were made with hydrogen acceptors. As an example, yeast RNA treated with methylene blue, which has an E0 of + .011 volts at pH 7.0, produced dramatic benefit in incipient colds. In many cases, the symptoms disappeared completely in less than 20 mins. The results were unsatisfactory when the cold was more then a few hours old.

    The proportion of excellent results was higher when resazurin, which has an E0 of + .051 volts at pH 7.0, was used and was still higher when thionin, which has an E0 of + .062 volts at pH 7.0 was used. These preparations are undesirable because they tend to produce methemoglobinemia and hyperuricacidemia.

    Therefore, I studied other possible oxidizing methods, such as quinone-hydroquinone resin, oxygen donors, and electron acceptors. The O/R potential of yeast RNA could be raised as high as Eh + .196 volts at pH 7.0. Although many of the results were dramatic, many persons could not tolerate a daily dose of 50 nanograms. Insomnia was the common complaint.

    This presented the possibility that an excessively high potential is just as toxic for nucleic acid as is an excessively low potential.

    One curious observation, which is being studied extensively, is that the excessively high potential RNA causes reduction rather then oxidation, of the urine. This phenomenon is similar to the finding of reduced urine in hyperthyroidism mentioned previously.

    These findings suggest that optimum Eh lies within as narrow a range as does pH. The toxic effect of hyperventilation of the body may be a similar phenomenon.

    The toxic effects of radiation which are beneficial in small doses may be similar also. So, the urine methods should provide as useful a guide to optimum dosage as they are to optimum dosage of streptococcal antigen.

    The different O/R potentials in different people preclude the possibility of using uniformly optimally oxidized nucleic acid for therapy. Moreover, optimum oxidation is only one phase of nucleic acid assimilation.

    In studying the effects of vitamins and minerals on the assimilability of yeast RNA, I observed that all of them produced unfavorable effects, but in different degrees. The unfavorable effects were identical with those produced by ingestion of hyper- or hypo- oxidized RNA and by the reductogenic streptococci.

    [Is it possible that vitamins, minerals, or any nutrient may have adverse effects if they are ingested by a sick person? Is this a reason for fasting: 'One man's meat is another man's poison'? For example, with the use of aminoguanidine to counter the Maillard reaction (linking glucose to proteins), normal rats apparently benefit because the crosslinking of their skin is slowed down, but diabetic rats may be harmed because the crosslinking of their skin seems to be accelerated.]

    It appears, therefore, that any substance or principle that interferes with nucleic acid assimilation, or renders it unassimilable, creates a low O/R potential within the body.

    Since the brown gummy substance is produced in such reactions, it is possible that this substance is a hydrolytic product of reduced nucleic acid. The substance appears to be identical, regardless of the nature of the interfering agent.

    [Does this mean that a wide variety of microbes can be an "interfering agent" that results in the production of the gummy substance?]

    Since the brown gummy substance is related to severity of symptoms, it may be the basic toxic principle in chronic diseases.

    I swallowed a spoonful of the gummy substance to test this point. I developed severe malaise, excessive tirednes, lassitude, acute sinusitis severe colitis and severe headache, which lasted several days.

    These observations coupled with the findings discussed previously lead me to conclude that the brown gummy substance is the major toxic principle in chronic diseases; that it usually originates from the reductogenic streptococci, but that many other substances and principles may produce it or may increase the amount produced by the reductogenic streptococci.

    The conclusion that the commonest and most potent source is the reductogenic streptococci is based on several considerations:

    1. "Vaccines" prepared from autogenous coliforms in genitourinary tract infections have little therapeutic value, but minute doses of reductogenic streptococcal antigen from the same person produce dramatic benefit.

    This indicates that the coliforms are not primary invaders, and suggests that the products of reductogenic streptococci may reduce local tissue resistance and permit proliferation of coliforms and other feebly pathogenic bacteria.

    If the reductogenic streptococci are responsible for primary tissue damage, the quat titer should be high, but it does not exceed 17 units.

    This presents the possibility that the brown substance is oxidized by the thyriod but that the streptococcal antigen, administered orally or parenterally, cause exaggeration of symptoms.

    A similar example is the failure to recover viable microorganisms from most eyes that have been enucleated because they showed marked evidence of severe inflammation.

    Griffiths and I (29) showed that organisms found in bile cultures are likely to be the result of contamination, e.g., from the duodenal tube.

