|Silicon dioxide||34,1 %|
|Sesame oil||30,0 %|
|Green tea extract (epigallo-catequin-gallate)||11,6 %|
|Zinc citrate||2,0 %|
|Fumaric acid||1,5 %|
Boosting by quantum dot technology:
1:34 for epigallo-catequin-gallate
Case study (Oelschlegel et al. 2014)
19 MS-patients received EGAA-6 for two months or more. 12 of them showed an improvement (3 of them a drastic improvement). In one case a little worsening occurred while in 6 patients no change could be found. But in 4 of these 6 it stayed unclear whether and when they had taken EGAA-6 at all. Without these 4 non-compliers the rate of improvement was 80%. Before 2 month no effects can be expected. Drastic improvements occurred after 6-12 months. In 2 of these three cases also lesions in brain were reduced. The third case has not been checked yet. Negative side effects could not be noted.
Pain had been treated successfully by using EGAA-1 a mixture of incense and curcumin as quantum dots. This mixture replaces cortisol (Hollmann 2010) without the side effect of cortisol.
In many cases weakening could be treated with EGAA-3 (Meyer, Mandel, Knapp 2011) a product against cancer. It opens membranes of mitochondria. After at least two days, weakening improved. Therefore the hypothesis of a mitochondropathy is not unlikely (Jennerich 2012). In cases of weakening also EGAA-2 (an anti-AIDS remedy) (Rohr, Gradl 2012) had been used successfully. Its action may be by improving immune response. Since depressions occur frequently in MS EGAA-4 had been used successfully. In this remedy sinigrin from onions being a serotonin-reuptake-inhibitor and coriander as degradation inhibitor of GABA and nutmeg and mugwort as degradation inhibitors of MAO are combined (all as quantum dots). Simultaneously it contains a guluronan-complex that improves oxygen supply of cells acting against exhaustions.
The used remedy and its functions
In the trials the herbal product EGAA-6 had been used.
It is mainly composed from three substances, the aminoacid glycine, fumaric acid and an extract of green tea (epigallocatequingallate) as quantum dots (Gradl 2008).
Mechanism of action
Glycine controls membrane permeability for chloride and hydrogen-carbonate ions in neurons of brainstem and spinal cord (Werman et al. 1968). It acts as an inhibiting neurotransmitter like GABA.
By binding to glycinergic receptors in brainstem and spinal cord balance between excitatory and inhibiting neurotransmitter systems is established (Baccei, Fitzgerald 2004) and rhythms of nerve cells are clocked (Gusev et al. 2000).
These effects could be used in treatment of
|ischemic strokes (Gusev et al. 2000; Zaslavskaja et al. 1999)|
|functional and organic brain traumata using glycine as a platelet-activating factor antagonists (Faden; Tzendzalian 1992)|
|limited intellectual performance and somnipathy (File et al. 1999; Hecht, Hecht-Savoley 2008)|
|clocking of brain rhythm in opium narcomania (Mashkova et al. 1996) and alcoholics (Sheveleva et al. 1996)|
By glycine breath rhythm in the respiratory centre of the brain is stabilized (Haji et al. 1990) and neuronal regulation of muscular tonus is controlled via brainstem and spinal cord (Waldegger, Jentsch 2000). Both mechanisms together with clocking of nerve rhythms (Gusev et al. 2000), its anti-spasmotic (Brune et al. 1996) and sedative (Shigemi et al. 2008) effect makes glycine a tool for stress protection and stress treatment (Goldstein et al. 1994).
The biggest part of amino acids in collagen is glycine. Collagen together with mucopolysaccharides and proteoglycans are an important part of extracellular matrix, the site of basic regulation in the body (Pischinger, Heine 2007). Glycine is enormous important for redox-homeostasis there.
Fumaric acid had been used for many years to treat psoriasis (Mrowietz, Christophers, Altmeyer 1999). It is able to act on dendritic cells. Recent experimental data point towards a skewing of the TH1-dominated T-cell response in psoriasis to a TH2-like pattern and inhibition of proliferation of keratinocytes.
