Monday, March 15, 2010

Double Blind Drug Trials (DBDT)not applicable to Homoeopathy

These tests are mandatory for every "scientific" allopathic medicine to pass before its release in the market for public prescription. But when applied to individual homoeomedicines in USA in the past, equivocal results were obtained. The then homoeopaths neither showed that DBDTs are inapplicable, nor suggested alternative controlled tests to establish the efficacy of homoeomedicines. Therefore, the science-conscious public demanded and government imposed ban on its practice, and some homoeopathic medical colleges were converted into allopathic ones. However, Dr. Sharma, for the first time in the world showed that DBDTs are not relevant or applicable to Homoeopathy mainly because : (i) A homoeophysician cannot remain "blind" but must know the total symptoms before and after every dose of known medicine to ensure that cure is occurring according Hering’s laws and aggravations , if any, are under control. (ii) The patients cannot be randomized into two groups for treatment with active medicine and similar looking inert placebo since no two patients with same pathology have identical symptom totality. (iii) The subjective symptoms, which have no diagnostic value for Allopathy, cannot be ignored but decide the selection of right homoeomedicine. The DBDTs , without these considerations will be, as in the past, inconclusive to bring bad name to Homoeopathy unjustly.

Dr RR Sharma

Homoeopathy explained through Xenobiology

Dr R.R. Sharma
Homoeopathy: Personal Experiences & Views

(6 March text revised 17 October 2001)

Copyright by the author Prof. Dr. Rati Ram Sharma- all rights reserved.

My attention had been drawn to the two critical web sites: "Homeopathy: The Ultimate Fake", by Stephen Barrett and "Homeopathy-a critique", by Rasmus Jansson. Since the main concern of these authors, in my opinion, was 'general well-being' I hoped they would gladly put my briefly described experiences & views (6 March text) on their web sites. A copy was emailed to Dr. Sahni, Mumbai who put it on the web site under 'articles/research papers' Jansson also put it on the web site: The Journal of Homoeopathy of Northern India published it in its 5 (1) Jan-March 2001 issue on pages 4-9. Barrett did not even acknowledge receipt of the article emailed on 6th March and on 28 August said he does not post "nonsense" on his web site. He has the support of, among others: (a) The Executive Director of the American Physical Society, which publishes several top class research journals of Physics. (b) The former Commissioner of the american Federal Drug Administration (FDA), who did not ban the homoeodrugs only because he was not sure of the Congress support. (c) Fortytwo prominant "critics of quackery & pseuduscience", who have petitioned the FDA against Homoeopathy.They all hold that homoeoremedies being "placebo" do not work and hence should be banned. In fact, Barrett, his group and such other "adherents of Scientific Medicine (Allopathy)" are where I was till mid 1960s. They all, like me, can take a U-turn to support Homoeopathy if they read my this article and the book Molecular Homoeopathy carefully. But the real change in their attitude will start if and when they themselves experience the advantage of Homoeopathy over Allopathy. In this article I answer some frequently asked questions, doubts and misconceptions about Homoeopathy.

According to Avogadro's law a gram-mole of a compound contains 6.02x1023 molecules. Therefore, "homoeopathic potencies of 12c centesimal or 24X decimal and higher, do not contain any molecule of the original drug and hence are placebos", is the perpetual criticism of Homoeopathy. The authors of the above two websites are not the first or alone to raise it. In future also it will be repeated if the "scientists" continue to ignore the underlying new scientific phenomena crying for recognition to enrich the sciences.

I, like any others knowing the Avogadro's law, used to discard and disregard Homoeopathy till mid 1960s when a homoeopath 'cured' my wife's eczema not amenable to Allopathy. This aroused my scientific curiosity to see these medicines act in my own hands and to explain the basic mechanisms within the framework of modern sciences. I studied widely, took a correspondence course and started treating my self, family members, associates and friends. After twelve years of hard work and thought I concluded, in the sick bed for jaundice, that Homoeopathy's art is valid but its science is nebulous. Some new concepts, phenomena, nay Sciences are needed to elucidate the science of Homoeopathy. This marked my search's end in one direction and beginning in another, culminating in the 1984-book 'Molecular Homoeopathy' / (Molecular Biophysics of the "Micro" Dose) and four dozen publications to date. Presently I am working on the new book "Scientific Homoeopathy / (The Rational Medicine)"

How to establish the efficacy of the homoeomedicines, is the big question. I first thought of the 'Double Blind Drug Trials' (DBDT) wherein both the doctor and the patient remain "blind" or unaware whether the given dose is active medicine or a similar looking inert placebo, and which every allopathic drug has to satisfy before coming to the market for public prescription. However, I found DBDT inapplicable to Homoeopathy mainly because patients with the same pathology cannot be randomized into two groups for treatment with active homoeomedicine and placebo. Because different patients usually have different symptom-totality, calling for different homoeomedicines. And the homoeophysician cannot remain "blind" but must know total symptoms before and after every dose of the known medicine to ensure that the cure is progressing according to the Herring's laws, the last symptoms are disappearing first, and medicines & dose frequency adjusted to the changing need.

I therefore got interested in those well worked out and firmly diagnosed cases, which served their own controls. These, for the Modern Scientific Medicine are: (a) incurable/fatal, (b) difficult-to-cure even with long, some times life long, medication, (c) requiring surgery, (d) viral infections where Allopathy offers nothing, (e) baby/child diseases where placebo does not work.

The cases of Indian Childhood Cirrhosis were diagnosed on liver biopsy, liver function tests, clinical history and physical examination. And were discharged from the referral hospital with hopeless prognosis, a week or ten days' short survival and with a whisper advice to take the child quick lest he should die on the way. But all showed definite signs of improvement within three days of the start of homoeopathic treatment with Ars, Phos etc. Since ICC is known to occur with a high frequency in the siblings, it is
interesting to report that the prophylactic treatment of the mother during pregnancy and then of the child after birth succeeded in three couples, one of whom had earlier lost five sons to ICC. President Radhakrishnan's ADC, who had suffered migraine for over 20 years and resigned in disgust when he did not get relief even with the treatments in Germany & U.K., was cured with Homoeopathy. A number of cases with confirmed diagnosis of psoriasis were homoeotreated satisfactorily. The MD in Pharmacology and Dean of Dharwar Medical College in Karnataka, after reading Molecular Homoeopathy (ref.1) came all the way by air for treatment of psoriasis and experienced relief within an hour of the homoeodose of Psor and "euphoria" on overnight crust shedding. An army Colonel had to fly in non-pressurized aircrafts during 1947-48 Indo-Pak war and developed labyrinth vertigo. Ever since he suffered giddiness & reeling sensation whenever he laid down in bed, turned on side or bent down. A number of E.N.T. experts were consulted and all sorts of tests were done without relief. He took homoeomedicine Nat. Sulph. From me in August 1974 on a Friday and did head-stand on Sunday. A four year girl child was treated in the advance institute of Scientific Medicine for acute Idiopathic Thrombocytopenic Purpura in 1993 with 2mg/Kg body weight prednisolone and later with 9 gm/day for 5 days immunoglobulin Ig G, then with Chinese medicine. In April 1997 she again had an acute episode, with platelet count <5000/mm3. The ITP is so serious a disease that a patient can internally bleed and collapse while talking! The homoeo-treatment with Lach raised the platelet count gradually from <5000 through <10000 on third day and then 33000, 79000 to 120000 tested weekly, rising later to 140000. The second case of ITP had platelet count <5000 five days after receiving 1500 mg prednisolone over 3 days in a referral Institute (PGI, Chandigarh). But showed a better response to homoeotreatment with Lach starting 1 September 2001. The platelet count rose to 14000, 25000, 1.3 lac and 3.39 lac after 4, 10, 24 and 45 days' treatment. A teacher in our Nursing College lost her fianc in the 1971 Indo-Pak war and developed Thyrotoxicosis complicated with Exophthalmos and Amenorrhea. The treating endocrinologist advised her to learn to live with it. But after homoeocure with Thyroidinum she married and had two children. Out of the several cases of arthritis and spondylosis the most striking one was that of general spondylosis threatening extremities and requiring urgent surgery. The homoeocure with Rhus and Calc carb gave her permanent relief. A senior executive was admitted for surgical removal of a solitary thyroid nodule. On learning of the possible homoeocure he left the hospital and was actually cured with Calc carb. The prolapse uterus in the third stage advised Thomson correction was rectified homoeopathically Lilium Trig. Several cases of renal stone and of viral hepatitis with jaundice were also homoeo-treated; the Australia antigen test undertaken in one case became negative after homoeotreatment. A case with confirmed diagnosis of active Idiopathic Ulcerative Procto-Colitis with ulcers in rectum and sigmoid colon and having passed blood with stools for years was cured with Phos and Merc. Earlier his sister had died in 1983 of ulcerative colitis and his family had given up hopes for his survival. Among several cases of asthma the most challenging was that of a young girl who had suffered for a decade with asthma, hives and remittent fever, occasionally spending the whole nights sitting. When Wysolone and Asthaline did not give adequate relief her treating allopathic physician sent her to me and she was cured with Ars, Ipec and Sepia. Potencies used for these homoeocures were 30c, 200c and 1000c, all far beyond the Avogadro's limit of 12c. But in my latest view (ref. 20) the 15c followed, if and when required
to change the potency, by (14c 16c) mixture suffice for clinical use, unnecessitating all other potencies. Presently clinical drug trials on these lines are under way at multiple centres with Ars, Bell, Bry, Calc, Rhus and Sulph.

These my few but convincing personal observations corroborate the overwhelmingly huge mass of persuasive evidence collected by innumerable, better homoeopaths than me, all over the world during the past over two centuries that homoeomedicines do cure even in high potencies with no molecule of the original drug in them. Clearly some molecules of diluent medium (lactose, water, ethanol) act curatively, suggesting a new scientific phenomenon bypassing the Avogadro's law. Really informative were the controlled animal experiments on Alloxan induced diabetes in rats and DMBA (Dimethyl-benz-anthracine) induced toxicity and cancer in mice. The dynamized 30c potency of the disease-causing chemical was curative but undynamized simple dilution of the same extent had no effect. These observations were confirmed by ref. 8. For preparing the 30c potency the mixture was vigorously agitated at each of 30 steps of 100 fold dilution but in simple dilutions the agitations were omitted. The mechanical dynamization processes of trituration or succussion at each step of dilution, which are unique only to Homoeopathy and not yet explored by "sciences", induce the diluent molecules (lactose, water, ethanol) to acquire and later mimic the chemical specificity of the original drug molecule so as to themselves act as the therapeutic agent. So, the homoeodose is not "micro placebo" but contains plenty of medicinally active
diluent molecules, removing for good the perennial conceptual impasse created by the Avogadro's law. Modern Physics recognizes the induction of magnetism and electric charge but not of the chemical specificity of one molecule into another which underlies the process of homoeopotentization.

The new science of Inductive Chemistry envisages studying the mechanism of preparation and properties of the 'induced molecules'. The other two new sciences of Xenobiology and Inductoxenopathy are elucidated elsewhere to explain the basis and operation of the Law of Similars etc.

All sciences are based on the real facts of observation and their theoretical explanation. If and when there is a conflict between theory and observation the former is revised to describe the latter faithfully. But if
the theory is so well established that the new observations seem anomalous the latter are again repeated in a different setting. If even then the observations get strengthened, there is a need for a deeper re-evaluation to discover new scientific phenomenon, which bypasses and yet is consistent with the old theory. This is exactly the challenge thrown up by the curative action of high potency homoeodrugs. The underlying new phenomenon is the induction of chemical specificity of the solute drug molecules into the molecules of solvent medium via dynamization processes. The modern sciences do not provide for it and hence will be enriched by recognizing and investigating it further. Revision of the physical basis of the 'chemical
specificity' of a molecule becomes necessary, however.

The chemical and biochemical discriminatory mechanisms recognize a molecule in two steps: first, of physical bonding via complementary 3-dimensional structures and second, of exchanging the energy dE
specifically characteristic of the recognizer-recognizee pair. The first step exercises a negative recognition and the second constitutes the positive recognition. The first step can be definite only to tell that the
molecule not binding to the receptor for A is not the molecule A. But the second step identifies A positively via the chemically exchanged energy dE, which is uniquely specific of the molecule A. If the molecule B is induced to carry the exchangeable energy of A, the discriminatory machinery is fooled to treat B as A, as here.

The molecules of lactose, water and ethanol have only -OH group in common. The oxygen atom in the -OH group, due to sp3 hybridization, has four equivalent valency orbitals. Two of these have bond pair electrons and the other two unshared lone pair electrons. The latter having no definite higher energy levels, can be raised, in small steps, to any desired energy level and hence play the basic role here. The organic solvent DMSO (Dimethyl-Sulphoxide) has lone pair electrons but no -OH group and does not serve as a diluent medium (ref. 21), emphasizing the role of the lone pair electrons of the -OH groups.

During forceful triturations and impacted succussions the outermost electron shell of the solute drug molecules comes repeatedly in close proximity with those of the diluent molecules. This induces resonant promotion of the lone pair electrons of the diluent -OH groups, in small steps, to energy levels of the chemically active electrons of drug molecules. The diluent molecules thus acquire the chemically exchangeable energy and hence the chemical specificity of the drug molecule to get "potentized" with the drug. During serial dilutions of potency preparation the original drug molecules get eliminated and the diluent molecules resonantly promoted by them take over the resonant promotion of the unpromoted diluent molecules. These considerations have experimental support (ref. 12, 13).

Smith & Boericke (ref.12) studied the CH3-, CH2- and -OH peaks in the Nuclear Magnetic Resonance spectra of ethanol, unsuccussed and succussed dilutions of sulphur in ethanol. Only the -OH peak of only the succussed potency spreads and reduces in area under the curve. No modern science can explain this observation which in our theory however, follows from the resonant promotion of lone pair electrons of -OH groups in the potentized ethanol.

The Laser Raman Spectral peak (ref. 13) of diluent alcohol disappears in succussed dilution of Potassium Dichromate and reduces in height in that of Ammonium Nitrate but a new peak of the solute appears in both cases. These results cannot be explained by modern sciences, but follow easily from the resonant promotion of the lone pair electrons of -OH groups of the potentized alcohol.

When a causative xenobiotic X' affects a strategic biomoleule M a molecular disease complex MX' is formed which alters the rates and/or routes of the basic biochemical reactions leading to some iochemical, biopotential and tissue changes and altered feelings and sensations. The allopathic treatments deal with these results and effects of disease leaving MX' alone. Attempts are made to keep the vital parameters only within the "normal range" since their normal value for the particular patient cannot be known.
Progressing basic disease requires gradually increasing dose creating side effects. Big molecules of allopathic drugs and low homoeopotencies in lactose cannot, but high potencies in ethanol can, cross the lipid and water channels in biological membranes to act inside and produce profound effects.
On this is based homoeopaths' belief that higher potency is more powerful. This is also why Homoeopathy, even in my own hands as mentioned above, could cure some cases not amenable to Allopathy. An adherent of the modern Scientific Medicine has to actually experience and (re)think to believe, like me, in the advantages of Homoeopathy over Allopathy. For instance, the recent large scale slaughter of animals with 'foot & mouth' disease in U.K. and Europe could be minimized if Homoeopathy were allowed its due role.

The symptom totality of a disease, in fact, indicates the total biological response of the healthy subject in the patient to the causative xenobiotic, as studied by the new science of Xenobiology. It has two components: activation of the defence mechanisms against the antigenic determinants on the xenobiotic molecule and the pathophysiology of the affected biomolecules, cells, tissues & organs. The potentized homoeomedicine Dx, prepared by resonantly promoting the diluent molecules D with the crude drug molecules X, contains a mixture of diluent molecules resonantly promoted with the antigenic and pathogenic determinants of X. The xenobiotic X', crude drug X and the homoeopotency Dx, carry similar chemically exchangeable energies and chemical specificities, hence elicit similar symptom totality. The homoeocure has two pronged effect: one of stimulating the immune response (aggravating some symptoms), and second of dislodging X' from the disease complex MX' formed with the biomolecule M (ameliorating some symptoms), through competitive chemical exchanges :
MX' Dx < > MD x X'
MD x < > M D x
X' is
biodegraded/ bioeliminated
D x is depromoted to D , the
diluent molecule D is metabolized .