    Coliforms and other weakly pathogenic bacteria usually occur outside the gastrointestinal tract only when a large number of strongly reductogenic streptococci is present. The coliforms in such cases do not possess increased pathogenicity. Therefore, they are likely to be secondary invaders.

    2. Reductogenic streptococci are found in increased number and in high proportion in all chronic invalids, but not in normal persons. Hence, they are an outstanding source of intoxication.

    3. The number and proportion of reductogenic streptococci is proportionate to the severity of the intoxication, as judged by the severity of symptoms, and to the quat titer of the gummy substance.

    4. An oral antigen prepared from the toxigenic streptococci had proved highly effective in a wide variety of diseases in doses that are close to the limit of Avagadro's number. Hence, this antigen must have a basic relationship to these diseases.

    These observations lead to the conclusion that the gummy substance of Fujita and Ging, and the substance isolated from the blood of schizophrenic and nonschizophrenic patients, is produced when a substance or principle interferes with nucleic acid assimilation and that the commonest origin of such interference is the reductogenic streptococci.

    On the basis of foregoing, it is concluded that the etiology of chronic diseases consists of the following sequence of events:

    • Initial entrance to the body is most likely to occur from a massive inoculation, such as close contact with parents, or from a severe debilitating disease.

    • This reduces nucleic acid assimilability, with consequent depletion of nucleic acids in cells and fluids.

    • Deficient erythropoietic and leukopoietic tissues gradually produce erythrocytes and leukocytes of inferior quality. The former deficiency lowers the oxygen carrying capacity to the tissues; the latter causes gradual reduction in the antibacterial power of leukocytes.

    • This, in turn, permits progressively greater activity of reductogenic streptococci, which are thus able to perpetuate themselves.
    Contrary to the general belief that these organisms are suspectible to antibiotics, administration of the antibiotic produces only temporary marked reduction in their number, particularly on exposed surfaces; the organisms, however, reappear and become just as numerous as before when the antibiotic is discontinued.

    Lack of competition from other pathogens permits even greater entrenchment of the reductogenic streptococci, and may account for the increased incidence of certain chronic diseases.

    Superficially, this concept might be confused with the "focal infection hypothesis," which is no longer tenable. However, there are several basic differences. The focal infection hypothesis assumes that "Streptococcus viridans" and "nonhemolytic streptococci," names that are based on unreliable taxonomic consideration, implant themselves in "primary foci."

    On the contrary, my work indicates that the pathological effects are produced by a toxic principle, which is produced by certain variants of S. salivarius and S. mitis, the nature of which has just been described.

    There is ample evidence that microbic dissociation is common in these two species, and to a certain extent they may able to gain or lose toxigenicity according to the enviroment. The undulating effect of changes in the weather is well known.

    Since the brown product of hydrolysis of reduced nucleic acid is the major toxic agent in chronic diseases,

    • is the intoxication caused by this reduced substance,

    • or is it caused by the negatively charged particles themselves?
    My experiments indicate the former, since an excessively high charge produces identical results.

    In short, this may explain pathological effects in oxidized tissues, such as thyroid and sex glands. Thus, the brown gummy substance is produced when nucleic acid is associated with an abnormal electrical charge.

    It is possible that metals, which have characteristic individual potentials, and vitamins, many of which are oxidizing agents, may act unfavorably on nucleic acid by producing an abnormal charge.

    It is possible, also, that a similar mechanism may hold for DNA, which could account for congenital defects. If DNA cannot be assimilated in adequate amounts, the DNA-deficient mother may not be able to produce a normal fetus.

    Top -- Home

    RNA, THE SECOND BIO INDICTATOR

    It has been established that DNA dictates heredity by directing it's replication in chromosomes. If interference with RNA assimilation is responsible for ill health, is the reverse true, viz., that RNA assimilation is essential for health?

    Is RNA the factor which controls and maintains orderly biochemical and biophysical relationships of the body? Progress already achieved in developing assimilable RNA indicates that this is true.

    When there is a deficient supply of RNA, the body cannot function normally, thus permitting development of a wide variety of biochemical, biophysical, cellular, hormonal, metabolic, and other abnormalities.

    When developing assimilable RNA, I was puzzled because many of the experimental lots were highly toxic even though some were physical mixtures of yeast RNA and common food ingredients.

