US 20130236563A1

(19) United States

(12) Patent Application Publication (10) Pub. No.: US 2013/0236563 A1
Samuelson et al. (43) Pub. Date: Sep. 12, 2013
(54) METHODS OF AMELIORATING OXIDATIVE is a continuation-in-part of application No. 12/383,
STRESS BY INCREASING THE EFFICIENCY 212, ?led on Mar. 20, 2009, noW Pat. No. 8,367,120,
OF GPX AND SOD ACTIVITY Which is a continuation-in-part of application No.
12/290,398, ?led on Oct. 30, 2008, noW abandoned.
71 A 1' t: REOXCYN DISCOVERIES GROUP
( ) pp lean INC Salt Lake City UT (Us) (60) Provisional application No. 61/001,010, ?led on Oct.
30, 2007.

(72) Inventors: Gary Samuelson, Sandy, UT (US);

Verdis Norton, Sandy, UT (US) Publication Classi?cation
(73) Assignee: Reoxcyn Discoveries Group, Inc, Salt (51) Int. Cl.
Lake City, UT (US) A61K 33/40 (2006.01)
(52) US. Cl.
(21) APPI- NO-I 13/857,225 CPC .................................... .. A61K 33/40 (2013.01)
(22) Filed: Apr- 5, 2013 USPC ......................................... .. 424/613; 435/375

Related US. Application Data (57) ABSTRACT

(63) Continuation-in-part of application No. 12/592,402, Methods of ameliorating cellular oxidative stress are dis
?led on Nov. 24, 2009, noW Pat. No. 8,455,010, Which closed.

1% A554

601711'0/ 30 min 60 mm 7 A 120 min seruwstarved

SAP}

NRFZ
; randomly q/c?ng
QAPI

Nut/ear NRFZ staining pr?/es in random/y cycling 3nd serumstarved

HMVEGL ce/ls.
Patent Application Publication Sep. 12, 2013 Sheet 1 0f 27 US 2013/0236563 A1

Reverse Osmios/s ./ 102

Distillation ./'1 04

Sa/t/ng _/'106

Chi/ling _/1 08

Y
Electra/yz/ng / 1 10

Storage/Testing _/ 1 12

Bottling _/114

FIG. 1
Patent Application Publication Sep. 12, 2013 Sheet 3 0f 27 US 2013/0236563 A1

202
g

258

FIG. 3
Patent Application Publication Sep. 12, 2013 Sheet 4 0f 27 US 2013/0236563 A1

Input
302 3

Output

FIG. 4
Patent Application Publication Sep. 12, 2013 Sheet 5 0f 27 US 2013/0236563 A1

FIG. 5
25

NaCl NaC/O A554

20

15

E 1 0..

5-

I
10
Patent Application Publication Sep. 12, 2013 Sheet 6 0f 27 US 2013/0236563 A1

{2r.e1?7l ed}

1.5

1,0

0.5
Patent Application Publication Sep. 12, 2013 Sheet 7 0f 27 US 2013/0236563 A1

20 16 I [pi/v}

FIG. 7
Patent Application Publication Sep. 12, 2013 Sheet 8 0f 27 US 2013/0236563 A1

:
___
i@E?$E3mm.6: $5a?g_pm.8v.9%
REA9%$3
EN.
9%..
gm.
Rm.
.a?
Rum.
mws?wmgwmmgg 43
.4
._
I-__._.
__
J.
_
:.I1a-w._.4

53m. wasgm.

mw 8 %
8HI

6E
%
Patent Application Publication Sep. 12, 2013 Sheet 9 0f 27 US 2013/0236563 A1

FIG. 9
0. 7000000
Oxygen / Nitrogen

0. 6000000

0. 5000000

0. 4000000

0.3000000

0.2000000

0. 1000000

0. 0000000
Water ASE/4 NaCIO

Ch/orine / Nitrogen
0. 00001 80
0. 0000160
0. 0000140
0. 0000120
0.0000100
0.0000080
0. 0000060
0. 0000040
0. 0000020
0. 0000000
Water ASEA IVaCIO

FIG. 10
Patent Application Publication Sep. 12, 2013 Sheet 10 0f 27 US 2013/0236563 A1

FIG. 11
Ozone / Nitrogen
0. 0002000
00001800
0. 0001600
0. 0001400
00001200
0. 0001000
0. 0000800
0, 0000600
00000400
0. 0000200
0.0000000
Water A554 NaCIO

Carbon Dioxide / N/tmgen

0, 0080000
1

0. 0070000
0. 0060000
0. 0050000 .,.

