CYAN EXHIBIT 1022
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`Weekly Report of Academy of Sciences Meetings
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`Published in accordance with a decision of the academy on July 13, 1835
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`By Mr. Perpetual Secretaries
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`Volume 223
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`July — December 1946
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`PARIS,
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`GAUTHIER-VILLARS, IMPRIMEUR-LIBRAIRE
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`Academy of Sciences Meeting Minutes
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`Quai cles Grands-Augustins, 55.
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`1946
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`

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`47
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`Meeting on July 1, 1946
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`Title: Antioxidant properties of carotenoids and their derivatives.
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`Author: Armand HERISSET
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`Journal: Comptes Rendus Hebdomadaires des Séances de l’Académie des
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`Sciences, v.253, pp. 47-49 (July — December) 1946.
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`Plant Biology: Antioxidant properties of carotenoids and their derivatives. Note
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`(1) by Mr. Armand Herisset, presented by Mr. Louis Blaringhem.
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`The plant cells of Trentepohlia Mart. and numerous chlorophyll cysts contain astaxanthin
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`dissolved in lipid droplets. It is traditionally thought that the carotenoid pigment is a protective screen
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`for chlorophyll, absorbing the short wavelengths of light; I have shown (2) that this complex appears as a
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`storage substance. Since lipid alone can fulfill this function, what is the role of the astaxanthin? The
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`frequent detection of carotenoids in the storage tissues leads us to think that these pigments must fulfill
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`a general function and, due to their high capability of acting as a reducer, we are also led to think that
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`they play a role in the preservation of the lipid or the storage substance with which they are associated.
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`l have, therefore, tried to examine the antioxidant properties of astaxanthin in algae, the carotene in
`carrots and vitamin A.
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`The first of these pigments was extracted from Trentepohlia and then purified; the two
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`remaining were provided to me, in their pure form, by industry. To characterize this function, I
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`examined the most sensitive reagent: the biological reagent.
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`It is known that mushrooms secrete oxido-
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`reduction enzymes; the mushrooms oxidize the phenols, giving them color, while the oxido-reduction
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`enzymes cause a more or less complete de-coloration of the methylene blue. An antioxidant slows the
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`oxidation reactions and, in contrast, accelerates the reduction reactions. Having studied several different
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`molds [fungi], l was able to determine that Mucor Rouxii Welm secretes oxidases when they are
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`cultivated in the following milieu, adjusted to a pH of 8.1: water 1000, glucose 30%, PO 4(NH4) 2H 1g,
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`N03NH4, 1g, SO4K2 0g,40,SO4Mg
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`(1) Meeting on June 24, 1946.
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`(2) Minutes, 221, 1945, pp. 707-708.
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`

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`48
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`7HZO 0.20g, SO4Fe.SO4 (NH4) 2%, 6HZO 0.10g, SO4Zn.7HZO 0.04g, SO4Mn.7HZO 0.05g, SO4Cu.5HZO
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`0.02g, agar 25g.
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`I added a drop of guaiacol solution, 1% per 5 ml of milieu, which is then cooled in an inclined
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`position. The difficulty comes from the fact that carotenoids are not easily soluble except in lipids: I
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`added the oily solution just before the solidification of the milieu and I energetically add it in order to
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`emulsify the mix. The control tubes must be shaken in the same manner because this operation
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`introduces air into the mass and it is necessary to operate under identical conditions (especially with
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`regards to the oxygen).
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`I prepared an equal number of control tubes and tubes containing 1/2500 carotene or 1/2500
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`astaxanthin or 1 drop of a vitamin A solution at 1/120,000 per gram.
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`These tubes were divided into 2 identical series, one placed in the light and the other in darkness, while
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`the temperature is kept at 25° in both cases.
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`At the end of3 to 4 days, the agar in the control tubes turned red, in darkness as well as in the
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`light. In the following days, the color increased then slowly diminished (perhaps a little more quickly in
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`the tubes exposed to light). During this time, the tubes containing carotenoids remained colorless or,
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`sometimes very lightly colored pink but this light coloration is slower and disappears very quickly. I
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`believe that it is produced in tubes in which the emulsification of the oily substance is less complete.
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`The substances studied completely prevent oxidizing catalysis caused by mushrooms or, make it more or
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`less minimal and transitory.
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`Although, among the species used, M. Rouxii alone secretes oxidases in a regular and consistent
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`manner, it is not the same with reduction enzymes: all of the stem cells produce it in abundance. I,
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`therefore, examined Mucor mucedo L., which develops quickly and rapidly. Mushrooms are cultivated in
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`an agarized Sabouraud milieu adjusted to a pH of 8.
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`I added a drop of 0.50% methylene blue solution per milieu tube which was then cooled in a
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`vertical position; the mold was then well-sown into it. Carotenoids were used, at the dosages indicated
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`in the first test and added by taking identical precautions. Two parallel series were prepared, one was
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`placed in the light and the other in the darkness.
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`After 24 hours ofculture, the tubes containing the vitamin began to lose color in the lower part
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`(in the light and in the dark).
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`At the end of36 hours of culture, I was able to create the following table:
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`— Carotene
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`Astaxanthin
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`Vitamin A
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`Light
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`the identical tubes placed in the light
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`The tubes have not
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`moved.
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`.
`Tubes are discolored
`_
`in the upper 1/2, green
`.
`in the lower 1/2.
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`The tubes were
`_
`.
`discolored In the
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`.
`lower 3A and green in
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`the upper %.
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`The tubes were
`.
`discolored almost
`.
`completely in the
`.
`upper portion.
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`Darkness
`
`.
`Identical
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`.
`Identical
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`Slightly more
`discoloration than in
`.
`.
`
`.
`Identical
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`

`

`Meeting on July 1, 1946
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`After 2 days ofculture, the controls began to become discolored in the lower parts. In general,
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`the tubes developed quickly to a more or less advanced reduction stage and, later, the coloration did
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`not vary much.
`At the end of 10 culture days, the situation was as follows:
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`49
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`————
`Light
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`The tubes turned
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`blue on the surface,
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`+/-clear green in the
`rest of the tube.
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`Tubes are discolored
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`were completely
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`All of the tubes were
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`Most of the tubes
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`in the upper 1/2, clear
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`discolored, some
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`green in the lower 1/2.
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`slightly green at the
`surface.
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`completely
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`discolored.
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`Identical
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`Identical
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`Identical
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`Darkness
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`Many of the tubes
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`were completely
`discolored, the others
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`were almost entirely
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`discolored; they only
`had a few small
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`patches of clear
`green.
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`The antioxidant properties of the carotenoids which were studied were associated with a more rapid
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`and complete discoloration (never observed in the control tubes).
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`Astaxanthin and vitamin A are powerful anti-oxidants; the carotene used was a little less active. The
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`action of the two latter is identical in the light and in the darkness; the former seems to be a little higher
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`in the light: under natural conditions, it is the only one of the 3 substances studied which is located in
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`cells exposed to the light.
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`

`

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`accurate translation from French to English of the French language original and was performed by a
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`Date article was
`Author. Title and Journal
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`emailed to us
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`June 6, 2013
`Title: Antioxidant properties of carotenoids and their derivatives.
`
`
`Author: Armand HERISSET
`
`
` Journal: Comptes Rendus Hebdomadaires des Séances de I‘Académie des
`Sciences. V253, pp. 4749 (July - December) 1946.
`
`
`
`
`
`Herl’sset C. RAc. Sc. v223 p.47
`Antiox_Eng Final
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