    When I swallowed such mixtures I usually developed a variety of toxic symptoms, such as excessive tiredness, lassitude, scanty urine, headache, heartburn, lack of mental alertness and clarity, foul light tan or spastic stolls, and even clay colored, mostly liquid stools, and a very high quat titer.

    The difficulty of correcting biochemical abnormalities by dietary restrictions or supplements is that the basic disturbance in nucleic acid continues to cause the abnormalities. In fact, such dietary attempts may aggravate the abnormality by increasing the interfering mechanism.

    When I began to find out how to make RNA assimilable, I observed that it no longer produced toxic effects, but favorable ones.

    As the method of preparing it was improved, there was better correction of the subjective and objective abnormalities.

    Not only did it include correction of abnormal pH relationships, improved thyroid function, reduction in gout symptoms (which had been brought on by earlier experiments), release of retained water, increase in mental alertness, and improvement in word blindness; but there was also dramatic improvement in allergy.

    During the early experiments, I had developed violent inhalent allergy to both RNA and DNA, para substituted benzenes, such as PABA and indophenols, and even to almost invisible traces of tobacco smoke.

    I now manipulate these chemicals without the slightest discomfort even through the air may be cloudy from nucleic acid dust.

    The effect of taking a dose of assimilable RNA is dramatic. Even when symptoms are severe and there are large amounts of derivatives of reduced nucleic acid in the urine, subjective and objective improvement begins in about 20 minuntes, even after a dose of only 1 mg. This amount is sufficient to maintain excellent health for about 24 hours.

    This indicates that RNA is essential for robust health, since adminintration of assimilable RNA corrects biochemical and other abnormalities and maintains their normal relationships. Soon after taking 1 mg. of assimilable RNA, the urine titer drops to less then 10 units and remains low for about 24 hours.

    Ill health follows when RNA cannot be assimilated. It is a dictator because the effective daily dose is about 1 mg., a ratio of about 1:70,000,000 for a 150 lb. person. It seems reasonable to conclude that RNA controls the basic physicochemical relationships of the body and, through them, normal physiological, cellular, and other relationships and functions.

    Assimilable RNA is a highly precise and easily distorted molecule. Even moisture ruins it in about 15 minutes. This precise configuration is necessary to overcome interference with normal nucleic acid assimilation. It is unnecessary in healthy persons because they are able to assimilate dietary nucleoproteins without interference.

    Top -- Home

    POSSIBLE RELATIONSHIP BETWEEN THE NUCLEIC ACID INTERFERING MECHANISM AND MALIGNANCY AND BLOOD DYSCRASIAS

    Earlier, physicians had reported favorable results in neoplastic diseases and blood dyscrasias by use of specific streptococcal antigen; but clinical evidence is inconclusive, no matter how dramatic it is, unless the experiments are conducted under well controlled circumstances, on a sufficiently large number of cases, and over a sufficiently long period of time to eliminate enthusiasm and to be sure that the results are really based on improvement in the basic disturbed relationship, and are not merely a temporary drug effect.

    [What is "specific streptococcal antigen"? See questions regarding bacterial antigens.]

    My experiments indicate that the basic disturbance which cause neoplastic diseases and blood dyscrasias is similar to that in other chronic diseases.

    The term "nucleic acid starvation" has been used in cancer research and is very appropriate. Warburg (30) maintained that cancer is produced by irreversible reduction of cellular respiration beyond a critical point.

    Use of the term "respiration" has led to rejection of his ideas in many quarters. But if his view were modified to indicate that cancer results when the O/R potential (and possibly the H-ion concentration) of the cell exceeds a critical limit or when the normal nucleic acid falls below a critical limit, it would approach the concept which I have described.

    My view that neoplastic diseases and blood dyscrasias are caused by interference with nucleic acid assimilation is supported by the following considerations:

    • 1. If cancer and blood dyscrasias have a similar basic etiology to other chronic diseases, then the basis is distortion of the nucleic acid molecule by reductogenic variants of S. salivarius and S. mitis and by other chemical and physical agents [including other microbes?].

    • 2. These streptococci are only temporarily affected by antibiotics, as described previously. Only superficial or exposed cells are suspectible to their action. With few exceptions, cancer is unaffected by antibiotics.