Competitive chemical exchanges between the pathogenic and curative
xenobiotics are thus basic to the homoeopathic drug action on the Law of
Similars. For this, the tiny amount of homoeomedicine as potentized ethanol
adsorbed/absorbed and retained for long via hydrogen bonds on sugar pills is
sufficient. For an 'incurable disease' this chemical exchange fails. Wurmser
(ref. 15) found that dynamized potencies of Arsenic and Bismuth increased
their elimination from animal tissues. I have treated a case each of
Arsenic toxicity with Arsenic album 200c, Opium toxicity with Opium 1M and
Belladonna toxicity with Bell 10M. The control of Alloxan induced diabetes
in rats with 30c Alloxan and of DMBA toxicity in mice with 30c DMBA,
presented above, are also supportive. The new science of Inductoxenopathy
envisages use of induced xenobiotics as medicines according to the Law of
Similars. Immunisation operates via the mediation of specific antibodies and as such is different from Homoeopathy.

Allopathy like Ayurved, as against Homoeopathy, operates on the 'Principle
of Opposites'. For instance digitalis, knwon for lowering the heart rate, is
given in large doses to control 'tachy cardia' (high heart rate) but is a
homoeomedicine for 'brady cardia' (low heart rate). Since the
allopathic/ayurvedic and homoeopathic medicines differ in their planes and
modes of action, the two can go together with advantage to the patient. A
judicious combination of Allopathy/Ayurved, Surgery, Homoeopathy and Yoga
offers a new Unified Therapeutics 'Navayurveda', the new Ayurved, to realize
the Holistic Health "connoting harmonious normalcy at all the four planes
of personality manifestation: social, physical, mental, supramental". Here
allopathic drug, as in emergencies, homoeopathic aggravations and advanced
stages of disease, keeps the symptoms within tolerance of the patient while
homoeomedicine effects the basic cure. The patient does not stop allopathic
drug suddenly before or on starting the homoeotreatment but tapers it off
with the progress of homoeocure and increasing relief.

According to World Health Organization "Health is a state of complete
physical, mental and social well-being and not merely an absence of disease
or infirmity". Here 'absence of disease is NOT health'. This internal
inconsistency or self contradiction arises because for diagnosis under the
Scientific Medicine a disease has to be advanced enough to create laboratory
detectable biochemical abnormalities and/or biopotential variations and/or
pathologic tissue changes out side the "normal range", since there is no way
of ascertaining the normal value of the diagnostic parameter for the
particular patient. In the pre- or sub-clinical stage, the clinical
laboratories report N.A.D. (no abnormality detected or no appreciable
disease) but the patient suffers some subjective/mental and physical
symptoms on which a curative homoeomedicine can be given to nip or abort the

disease. Therefore, Homoeopathy can and does serve as an effective
Preventive Community Medicine for preventing diseases from progressing to
advanced dangerous stages.

The other deficiency in WHO's definition is that it does not recognize the
existence or achievability of the "supra mental health". However the Yoga
practices of pranayam (breath control) and dhyan (meditation) have been
shown, as in my own experience, to induce the rare abilities like intuition,
self control on thought, assuming alpha and theta brain states at will, etc.
Homoeopathy and Yoga help attenuate anxiety and other psychological
components of disease.

Homoeopathy has always been and still is beyond the contemporary sciences
but provides the bases, and needs recognition, of three new sciences:
Inductive Chemistry, Xenobiology & Inductoxenopathy. It has vast
therapeutic potentials, both curative and preventive, yet is cheap and free
from the side effects of Allopathy. It is NOT a "Quackery" or "Ultimate
Fake", but the main (not mere alternative) Medicine of the new
century/millennium. An open minded approach, keeping the general well-being
upper most, is needed to develop Navayurveda based Medical Education and
Health Care Delivery System.

Saturday, March 13, 2010

Evidence of clinical efficacy of homeopathy A meta-analysis of clinical trials

M. Cucherat1, M. C. Haugh1, M. Gooch2, J. -P. Boissel1 and for the HMRAG group

(1) Department of Clinical Pharmacology, Hospitals of Lyon and University Claude Bernard, Faculté RTH Laennec, BP 8071, F-69376 Lyon, France e-mail: Tel.: +33-478-785757; Fax: +33-478-776917, FR
(2) Academic Departments, Glasgow Homeopathic Hospital, Glasgow, Scotland, UK, GB
: To establish, using a systematic review and meta-analysis, whether there is any evidence from randomised controlled clinical trials of the efficacy of homeopathic treatment in patients with any disease.
Data sources: Published and unpublished reports of controlled clinical trials available up to June 1998, identified by searching bibliographic databases (Medline, Embase, Biosis, PsychInfo, Cinahl, British Library Stock Alert Service, SIGLE, Amed), references lists of selected papers, hand searching homeopathic journals and conference abstracts, and contacting pharmaceutical companies.
Trials selection: Trials were selected using an unblinded process by two reviewers. The selection criteria were randomised, controlled trials in which the efficacy of homeopathic treatment was assessed relative to placebo in patients using clinical or surrogate endpoints. Prevention trials or those evaluating only biological effects were excluded. One hundred and eighteen randomised trials were identified and evaluated for inclusion. Sixteen trials, representing 17 comparisons and including a total of 2617 evaluated patients, fulfilled the inclusion criteria.
Data extraction: Data were extracted by two reviewers independently, using a summary form. Disagreements were resolved by a third person.
Data synthesis: The evidence was synthesised by combining the significance levels (P values) for the primary outcomes from the individual trials. The combined P value for the 17 comparisons was highly significant P=0.000036. However, sensitivity analysis showed that the P value tended towards a non-significant value (P=0.08) as trials were excluded in a stepwise manner based on their level of quality.
Conclusions: There is some evidence that homeopathic treatments are more effective than placebo; however, the strength of this evidence is low because of the low methodological quality of the trials. Studies of high methodological quality were more likely to be negative than the lower quality studies. Further high quality studies are needed to confirm these results.

Key words Homeopathy - Meta-analysis - Randomised clinical trial

Received: 19 August 1999 / Accepted in revised form: 29 December 1999

A meta-analysis of homeopathic treatment of pollinosis with Galphimia glauca

Wien Med Wochenschr. 1997;147(14):323-7.
[Article in German]

Lüdtke R, Wiesenauer M.

Institut für Medizinische Informationsverarbeitung, Tübingen, Deutschland.
OBJECTIVE: 1) To assess the efficacy of homeopathic prepared Galphimia glauca compared to placebo in the treatment of pollinosis. 2) To estimate the corresponding overall success rate of Galphimia glauca. Meta-analysis of clinical trials.
STUDY SELECTION: 7 randomized double-blind placebo-controlled trials and 4 not placebo-controlled trials (1 randomized and controlled, 1 prospective uncontrolled, 2 retrospective uncontrolled) performed by our study group between 1980 and 1989. An additional MEDLINE search revealed no further trials on this topic. Exclusion and inclusion criteria were identical over all trials. In total, 1038 ambulatory patients that suffered from acute pollinosis (752 in placebo-controlled trials) entered the analysis.
MAIN OUTCOME MEASURES: Relative frequency and relative risk for showing noticeable and soothing relief in ocular symptoms as assessed by the patient. The overall rate of improved eye-symptoms is about 1.25 (CI: 1.09 to 1.43) times higher in the verum than in the placebo group. Verum success rate is estimated by 79.3% (CI: 74.1% to 85.0%). Across the single studies the results were highly comparable except for the study run in 1985. A significant superiority of Galphimia glauca over placebo is demonstrated. Estimates of verum success rates are comparable with those of conventional antihistaminics, but no side effects occurred. As not all of the single studies were analyzed by intention to treat analysis the results may be biased.

PMID: 9381725 [PubMed - indexed for MEDLINE]

Homeopathic vs Conventional Treatment of Vertigo

A Randomized Double-blind Controlled Clinical Study

Michael Weiser, MBChB; Wolfgang Strösser, MD, MBChB; Peter Klein, MSc
Arch Otolaryngol Head Neck Surg. 1998;124:879-885.

Objective To compare the efficacy and safety of a homeopathic remedy (Vertigoheel, Heel Inc, Albuquerque, NM) vs betahistine hydrochloride (active control) in the treatment of patients with vertigo of various origins in a confirmative equivalence trial.

Design Randomized (1:1) double-blind controlled clinical trial.

Setting Fifteen study centers (general practice) in Germany between November 1995 and November 1996.

Subjects A total of 119 patients with vertigo of various origins (from whom 105 patients could be analyzed as intended per protocol).

Main Outcome Measures Frequency, duration, and intensity of vertigo attacks.

Results Both homeopathic and conventional treatments showed a clinically relevant reduction in the mean frequency, duration, and intensity of the vertigo attacks. The therapeutic equivalence of the homeopathic remedy and betahistine was established statistically.

Conclusions Concerning the main efficacy variable, therapeutic equivalence between the homeopathic remedy and betahistine could be shown with statistical significance (confirmative analysis). Both remedies reduced the frequency, duration, and intensity of vertigo attacks during a 6-week treatment period. Also, vertigo-specific complaints were significantly reduced in both treatment groups.

From the Biologische Heilmittel Heel GmbH, Baden-Baden (Dr Weiser), Clinical Research and Pharma Consulting, Bergisch Gladbach (Dr Strösser), and Datenservice Eva Hönig GmbH, Rohrbach (Mr Klein), Germany.

Monday, March 8, 2010

Intrathecal baclofen and homeopathy for the treatment of painful muscle spasms associated with malignant spinal cord compression

Elizabeth Thompson
Palliative Medicine, St Margaret's Hospice, Clydebank, Strathclyde

Fiona Hicks

In this case study we describe a dual approach to the palliation of difficult muscle spasms using intrathecal baclofen via a fully implanted system, together with the homeopathic approach to symptom control. The homeopathy is seen to complement rather than to replace conventional prescribing and using both approaches together appears to have avoided the necessity for increasing drug doses and to have minimized side-effects. As well as encouraging us to take on experience from other disciplines, this case study also suggests that palliative care could be a forum for evaluating the effectiveness of the homeopathic approach in symptom control in carefully designed studies.

Palliative Medicine, Vol. 12, No. 2, 119-121 (1998)
DOI: 10.1191/026921698666737949

Animal models for studying homeopathy and high dilutions: Conceptual critical review

Leoni Villano Bonamin1, 2, , and Peter Christian Endler3
1 Laboratory of Cellular and Molecular Biology, University Paulista, Brazil
2 Laboratory of Pathology, University of Santo Amaro, Brazil
3 Interuniversity College for Health and Development Graz, Austria

This is a systematic review of the animal models used in studies of high dilutions. The objectives are to analyze methodological quality of papers and reported results, and to highlight key conceptual aspects of high dilution to suggest clues concerning putative mechanisms of action.
Papers for inclusion were identified systematically, from the Pubmed-Medline database, using ‘Homeopathy’ and ‘Animal’ as keywords. Only original full papers in English published between January 1999 and June 2009 were included, reviews, scientific reports, thesis, older papers, papers extracted from Medline using similar keywords, papers about mixed commercial formulas and books were also considered for discussion only. 31 papers describing 33 experiments were identified for the main analysis and a total of 89 items cited.
Systematic analysis of the selected papers yielded evidence of some important intrinsic features of high dilution studies performed in animal models: a) methodological quality was generally adequate, some aspects could be improved; b) convergence between results and materia medica is seen in some studies, pointing toward to the possibility of systematic study of the Similia principle c) both isopathic and Similia models seem useful to understand some complex biological phenomena, such as parasite–host interactions; d) the effects of high dilutions seem to stimulate restoration of a ‘stable state’, as seen in several experimental models from both descriptive and mathematical points of view.

Effects of Potentised Substances on Growth Kinetics of Saccharomyces cerevisiae and Schizosaccharomyces pombe

C. Scherra,c; S. Baumgartnera,b; J. Sprangerc,e; M. Simond

aVerein für Krebsforschung, Institut Hiscia, Arlesheim, Schweiz bKollegiale Instanz für Komplementärmedizin (KIKOM), Universität Bern, Insel-Spital, Imhoof-Pavillon, Bern, Schweiz cForschungsinstitut für biologischen Landbau, Frick, Schweiz dInstitut für Chemie und Biologie des Meeres (ICBM), Universität Oldenburg, Deutschland eInstitut für anthroposophische Veterinärmedizin, Frick, Schweiz

Forsch Komplementmed 2006;13:298-306
(DOI: 10.1159/000095302)


Homeopathic potencies are used as specific remedies in complementary medicine. Since the mode of action is unknown, the presumed specificity is discussed controversially. Objective: This study investigated the effects of potentised substances on two yeast species, Saccharomyces cerevisiae and Schizosaccharomyces pombe, in a stable and reliable test system with systematic negative controls. Materials and Methods: Yeast cells were cultivated in either potentised substances or water controls in microplates and their growth kinetics were measured photometrically. Water control runs were performed repeatedly to investigate the stability of the experimental set-up (systematic negative controls). Results: 4 out of 14 screened substances seem to have affected the growth curve parameters slope or yield. Out of these substances, azoxystrobin and phosphorus were chosen for 8 further replication experiments, which partly confirmed the results of the screening. On the average of all experiments, azoxystrobin affected the slope of the growth curve of Saccharomyces cerevisiae (p < 0.05), and phosphorus affected the slope of the growth curve of Schizosaccharomyces pombe (p < 0.05). No effects were seen in the water control runs. In addition, significant interactions between treatment with potentised substances and experiment number were observed in all experiments with potentised substances (p < 0.01), but not in the water control runs. Conclusions: Both yeast species reacted to certain potentised substances by changing their growth kinetics. However, the interactions found point to additional factors of still unknown nature, that modulate the effects of potentised substances. This stable test system with yeasts may be suitable for further studies regarding the efficacy of homeopathic potencies.

Copyright © 2006 S. Karger GmbH, Freiburg


The present invention relates to a method of treatment, in particular to a method of treating an effect of glutamate or glutamic acid by administering a dilution or an ultra-high dilution or potentised preparation of glutamic acid or glutamate.

The reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that that prior art forms part of the common general knowledge in Australia or elsewhere.


Homoeopathy employs minute doses of usually harmful or toxic agents to stimulate organisms back to health. The agents used in homoeopathy are selected precisely on the basis of their ability to induce disease-like symptoms and signs in healthy people when administered in toxic doses, or one or more times in sub-harmful doses. These agents will, in properly diluted form, cure a sick person with similar symptoms. While microdose effects are now well accepted, for example in the phenomenon known as hormesis, some homoeopathic solutions are attenuated beyond Avogadro's constant, i.e. in theory none of the original agent remains. There have been a great number of reported experiments that demonstrate that homoeopathic remedies are effective in treating a variety of symptoms.

' Many chemical agents induce undesirable effects on organisms such as mammals. The effects which result from chemical agents having one or more chiral centres often result from a specific stereoisomer of the chemical agent. For example, (-)-adrenaline is the isomer which is found in humans and is the chemically active agent. It is about 15 times more active than (+)-adrenaline physiologically. Although chemically hard to differentiate in vitro, in vivo they are readily differentiated by the stereo-specificity of enzymes.