In a multi centred study (Schilling, et al. 2006) it turned out that methyl hydrogen fumarate (MHF) and dimethyl fumarate (DMF) in chronic experimental autoimmune encephalomyelitis (EAE) induced by immunization of C57BL/6 mice with MOG peptide aa 35-55 was preventive. Fumaric acid esters were delivered twice a day by oral gavage. Both esters had a significant therapeutic effect on the disease course and histology showed a strongly reduced macrophage inflammation in the spinal cord. Multiparameter cytokine analysis from blood detected an increase of IL-10 in the treated animals. Linker et al. (2011) could elucidate the mechanism via activation of Nrf2 pathway. The protein encoded by this gene is a polypeptide hormone and nerve growth factor whose actions have mainly been studied in the nervous system where it promotes neurotransmitter synthesis and neurite outgrowth in certain neuronal populations including astrocytes. The protein is a potent survival factor for neurons and oligodendrocytes and may be relevant in reducing tissue destruction during inflammatory attacks. A mutation in this gene, which results in aberrant splicing, leads to ciliary neurotrophic factor deficiency, but this phenotype is not causally related to neurologic disease. In addition to the predominant monocistronic transcript originating from this locus, the gene is also co-transcribed with the upstream ZFP91 gene. Co-transcription from the two loci results in a transcript that contains a complete coding region for the zinc finger protein but lacks a complete coding region for ciliary neurotrophic factor.[ CNTF has also been shown to be expressed by cells on the bone surface, and to reduce the activity of bone forming cells, osteoblasts. Beurrier et al. (2010) could proof that a main action of CNTF are enhanced glutamate transporters.
One third of green tea dry matter consists of epigallocatechin gallat (EGCG), an antioxidant with many positive effects on health. In black tea it is reduced by fermentation to theaflavines.
Neuro-degenerative diseases like Alzheimer and Parkinson’s disease are caused by amyloid fibres or by wrong folding of proteins. EGCG binds to yet unfolded polypeptides. Instead of toxic fibres spheric oligomers are formed (large, mature α-synuclein and amyloid-β fibrils form smaller, amorphous protein) (Ehrnhoefer et al. 2008, Bieschke 2010). EGCG is able to disintegrate already existing plaques. In model-mice plaques in cortex, hippocampus and entorhinal cortex could be reduced by 54, 43 and 58 % after a six month treatment (Rezai-Zadeh et al. 2008).
In multiple sclerosis EGCG can protect nerves of the CNS and can control T-lymphocytes being responsible for the disease. EAE (the animal model for MS) had been significantly less severe in animals who received EGCG (Aktas et al. 2004). EGCG neutralizes TNF- and reduces production of IL-6 and IL-8, the reason for its immuno-suppressive action.
Others (Sun et al. 2013) could find reduced disease severity in EAE by decreasing brain inflammation and demyelination damage, accompanied by decreased encephalitogenic T-cell responses and reduced expression of inflammatory cytokines and chemokines. The effect of EGCG was attributable to its selective inhibition of interferon-gamma and interleukin-17 production in CD4+ T-cells, mediated via alteration of the STAT-pathway and the transcription factors T-bet and retinoid-related orphan receptor (ROR) gammat/ROR-alpha. More important, EGCG has been found novel properties of directly inhibiting TH1 and TH17 cell-differentiation in this study. On the other hand, EGCG-treated antigen presenting cells (APC) exhibited reduced co-stimulatory function as a result of altered expression of CD80 and CD86.
Wang et al. (2012) proposed that EGCG can improve cognitive function by impacting the generation of neuron cells, a process known as neurogenesis. They focused on the hippocampus, the part of the brain which processes information from short-term to long-term memory. They could proof, that the production of neural progenitor cells, which like stem cells can adapt, or differentiate, into various types of cells is enhanced by EGCG. In mice this increased cell production gave an advantage to memory or spatial learning. Like other catechines EGCG is a radical scavenge for ROS and RNS being responsible for DNA-damage (Lee, Lee 2006).
Material and Methods
Treatment of mice and rats fed with triethyltin (TET)
All animals are fed ad libitum with 10 mg/l TET (p. a. as chloride) and with pelleted standard feed (Altromin). 10 animals each received i. p. an injection of EGAA-6 in 0.15 M NaCl. 10 animals were not treated (only TET).
Mice were female NMRI1 22-24 g. On days 6, 8 and 10 0.5 ml of EGAA-6 was administered i. p. (1% except P2, which was administered 0.1%). The animals were killed at day 12. The health status was scored in a 10-scaled score daily.
Rats were male Lewis 180-220 g. The animals received EGAA-6 on days 6, 8, 10 and 12 i. p. (1 ml of a 14% solution). All animals were scored daily for their health status.
Experimental Autoallergic Encephalitis (EAE) in guinea pigs
EGAA-6 was administered to 10 female guinea pigs (Hartley 560-720 g). 10 animals were taken as a control. All animals were vaccinated using 500 µg basic myelin protein (BP) together with 2.4 mg mycobacterium tuberculosis (H37Rv) in an emulsion of 140 µl 0.15 M NaCl and 260 µl complete Freunds’ adjuvant i. c..