    • 3. Interference with assimilation of nucleic acids by the mechanism described previously could account for the "nucleic acid starved cells" and the abnormal nucleic acid metabloism which occurs in cancer. It is possible that the body is compelled to accept some of the distorted nucleic acid molecule, resulting in an abnormal nucleic acid pattern.

    • 4. Reductogenic streptococci are able to produce genetic mutations, as described previously. Mutation is regarded as an integral part of carcinogenesis.

    • 5. Although many cancer tissues reduce TTC and cancer tetrazoliums, the phenonenon is inconstant. Cancer may occur in highly oxidized tissues, such as the thyroid and sex glands. Also, cancer can be caused by highly oxidized hormones.

      Huggins and Yang (31) showed that cancer can be induced both by strong electron donors and acceptors. These apparently paradoxical relationships are explained by the finding that any interference with nucleic acid assimilation may prevent or retard cellular maturity, with consequent inability of the cell to assume its normal functions.

      Because of this impaired nourishment, the cell is relatively undifferentiated, which is characteristic of many cancer cells.

    • 6. The suspected relationship of free radicals could be a function of their unpaired electron.

    Top -- Home

    SUMMARY AND CONCLUSIONS

    Streptococcus salivarius and S. mitis dissociate into pathogenic and nonpathogenic variants. The pathogenic variants are pathogenic for rabbits. The variants are found constantly in chronic invalids, the number and proportion to total streptococci fluctuating with severity of symptoms, provided that "minor ailments" are considered disease processes.

    The pathogenic variants are uncommon in persons with robust health. In addition to being resistant to a number of chemical agents, they possess strong reducing power which lowers oxidation-reduction potentials in the body. The reduced enviroment affects ingested nucleoproteins and nucleic acids, making them relatively unavailable to the body.

    A number of other principles and substances also interfere with nucleic acid assimilation. The unassimilable nucleic acid, which becomes the gummy substance of Fujita and Ging, produces highly toxic effects.

    Evidence is produced which indicates that the gummy substance is the common toxic principle in chronic diseases.

    Hydrolytic products of unassimilable nucleic acid can be measured in the urine, which provides data about interference with nucleic acid assimilability.

    The poor acceptance of nucleic acids in chronic diseases leads to "cell starvation" that, after it reaches a critical point, may cause the cells to become neoplastic. Blood-forming organs become malnourished and have difficulty in producing normal blood cells.

    Attempts to introduce RNA into the body without interference indicate that RNA dictates basic health relationships and that disease follows any interference with this normal process.

    LETTER to SCIENTIFIC RESEARCH (now Science News), 9-15-69, by George H. Chapman

    ... I presented substantial evidence:

    • (1) that reductogenic (i.e., electron donor) dissociates of two species of streptococci are the common denominator of chronic disease;

    • (2) that the acid - reduced state that they produce causes "rejection," which I now recognize as precipitation of RNA;

    • (3) that the rejected RNA is hydrolyzed and metabolized and the products are excreted in the urine; and

    • (4) that one of these hydrolytic products is the straw or amber coloring matter of urine which can be titrated with benzalkonium chloride.

    Loss of RNA by this process is slight in normal persons, but increases parallel to the severity of the disease process and with many subjective and objective symptoms that I described.

    It is almost impossible to maintain strictly optimum pH and E0 at all times, even in persons with robust health. Therefore there is constant but slight loss of RNA, even in normal persons. This is the basis of the aging process.

    Persons who have had only slight loss are young at 75, whereas those who have had heavy loss are old at 50.

    Consequently I differentiate between aging (slight loss) and accelerated aging (moderate to severe loss) ...

    ["Aging" is probably the result of numerous influences, some of which, when acting at the same time, may have multiplicative (interacting) effects. Dr. Chapman has presented strong evidence that chronic microbial infections are among these influences.