The inventor has now found that the effects of glutamate within the nervous system or within the organism generally can be treated by administering a dilution or an ultra high dilution or potentised dilution of a stereoisomer of glutamate or glutamic acid.

hi the context of the present invention it will be understood by those skilled in the art that the term " glutamic acid or glutamate " refers to one or more stereoisomers which induce the effect to be treated in the organism. The stereoisomer of glutamic acid or glutamate which may be used to treat the effects of the glutamic acid or glutamate within the organism or nervous system will be the enantiomer of the form of glutamate or glutamic acid naturally occurring within the organism. Therefore, a potency or dilution of D-GA may be used to modulate the toxicity of L-GA. Similarly a potency or dilution of L-GA may be used to modulate the toxicity of D-GA. (Note that D-GA = D-Glutamic Acid = D-GLU and L-GA = L-Glutamic Acid = L-GLU. Glutamic Acid = GA)

Accordingly, there is provided a method of treatment of an organism suffering from the effects of glutamic acid or glutamate, said method comprising the steps of potentising a stereoisomer of glutamic acid or glutamate, and administering said potentised glutamic acid or glutamate to the organism.

Another aspect of the invention provides a method of treatment of an organism suffering • from the effects of glutamic acid or glutamate, said method comprising the steps of diluting a stereoisomer of glutamic acid or glutamate, and administering said diluted glutamic acid or glutamate to the organism.

Still yet another aspect of the invention provides a method of treatment of an organism suffering from the effects glutamic acid or glutamate, said method comprising the steps of diluting a stereoisomer of glutamic acid or glutamate to an ultra-high dilution of said stereoisomer, and administering said ultra-high diluted stereoisomer to the organism.

However, the same effect may be obtained by using dilutions such as those used in

investigations involving hormesis. Such dilutions exist below the toxic range of a given compound, substance or molecule. Such dilutions below the toxic range are stimulatory rather than toxic, but may be bilphasic or multiphasic in their effects, i.e., stimulatory or inhibitory in their effects, depending on the concentration below the NOAEL which is being used. Usually this phenomenon exists in a narrow range of concentrations just below the toxic range. Put another way, the phenomenon exists just below the 'no observed adverse effect level' or 'NOAEL'.

The method of the present invention may be used for the treatment of any organism, including for example, animals, plants, microorganisms and in particular, humans.

Another aspect of the invention is the use of a dilution or an ultra-high dilution or potentised preparation of glutamate or glutamic acid, for the preparation of a medicament for the treatment of the toxic, physiological and/or pathological effects of said glutamate or glutamic acid.

Still yet another aspect of the invention is the use of a dilution or an ultra-high dilution or potentised preparation of glutamate or glutamic acid, for the preparation of a medicament for the treatment of the addictive and other undesirable effects of drugs of addiction.

Still yet another aspect of the invention is the use of a dilution or an ultra-high dilution or potentised preparation of glutamate or glutamic acid, for the preparation of a medicament for the alleviation of the physical and psychological effects of drugs of addiction.

The present invention may be used for the following non-limiting examples:

1. Increase or decrease neurotransmission

2. decrease apoptosis

3. increase apoptosis, e.g. in the treatment of malignancy

4. treat neuronal degeneration

5. treat Alzheimer's disease

6. treat Parkinson's disease

7. treat Huntington's chorea

8. treat HIV-associated dementia

9. treat multiple sclerosis

10. treat amyotrophic lateral sclerosis

11. treat glaucoma

12. treat obsessive-compulsive disorder

13. treat stroke

14. treat dementia 15. treat neuropathic pain

16. treat chronic pain

17. treat anxiety

18. treat depression

19. treat bipolar disorder

20. treat post-traumatic stress disorder

21. treat retinal degeneration of whatever cause

22. treat nicotine addiction or withdrawal

23. treat cocaine addiction or withdrawal

24. treat amphetamine addiction or withdrawal

25. treat morphine or opiate addiction or withdrawal

26. treat addiction or withdrawal due to licit or illicit drugs

27. treat cerebral ischaemia

28. treat neuronal injury in situations like head injury or trauma

29. treat epilepsy

30. treat schizophrenia

31. treat psychosis

32. treat Chronic Fatigue Syndrome ("CFS") - CFS has a common physiological mechanism compared with depression, anxiety and stress.

33. treat addiction in general

34. treat alcohol addiction or withdrawal

Alternatively, the invention may be used for the preparation of a medicament for the alleviation of the physical and psychological effects of any of the conditions listed above and numbered from 1-34 inclusive.

It has also been found that the enantiomer of the endogenously occurring isomer of glutamate or glutamic acid, used to treat the organism, may be derived from other salts of glutamate or glutamic acid, e.g., instead of glutamic acid being used, sodium glutamate may provide a suitable isomer of glutamate.

It should be noted that (-)-glutamate (or L-glutamate) is the naturally occurring isomer in the human. Therefore a preparation of (+)-glutamate (or D-glutamate) would be used to treat the human in the context of this method, when the effects of endogenous, naturally occurring glutamate, are being treated.

Potentization of the stereoisomer, glutamate or glutamic acid, may be according to the practices used in homoeopathy, homotoxicology or any other system of medicine or
treatment which uses potencies, for example, anthroposophical medicine
. For a discussion of the method of preparation of stereoisomers in general, and this includes glutamate, into potency, refer to previous patents by Kuzeff as inventor, especially 'Spacetime modification of homeopathic medicinal action',22'23 and also other patents, including AU 2003208170, AU 2005100336, AU 2005100337 and AU 2007100082. The description of manufacture of potencies in the following pages applies to any stereoisomer including Glutamic Acid. Preferably the stereoisomer is potentised by succussion or trituration. Attenuation or dilution of the medicinal substance is, in homoeopathy, usually performed in the decimal, centesimal and fifty millesimal (LM) systems as the standard scales of attenuation, under which each successive attenuation contains just 1/10, 1/100 or 1/50,000 as much of the medicinal substance as the preceding attenuation. It is preferred that after each attenuation, the attenuated medicinal substance is succussed (typically between 10 to 100 times at each stage of attenuation) or triturated. Generally, soluble substances may be subjected to succussion and insoluble or solid substances may be subjected to trituration. Other forms of agitation may be used instead of succussion or trituration, e.g., sonication, Lab dancer, etc.

In order to prepare the dilutions or potencies A ml of tincture or B grams of medicinal substance are added to C ml or D grams or E parts of vehicle. Subsequent liquid or solid attenuations are made by serial progression, succussing or triturating one part of the preceding attenuation to C ml, D grams or E parts of the vehicle respectively. A, B, C, D, and E, are any numbers greater than zero. When preparing consecutive attenuations, it is not necessary for A, B, C, D or E, to be kept constant. For example for the first, second, third, fourth, fifth, etc. attenuation, the values of A could be Ai, A2, A3, A4, A5 ... etc., where A1, A2, A3, A4, A5 ... etc. represent any numbers greater than zero. The same principle applies for values of B, C, D, and E.

hi the decimal scale of attenuation is generally practised one millilitre (1.0ml) of tincture, one millilitre of IX aqueous solution, or one gram (1.Og) of IX trituration represents 0.10 gram of dry crude medicinal substance. One millilitre of 2X attenuation, or one-gram (1.Og) of 2nd trituration contains 0.01 gram of the dry crude medicinal substance. Subsequent liquid or solid attenuations are made by serial progression, succussing or triturating one (1) part of the preceding attenuation to nine (9) parts of the vehicle, and represent the following proportions of active principle (i.e. dried medicinal substance):

2X = IO-2 6X = IO-6

3X = IO'3 7X = IO"7

4X = IO"4 8X = IO"8 5X = IO"5 (and so on ....)

In decimal attenuations nX = 10"n where n is an integer greater than 0. In the case of centesimal attenuations, each attenuation contains just one hundredth of the medicinal substance of the one before, nC = 10'2n. In the case of fifty millesimal attenuations one millilitre (1 ml) of the first fifty millesimal attenuation (ILM) represents 4.OxIO"9 gram of dry crude medicinal substance. One millilitre (1 ml) of the second fifty millesimal attenuation (2LM) represents 8.OxIO'14 gram of dry crude medicinal substance. Each subsequent attenuation represents a further decrease in concentration of dry crude medicinal substance by a factor of 2xlO"5. Each attenuation such as 2X, 3X or nX (2C, 3C or nC) (2LM, 3LM etc) is generally referred to as a potency.

hi order to prepare the solid or liquid stereoisomers of chemical agents, it is effective to add 2 or more different potencies or attenuations together. A potency refers to a solution, which has undergone serial dilution and succussion and/or agitation whereas attenuation refers to a ' process of dilution, which may or may not involve succussion or agitation. The term potency also refers to solid attenuations as described herein. The term 'different potencies or attenuations' encompasses 2 potencies or attenuations of different dilutions as well as solutions which have undergone a different number of steps of serial dilution or attenuation with succussion, or, in the case of solid attenuations, a different number of steps of serial trituration as described herein. For example, a person skilled in the art could add the fourth and twelfth potencies or attenuations together in equal or unequal quantities. The solution may then be succussed (shaken) N times, where N is any integer greater than zero.

Alternatively, the solution is not succussed. Similarly, one could add the fourth, twelfth, or thirtieth potencies or attenuations together, or any number of combinations of potencies or attenuations. It is also common practice in homeopathy to make mixtures of different potencies of the same medicinal substance or mixtures of potencies of different substances in various combinations. Such mixtures are called complexes and are have been commercially available for many years.

Accordingly, one or more potencies of glutamate or GA could be included in such mixtures. It would be possible to add GA potencies to existing available commercially available complexes. Note that the term 'different potencies or attenuations' also encompasses the situation where the same or different potencies, made from 2 or more different medicinal substances, are added or mixed together. Such mixtures are common practice in homeopathy and are called 'complexes'.

Also, there is the situation where one could treat the effects of a racemic mixture or of specific stereoisomers (or optical isomers) with both the (+)- and (-)- enantiomers contemporaneously, either simultaneously or within the same course of treatment. This could be with a mixture of equal or unequal volumes of potencies or attenuations or dilutions or a combination of one or more of same or different potencies or attenuations of each enantiomer. This is illustrated in the discussion and non-limiting examples which follow:

For instance, one could mix one, two, three, four, five or more potencies, attenuations or dilutions of the (+)- enantiomer to one, two, three, four, five or more potencies of the (-)- enantiomer or vice versa. One or more potencies, attenuations or dilutions of an enantiomer may be prepared in one mixture, and added to an equal or unequal number of the same or different or any combination of potencies or attenuations of the other enantiomer prepared in a separate mixture. Alternatively, all potencies, attenuations or dilutions could be added to the same mixture.

(+)- and (-)- enantiomers may be mixed 50:50 or 25:75 or in any proportion. Thus, a mixture could be prepared by adding 2ml of 4th, 12th and 30th potencies of (+)- enantiomer to 2ml of 4th, 12th and 30th potencies of (-)- enantiomer, or vice versa. Alternatively 0.5ml of 4th, 2ml of 12th and 3.4ml of 30th potencies could be used, and in fact the numbers 0.5, 2 and 3.4 could be replaced by any numbers greater than zero or equal to zero. Also, the 4th, 12th and 30th potencies could be replaced by any potencies represented by integers greater than or equal to 1. The mixtures of (+)- and (-)-enantiomers prepared separately, could then be administered separately or mixed together. If mixed together, the resulting solutions could be succussed or not succussed, and subsequently serially diluted or attenuated or potentised or not. Alternatively, the (+)- and (-)-enantiomers may not be prepared

separately, and mixing could occur in the one container.

Also, in the spirit of the above description, 0.4ml of 3rd potency, attenuation, or dilution of the (-)-enantiomer, may be added to 1.2 ml of the 13th, 5.2 ml of the 41st, 4.5 ml of the 200th' and 3.7 ml of the 1000th potency, attenuation or dilution. This may then be succussed or not. In turn, this may then be added to a mixture of 5.1 ml of the 5th potency, attenuation or dilution, 0.3 ml of the 37th, and 6.3ml of the 105th potency, attenuation or dilution of the (+)- enantiomer. This latter mixture may have been succussed or not. The resulting combination of mixtures may then be serially diluted, attenuated or potentised or not. In this paragraph the number denoting potencies, attenuations or dilutions can be replaced by any integers greater than zero. Numbers representing millilitres of potency, attenuation or dilution, can be replaced by any numbers greater than or equal to zero.

(+)- and (-)- enantiomers may be mixed 50:50 or 25:75 or in any proportion. Thus, a mixture could be prepared by adding 2g of 4th, 12th and 30th potencies of (+)- enantiomer to 2g of 4th, 12th and 30th potencies of (-)- enantiomer, or vice versa. Alternatively 0.5g of 4th, 2g of 12th and 3.4g of 30th potencies could be used, and in fact the numbers 0.5, 2 and 3.4 could be replaced by any numbers greater than zero or equal to zero. Also, the 4th, 12th and 30th potencies could be replaced by any potencies represented by integers greater than or equal to 1. The mixtures of (+)- and (-)-enantiomers prepared separately, could then be administered separately or mixed together. If mixed together, the resulting solutions could be succussed or not succussed, and subsequently serially diluted or attenuated or potentised or not. Alternatively, the (+)- and (-)-enantiomers may not be prepared separately, and mixing could occur in the one container.

Also, in the spirit of the above description, 0.4ml of 3rd potency, attenuation, or dilution of the (-)-enantiomer, may be added to 1.2g of the 13th, 5.2g of the 41st, 4.5g of the 200th' and 3.7g of the 1000th potency, attenuation or dilution. This may then be succussed or not. In turn, this may then be added to a mixture of 5.1g of the 5th potency, attenuation or dilution, 0.3g of the 37th, and 6.3g of the 105th potency, attenuation or dilution of the (+)-enantiomer.

This latter mixture may have been succussed or not. The resulting combination of mixtures may then be serially diluted, attenuated or potentised or not. In this paragraph the number

denoting potencies, attenuations or dilutions can be replaced by any integers greater than zero. Numbers representing grams of potency, attenuation or dilution, can be replaced by any numbers greater than or equal to zero.

The same procedures as described above could be used for mixtures of stereoisomers in general, including diastereoisomers or mixtures of diastereoisomers and enantiomers.

The administration of the potentised stereoisomer is typically by an oral route but may be administered intravenously, intramuscularly, transdermally, subcutaneously, topically, intrathecally, intraperitoneally or via any mucous membrane (typically sublingually). It is particularly preferable to administer the potentised stereoisomer orally or sublingually.

Specific examples of administration of the potentised stereoisomer include tablets, globuli, liquid dilutions for injection and liquid external preparations. Administering potencies via different routes is part of routine optimization of therapy in homeopathy; i.e., one uses whichever route is most expedient.

Another method of administering the potentised or attenuated stereoisomer is to use devices such as the MORA machine, Listen Machine or Vega Select Machine or other bioresonance or electrodermal testing devices to detect an electromagnetic or bioresonance signal from the potency or attenuation and then administer the signal in an unchanged, modified or inverted form to the organism to be treated. These devices which are commercially available, claim to be able to copy the effects of medicines, dilution or potency and pass the attributes of a medicine, dilution or potency onto a heretofore placebo or medicinally inactive vehicle. The terms "modification" or "inversion" of the signal includes changing the polarity of the signal.

hi order to determine the appropriate potency of the stereoisomer in order to achieve the most effective treatment of the undesirable effect of the chemical agent it is usually preferred to commence by administering a low potency of the stereoisomer, say between 1C- 1OC inclusive (or IX - 10X) and gradually incrementally increasing the potency until the treatment is optimised. Experience may show that 6C, 15C, 30C, 200C are appropriate attenuations in most cases. Attenuation of lOOOC, 10,000C or higher may also produce desirable results. This also applies for non-decimal and non-centesimal potencies. Trying different potencies to determine the optimal effect in a patient is part of routine optimization of therapty in homeopathy.

The vehicles used to attenuate the stereoisomer may be selected from the group consisting of water, such as water for injection B.P. or U.S.P., lactose B.P. or U.S.P., sucrose B.P. or U.S.P. ethanol typically in suitable concentrations (e.g. 15 - 95%). Also absolute ethanol, purified water, glycerol 85% or other ethanol/water mixture or dilution of glycerol may be used. Other vehicles will also be apparent to those skilled in the art of homoeopathy.