0. 0040000
0. 0030000
0. 0020000 I
0.0010000

0. 0000000
Water ASEA NaCIO

FIG. 12
Patent Application Publication Sep. 12, 2013 Sheet 11 0f 27 US 2013/0236563 A1

FIG.13

Q0 0 80 9 40 0
Patent Application Publication Sep. 12, 2013 Sheet 12 0f 27 US 2013/0236563 A1

FIG. 14

10 Li: :| 2

#3

Power Source

FIG. I 5
"WV "w 1

10 WW Mi 2
W- 2- ME :- WW1

PavarSoum
Patent Application Publication Sep. 12, 2013 Sheet 13 0f 27 US 2013/0236563 A1

FIG. 16

Wall Terminal Strip

Potentiometer
EI
i _' Transformer
E
E l Recti?er

FlG. l 7

El\> Ground
I
Wall Terminal Strip Current Transformer

@ l \ @
Potentiometer El

Transformer < E

Recti?er _ [II
I
DC Shunt <__ E
Patent Application Publication Sep. 12, 2013 Sheet 14 0f 27 US 2013/0236563 A1

0.25

02
:5: T Oneway 000m, 0010.002;
9. T 00 P<0.015 vs. Sad-PL
g, 0.15
g 0k
8 0.1 T 9%
in T
g
-~l
0.05 _

0
Sad-PL Sad-A554 Run-PL Run-A554

FIG, 18
Patent Application Publication Sep. 12, 2013 Sheet 15 0f 27 US 2013/0236563 Al

w%

2. 00

GOqplix1urtacdho?nze(mp0Is/g} 1.50

1.00
__

0,50 _

Sad-P1. 56d~A$EA Run-P1. Run-ASEA

FIG. 19A

Oxidized G/utathione (6556) in muscle

3 7 9k

FIG. 19B CRonutrnol RunASEA

sCeodntra!y ASe5dni4ary
Patent Application Publication Sep. 12, 2013 Sheet 16 0f 27 US 2013/0236563 A1

FIG. 20
Patent Application Publication Sep. 12, 2013 Sheet 17 0f 27 US 2013/0236563 A1

FIG. 2]

120

6O _ V, i , Seriesi

W Expon.(Series1)

40 i i v i i , , i i \ 2, ~ y:98.615e0,0311l

20

0 ' t 3 1
Patent Application Publication Sep. 12, 2013 Sheet 18 0f 27 US 2013/0236563 A1

F]G. 22

Bottle versus Pouch

120
100 0 Bottle Decay

80
I Pouch Decay
6O
4O -Expon. (Bottle Decay)

2O --Expon. (Pouch Decay)

O y = 94.731601)X
0 5 10 15
Month y = 994959003X
Patent Application Publication Sep. 12, 2013 Sheet 19 0f 27 US 2013/0236563 A1

FIG. 23

Expt. 5f07 ROS Assay -- using data from 0-250 min

18000

11.

Ohm6ocD.om<5BsE 4862o 0o 0o o0
_
_

nnnnuilununu
IIIIIIIIIIIIII
m n

0.01 0.1

AAPH Concentration [mM]

FIG. 24
Determination of lntraassay Variation Using
Two Levels of AAPH

_.L"IZY
0.369E3 uMIL

buffer control

O1. 6.
0 4.
5.
7.
8.
3
D

3.69E3 uMIL

200 400 600 800 1000 1200 1400

Time [mins]