    For more on the impact of infection on human aging, see "Are there spirochetes in the brain in more than 90 percent of Alzheimer's Disease cases?"

    www.avonhistory.org/bug/alz11.htm]

    Top -- Home

    REFERENCES

    1. FUJITA, S. & N. S. GING. 1961. SCIENCE. 134: 1687.

    2. CHAPMAN, G. H. 1947. TRANS. N.Y. ACAD. SCI. 10: 45.

    3. SHERMAN, J. M. 1937. BACTERIOL REV. 1: 3.

    4. KOPPER, P. H. 1952. J. BACTERIOL. 63: 639.

    5. RAWLS, W. B. & G. H. CHAPMAN. 1935. J. LAB. CLIN. MED. 21: 437.

    6. CHAPMAN, G. H., C. BERENS, & E. L. NILSON. 1936. J. BACTERIOL. 31: 339.

    7. RUGGIER, J. C., & G. H. CHAPMAN. 1941. J. TENN. STATE DENT. ASSOC. 21: 149.

    8. CHAPMAN, G. H. 1943. J. AM. COLL. DENTISTS. 10: 70

    9. ALBERT, L. 1942. MED. TIMES. 70: 118.

    10. STILES, M. H. & G. H. CHAPMAN. 1940. ARCH. OTOLARYNGOL. 31: 458.

    11. LAROCHE, G., ET AL. 1962. FEDERATION PROC. 21: 300.

    12. TRAXLER, R. W. & C. E. LANKFORD. 1957. APPL. MICROBIOL. 5: 70.

    13. GOEB, F.. 1947. J. EXP. MED. 85: 499.

    14. GRIFFITHS, A. F. & G. H. CHAPMAN. 1950. MILITARY SURGEON. 106: 53.

    15. CHINARD, F. P. 1948. J. BIO;. CHEM. 176: 1439.

    16. PENNELL, R. B. & C. A. SARAVIS. 1962. ANN. N.Y. ACAD. SCI. 96: 462.

    17. BERGEN, J. R., W. P. KOELLA, H. FREEMAN & H. HOAGLAND. 1962. ANN. N.Y. ACAD. SCI. 96: 469

    18. FROHMAN, C. E., E. GOODMAN, P. G. S. BECKETT, L. K. LATHAM, R. SENF & J. S. GOTTLIEB. 1962. ANN. N.Y. ACAD. SCI. 96: 438.

    19. SANDERS, B. E., E. V. C. SMITH, L. FLATAKER & C. A. WINTEN. 1962. ANN. N.Y. ACAD. SCI. 96: 448.

    20. BERGEN, J. R. 1962. CITED BY PENNELL, R. B. & C. A. SARAVIS. REFERENCE 16.

    21. SCHREK, R. 1939. J. LAB CLIN. MED. 25: 180.

    22. STILES, M. H. 1941. AM. J. CLIN. PATHOL. 11: 871.

    23. BERGEIM, O. 1924. J. BIOL. CHEM. 62: 45.

    24. CROCKER, W. J. & E. H. VALENTINE. 1934. J. LAB. CLIN. MED. 20: 172.

    25. CHAPMAN, G. H., M. H. STILES & C. BERENS. 1939. AM. J. CLIN. MED PATHOL, TECH. SUPPL. 9: 20.

    26. RAWLS, W. B. & G. H. CHAPMAN. 1939. REV. GASTROENTEROL. 6: 317.

    27. CHAPMAN, G. H. 1951. AM. J. PUBLIC HEALTH. 41: 1381.

    28. LARA, F. J. S. 1952. J. BACTIOL. 64: 271.

    29. CHAPMAN, G. H. & A. F. GRIFFITHS. 1939. REV. GASTROENTEROL. 6: 243.

    30. WARBURG, O. 1956. SCIENCE. 123: 309.

    31. HUGGINS, C. & N. C. YANG. 1962. SCIENCE. 137: 257.

    Top -- Home

    www.washingtonpost.com/national/health-science/new-studies-show-that-young-blood-reverses-the-effects-of-aging-when-put-into-older-mice/2014/05/04/1346baac-d2eb-11e3-8a78-8fe50322a72c_story.html

    Article from the Washington Post

    [Young blood proves good for old brains]

    By Meeri Kim, May 4, 2014

    New studies show that young blood reverses the effects of aging when put into older mice

    A blood-based protein that can rejuvenate the hearts of aging mice, has a similar effect on the mice's brain and skeletal muscle function, according to scientists at Harvard University ...

    A trio of new studies has discovered that the blood of young mice appears to reverse some of the effects of aging when put into the circulatory systems of elderly mice.

    After combining the blood circulations of two mice by conjoining them -- one old, the other young -- researchers found dramatic improvements in the older mouse's muscle and brain. After four weeks, stem cells in both those areas got a boost of activity and were better able to produce neurons and muscle tissue.

    They later discovered that injections of a special protein found abundantly in young blood -- or even transfusions of whole young blood -- give the same advantages as sharing a blood supply.