The methods of preparation of solid or liquid stereoisomers of chemical agents into potentised attenuations include where water-soluble or alcohol-soluble isomers are to be prepared into potencies the use of water B.P. or purified water alone, or in a mixture of water and ethanol, say, 30-45% ethanol. Ethanol-soluble stereoisomers may be prepared using higher concentration ethanol solutions, say 55-95% ethanol or absolute ethanol. It is possible to start using lower and incrementally lower ethanol concentrations as the potency reaches 3 to 5X or 3 to 5C. Final homeopathic liquids often contain 30-40% ethanol.

The stereoisomer may be prepared by a process of trituration. The process of trituration is particularly advantageous when the stereoisomer is not readily soluble in water, ethanol or water/ethanol mixes. At potencies beyond 3C or 6X it is possible to convert from trituration to liquid dilutions or vice versa. For example, liquid dilutions may be prepared by first making a trituration and then diluting it in liquid such as water for injection.

In trituration one part of the medicinal substance, when preparing the first potency, or one part of the preceding attenuation when preparing the second or subsequent potencies, is added to one third of the total vehicle (e.g. lactose B.P.) used for that potency. The process of trituration is typically performed with mortar and pestle for 15 to 20 minutes. The side of the mortar is then scraped for five minutes to dislodge any attenuated substance with the pestle or with a spatula. Then the second third of the vehicle for attenuation is added to the mortar and the contents subjected to a further 15 to 20 minutes trituration prior to scraping the sides of the mortar for a further five minutes to dislodge attenuated substance. The remaining one third of the vehicle is added to the mortar and the combined mixture is subjected to trituration for a further 15 to 20 minutes to complete the trituration for that
potency. Alternatively, the total vehicle may be added to the medicinal substance or preceding attenuation at each successive stage of potency preparation and subjected to 60 minutes of trituration. Each successive level of attenuation is called a potency.

The process of the present invention may be used to reverse, or in another embodiment enhance, the effects in vivo, and in vitro, of glutamate or any of its compounds. Another aspect of the invention provides for the use of the method of the present invention to enhance or reverse the effects in vivo and in vitro of (-)-glutamate or any of its compounds or to enhance or reverse the effects in vivo and in vitro of (+)-glutamate or any of its compounds.

It is to be understood that in this specification the terms enantiomer, stereoisomer and optical isomer refer to the relevant isomers of glutamic acid.


The present invention is further described by the following non-limiting examples.

Use of Enantiomeric Glutamate in the treatment of Depression, Anxiety and Stress - a placebo controlled randomized study


Previous papers describe the modulation of toxic effects of optical isomers by use of their stereoisomers.23"26 To abbreviate the current concept, it could be said that the toxicity of optical isomers can be counteracted by administration of a potency of its enantiomer, however, this can be extended to stereoisomers in general and also can be used to increase isomer activity as well as inhibit it. The present study entails the use of homeopathic potencies of (+)-glutamic acid for the treatment of depression, anxiety and stress as measured by DASS questionnaire.


To paraphrase Hyman, major depression has a lifetime prevalence of 1 in 10,9 attention deficit hyperactivity disorder is attributed to 8.7% of children between 8 and 15,12 eight million or 1 in 10 children in the United States take stimulants like Ritalin, where the use of such medications increased 400% in the decade from 1986.53 Autism affects 1 in 166 children, which is an increase of 11 -fold in the last 10 years.52 Learning disabilities affect 5- 10% of school age children.14 Alzheimer's disease affects 30% of people over 85 years and is expected to increase 3 times by 2050, costing tens of billions of dollars annually.7 Psychotropic medications are the number 2 selling class of prescription drugs.4 With this impressive list of facts it would appear that modern society is in the grip of an epidemic of psychiatric disorders. Hyman describes how such conditions may be viewed as systemic disorders affecting the brain rather than primarily brain disorders, and describes how the problem may be approached from the perspective of nutritional and environmental medicine using a systems approach, and presents a case history with what was a well-demonstrated and obviously very satisfying outcome for patient, parents and practitioner.18

It is noted that a number of the above conditions have been associated with increased action of the neurotransmitter glutamate at NMDA receptors. Glutamate is the major excitatory synaptic neurotransmitter in the brain and is found in 80% of neurons and there is increasing" evidence that antagonists of glutamate action at NMDA receptors have antidepressant-like action,32 and anxiolytic action.19'32 Glutamate is characterized by a number of experts to be largely responsible for the ability of the nervous system to rapidly transmit information from one part of the body to another, and to be important in thought formation and memories.27

Excitotoxicity is a term applied to glutamate and is the excessive exposure to the neurotransmitter glutamate or to stimulation of its membrane receptors, and is considered a main contributor to neuronal injury and death in numerous conditions.27 Excitotoxicity in this context was first described in 1969 35 (in Lipton). hi the case of the NMDA receptors these include Alzheimer's disease, Parkinson's disease, Huntington's disease, HTV- associated dementia, multiple sclerosis, amyotrophic lateral sclerosis, and glaucoma, obsessive-compulsive disorder, stroke, dementia and neuropathic pain,3 and also anxiety and depression.19 Of note is that the role of glutamate in nervous system functioning and its excitotoxic and apoptotic effect draw interesting parallels with comments by Rudolf Steiner, the founder of Waldorf Schools, in the early 20th century, about 1913. He mentions that the process of normal thinking, as opposed to what he calls 'sense free thinking', exerts an effect on the organism in general and the nervous system in particular, contributing gradually to death and cell death. GA is a molecule considered to be central to nervous system functioning, and has an effect leading to cell death. It therefore seems to fulfill important criteria in the context of Rudolf Steiner's claims.1'2 This is really interesting, since the same author, who has a tendency to startle some people with his firm beliefs, nonetheless has a record of being quite prescient.

The situation where one specific physiological portal, as represented by GA, may lead to wide ranging and multiple pathologies the inventor calls a devils' door phenomenon. The overall impression is that modulation of GA activity may lead to significant CNS effects.

Another group of glutamate receptors are the metabotropic glutamate receptors. Evidence suggests that inhibition of some receptors belonging to this category may play a part in counteracting nicotine addiction; other receptors in the same category may assist with depression occurring in early nicotine withdrawal and may be useful in treatment of depression generally.31 Metabotropic glutamate receptors may also be promising targets in the treatment for neurologic disorders derived from abused drugs such as cocaine, morphine and amphetamines, and may also play a part in the regulation of several neurodenerative disorders, epilepsy, and ischemia.51 Some metabotropic receptor agonists may be useful in the treatment of psychotic disorders including schizophrenia.4'5 Note that this sentence refer to agonism on a glutamate receptor function. On the basis of available evidence and consistent with the well-known homeopathic principle of aggravation, it is also possible to increase toxicity of a compound using stereoisomers in potency, and accordingly, future experiments could investigate increase of optical isomer, and in the case of the present paper, glutamate, activity.13'23'32'50

There are 3 classes of glutamate-gated ion (or ionotropic.) channels, known as AMPA, kainite, and NMDA receptors. Excessive activation of the NMDA receptor leads to production of damaging free radicals and other enzymatic processes contributing to cell death.28'29'33 In addition the are at least 10 types of metabotropic glutamate receptors. It should be noted that the functions and interactions of receptors in the brain do not act in isolation. GABA receptors for instance balance the actions of glutamate to prevent

1. Steiner, R. The Incarnation of Ahriman: The Embodiment of Evil on Earth. Rudolf Steiner Press. Forest Row. ISBN 1 85584 187 9, pages 101-103, 2006.

2. Steiner, R. Approaching the Mystery of Golgotha, Anthroposophic Press, Massachusetts, 2006. page 12. ISBN 0-88010-606-9

hyperexcitation of neurons, and abnormal GABA-glutamate tone in the brain may contribute to panic prone or anxiety states.19

Similarly it should be noted that fear extinction involves the new learning of fear inhibition and is considered crucial for effective anti-anxiety treatment.49 Although the same authors note that NMDA antagonists may have anti-anxiety action, they also note that a partial NMDA receptor agonist may facilitate extinction.13 It should be noted that clinical experience using homeopathic potencies of enantiomeric glutamate indicate a likely anxiolytic, antidepressant and anti-stress action measurable after 4-6 weeks treatment, as measured by DASS questionnaire.34 This has even been noted in patients with severe psychosocial problems and chronic pain. The effect on pain has not been measured at the present time, but it may be a good idea in such patients to record pain scales at baseline and at 2 weekly intervals together with the DASS questionnaires.6'16

Similarly it could be noted that other glutamate receptors have relevance in other areas. AMPA potentiators may be of benefit in enhancement of cognitive function.32

Other agonists of the NMDA receptor such as aspartate and D-serine are known. Glycine is a well known co-agonist of the NMDA receptor with glutamate. Accordingly, potencies of D-serine, aspartate and glycine may be used to modulate the activity of these agonists of the NMDA receptor, and could be utilized in combination either as a complex, or separately as simplexes, to modulate NMDA receptor function.

One of the best opportunities for studying the effects of similar substances, which are almost identical but are in fact not identical, is in the case of optical isomers. Stereoisomers, which are enantiomers of each other, are almost identical chemically. Stereoisomers have identical molecular formulae and structural formulae, but different configurational formulae, i.e., the molecules are identical to each other except for their spatial orientation. Enantiomers have the peculiarity that they are mirror images of each other. The 2 mirror images are typically differentiated with the notations (+)- and (-)- referring to the ability of the compound to rotate polarized light in a polarimeter, and (R)- and (S)- which refers to the Cahn-Ingold- Prelog convention specifying the orientation of the molecular weights of moieties attached to a chiral centre in the molecule. Their main identifying difference is that they rotate polarized light differently when analysed using a polarimeter. Furthermore, enantiomers rotate polarized light by an approximately equal number of degrees in a polarimeter, but in opposite directions. ' ' 10

The profile of physiological actions of optical isomers and their enantiomers or diastereoisomers are typically very similar, however, the potency of each form of stereoisomer in terms of a given physiological action may vary widely, and the actual physiological actions are very different. For example adrenaline found in humans is the minus isomer and is about 15 times more active than the plus isomer physiologically.

Similarly, the β-receptor blocker (-)-propranolol is about 60-100 times more active than (+)- propranolol in blocking the inotropic, chronotropic, and vasodepressor actions of the β- receptor stimulant isoprenaline. However, the (+)-isomer is more effective at inhibiting oubain-induced arrhythmias in dogs.17

Previous studies have described how it was sought to treat the toxic or physiological effects of optically active compounds by using their stereoisomers, and more specifically, their enantiomers, in potentized form.23"26 This is particularly of interest since in nature since a large proportion of biomolecules are stereoisomers. By so doing, the attempt was made to mimic the simillimum principle of homeopathy. However, rather than focus on the actual symptoms and signs as is done in homeopathy, the method of the instant application focuses on the profile of physiological actions of stereoisomers and specifically glutamic acid.

Looked at in its simplest manifestation, the simillimum principle says that to treat, a sickness one must administer a medicinal preparation which in suitably attenuated form, is capable of producing the same symptoms and signs which are being exhibited and experienced by the patient. Stereoisomer symptoms are different.

Experiments using isopathy have been previously reported3'4'5'6'7'8' '' >u Isopathy is not the same

3. Cazin J.C., Cazin M., Gaborit J.L., Chaoui A., Boiron J., Belon P.. Cherruault Y., Papapanavotou C. A study of the effect of decimal and centesimal dilutions of arsenic on the retention and mobilization of arsenic in the rat. Hum Toxicol 6, 315-20 (1987).

4. Fisher, P., House, I., Belon, P., and Turner, P. The influence of the homoeopathic

as homeopathy. In homeopathy something similar is used and not something identical to the pathological state (as is the case in isopathy) or to that which causes the pathological state. In isopathy the effects of a morbific agent are treated by a dilution or potency of the same morbific agent. Therefore, to treat the toxicity of intraperitoneal injections of (-)-U-50488 in an isopathic way, one would use potencies of (-)-U-50488. However, to use a potency of an enantiomer or diastereoisomer, is not isopathy, since we are not administering a potency of the identical substance, but rather a configurationally and functionally different molecule. When the enantiomer is used, we call this enantiomeric treatment or therapy. More generally, it can be called stereoisomeric treatment therapy. Note that the term 'treatment' has been used in a different context in the bioassay section, describing experiments with microorganisms, which occupies a later section of this specification.

Although interesting and significant effects have been reported in previous studies involving isopathy, these have tended not to be of the magnitude one seems to observe in the clinic. This suggested that the effects seen with isopathy to date are not representative of true homeopathic effect, where effects seem to be more dramatic.

remedy plumbum metallicum on the excretion kinetics of lead in rats. Hum Toxicol 6, 321-4 (1987).

5. Jonas, W., Lin, Y., and Tortella, F. Neuroprotection from glutamate toxicity with ultra-low dose glutamate. Neuroreport 12, 335-9 (2001).

6. Jonas, W.B. Do homeopathic nosodes protect against infection? An experimental test. Altern Ther Health Med 5, 36-40 (1999).

7. Aabel, S., Laerum E., Dølvic S., Djupesland, P. Is homeopathic 'immunotherapy' effective? A double-blind, placebo-controlled trial with the isopathic remedy Betula 30c for patients with birch pollen allergy. Br Homeopath J. 2000 Oct;89(4):161-8

8. Aabel, S. No beneficial effect of isopathic prophylactic treatment for birch pollen allergy during a low-pollen season: a double-blind, placebo-controlled clinical trial of homeopathic Betula 30c. Br Homeopath J. 2000 Oct;89(4):159-160.

9. . Berchieri A. Jr., Turco W.C., Paiva, J.B., Oliveira G.H., Sterzo, E.V. Evaluation of isopathic treatment of Salmonella enteritidis in poultry. Homeopathy. 2006 Apr;95(2):94-97.

10. Velkers, F.C., te Loo, A. J., Madin, F.,van Eck, J.H. Isopathic and pluralist homeopathic treatment of commercial broilers with experimentally induced colibacillosis. Res Vet Sci, 2005, Feb;78(l):77-83.

11 . de Almeida, L.R., Campos, M.C., Herrera, H.M., Bonamin, L.V.,da Fonseca, A.H. Effects of homeopathy in mice experimentally infected with Trypanosoma cruzi. Homeopathy. 2008 Apr;97(2):65-9.

The basic principle of homeopathy is "like cures like". This is also called the "law of similars". Note that it is not "the law of identicals", and in homeopathy one does not say "identical cures identical". Isopathy represents the latter approach rather than the former. In homeopathy minute doses of (usually) harmful or toxic agents are used to stimulate a person back to health. The agents used in homeopathy are selected precisely on the basis of their ability to create the noxious symptoms and signs experienced by the patient, if given to healthy people in toxic doses, or repeatedly in sub harmful doses.15 In homeopathy administration of substances in potentized form is believed to accompany the most dramatic effects clinically. It is hypothesized that the enantiomeric treatment of optical isomer effects will demonstrate larger effects than isopathic treatment, if the law of similars is being more faithfully mimicked.

In previous experiments we used enantiomers of propranolol, U-50488, isocyanate to provide an experimental model which attempted to mimic the homeopathic law of similars, rather than some notion of a "law of identicals" 26

In this study we use the amino acid glutamic acid. However, it should be noted that other amino acids could also be of use. These include aspartate and glycine, hi fact aspartate, glutamate and glycine isomers could be used to perform experiments to modify plant metabolism, e.g., carbon fixation. Glutamine isomers could be used potentially to influence nitrogen fixation in plants.

Materials and Methods

This study compares the ability of a (+)-Glutamic acid homeopathic complex with placebo, in terms of their respective abilities to improve DASS questionnaire scores over a 6-week period, in patients scoring an average of 14 or more points on the DASS questionnaire. Patients eligible for admission would be those with depression, anxiety or stress. Patients with bipolar disorder, PTSD, OCD and panic attacks would also be admissible. Patients with psychotic disorders or a history of psychosis would not be eligible.

