Microcurrent Experimental Results

MicrocurrentExperimentalResults

Microcurrent Experimental Results.
Research Conducted by Dr Vivienne Reeve, University of Sydney and Wayne Reilly,
Health World Ltd. Feb 12.03
Hypothesis 1. FSM alters LOX induced inflammation using an accepted model of inflammation?
Table 1.
4 minutes of frequency 40/116
at (200microamps) immediately
after painting of AA gives a
70% reduction in ear swelling.
4 minutes of frequency 40/355
at (200microamps) immediately
after painting AA gives a 70%
reduction in ear swelling
Test hypothesis using an ear inflammation model – Arachadonic acid induced swelling.
Male SKh-1 albino hairless mice aged 6 weeks. Ears were measured with a micrometer, 5l of
25mg/ml of Arachadonic acid in ethanol, was painted on both sides of the ear and both ears were
treated. FSM was applied immediately after it had dried and measurements were taken at 1 hour
post application. The mean ear swelling for the change in thickness was determined and presented
in a table.
The response is time dependent as 1min gives no protection, 2 mins gives a 50%
protection and 4 mins gives 100% protection. Data Appendix 1.
Conclusion.
Frequency Specific Microcurrent can effectively reduce AA induced ear swelling
demonstrating its utility in applications for the treatment of acute inflammation, (Table 1.)
Experiment 1
0
5
10
15
20
25
30
35
40
1 2 3
Mean Ear Swelling
AA
40/116
40/355
2
Repeat of 1st experimental results using 40/116 (Inflammation in the immune system)
R e p e a t o f 4 0 /1 1 6 E x p e r im e n t
0
2
4
6
8
1 0
1 2
1 4
1 6
A A 4 0 /1 1 6 0 /0
M e a n e a r S w e llin g
A A
4 0 /1 1 6
4 0 /0
Test hypothesis using a ear inflammation model – Arachadonic acid induced swelling.
Male SKh-1 albino hairless mice aged 8 weeks. Ears were measured with a micrometer, 5l of
25mg/ml of Arachadonic acid in ethanol, was painted on both sides of the ear and both ears were
treated. FSM was applied immediately after it had dried and measurements were taken at 1 hour post
application. The mean ear swelling for the change in thickness was determined and presented in a table.
3
Table 2.
Hypothesis 2. a. Frequency is an absolute requirement for the observed anti-inflammatory
response., b. the frequency is specific for target and tissue.
4 minutes of frequency
0.1/0.1 at (200microamps)
immediately after painting of
AA gives no reduction in
ear swelling
4 minutes of frequency
Mineral/Bone at
(200microamps) immediately
after painting of AA gives no
reduction in ear swelling
4 minutes of partial
Intermediate protocol
frequency at (200microamps)
immediately after painting of
AA gives no reduction in
ear swelling
Results from 23.1.03
0
5
10
15
20
25
30
35
40
45
50
Group1 Group 2 Group 3
Mean Ear Swelling
AA
Min/Bone
Int/Injury
Conclusion.
Current alone was found to have no effect on inflammation, the mean ear swelling was the same
as for AA. Frequency was found to have specificities in both A/B channels and as seen in Table
1, ie particular frequencies are required for the reduction of inflammation in the model. (see
Table 2).
4
Hypothesis 3. FSM can alter the UV induced skin fold oedema associated with sunburn.
Table 3.
1 2 3
0
20
40
60
80
100
21h
23h
25h
27h
SSUV
SSUV+MICRO t0
SSUV+MICRO t2
*
*
**
**
#
#
@
@
*, #, @ P<0.05
** P<0.01
Skinfold
thickness
increase +/-
SEM
(mm x 0.01)
Development of Erythema/Oedema following 2MED of SSUV,
with microcurrent immediately after (to) or at 2h
Female albino SKh-1 mice were irradiated for 28.95 minutes under a solar simulated light
source. This was equivalent to two MED’s of UV (This is enough UV to produce a sunburn
response as measured by increased skin fold thickness (skin oedema) which reaches a maximum
at 24 hours after UV exposure. Groups of 4 mice were exposed to UV and treated with FSM
immediately after and 2 hours after UV exposure. Mice were treated simultaneously using a box
modified for FSM treatment. The frequencies used were 40/116 for 4 minutes at 200amp;
40/355 for 2 minutes at 200amp and 40/103 for 2 minutes at 200amp. The results are
presented in Table 3.