    Old mice who were injected with the protein or who received a blood transfusion navigated mazes faster and ran longer on treadmills. They easily outperformed their control peers, who were given only saline.

    But for the young mice, getting old blood was a definite setback. When conjoined to an older mouse, the creation of new cells in the young mouse slowed. Old blood seemed to cause premature aging.

    Two of the studies, both published online Sunday [5-4-14] in the journal Science, came out of collaborations at the Harvard Stem Cell Institute that shared specimens of mice -- one focused on muscle changes and the other specialized in the brain. The third, published Sunday in Nature Medicine, came from a group of researchers from Stanford University and the University of California at San Francisco.

    "The Stanford group has been working in this area for a while, but we weren't involved in their study," said Science study author and biologist Amy Wagers of the Harvard institute ...

    Nature Medicine study author and neuroscientist Tony Wyss-Coray of Stanford said he hopes to dive into human studies immediately. His new start-up company, Alkahest, is planning the first young-blood clinical trial at Stanford this year. Patients with Alzheimer's disease will be given young blood, with researchers measuring their cognitive condition before and after ...

    "Most diseases that affect industrialized nations have a very strong aging component, and these are currently studied in isolation," Wyss-Coray said. "But age is the key risk factor for all these diseases."

    Judith Campisi, a biochemist at the Buck Institute for Research on Aging who was not involved in the studies, agreed that fundamental aging research is necessary and can have wide benefits ...

    "Even in the best of circumstances of being completely disease free, things just can't be maintained with age," said Ronald Kohanski, the deputy director of the National Institute on Aging, who was not involved in the research.

    The studies started with a ... setup called parabiosis. Small flaps of skin from the sides of two genetically identical mice are cut and sewn together. As the wounds heal, their tissue begins to fuse. The mice, now conjoined, share a single blood supply. Pairing old and young mice, or heterochronic parabiosis, has become an unexpectedly insightful tool for age research ...

    A few recent animal studies have claimed to increase longevity. In 2009, a drug called rapamycin was shown to extend the life span of mice by about 10 percent. Also, a calorie-restricted diet received much attention for its proven health benefits for monkeys. However, nothing has been proven to reverse the adverse effects of aging -- something that young blood appears to do.

    In particular, the two studies published in Science focused on a specific protein in young blood, called growth differentiation factor 11. GDF11 circulates at high levels in the bloodstreams of youthful mice but declines with age. Last year, a study showed that injections of GDF11 appeared to rejuvenate the toughened heart muscle of elderly mice.

    "(Rapamycin) and caloric restriction seemed to slow the aging process, not necessarily stop or even reverse it," Campisi said. "But GDF11 seems to reverse it."

    In the new experiments, GDF11 treatment had a similar turn-back-the-clock effect on both skeletal muscle and the brain.

    "It could have appeared that the GDF11 effects were limited to the heart," said Wagers, who authored both the heart and muscle studies. "These new studies extend the impact to other types of tissues."

    The Science study found that after four to five weeks of heterochronic parabiosis, the muscle stem cells from the older partners had less DNA damage compared with controls. Their neural stem cells got a boost of activity as well, and they had a greater amount of blood flow in their brains.

    Then the researchers switched to pure GDF11 injections. When they gave a new group of aged mice four weeks of treatment, they found that the protein itself gave similar enhancements as shared circulation. There were more stem cells in their muscles to create new tissue, and they performed better on strength and endurance tests than controls given saline. GDF11 treatment also increased the number of blood vessels in their brains.

    The research detailed online in Nature Medicine did not distinguish a specific protein but instead injected whole, young blood. Scientists focused on the hippocampus, a region of the brain important for memory and spatial navigation, where neurons are created from neural stem cells.

    The lab's previous study found a reactivation of neural stem cells using young blood -- but this time, the team wanted to see whether it really meant a smarter mouse. The researchers found that it did. The old mice treated with young blood could navigate mazes and recall fear memories better than controls ...

    In a small group of human subjects, Wagers has found that GDF11 levels are consistent with what she saw in mice -- higher in the young and lower in the old ...

    [It is possible that a reduction of harmful ingredients in old blood could be important, for example inflammatory molecules that rise with age. Also, it would be interesting to test the effects of plasma from young donors on people with Alzheimer's disease.]

    Bug War -- Top -- Home -- What's New