(+)-Glutamic acid homeopathic complex or simplex will be administered to 90 patients in the course of their normal treatment in a prospective randomized placebo controlled study. 45 patients will be in each arm. This sample is sufficient to detect a difference of 10 points in the

DASS questionnaire score with power 0.95 at the 2.5% level.

Patients must have average DASS questionnaire scores above 14 will be suitable for admission to the study. Randomization and blinding of Glutamic Acid potencies and indistinguishable succussed placebos will be performed by Brauer Biotherapies, Tanunda, S. A. Bottles will be coded IA and IB up to 82 A and 82B or further depending on sample size. Either A or B will be medicine or placebo according to randomization and blinding. Consecutive patients will receive bottles 1 A,B up to 82A,B. Patient will take the 'A' bottles for 6 weeks. Alternatively, bottles will be randomized by coin toss to medicine or placebo groups and numbered consecutively 1 ,2,3,4. This latter method will be more practical in the performance of the study in view of the next paragraph, since each sequential bottle is randomized and has a 50% chance of being medicine or placebo.

When data for each patient is collected, results will be sent to Brauer who will record the result ' and then break the blinding for that patient. If the patient received placebo he will then be offered treatment with the active bottle if he so chooses. If the patient received the active treatment in bottle A, then he will be offered the chance of continuing treatment if he is favorably disposed. The minimal sample size needed for a plausible study would be 31 in each arm - see below.

The DASS questionnaire consists of 42 questions divided into 3 groups of 14 questions relating to 3 areas: Depression (D), Anxiety (A) and Stress (S). The investigator can score for the individual items or take an average of the entire score. Scores for each of the D, A and S data range from a minimum of zero, to a maximum of 42. Up to approximately 14 is considered a normal score for all 3 items - depression, anxiety and stress. It is proposed to take an average score of all the items as the primary end-points in the study at baseline, 2 weeks, 4 weeks and 6 weeks, however, at the end of the study analyses will be performed on each of the 3 areas of Depression, anxiety and stress at the baseline, 2 week, 4 week and 6 week milestones. Therefore this will be a repeated measures study. The study will not be stratified to give a definitive answer for each individual outcome - D, A and S - since this will require a trebling of the sample size. Also, we will not restrict the study to just D, A or S, since this will limit the number of eligible patients for admission to the study, since only one clinic is involved in the recruitment.

To detect a difference of 5 points in the DASS questionnaire we require 82 patients in each arm for power of 0.95 and p=0.05. To detect a difference of 10 points in the DASS questionnaire we require 45 patients in each arm for power of 0.95 and p=0.025. For power of 0.8 and p=0.05 and to detect a difference of 10 points on the DASS questionnaire we require a sample size of 31 in each arm.8 Normative data available for the DASS questionnaire were used in the generation of the table below:6' 8;3°

The Table, below, shows values for power calculation:

Power calculation table for Glutamic Acid study using DASS questionnaire based on published normative data.6' 8 30

Average (Pre Test - Post Treatment Stress Score)

VAR(D) AlphaZ-AlphaPowerZ-Beta 1 4 5 6 7 8 9 10

0.050 1.64 0.80 0.8

0.025 1.96 0.80 0.8

125.140.050 1.64 0.90 1.28

0.025 1.96 0.90 1.28

0.050 1.64 0.95 1.64

0.025 1.96 0.95 1.64

These calculations give the sample size required to detect significant differences between Pre and Post test stress scores. They test the null hypothesis that expected Pre Test Scores = expected Post Test score. For example, under the null hypothesis a difference of 5 is significant at the 5% level (one sided) with a power of 0.8 with a sample size of 62. These calculations assume that the SD for high Stress scores is the same for low scores (specifically I've used the Normative value of 7.91 throughout). Furthermore, I have assumed that the pairs of scores are uncorrelated; this gives an upper bound for the sample size for a given Alpha level and power.

Clinical Cases Persons presenting to the inventor with chronic depression, anxiety or stress according to DASS questionnaire, were offered treatment, free of charge, with potentized D-Glutamic Acid (D-Glutamic Acid = D-GA, L-Glutamic Acid = L-GA). Chronic in the context of the following case data means more than 6 months duration, but most cases had a history of years of distress not adequately responding to treatment with usual medical therapy. Note that prior to days on which patients were recruited which are reported below, the invention had not been discussed with any of the patients. Patients were recruited and given D- Glutamic acid potencies after giving verbal informed consent.

The first person to be recruited and to receive potentized glutamic acid was case 1. The second was case 2 and so on. No cases are omitted. Data was extracted from the computerized records between approximately 17/7/2008 and 27/7/2008. The first case was recruited and commenced on potentized D-Glutamic acid therapy on 9th November 2007. Patients were asked to perform DASS scores at a two weekly intervals for 6-8 weeks at least.

Failing this, the data was collected opportunistically upon representation for consultation. For each patient, the data collected is included in the graphs below, with intervening measurements not omitted. Measurements were supposed to be collected every 2 weeks preferably and not sooner. Advice for taking potencies is below after Case 21 of the clinical cases.

Potencies were manufactured according to the Homeopathic Pharmacopeia of the United States, HPUS Revision Service General Pharmacy, 2004. 100 succussions occurred at each stage of potency preparation. Adding equal volumes of the individual constituent potencies together made final potencies, e.g., to make the LM 4/12/30 potency, equal volumes of the LM 4, 12 and 30 potencies were added together. No succession occurred. Potencies were prepared in 30% ethanol. All potencies given to patients were in drop form in 30% ethanol, unless otherwise stated. The results are presented in the attached Case Tables.

Case l A patient presented to the clinic, which he/she had attended since August 2007, who was taking regular large doses of intramuscular narcotic for pain control. Chronic pain has been a problem for years. Patient was recruited on 9th November 2007 (9/11/07 or 9.11.07 according to Australian notation) and commenced treatment with D-Glutamic acid potencies. The patient was asked to alternate a potency chord of D-Glutamic Acid LM 4/12/30 daily with a potency chord of D-Glutamic Acid 4/12/30 C every 2 days. Thus the first potency chord referred to is a mixture of LM 4, LM 12 and LM 30 potencies of D- Glutamic acid. The LM notation refers to the 50th millesimal scale of attenuation in homeopathy. The second potency referred to is a mixture of 4th, 12th and 30th centesimal potencies. 'C chord' and 'LM chord' in the following text means the 4/12/30 C potency chord of D-Glutamic acid and the LM 4/12/30 potency chord of D-Glutamic acid respectively. The reason for alternating potencies, which is unusual if not unknown in homeopathic practice, was in order for the practitioner to obtain an impression of whether the two potency chords differed in their clinical efficacy, or indeed if they would work synergistically. If it transpired that one of the potency chords was more efficacious, then it was anticipated putting forward a proposal to conduct a double blind randomized controlled trial using the more efficacious potency chord. If both of the chords were not effective in clinical use, then another potency or combination would be suggested after a period of clinical use. The full 42 question DASS questionnaire was used for this patient.

Patient reported a marked improvement in DASS score and cessation of intramuscular narcotic use. MK has it on good authority that the patent is not a drug seeker. Patient recommenced narcotic use for pain, but at a much lower dose of methadone lOmg twice daily orally.

Unfortunately the DASS question sheet for 9/11/07, which is the date of recruitment to the study, has been misplaced in the files, or perhaps the patient did not hand one in on the day. However, a DASS questionnaire completed on 1/11/07 prior to recruitment produced the scores used in the graph above.

These data are consistent with efficacy of alternating LM and C glutamic acid potency chords.

Case 2

New patient presents for consultation for first time on 9th November 2007. Usually attends another practitioner. Also attends a psychiatrist for anxiety. Recruited and commenced on D-Glutamic Acid LM 4/12/30 and D-Glutamic Acid 4/12/30 C potency chords, alternating them every 2 days.

Score of first page of 42 question DASS questionnaire was: Depression = 24, Anxiety = 34 and Stress = 30. Patient did not re-present. Thus, no conclusion is possible.

Case 3

Patient with long history of recurrent depression who had been attending the practice since 5/11/07. Recruited to the study on 16 November 2007 with LM 4/12/30 and 4/12/30 C potency chords of D-Glutamic Acid alternating every day, instead of every 2 days, i.e., LM chord taken on day one, and C chord taken on day 2 and so on, with the chords alternating each other on a daily basis. The full 42 question DASS questionnaire was used for this case. 7.12.07: Patient had already been taking long-term antidepressant at presentation.

Complained of neurological symptoms. Pre-existing hypertensive. Neurological examination normal. CT brain normal. A new antihypertensive medication was started in view of poorly controlled hypertension. Increased anxiety score on this date consistent with presentation. Other scores lower. Patient stopped his antidepressant medication at his/her own insistence. Patient is reluctant to take antidepressant medication.

17.1.08: Patient presented to locum. Patient reported feeling better on treatment. 31.1.08: Patient stopped glutamic acid treatment. Patient is reluctant to take any antidepressant treatment including D-Glutamic acid. Scores noted to gradually deteriorate over the next 3 months. It is noted that at the time of cessation of glutamic acid treatment, all readings were less than baseline. Therefore, the results are consistent with beneficial effect of alternating LM and C glutamic acid potency chords.

NOTE; For the following cases, DASS 21 scores were used:

Case 4 Case 4 has many years history of panic attacks and takes regular medication for this.

Given full 42-question DASS questionnaire on 7/11/08, recruited to glutamic acid treatment on 16/11/07, and commenced D-Glutamic acid alternating LM 4/12/30 potency chord with 4/12/30 C potency chord thereof on 16/11/07. Potency chords were alternated daily. Results were collected opportunistically for this patient.

Results are consistent with beneficial effect of alternating LM and C glutamic acid potency chords.

Case 5.

A narcotic and anxiolytic using patient with many years history of fluctuating mood and anxiety was recruited on 16/11/07 at a time when he/she was relatively well, to see if alternating D-Glutamic potency chords could help maintain his/her low scores. Measurements given by the patient were far between. Also, patient did not have enough potency to last 6 months. Alternating drops daily, the supply would last at most 4 months.

Results are not consistent with an effect of D-Glutamic Acid potency chords, but in the context are difficult to interpret. Also, compliance is an issue. If the patient was not improving, then the usual practice would have been to modify the treatment after about 1 month from recruitment and commencement of potencies, e.g., change to exclusive use of the LM potency chord.

Case 6 Case 6 was recruited on 16/11/07. DASS was not done on that day. A questionnaire had been completed on 8/11/07 and this was used as baseline. Long term chronic back pain, anxiety and depression. Commenced on daily alternating LM 4/12/30 and 4/12/30 C D- Glutamic Acid potency chords.

5.12.07 Administration of alternating potency chords increased from every 2 days to daily alternation. Date on question sheet was 6/12/08, but patient presented it on 5/12/08.

2.1.08 Patient's opinion is that LM potency chord helps the most. Changed to daily LM 4/12/30 potency chord. 4/12/30 C potency chord stopped.

12.2.08 Patient deteriorated after changing to daily LM potency from alternating LM/C potencies, however, anxiety and stress scores remained lower than at baseline.

Not a big change overall, but results are consistent with beneficial effect of the LM and C glutamic acid potency chords.

Case 7

Patient was recruited on 12/12/07. Many years anxiety and panic attacks. Severe 'white coat hypertension'. Commenced on D-Glutamic acid LM 4/12/30 potency chord daily. Patient had relatively low scores to start with, however, it was determined to see if the stress

score could be decreased further. Results of DASS 21 questionnaire are in Table 7. Relevant comments from the notes are included below the graph, but are not necessarily made on the same dates as the questionnaires.

14.3.08 All scores had decreased. LM potency chord was stopped. Patient changed to 4/12/30 C potency chord daily. DASS sheet not available for this day.

Patient was changed to C chord to see if lower scores would be maintained. 1.4.08 Scores virtually identical since changing to 4/12/30 C potency. Improvement in all scores since baseline. Patient changed to daily LM 4/12/30 potency.

4/12/30 C potency stopped. 18.4.08 From this time patient's scores gradually deteriorated. It transpired that contrary to instructions the patient was taking the potency at night and was having difficulty with sleep. Results from 18.4.08 are consistent with homeopathic aggravation. Patient needed to stop potency. It would be suggested to restart with a potency chord composed of lower D-Glutamic acid potencies, e.g, LM 2/3/6, after a period of abstinence from D-glutamic acid potency, or perhaps to restart potency at less frequent rate of administration.

Potency should then be taken during the day and not at bedtime. 16.6.08 LM 4/12/30 administration increased to twice daily, morning and mid afternoon. Contrary to instructions patient continued taking second dose at bedtime. Improvement after increased rate of administration of LM potency was noted.

For the first 4 months to 18.4.08 results were consistent with action of D-glutamic acid potencies. For the last 3 months results were consistent with homeopathic aggravation and/or incorrect time of dosing contrary to instructions. Scores improved after increased potency administration to twice daily.

Case 8

Patient with years of chronic pain and narcotic use was recruited on 2/1/08 and commenced on LM 4/12/30 and 4/12/30 C D-Glutamic Acid potencies alternating every 2 days. Results are shown in Table 8.

18.2.08: Patient changed to D-Glutamic acid 4/12/30 C potency daily, in view of deterioration in scores on 2.2.08 compared with previously.

27.3.08 Improved scores but some deterioration since last time. Patient.changed to

LM 4/12/30 potency. C potency chord stopped. Deterioration since last visit. Today this patient was given LM 4/12/30 D-

Glutamic acid potency in globule (round pill) form. To do this 5 drops (20 drops = ImI) of the LM 4/12/30 D-Glutamic acid potency was added to 30 gm sucrose placebo globules available from Brauer Biotherapies, Tanunda, South Australia, product code D3770 in a 50ml Brown tinted bottle with plastic screw top. The contents were immediately given 20 forceful downward succussions at approximately 45 succussions per 60 seconds or 1- 2 Hz.

2.5.08 Deterioration compared with last visit but still improved since baseline.

Has been placing pills under the tongue contrary to instructions. Suggest put in water first and mix before placing contents under the tongue.

16.5.08 Overall improvement. LM pill dose increased to twice daily, morning and mid afternoon.

27.5.08 Marked improvement over the course of treatment. Average DASS score decreased from 25 to 5.3. The trend of the 3 graphs has been downward throughout the course of treatment. Results are consistent with action of D- Glutamic acid.

Case 9

Patient with long history of depression over years was recruited on 5/2/08 and started D-

Glutamic acid LM 4/12/30 and 4/12/30 C potency chords alternating daily doses. Patient had been using anti-depressant medication since at least 2003. 25.3.08: There had been no change over 6 weeks of treatment with alternating potency, therefore, C potency was stopped and the patient continued with daily D-Glutamic acid LM 4/12/30 potency.

3.4.08: Patient was changed from the drops to globules to see if there were any effects on scores subsequently. Globules were prepared as described above in Case 8.

Results are consistent with effect of D-Glutamic acid potency, in particular the LM potency.

Case 10

Long history anxiety. Commenced on daily alternating 4/12/30 C D-Glutamic Acid and LM 4/12/30 D-Glutamic acid potency chords. Changed to daily LM potency chord on 18/4/08. Consistent with action of D-Glutamic acid potencies.

Case 11

Patient with long history of chronic pain was recruited on 11.2.08 and commenced on daily alternating D-Glutamic acid potencies - alternating 4/12/30 C and LM 4/12/30 potency chords on a daily basis. 10.3.08: Scores had not improved. Alternating doses were ceased and patient was changed daily LM potency chord. 10.4.08: Scores improved.

9.5.08: Ran out of glutamic acid LM drops 4 days previously. Changed to LM potency chord globules produced as described above - 7 globules daily mixed for 10-20 seconds with 1A-I teaspoon water in a glass and placed under the tongue for 10-

20 seconds before swallowing the liquid - remainder of pills were left under the tongue to dissolve. 22.5.08: Patient perception is that globules are better than drops, and indeed scores did drop further compared with the previous visit when globules were started. 29.5.08: Slight deterioration in scores. LM globules increased to twice daily.