On days 6, 8, 12, 14 and 16 after vaccinations the animals received 2 g of EGAA-6 (1% in a trituration of bentonite containing 2% Vaseline and 0.5% castor triglyceride-polyglycolether by feed.
Animals were weighed daily. The lameness was scored in a 4-scaled score (0= no signs of lameness to 3 = severe lameness). All surviving animals were killed on day 20. At this time serum as well as spleen cells were gained under sterile conditions.
Globulin fraction of serum was gained by salting out (using ammonium sulphate) and a dialysis, which followed. In a precipitation test (Ouchterlony), 5 µl diffunded against a dilution series (1:2) in two parallels for 20 hours against 5 µl of a bovine BP-solution (5 mg/ml). After staining with Coomassie Blue titres were determined. It is necessary to use globulin fraction, because albumin precipitates with BP as well.
Gliotoxic serum activity
Glioblastoma cells of mice (107 cell/ml) in Dulbeccos MEM supplemented with 10% inactivated foetal calf serum were filled 100 ml each in micro plates and incubated for 24 hours (37°C; 5% CO2, 100% moisture). Afterwards 100 µl of inactivated serum respectively a dilution series (1:2) are added in parallels and incubated for 30 minutes. Afterwards
10 µl guinea pig complement was added (diluted 1:4). After another incubation period of 90 minutes under a reverse microscope, the percentage of damaged cells (cell extensions withdrawn) was assessed. 3 troughs were used as a control (without serum). 50% damage was used as the titre-limit.
Cellular immunity against BP
Lymphocyte transformation was determined by incorporation of 3H-thymidin in spleen cells of guinea pigs. To 4 x 106 cells in 4 ml medium (TC 199 ) + 10% inactivated foetal calf serum in 3 parallels 8 µl BP-solution (5 mg/ml in 0.15 M NaCl) respectively 10 µl PHA-P (Difco 1.50 diluted with medium) were added. Three troughs were used as a control. After 48 hours (37°C) 80 µl 3H-thymidin (20 µc/ml; 2000 mCi/mmol,) were added for 12 hours. Cells were filtered, washed and dried and than measured twice in scintillation tubes. From the means counts of controls and stimulated cells a stimulation factor (quotient) was determined.
At day 18 guinea pigs were injected i. c.: 50 µl of a BP-solution (0.1% in 0.25 M NaCl) and its dilution series (1:2; 1:4; 1.8). The same concentration of bovine albumin was used as a control. After 24 hours and 48 hours reddening and hardening were determined.
Histological scoring: Brain and medulla were taken, fixed in buffered formaldehyde, embedded in paraffin and cut. Cuts were stained (HE and Kluever-Barrerra) and assessed under a microscope (no lesions = 0; to severe demyelination = 3). Four cuts per animal were scored as a minimum.
Effect on TET-activity in mice and rats
Mice: In all cases the clinical data of mice improved. Figure 6 shows the change of the protective index from day 6 to the end of the trial. The biggest difference could be seen at the end of the trial (days 11-13) when the not protected animals became worse steadily. The maximum protective index was 6.03
Rats: The clinical data for rats are shown in fig. 7. Here also the difference between treated and not treated was highest at the end of the trial.
Effect on EAE in guinea pigs
All not treated animals died within 20 days (control) with signs of very sever lameness. In the group treated with EGAA-6 one out of 10 animals died. All survivors had no or only little signs of being sick.
In the group treated with EGAA-6 in nine out of ten animals clinical signs of EAE were suppressed. In these animals the degree of lesions of the brain was less than in the control group. Also in this group the highest score (3) never occurred, while this score was most frequent in the control group (fig. 8).
Precipitating BP-antibodies occurred in high titres in guinea pigs treated with IMMUNO-SHIFT while they did not occur in the control group. Guinea pigs, which became sick temporarily, had lower titres than treated animals with no signs of being sick (fig. 9). Gliotoxic serum activity was lower in treated animals.
None of the animals whether treated or not showed a reaction against BP in the skin irritation test or in the transformation test. No difference between the groups could be found thereby.
The effect of EGAA-6 on EAE showed: Cellular immunity, the pathogenetic mechanism of EAE did not change, while clinical and histological signs improved by treatment. Gliotoxic antibodies (IgG2-antibodies) were reduced while precipitating BP-antibodies increased in treated guinea pigs. These antibodies also could be found after vaccination of EAE-resistant guinea pigs in comparison to non-resistant strains.