Conclusion.
FSM at the frequencies tested was shown to have a partial effect on Skin Fold Thickness
indicating some protection against the UV induced sunburn response. The result at 2
hours post UV was statistically significant to P=0.01. It is highly probable that other
specific frequencies could have even greater effects on this model of inflammation.
5
Hypothesis 4.
FSM can alter UV induced systemic immunosuppression. Demonstrating the
immunomodulatory effect of FSM on Th1 immunity.
Table 4.
Experimental Conclusion.
These experiments clearly demonstrate the efficacy of FSM in these well characterised models of
inflammation and immunity. The key findings are as follows.
1. Four (4) mins of 40/116 or 40/355 will produce a greater than 60% reduction in ear
swelling in a recognised model of inflammation.
2. The effect is repeatable with 40/116 and the result was determined blinded demonstrating
efficacy.
3. Frequency was shown to be required to obtain a result and frequencies appear to be
specific.
4. A statistical significant reduction in inflammation was also observed in a second
inflammation model.
5. A systemic immunomodulatory effect was demonstrated in relation to FSM’s ability to
reverse immunosuppression to UV as measured by CHS responses to oxazalone.
Effect of Microcurrent on the CHS Responses of Mice to
Oxazalone
0.00 5.00 10.00 15.00 20.00 25.00 30.00 35.00
1
Groups
Ear Swelling(0.01xmm)
IS 63.4%
IS 57.48%
IS 31.05%
UV Group
UV+2hr Micro-current
UV+Immed Micro-Current
Non-UV Group
The mice that had been used in the sunburn experiment, Hypothesis 3, were then sensitised with
Oxazalone a well known chemical sensitiser. This method measures the ability of an animal to produce
an immune response to the contact sensitiser (CHS response) and is a measure of systemic immunity as
challenge with Oxazalone, at a site away from the sensitisation site, will induce an inflammatory
response. UV is a known immune suppressor of this ability to mount an immune response on subsequent
challenge to a chemical sensitiser.
If FSM is able to overcome UV induced immune suppression it is proof of its activity on the systemic
immune system. It also suggests that a Th1 response has been induced and that a systemic memory to the
Oxazalone has developed. The results are in Table 4.
Conclusion.
FSM when given immediately after UV was shown to have a protective effect on UV induced immune
suppression. This clearly demonstrates systemic immune modulation.
6
References to validate models.
1. Kim SY, Son KH, Chang HW, Kang SS, Kim HP . Inhibition of mouse ear edema by steroidal
and triterpenoid saponins. Arch Pharm Res 1999 Jun;22(3):313-6.
Abstract
Certain steroids and triterpenoids isolated from diverse plant families were
known to possess anti-inflammatory activity. In the course of finding new
anti-inflammatory natural products, some steroidal and triterpenoid saponins
were isolated and evaluated for their anti-inflammatory activity using in vivo
mouse ear edema test. At the oral dose of 100 mg/kg, several steroidal saponins
and triterpenoid saponins such as hederagenin glycosides showed significant
inhibition of ear edema (20-37% inhibition), though less potent than
indomethacin and hydrocortisone.