9.6.08: Increase in scores noted on this day, but had decreased again by one month later. Consistent with action of D-Glutamic acid potency.

Case 12

Patient admitted to study on 19 March 2008 and was given D-Glutamic Acid 4/12/30 C potency chord. DASS21 : D = IO, A= 6, S= 26. Patient did not complete subsequent DASS questionnaires. However, at presentation on 30/6/08 the patient was noted to have a low KlO (Kessler Psychological Distress Scale) score of 12. No conclusion can be drawn.

Case 13

Patient admitted on 19 March 2008 who usually attended another practitioner in the practice. DASS 21 scores: D=14, A=12, S=34. Patient did not represent. No conclusion can be drawn.

Case 14

Admitted 3 April 2008. Years of psychological distress. Past diagnoses include Autism and Schizophrenia. Started on LM 4/12/30 D-Glutamic Acid potency chord. Compliance was an issue and a 3rd party undertook to administer the D-GA.

Results consistent with effect of D-Glutamic Acid. Data only available for 2 weeks after recruitment.

Case 15 Years of depression and anxiety recruited on 8/4/08. Started on LM D-Glutamic acid (D- GA) 4/12/30 potency chord, 10 globules daily.

21/5/08: Patient presented for the first time since recruitment and presented all DASS

21 questionnaires simultaneously, the preferred procedure being to complete and forward a questionnaire to the investigator every 2 weeks, thereby allowing dose and potency adjustment. No discernible effect D-GA at this stage. D-GA potency was increased to twice daily - morning and mid afternoon preferred.

5/6/08: Substantial improvement in DASS 21 scores.

3/7/08: Further improvement in DASS 21 scores. 17/7/08: Some deterioration in DASS 21 score, but still substantially improved compared with baseline. Consistent with effect of D-GA potency.

Case 16

Several months depression and anxiety. Recruited 9/4/08. Started on D-GA 4/12/30 C potency 7-10 drops daily.

No improvement in this patient. This patient did not respond to concurrent treatment with a substantial dose of a common anti-depressant either, and was subsequently admitted for psychiatric evaluation and management. Not consistent with efficacy of D-GA.

Case 17

Recruited on 22/5/08 and started on D-GA LM4/12/30 potency chord drops, 7-10 drops daily.

30/5/08: Substantial improvement in scores consistent with efficacy of D-GA.

13/6/08: Slight deterioration in anxiety and stress score since last record and substantial deterioration in depression score. Patient reported that family member had experienced a misfortune since the last DASS questionnaire result. Anxiety and stress score remain less than half of baseline.

Case 18 Recruited on 30/4/08.

1/5/08: Patient started D-GA LM pills 7 daily. Consistent with effect of D-GA potency

Case 19 Recruited 7/5/08. History of depression and fibromyalgia. Consistent with effect of D-GA potency

Case 20

Recruited 6/5/08. Did not take D-GA potency daily and hand in all completed DASS questionnaires at one time on 11/6/08, without any consideration of the 2 weekly dates written on the sheets. Order of completion of questionnaires not known. Hence, it was impossible to know when or in what order the DASS 21 questionnaires were completed. Poor compliance. Unable to interpret data.

Case 21

Many years of anxiety and depression. Long term major tranquilizer and antidepressant medication use.

13/06/08: Recruited to study. Consistent with effect of D-GA potency

Instructions for drops and pills:

Drops: Place one-teaspoon water in a clean glass. Add about 7 drops of the homeopathy bottle, which has a dripulator (20 drops= ImI). Put the liquid under your tongue and leave it there for about 10 seconds before swallowing. Take the drops every day. Take the drops at a time of day when you feel relatively well. For example, if you tend to feel worse in the morning, then take the drops later in the day. Taking drops at night may interfere with sleep.

Store on a shelf away from direct sunlight. Do not store near electrical appliances, computers, TV etc. Do not store in the pantry, medicine cabinet or fridge. Do not store near strong smelling things like perfume, vicks, camphor, tiger balm etc.

Pills (globules):

" Place one-teaspoon water in a clean glass. Add about 7 pills to the water. Swirl the contents in the glass for about 20 seconds backwards and forwards. The pills will not completely dissolve. Put the liquid and any undissolved pills under you tongue and leave it there for about 20 seconds before swallowing. Allow the pills to dissolve under the tongue.

Take this dose of pills every day. The best time to take them is at a time of day when you feel relatively well. For example, if you tend to feel worse in the morning, then take the drops later in the afternoon, or if you feel worse in the evening, then take them in the morning. Taking drops at night may interfere with sleep.

Store on a shelf away from direct sunlight. Do not store near electrical appliances, computers, TV etc. Do not store in the pantry, medicine cabinet or fridge. Do not store near strong smelling things like perfume, Vicks™, camphor, Tiger Balm™ etc.

Toxicity testing of L- and D-glutamic acid with different test organisms

Vibrio fischeri bioluminescence inhibition test

Testing L- and D-glutamic acid

Different masses of glutamic acid (GA), ranging from 0.4 mg to 40 mg, were measured into test-cuvettes. 200 μl bacterial cell-suspension was poured into each cuvette. Luminescence was measured after 30 minutes contact time. GA demonstrated inhibition of Vibrio fischeri.

The calculated inhibition rate values, expressed as H% (H% = (Icontrol - Isample) / Icontrol) are shown in the Table below. (Icontrol = Inhibition by Control; Isample= inhibition by GA sample.)

Table: Light intensity inhibition of L-glutamic acid

*- pH vas measured with paper-stripes, immediately after testing

A sudden inhibition-increase was measured on the effect of L-GA between 5 and 10 mg/mL. From the dose-response curve: (with the software: ORIGIN 6.0):

L-GA EC20 — 4.8 mg/mL EC so = 6.7 mg/mL

This means that 4.8 mg/mL L-glutamic acid resulted in 20%, and 6.7 mg/mL L-glutamic acid in 50% reduction in the light emission of Vibrio fisheri test-bacterium.

The same was measured for D-glutamic acid, the Table below shows the results.

Table: Li ht intensit inhibition of D- lutamic acid

From the dose-response curve: (with the software: ORIGIN 6.0): D-GA EC20 = 5,0 mg/mL EC50 = 6,3 mg/mL

Vibrio fisher i is a suitable testorganism for testing potency.

The results indicate that D-GA is a little more toxic in Vibrio fischeri than L-GA.

(In comparison, the toxicity of D-GA expressed in a copper equivalent. It is a fictive scale of toxicity: if the same inhibition were caused by copper.

EC20 cu = 11 -2 mg/L EC50 Cu = 17.6 mg/L)

Bacterial growth inhibition Two bacteria, Escherichia coli and Pseudomonas fluorescens were used as test organisms to measure the growth inhibitory effect of L- and D-glutamic acids.

We used a 2-fold dilution from the optimal nutrient medium, because our experience is, that bacteria are more sensitive, when the life conditions are not optimal, e.g. nutrients are lacking.

The cell growth was followed by measuring optical density, which is proportional to the cell- concentration. Optical density does not differentiate between living and dead cells, and reflects cell number. The values in the Tables below were measured after 42 hours cultivating with and without L-glutamic acid.

Table: Growth inhibition of L-GA on bacteria

5 mg/mL L-glutamic acid results in close to 90% reduction in the optical density of the culture in the nutrient medium. It means that the inhibition of cell growth is close to 100%. L-GA is a little-bit more toxic for E.coli, than D-GA. EC so of L-GA is: 2 mg/mL. E. coli is a suitable microorganism for testing potency.

Pseudomonas is even more sensitive than E. coli, and both GA-s are equally toxic.

EC so of L-GA is 0.5 mg/mL. The concentration range between 0 and 2 mg/mL should be used for the potency-testing. These findings also indicate that glutamic acid may be suitable, as a therapeutic agent, for topical administration in clinical cases of Pseudomonas infection, particularly with antibiotic-resistant strains, and perhaps also useful for similar treatment of E.coli. This species of bacteria is recommended for toxicity testing and testing of potencies.

Yeast growth inhibition

Yeast growth inhibition of L- and D-GA

This yeast species is less sensitive, than the bacteria, and its inhibition curve has two stairs, what means that probably two different concentration-dependent mechanisms play role in the effect (see Figure 4.).

Pink and blue H% are based on the optical density, yellow is based on cell count.

Note that the pink, blue and yellow colours are not visible on the PCT specification which is in black and white, however, the context of the lines in the graph makes it obvious which lines are being referred to.

Tetrahymena Growth Inhibition

The growth curve was taken up and the specific growth rate was calculated and used for the calculation of the H%. The results are summarised in the Table below.

Table; Growth of Tetrah mena on the effect of increasin L- lutamic acid concentration

The effect testing was carried out in a nutrient medium, which is a 2-fold dilution of the normal one (Control 1A), to see better the acute effect of the GA on the cells propagation. 2 mg/mL GA concentration caused 100% growth inhibition and depletion in the cell number after two days, 5 mg/mL caused depletion already after 20 hours. The concentration range between 0 and 2 mg/mL should be used for potency testing.

The results show that the single cell animal test-organism is highly sensitive to the L-GA, the concentration of 0,01 g/litre (=mg/mL) caused 100% inhibition in the growth.

Tetrahymena could be used as test-organism for testing potency.

The test in this form is too complicate and durable, with a lot of labour requirement, so I would simplify the test for measuring not the whole growth curve, but only one point of time, e.g. after 68 hours growth.

Acute and Chronic Toxicity testing of L- and D-glutamic acid with Folsomia Candida animal test organism

The goal of the study was to determine the effect of L- and D-glutamic acid on the survival and reproduction of Folsomia Candida (Collembola) in feeding experiments.

The Collembolans, commonly known as springtails, are the most numerous and widely occurring insects in terrestrial ecosystems.

Microarthropods as e.g. springtails are said to have an important function regarding the maintenance of soil functions. Due to their short life cycles, high number of species and their

high density, important requirements for using them as indicator organisms is fulfilled. Folsomia Candida is a little 3—4 mm long white animal.

Test organism: Folsomia Candida (Collembola: Isotomidae) obtained from synchronized culture

Test endpoint: acute test: survival (mortality) chronic test: number of newly born animals (inhibition of reproduction)

Duration of the test: acute test: 2 weeks chronic test: 4 weeks

Guideline: ISO/FDIS 11267, 1998

The acute and chronic tests were performed simultaneously. The first assay (survival test) was terminated after 2 weeks and the second assay (reproduction test) will be terminated after 4 weeks.

1. Folsomia Candida mortality test

Effects of L- and D-glutamic acid on Collembola (Folsomia Candida) survival

Tightly closed glass containers (200 ml of volume), containing charcoal-gypsum at the bottom were prepared for the experiments.

Before starting the experiment this charcoal-gypsum underlay was wetted with 1 ml of water.

Twenty (20-20) 12 days old juvenile springtails (obtained from synchronized culture), were introduced in each test containers.

As a food source for the Collembola, granulated yeast was added.

Tested glutamic acid was thoroughly mixed in the yeast. Different doses of L- and D- glutamic acid, mixed into the yeast granules were tested as shown in the Table below.

Table: Doses of L- and D-glutamic acid mixed with yeasts

The pots were incubated for 14 days at a temperature of 25 0C in the dark. The test containers were opened once a week for aeration. After 2 weeks surviving juveniles F. Candida were counted.

Results and evaluation: After two weeks all of the animals were survived; there was no toxic effect observed.

2. Inhibition of reproduction of Folsomia Candida — effect of L- and D-glutamic acid

The experimental conditions, excepting the duration, were the same as those used for the acute test. After 4 weeks the reproduction was characterised by the number of animals. The number of the second generation animals was the same in all of the pots, no inhibition was observed.

Treatment of the toxic effects of L-glutamic acid with D-glutamic acid potencies Preventive treatment and curing

Testing bacterial growth inhibition Concepts of testings

Two bacteria, Escherichia coli and Pseudomonas fluorescens were used as testorganisms to measure the growth inhibitory effect of L-glutamic acids in the presence of placebo and D- glutamic acid potencies.

We tried to find the best way to test the efficacy of the potency, so we applied preventive and curative treatments. By 'preventative treatment' we mean that D-GA potency (D-Glutamic Acid potency) was administered prior to addition of L-GA (L-Glutamic Acid) to cell cultures. By 'curative treatment', we mean that D-GA potency was administered after addition of L-GA to cell cultures.

Step 1 & 2: 1. In the case of preventive treatment (= 'pre-treated') the mixture of 4th +12* +3Q^ potencies of D-glutamic acid was added to the bacterial cell culture 24 hours before administering the L-GA toxin. The comparison happened between non-pre-treated and pre-treated bacterial cultures. The growth of both were followed with L-GLU and with placebo. 2. Curing is different from prevention, because in this case the potency was applied directly after the addition of L-GLU. Potencies of 5*, 13th, 31st and the mixture of 4th + 12th + 30th potencies (H2) of D-glutamic acid were added to the bacterial cell culture 10 minutes after the application of L-GA to the bacterial cell culture. It is reiterated that 'curing' is also referred to in the bioassay section of this specification as 'treatment' and means that potencies were only administered after L-GA was added to cultures (10 minutes after) and not before. The bioassay section starts with the section called 'Toxicity testing of L- and D-glutamic acid with different test organisms.' The term 'pre-treatment' means that H2 was administered 24 hours before addition of L-GA to cultures. 'Pre-treatment' is also referred to as 'prevention'.

Materials and methods General conditions

The potencies were prepared in glass tubes, taking care not to contact or being close to metal objects or electric wires. (This is possibly being over cautious in the case of electric wires, since it must be admitted that it has not been the experience of homeopaths in the clinic, that potencies lose efficacy, when they are stored in quite strong electric fields near power sockets.)

The propagation of the microbes, inoculation and other microbiological manipulation was executed in an alternative way compared to usual, excluding contact with metal tools, surfaces and equipments. Potencies in ethanol were stored in wooden cupboard (far from electric cables) in dark.


Pseudomonas fluorescens (historical strain from the department's culture collection) Escherichia coli (ATCC 25922) Materials L-Glutamic acid, 99 % (CAS : 56-86-0)

Cat.: 12,843-0 (ALDRICH®, Sigma- Aldrich Chemie GmbH, Germany) Lot.: S45561-108, molecular weight 147.13 D-Glutamic acid, minimum 99 % TLC (CAS:6893-26-l)

085Kl 083 SIGMA®, Sigma- Aldrich Chemie GmbH, Germany Ethanol 96 % puriss. (Spektrum 3D® 64-17-5), molecular weight 147.13

Nutrient medium I

Nutrient liquid media used for cultivation of Pseudomonas fluorescens and Eschericia coli L

2.5 g peptone from meat (trypsin digest) (REANAL® 25773-1-99-33, CAS:73049-73-7) 2.5 g D-glucose (Spektrum 3D® CAS:5996-10-1)

1.5 g meat extract (REANAL® 02117-37-33)

0.25 g NaCl (REANAL® CAS:7647-14-5)

1000 cm3 distilled water pH=7

Sterilised by autoclaving at 121 0C, for 10 minutes.

Nutrient medium II

Nutrient agar media used for counting (spread plate counting technique) of bacteria II.

5 g peptone from meat (trypsin digest) (REANAL® 25773-1-99-33, CAS:73049-73-7)

5 g D-glucose (Spektrum 3D® CAS:5996-10-l)

3 g meat extract (REANAL® 02117-37-33)

0.5 g NaCl (REANAL® CAS:7647-14-5) 17 g agar-agar OXOID® m CAS:9002-l 8-0 1000 cm3 distilled water pH=7

Sterilised by autoclaving at 121 0C, for 10 minutes.

Sterile glass tubes were used for the propagation of the bacterial cells.

For characterising optical density of the bacterial culture Kartell Art.01937-00 Cuvette (2 clear side) were applied. Absorbance was measured by SANYO SP55 UV/Vis



Preparation of potencies

Thirty round bottom 10 ml of volume calibrated glass test tubes with glass stoppers (CLS842450 Aldrich Pyrex® round-bottom centrifuge tube) were used for the D-GA potency preparation and were numbered from 1 to 30.

Potencies were all prepared and stored in 35% ethanol, except the final potencies, which were administered to organisms, because they were prepared in distilled water.

Into the first tube 10 mg D-glutamic acid was placed and then 1 ml 35% ethanol. The test tube was sealed and given 20 forceful downward successions at 0.75-1 Hz. The same ethanol stock bottle was used for all of the test tubes described below. The next 29 tubes, i.e. tube numbers 2- 30, each contained 5 ml 35% ethanol in water.

One drop (20 μl) was removed from the 1 st test tube using automatic pipette, and was added to the second test tube. This was given 20 succussions as shown by the video. 20 μl from the second test tube was removed and added to the third test tube and so on until the 30th potency. All potencies were succussed by hand.

In the experiments the 5th , 13th and 31 st potencies separately and the mixture of 4th , 12th and 30th potencies of D-glutamic acid were tested and indistinguishable placebos.

To make the separate 5th, 13th and 31st potencies, 20 μl (one drop) of the 4*, 12th and 30th potencies were added to 5 ml distilled water in a 100 ml glass flask with plastic screw stopper (SIMAX Reagent bottles with screw GL 45 ace. to DIN 100 ml Code: 414 321 100).

These were given 20 succussions by agitating the freshly diluted solutions by rapping the glass reagent bottles hard against a hard and elastic object such as a leather-bound book) to produce the potencies in distilled water.

To prepare the mixture of 4th, 12th and 30th potencies the total contents of test tubes 4, 12 and 30 (all prepared in 35% ethanol) were added to a 100 ml glass flask with plastic screw stopper. The content was given 20 forceful downward successions at 0.75-1 Hz. 20 μl of the content was removed using an automatic pipette and added to a 100 ml glass flask with plastic screw stopper containing 5 ml distilled water. This test flask was then succussed (by agitating the freshly diluted solution by rapping its container hard against a hard and elastic object such as a leather- bound book) 20 times.

The placebo was prepared on the same way but without D-glutamic acid.

These final homeopathic D-GA potencies in distilled water were prepared immediately before application (in preventive treatment and also in curing tests). These were applied within 5-10 minutes after preparation.

Preparation of bacterial inoculum

The suspensions of test-organisms (bacteria) were prepared in two subsequent steps.

1) Two different shaken flasks containing inocula of bacteria were prepared: each of these contained 25 ml of nutrient liquid medium I, noted above, inoculated with bacteria. One of these was complemented with 2.5 ml of sterilised water (non-treated) the second one was complemented with 2.5 ml of freshly prepared mixture of 4th, 12th and 30th potencies (H2 pre-treated bacteria, preventi ve treatment). After the treatment we cultured bacterial cells for 24 hours at 28 0C.

2) 5 ml of step 1) cell suspensions was added to 350 ml fresh nutrient liquid medium L, so we have got a. Flask one with 355 ml non pre-treate d bacterial inoculum, b. Flask two with 355 ml one-day long H2 potentised bacterial inoculum.

The inocula were used immediately in the experiments: treatment by L-GLU and the potency.

Treatment of the bacterial cells with L-Glutamic acid

1. 1. 0, 2.5, 5.0, 10.0 and 25.0 mg L-glutamic acid (L-GLU) were measured and placed into empty glass test tubes with plastic cups (sterilised by autoclaving at 121 0C, for

10 minutes).

2. 5 ml from the 355 ml non-treated and parallel pre-treated bacterial inocula were added to the test tubes containing L-GLU. These two steps resulted in bacterial cell culture series containing increasing L-GLU concentration: 9 pre-treated with potency and 9 non-pre-treated ones.

Application of placebo and D-glutamic acid potencies on test-bacteria

3. 500 μl of freshly prepared potencies were added to a pre-treated and a non-pre- treated test tube series (resulted in step 1 and 2) within 20 minutes: the effect of 5th, 13th and 31st potencies (separately) and the mixture of the 5th, 13th and 31st potencies were tested. This means 2x4 series.

4. A second 2x4 series were treated with freshly prepared placebo: 500 μl placebo was added into each tube.

5. An additional L-GLU concentration series was treated by 500 μl of sterile water: one pre-treated and one non-pre-treated. Growing the cells

6. The treatments were continued with a 24 hour bacterial growth period at 28 0C, in dark (in non-metal conditions), by moderate aeration. Instead of the whole growth curve, we only measured the cell concentration after 24 hours growing of the bacteria. Measuring cell concentration in the test tubes

7. The cell growth in all of the test tubes was measured by the optical density at the 600 nm maximal absorption wavelength, which is proportional to the cell-concentration within a certain range. The optical density unit is not able to distinguish between

dead and living cells. This means that dead cells or suspended material in the culture may cause light absorption. The results are given in ODU (Optical Density Unit). 8. Some members of the L-GLU concentration series were evaluated based on live cell counts. In the case of samples containing 25 mg L-GLU {Pseudomonas fluorescens) and 10 mg L-GLU {Escherichia colϊ) cell counting by spread plate counting technique was also carried out. This technique is based on the cultivation of the cells and gives only the number/concentration of the living cells. The results were given as Colony Forming Unit (CFU/ml).

Evaluating measured data

Primary (measured) data were placed into an excel table and the comparative evaluation was executed with them. We always compared placebo and potency, pre-treated and non-pre-treated, etc. Instead of parallels with the same L-GLU-concentration we applied the L-GLU concentration series, which makes the fitting of a dose-response curve possible. We can suppose that this curve is always an "S"-shaped curve, which we can read (i.e., attribute) EC50 or similar values too, so we can interpret toxicity data better.


The optical density values (Tables \—i, below) were measured (600 nm) after 24 hours culturing of potentised and non-treated Pseudomonas fluorescens with and without L-GA, in the presence of water, placebo and D-glutamic acid potencies.

The optical density values (Tables 5-8, below) were measured (600 nm) after 24 hours culturing of potentised and non-treated Escherichia coli with and without L-GA, in the presence of water, placebo and D-glutamic acid potencies.

The experimental design is further clarified in the tables below. These show that 0, 2.5, 5, 10 and 25 mg L-GA were added to tubes in the non-treated (i.e., non-pretreated) arms containing bacterial culture (The L-GLU concentrations in the tubes are: 0, 0.45, 0.9, 1.8 and 4.5 mg/ml.) Then either placebo or potency were added 10 minutes later. The effect on bacterial growth of the intervention vs. placebo was then determined by measuring optical density 24 hours later. In another series shown in each table, bacterial cultures were administered a complex of D-GA 24 hours prior to administration of L-GA (L-GLU or L-Glutamic acid) at the doses shown in this

paragraph already. These cultures were then administered either placebo or D-GA (D-GLU or D-Glutamic acid) potency. The effect on bacterial growth was then determined by optical density measurement 24 hours later.

Note that in the tables which follow, the number in the placebo columns mean that these were the placebos made for the potencies indicated by the number, and it does not mean that a potency of glutamic acid was administered. The same interpretation applies to all tables 1-8 inclusive. Hence, using table 1 as an example, the first 5 samples, starting from the top of the table and going down, received just placebo 10 minutes after L-GA was added to the cultures. The next 5 samples received 5th potency 10 minutes after L-GA was added to the cultures. The next 5 samples, received the 4/12/30 complex of D-GA pre-treatment 24 hours before addition of L-GA to the cultures, and placebo 10 minutes after addition of L-GA. The final 5 samples in table 1 received the 4/12/30 complex of D-GA 24 hours before addition of L-GA and the 5th potency of D-GA 10 minutes after addition of L-GA to the cultures.

Table 1. The effect of the 5 th potency on the growth of H2 pre-treated and non-pre treated Pseudomonas fluorescens: optical density (ODU) and living cell concentration (CFU)

The optical density is proportional to the cell concentration within a certain range. The difference between optical density and living cell number originates mainly from the dead cells: which are already naturally dead (at the end of the growth curve) or killed before time by the toxic agent. These two possibilities cannot be differentiated from each other.

The curing effect of the 5 th potency is significant, compared with placebo. Pre-treatment with the mixture of 4th, 12th and 30th potencies (H2) shows little effect.

The living cell concentrations shown in the table give a better picture: cell number is 1x105 cell/ml in the placebo grown culture, 18x105 cell/ml in the H2-pre-treated. 5 th potency resulted in 500-fold cell growth compared to the culture with placebo.

The H2 pre-treated culture's cell number is 13 times higher in the 5 th potency treated arm compared with the placebo treated arm, i.e., 18xlO5 cell/ml cf. 253xlO5 cell/ml.

NOTE that in all the graphs below, the 1-glu concentration is in units of 0, 2.5, 5, 10 and 25mg per tube and not, as it appears sometimes 0, 2,500, 5,000, 10,000 and 25,000 mg per tube.

The growth of Pseudomonas fluorescens with placebo is highly inhibited by L-GLU at a concentration higher than 5 mg/tube. The same L-GLU concentration does not effect the culture treated by 5th potency.

H2 pre-treatment had minimal effect on the growth of Ps. fluorescens compared with the non-pre-treated arm shown in graph 1. The L-GLU toxicity shows inhibition at a higher concentration. The 5th potency treatment of the pre-treated culture looks less effective compared with the non-pre-treated culture (previous graph).

Table 2. The effect of the 13th potency on the growth of H2 pre-treated and non-pre treated Pseudomonas fluorescens: optical density (ODU) and living cell concentration(CFU).

The curing effect of the 13th potency is also significant, compared with placebo. Pre-treatment with H2 shows practically no effect.

The living cell concentrations show a similar tendency, than in the case of the 5th potency: cell number is 16x105 cell/ml in the placebo grown culture and 105x105 cell/ml in the H2-pre- treated. It is an 8-fold higher value.

13th potency resulted in 50-fold cell growth compared to the culture with placebo.

The H2 pre-treated culture's cell number on the effect of the 13th potency (166x105 cell/ml) is similar to the placebo-treated culture (105x105 cell/ml) and is significantly higher, than the same for the placebo treated culture.

Placebo treated Ps. fluorescens culture shows a curve similar to the culture treated with the 5th potency. The L-GLU concentrations higher than 5 g/tube inhibit the growth considerably. 13th potency is able to compensate this effect of the L-glutamic acid. The sudden drop in the growth in the tube of 10 mg L-GLU/tube is strange, and looks like a mistake on first impression.

Pre-treatment of the culture with H2 resulted in lower sensitivity of the bacterial cell culture to the toxicity of L-GLU. Pre-treatment is able to defend the cells from the toxic effect of L-GLU even in the highest concentration. (This was not the case in the previous, practically parallel experiment.) The 13th potency line in Graph 4 represents a sample which received H2 potency 24 hours before addition of L-GA to the sample, and then received 13th potency 10 minutes after addition of L-GA. The placebo line indicates a sample which also received H2 potency 24 hours before addition of L-GA, but 10 minutes after addition of L-GA, placebo was added to the culture and not potency. Thus, both the arms had pre-treatment with H2 24 hours before addition of L-GA as toxin. It is reiterated that all potencies used in the examples in this specification were made from D-Glutamic acid, according to the method described in the specification.

It is possible that more frequent administration of 13th potency may confer greater protection against L-GLU toxicity. It is possible that more frequent administration of 12th potency may confer greater protection against L-GLU toxicity. We have administered minimal doses of potency in these experiments. The usual practice in homeopathy in a situation of acute toxicity as in these experiments would be to preferably administer potency much more frequently. This could be daily, twice daily, three times daily or more often, hi some instances of acute pathology in, say, humans, potency may be administered every few minutes. This is illustrated in previous patent applications by the inventor R.M. Kuzeff.

Table 3. The effect of the 31st potency on the growth of H2 pre-treated and non-pre treated Pseudomonas fluorescens: optical density (ODU) and living cell concentration (CFU)

Potency pre-treated 2,5 31 0.292

Potency pre-treated 31 0.245

Potency pre-treated 10 31 0.243

Potency pre-treated 25 31 ι?J 0.066 ■ 1

Potency pre-treated 31 0.324

Potency pre-treated 2,5 31 0.314

Potency pre-treated 31 0.310

Potency pre-treated 10 31 0.327

Potency pre-treated 25 31 0.323 3190

The toxicity inhibiting effect of the 31st potency compared to the placebo-treated culture is weaker than that of the 5 or 13 . When looking at the H2 pre-treated culture a significant effect of 31st potency can be seen.

The living cell concentrations show similar tendency, cell number with only placebo is 153 xl O5 cell/ml (which is an uncertain value again: probably a mistake) and only 1x105 cell/ml in the

H2-pre-treated culture. The last one is more realistic, but we have to accept, that every tube with a bacterial culture is a microcosm, which may have its individual fate and evolution.

31 st potency resulted in 3000-fold cell growth compared to the culture with placebo in the case of the H2 pre-treated culture, which is a considerable difference.

The results are shown on Graph 5 and Graph 6 below.

The 31st potency was able to effectively increase or, alternatively, not allow a decrease in the growth rate, even in the case of treatment with 25.0 mg/tube L-GLU. 31st potency was effective in both cases: with and without H2 pre-treatment.

Both placebo and the treatment with potency shows the formerly experienced effect: without potency (placebo), H2 pre-treatment lowers L-GLU toxic effect, 31st potency keeps high cell concentration, no damage of the cells or inhibition of the growth can be measured.

Table 4. The effect of the H2 potency on the growth of H2 pre-treated and non-pre treated Pseudomonas fluorescens: optical density (ODU) and living cell concentration (CFU) (H2: the mixture of 4th + 12th +30th potencies)

Pseudomonas L-GLU Placebo Potencies Absorbance Living cell fluorescens [mg/tube] (ODU) CFU/ml*105

Non-treated 0 4+12+30 0.310

Non-treated 2,5 4+12+30 0.311

Curing with H2 was not successful, but the preventive application (pre-treatment) resulted in high cell concentration, no inhibition by L-GLU.

The growth of the microbes on the effect of H2 gave irregular values in some cases, e.g. the cell growth is strongly inhibited by 10 mg L-GLU/tube both in case placebo and potency treatments. (To consider the ODU 0.035 result as a mistake, is not reasonable, because the living cell number is also low in this case.) The relatively good resistance of the pre-treated culture to L-GLU (799x105 cell/ml) cannot be increased when repeatedly treated with H2 after the administration of L-GLU toxin.

Placebo and H2 treatment do not show much difference. The drop at 10 mg of L-GLU maybe interesting, because it is not the only case, when this L-GLU concentration resulted in extremely low growth.

The pre-treatment with potency H2 gives similar results both for H2 and potency post- treatment: the preventive treatment was effective, it was able to eliminate the growth inhibitory effect of L-GIu - compare in this regard graphs 8 and 9. In the next 2 Graphs showing effect of placebo in the non-pretreated (with H2) and pre- treated groups, PI refers to data from Graph 1 , PII from Graph 2 and so on.

In Graph 9 we show all the placebo treated tubes results together, to see the trends and the deviation better. We can see that there is a proportional growth inhibition: higher concentration of L-GLU results in higher inhibition until the concentration of 10 mg/tube L- GLU. The highest concentration of L-GLU cannot inhibit cell growth in two cases. It is a known mechanism in toxicology: too many toxin-molecules cannot reach the receptors due to steric inhibition. Comparing the placebos in graph 9, with the different treatment and pre- treatment combinations in graphs 10-12 inclusive, one can see that potency administration produces a different effect on Pseudomonas flourescens depicted in these graphs.

Pre-treatment with H2 is able to stop or hinder the toxic effect of L-GLU. The differences cannot be explained easily, but knowing the statistics of microbial processes, the trend is the relevant issue; and the trend of the pre-treated and not pre-treated is significantly different.