2. Qian C, Hwang SB, Libertine-Garahan L, Eckman JB, Cai X, Scannell RT, Yeh CG .
Anti-inflammatory activities of LDP-392, a dual PAF receptor antagonist and
5-lipoxygenase inhibitor. Pharmacol Res 2001 Sep;44(3):213-20
Abstract
Leukotrienes (LTs) and platelet-activating factor (PAF) are important mediators
of inflammation and allergy. LDP-392, a novel dual PAF receptor antagonist and
5-lipoxygenase (5-LO) inhibitor, has been identified. LDP-392 is 17.9-fold more
potent than zileuton (5-LO inhibitor) in the RBL cytosolic 5-LO assay, and
equally potent as MK 287 (PAF receptor antagonist) in the human platelet PAF
receptor binding assay. The in vivo dual activities of LDP-392 were confirmed by
measuring the inhibition of ex vivo LTB(4)production in rats and PAF-induced
hemoconcentration in mice. Intravenous administration of LDP-392 demonstrated
greater inhibition than zileuton, BN 50739 or MK 287 on arachidonic acid-induced
ear edema and protected mice from LPS-induced lethality. Topical administration
of LDP-392, in a dose-dependent manner, inhibited TPA-induced ear edema in mice
and UVB-induced erythema in guinea-pigs. These data suggest that LDP-392, as a
dual PAF receptor antagonist and 5-LO inhibitor, may be of greater clinical
effectiveness. Copyright 2001 Academic Press.
3. Ueda H, Yamazaki M Anti-inflammatory and anti-allergic actions by oral administration of a
perilla leaf extract in mice. Biosci Biotechnol Biochem 2001 Jul;65(7):1673-5
Abstract
The anti-inflammatory and anti-allergic activity of perilla leaf extract was
investigated. The oral administration of perilla leaf extract to mice inhibited
two types of acute inflammatory models, arachidonic acid-induced ear edema and
12-o-tetradecanoylphorbol-13-acetate-induced ear edema. Oral administration of
perilla leaf extract also inhibited the contact dermatitis model,
oxazolone-induced ear edema, by affecting sensitization.
4. Danno K, Ikai K, Imamura S.Anti-inflammatory effects of eicosapentaenoic acid on experimental
skin inflammation models. Arch Dermatol Res 1993;285(7):432-5
Abstract
Anti-inflammatory effects of eicosapentaenoic (EPA) and docosahexaenoic acids
(DHA) were examined on three models of skin inflammation induced in mice by
topical application of an arachidonic acid (AA) solution, ultraviolet-B (UVB)
irradiation, and contact sensitization with dinitrofluorobenzene. Ear oedema
reactions induced by AA and UVB irradiation were significantly suppressed in
mice fed a daily dose of 300 mg/kg EPA for 2 weeks. The contact hypersensitivity
reaction was not impaired by EPA. None of the skin reactions was significantly
7
inhibited in mice fed DHA or safflower oil. The results suggest that EPA, but
not DHA, has anti-inflammatory effects on AA- and UVB-induced acute inflammation
reactions.
5. Kotyuk B, Raychaudhuri A, DiPasquale G.
Effect of anti-inflammatory compounds on edema formation and myeloperoxidase
activity in the arachidonic acid-induced ear model in the mouse. Agents Actions 1993;39 Spec
No:C46-8
Abstract
The arachidonic acid (AA)-induced ear edema model in the mouse has been
demonstrated as an effective in vivo experimental tool to screen compounds
showing anti-inflammatory activity. Since neutrophil influx is a component of
the inflammatory reaction, we have modified this assay by quantitating
myeloperoxidase (MPO) levels which reflect neutrophil accumulation in the
edematous biopsies of the mouse ear. Our work has shown that orally administered
5-lipoxygenase inhibitors, dual inhibitors (CO/LO), and steroids
dose-dependently inhibit both edema formation and MPO activity, whereas oral
activity is not seen with NSAID’s. There is a good correlation between the
inhibition of edema formation and of MPO activity by these compounds. Thus,
measurement of MPO, in addition to the AA-induced edema in the mouse ear, can
provide another parameter to profile potential anti-inflammatory compounds.

 

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