The treatment with the different potencies resulted in lower toxicity and with the exception of the 10 mg/tube L-GLU concentration in Graphs 3 and 7, potency was able to prevent the Pseudomonas cells from the toxic effect of L-GLU. It is reiterated that 'treatment' in the bioassay section of this specification means that potency and placebo were only administered 10 minutes after administration of L-GA to cultures, and not before.

From this comparative graph we can see, that treatment administered after L-GLU was added to cultures was effective in all cases. But the most efficient was in case of H2, when the pre-treatment happened with H2.

Escherichia coli

The same experiment as in case of Pseudomonas fluorescens was executed by using E. coli. As a general conclusion, we can state, that absorbance is not a proper endpoint for following the differences in E. coli cultures. The optical density values and the cell-numbers are not proportional with each other. Living cell concentrations were measured only at 10 mg/tube L-GLU concentration. These cell concentration values in some cases prove the efficiency of potency compared to placebo.

The treatment with 5 potency resulted in a 5-fold increase in cell concentration compared to

placebo. H2 pre-treatment has weaker but still significant effect against L-GLU (2.5-fold). The pre-treatment with H2 together with post-treatment with the 5th potency was the most effective, as it is shown in Table 5.

Table 5. The effect of the 5th potency on the growth of H2 pre-treated and non-pre treated Escherichia coli: optical density (ODU) and living cell concentration (CFU)

The cell numbers in Table 6. shows some increase in the cell concentration on the effect of potency treatment compared to placebo. The H2 pre-treatment itself did not proved to be efficient against L-GLU.

Table 7. The effect of the 31st potency on the growth of H2 pre-treated and non-pre treated Escherichia coli: optical density (ODU) and living cell concentration (CFU)

Potency pre-treated 25 31 0.389

Potency pre-treated 31 0.520

Potency pre-treated 2,5 31 0.468

Potency pre-treated 31 0.458

Potency pre-treated 10 31 0.396 2720

Potency pre-treated 25 31 0.315

The treatment with 31st potency resulted in non-significant increase in cell concentration compared with the placebo treated culture. H2 pre-treatment's effect is also not significant.

Table 8. The effect of the H2 potency on the growth of H2 pre-treated and non-pre treated Escherichia colϊ optical density (ODU) and living cell concentration (CFU)

The differences between potency and placebo treated cultures are not significant, but the culture which was pre-treated and repeatedly treated with H2 shows higher cell concentration.

Tests with Tetrahymena

In these tests we investigated the effect of D-Glutamic Acid 13th potency prepared as described above on inhibition of L-Glutamic acid toxicity in Tetrahymena.

First trial with Tetrahymena'. propagation in test-tubes; curing type treatment by 1/10 volume ratio of 13th potency or placebo, i.e., 1 part potency or placebo in 10 parts culture.

In graphs 13 - 17 the notation ' 15 GIu' means that the particular arm of the study, indicated by the line to the left of the ' 15 Glu'notation, in the key on each graph, received L-GIu added to the culture as a toxin, at a concentration of 0.5 mg/ml or 15mg/30ml in each test. The absence of 15 GIu next to the description of a line in the key on each graph, indicates that L- GIu was not added to the culture. Hence, 'Graph 13- Tetrahymena' shows that the 13th potency of D-GIu added to culture resulted in substantially less inhibition in Tetrahymena growth after 60 hours of incubation.

As in all the bioassay experiments in this specification, once again 'treatment' means that D-GIu potency was added 10 minutes after addition of L-GIu (as toxin) to cultures. 'Pre- treatment' means that H2 potency was added 24 hours prior to addition of L-GIu to cultures.

2nd trial: propagation in 30 ml flasks, prevention type pre-treatment with 13th potency in 1/10 volume ratio, curative treatment with 13th potency in 1/60 volume ratio: all combinations, i.e., 1 part potency or placebo in 60 parts culture.

In 'graph 14 - Tetrahymena' the first 2 lines in the key on the graph have 15 GIu next to them. This means that these 2 arms had L-GIu added to them as a toxin. The other 3 arms in the key did not have addition of L-GIu. The arm which received 13th potency D-GIu 10- 20 minutes after addition of L-GIu, and also at 24 hours before addition of L-GIu, showed less inhibition of growth due to L-GIu compared with the identical placebo depicted with squares along the growth curve. The 4th and 5th lines on this graph show 2 types of placebos with very similar growth curves. They show the growth curve unaffected by L-GIu toxicity.

The 3rd line shows the growth curve in the absence of L-GIu toxicity, but with pre-treatment with 13th potency administered 24 hours prior to addition of L-GIu toxin.

The same without potency pre-treatment, only curative treatment is shown in Graph 15, which shows a similar trend to the 'Graph 14-Tetrahymena', but without any pre-treatment with potency or placebo at the 24 hour mark prior to addition of L-GIu to cultures at the

above mentioned dose. In graph 15, potencies and placebos were only added after addition of L GIu to the cultures.

Third trial: repetition of the 2nd: Tetrahymena was not co-operative in this case: its behaviour was not normal: much slower growth, etc. But still can be seen the effect of the potency compared to placebo.

In the context of the descriptions given above, the relevant interpretation of 'Graphs 16- and 17 tetraymena' below are obvious. They also show action of D-GIu potency inhibiting the toxic effect of L-GIu on growth of Tetrahymena, as represented by cell concentrations. In 'graph 16 — Tetrahymena' and 'graph 17 - Tetrahymena', the notation '15 GIu' appears next to all the arms depicted by lines in the key contained in each graph. Hence, L-GIu was added as toxin to all the cultures represented by these graphs.

Pre-treatment was not effective, but curative treatment yes, in spite of the abnormally slow growth. (Slow growth is not association with the potency or placebo treatment: growth was slow without L-GLU treatment as well). Note that all graph lines in 'Graph 17 - Tetrahymena' are clustered along the x-axis except for 'potency pre-treated + potency, 15 GIu'

In addition to the cell numbers, we prepared some photos about the cells, because we observed morphological differences between potency treated and placebo cultures. To demonstrate it, we show some of microscopic pictures in Figures 6, 7 and 8.

Normal cells in optimal culture medium are shown in Figure 6, without any treatment: the cells are big and healthy. Moving intensively with their hair-like filaments. The size can be seen from the lines under the cells, which are the engravings of a Buerker-chamber, and shown on most of the pictures.

Figure 7 shows L-GIu + placebo: the L-GIu toxicated cells are not only fewer in number but the still living ones are much smaller than the healthy ones. Their inner structure is also different, less structured morphology.

Figure 8 shows L-Glu+pre-treatment + treatment with potency and L-GIu + potency. The L- GLU toxicated cell with potency looks healthy, and their number is higher, than the L-GLU treated but not cured by potency sample. Only curative and preventive+curative are both very effective from the morphological point of view.

The pictures can be compared according to the size: the same magnification was used for each. The sizes of the pictures are comparable by the engraving of the chamber. If the two lines are an identical distance apart, the magnification of the picture is the same (when no engraving can be seen, it is, because the cells were not co-operative in being fixed on a certain scratch.

The observation of the morphology is also a good tool, but not quantitative like the number of cells. But sometimes it is better evidence, although the best is to consider the two together. Bacteria are too small to use this method on. hi Figure 9, the difference can be seen visually in the respective flasks, hi the photograph can be seen: left: normal growth (nothing added), in the middle: L-Glu+potency, right: L-Glu+placebo.

Reference List

1. Aboul-Enein, H. Y. Wainer I. W. The impact of stereochemistry on drug development and use. Aboul-Enein, H. Y. Basha L. I. Chirality and Drug Hazards. 1-21. 97. New York, John Wiley & Sons.

2. Cazin, J.C. et al. A study of the effect of decimal and centesimal dilutions of arsenic on the retention and mobilization of arsenic in the rat. Hum Toxicol 6, 315-20 (1987).

3. Chakrabarty, K., Bhattacharyya, S., Christopher, R., and Khanna, S. Glutamatergic dysfunction in OCD. Neuropsychopharmacology 30, 1735-40 (2005). 4. Chavaz-Noriega, L. E., Marino, M. J., Schaffhauser, H., Campbell, U. C, and Conn, P. J. Novel Potential Therapeutics for Schizophrenia: Focus on the Modulations of Metabotropic glutamate Receptor Function. Current Neuropharmacology 3, 9-34. 2005.

5. Chavez-Noriega, L.E., Schaffhauser, H., and Campbell, U.C. Metabotropic glutamate receptors: potential drug targets for the treatment of schizophrenia. Curr Drug Targets

CNS Neurol Disord 1, 261-81 (2002).

6. Crawford, J.R. and Henry, J.D. The Depression Anxiety Stress Scales (DASS): normative data and latent structure in a large non-clinical sample. Br J Clin Psychol 42, 111-31 (2003). 7. Cummings, J.L. Alzheimer's disease. N Engl J Med 351, 56-67 (2004).

8. Desu M.M. and Raghavarao D. Nonparametric Statistical Methods For. Complete and Censored Data. Chapman & Hall/CRC, Boca Raton, Florida (2004).

9. Eaton, W. W., Kalaydjian, A., Scharfstein, D.O., Mezuk, B., and Ding, Y. Prevalence and incidence of depressive disorder: the Baltimore ECA follow-up, 1981-2004. Acta Psychiatr Scand 116, 182-8 (2007).

10. Eliel, E.L. and Wilen, S.H. Stereochemistry of Organic Compounds. John Wiley & Sons, INC., New York (1994).

11. Fisher, P., House, L, Belon, P., and Turner, P. The influence of the homoeopathic remedy plumbum metallicum on the excretion kinetics of lead in rats. Hum Toxicol 6, 321-4 (1987).

12. Froehlich, T.E. et al. Prevalence, recognition, and treatment of attention- deficit/hyperactivity disorder in a national sample of US children. Arch Pediatr Adolesc Med 161, 857-64 (2007).

13. Garakani, A., Mathew, S.J., and Charney, D.S. Neurobiology of anxiety disorders and implications for treatment. Mt Sinai J Med 73, 941-9 (2006).

14. Grizzle, K.L. Developmental dyslexia. Pediatr Clin North Am 54, 507-23, vi (2007).

15. Hahneman, S. The Organon of Medicine. M.Bhattacharyya & Co., 8th Indian Edition (5th &6th Edition), Calcutta, 1984

16. Henry, J.D. and Crawford, J.R. The short-form version of the Depression Anxiety Stress Scales (DASS-21): construct validity and normative data in a large non-clinical sample. Br J Clin Psychol 44, 227-39 (2005).

17. Howe, R. and Shanks, R.G. Optical isomers of propranolol. Nature 210, 1336-8 (1966).

18. Hyman, M.A. Is the cure for brain disorders outside the brain? Altern Ther Health Med 13, 10-5 (2007). 19. Johnson, P.L. and Shekhar, A. Panic-prone state induced in rats with GABA dysfunction in the dorsomedial hypothalamus is mediated by NMDA receptors. J Neurosci 26, 7093-104 (2006).

20. Jonas, W., Lin, Y., and Tortella, F. Neuroprotection from glutamate toxicity with ultra- low dose glutamate. Neuroreport 12, 335-9 (2001). 21. Jonas, W.B. Do homeopathic nosodes protect against infection? An experimental test. Altern Ther Health Med 5, 36-40 (1999).

22. Kuzeff, R. M. Spacetime modification of homeopathic medicinal action. Australian patent application number 2007100082. 2007.

23. Kuzeff R.M. Treatment of effect of chemicals with their ultradilute stereoisomers. Sempach Pty Ltd. 2007.

24. Kuzeff, R.M., Mecheva, R.P., and Topashka-Ancheva, M.N. Inhibition of (-)- propranolol hydrochloride by its enantiomer in white mice— a placebo-controlled randomized study. Forsch Komplementarmed Klass Naturheilkd 11, 14-9 (2004).

25. Kuzeff, R.M., Topashka-Ancheva, M.N., and Mecheva, R.P. Inhibition of (-)- propranolol hydrochloride by its enantiomer in white mice. Forsch Komplementarmed

Klass Naturheilkd 10, 309-14 (2003).

26. Kuzeff, R.M., Topashka-Ancheva, M.N., and Mecheva, R.P. Inhibition of (-)-trans- (1S,2S)-U5O488 hydrochloride by its enantiomer in white mice — a placebo-controlled, randomized study. Forsch Komplementarmed Klass Naturheilkd 11, 144-9 (2004). 27. Lipton, S.A. Failures and successes of NMDA receptor antagonists: molecular basis for the use of open-channel blockers like memantine in the treatment of acute and chronic neurologic insults. NeuroRx 1, 101-10 (2004).

28. Lipton, S.A. and Nicotera, P. Calcium, free radicals and excitotoxins in neuronal apoptosis. Cell Calcium 23, 165-71 (1998). 29. Lipton, S.A. and Rosenberg, P.A. Excitatory amino acids as a final common pathway for neurologic disorders. N Engl J Med 330, 613-22 (1994).

• 30. Lovibond, S. H. and Lovibond, P. F. Manual for the Depression , Anxiety and Stress Scales. Psychology foundation monograph, School of Psychology, University of NSW. 95.

31. Markou, A. Metabotropic glutamate receptor antagonists: novel therapeutics for nicotine dependence and depression? Biol Psychiatry 61, 17-22 (2007).

32. Mathew, S.J., Keegan, K., and Smith, L. Glutamate modulators as novel interventions for mood disorders. Rev Bras Psiquiatr 27, 243-8 (2005).

33. Nicotera, P. and Lipton, S.A. Excitotoxins in neuronal apoptosis and necrosis. J Cereb Blood Flow Metab 19, 583-91 (1999). 34. Nieuwenhuijsen, K., de Boer, A.G., Verbeek, J.H., Blonk, R. W., and van Dijk, F.J. The Depression Anxiety Stress Scales (DASS): detecting anxiety disorder and depression in employees absent from work because of mental health problems. Occup Environ Med 60 Suppl 1, i77-82 (2003).

35. Olney, J. W. Glutamate-induced retinal degeneration in neonatal mice. Electron microscopy of the acutely evolving lesion. J Neuropathol Exp Neurol 28, 455-74


40. Paulose-Ram, R., Safran, M.A., Jonas, B. S., Gu, Q., and Orwig, D. Trends in psychotropic medication use among U.S. adults. Pharmacoepidemiol Drug Saf 16, 560-70 (2007). 41. Sankaran R. An Insight into Plants, Vol. 2. Homoeopathic Medical Publishers, Mumbai (2002).

42. Sankaran, S. The Substance of Homeopathy. Homoeopathic Medical Publishers, Bombay (1994).

43. Sankaran, S. The Spirit of Homeopathy. Homoeopathic Medical Publishers, Bombay (1995).

44. Sankaran, S. The Soul of the Remedies. Homoeopathic Medical Publishers, Bombay (1997).

45. Sankaran, S. The System of Homeopathy. Homeopathic Medical Publishers, Mumbai (2000). 46. Sankaran S. An Insight into Plants, Vol. 1. Homoeopathic Medical Publishers, Mumbai (2002).

49. Vianna, M.R.M., Cammarota, M.P., Coitinho, A.S., Medina, J.H., and Izquierdo, I. Pharmacological Studies of the Molecular Basis of Memory Extinction. Current Neuropharmacology 1, 89-98 (2003). 50. Wang J.Q. et al. Glutamate-regulated Behavior, Transmitter Release, Gene Expression and Addictive Plasticity in the Striatum: Roles of Metabotropic Glutamate Receptors. Current Neuropharmacology 1, 1-20 (2003).

51. Wang, J.Q. et al. Glutamate signaling to Ras-MAPK in striatal neurons: mechanisms for inducible gene expression and plasticity. MoI Neurobiol 29, 1-14 (2004).

52. Wazana, A., Bresnahan, M., and Kline, J. The autism epidemic: fact or artifact? J Am Acad Child Adolesc Psychiatry 46, 721-30 (2007).

53. Zuvekas, S.H., Vitiello, B., and Norquist, G.S. Recent trends in stimulant medication use among U.S. children. Am J Psychiatry 163, 579-85 (2006).