PROVINCE OF ONTARIO

CANADA

I, JORDANA ENIG STOCKHAMER, a Notary Public in and for the Province of Ontario, by Royal

Authority duly appointed, residing in the City of Vaughan, in the Regional Municipality of York, in

the Province of Ontario, DO HEREBY CERTIFY that the paper writing hereunto annexed, each

page of which bears an impression of my seal, is a true and correct copy of A CASE-CONTROL

STUDY OF THE EFFECTS OF PHYTOTHERAPY WITH THE ALVEO FORMULA ON

IMMTINITY IN CHILDREN signed by Josef Richter, MD, PhD, of the Department of

Immunology, Regional Public Health Institute, Usti nad Labem, Czech Republic, the same having

been compared by me with the.original document, an act whereof being requested I have granted

under my hand and notarial seal of office to serve and avail as occasion shall or may require.

DATED at Toronto, Ontario, this 26tr day of August, 2010

))))

Public in and for theProvince of Ontario

an health se1ices, treatment and health procedures worldwide are provided by the practices

of CAM (2).

The public's growing interest in botanical formulas is notable. There has been a wide

application of phytotherapy in the past decade (9, 10, 13, 15, 18, 19, 27'and 29). The general

public is becoming more concerned with the use of various herbal formulas, herbal extracts

and mixtures. Many clinical case studies based on a wide scale of herbal products have

already been published (1, 2,7,8, and l9).

This is what inspired us to conduct a study on the effects of the botanical formulation

ALVEO. We were mostly interested in the response of clinical changes and the changes in

seeretory immunity after the administration of Alveo. After studying its individual

components we suspected the possibility of positive effects. To observe its impact we selected

two groups of children: one group consisted of children being treated at our clinic for chronic

upper respiratory tract infections and the other was a control group of healthy children'

ALVEO is a botanical tonic manufactured by Akuna Health Products Inc. in Canada-

It was used in the recommended dose of 28 ml per day. The study was conducted between

September 2006 and September 2007. Following are the partial results of this study.

Study methodolory:

The case group (Group A) consiste d of 27 children ( 14 boys and I 3 girls) undergoing long-

term treatment at the Department of Microbiology and Immunology; the control group (Group

B) consisted of 35 children (18 boys and 17 girls) from an elementary school in Usti nad

Labem, Czech Republic. The children in Group A were between the ages of 6 and 13 with the

average age being 10.95 years; the children in Group B were between the ages of 9 and 14

with the average age being 10.74 years. Group A was given I ounce or 28 ml of ALVEO

daily on an empty stomach or half an hour after a meal. Every child consumed I bottle of

product in Bn average of 35 days. The children in Group B received a placebo which

resembled Alveo in appearance and contained the same amount (950 ml) of liquid but no

active ingredients. The length of application was averaged 34 days.

The clinical disposition of each child was recorded on a daily basis using a

questionnaire with an emphasis on the symptomatology of infections mainly of the upper

respiratory tract (cough, phlegm discharge, temperature, rhinitis, pharyngitis, and otitis).

Collected data was processed and duration of illness observed. Anthropometric measurements

were taken for each child and hiVher BMI was calculated'

At the same time, the saliva of each child was collected prior to and after application

using the standard method of gathering unstimulated saliva. These specimens were stored in

standard plastic test tubes at less than 70 o C until processing. The exact time of collection

was observed when the effects of circadian rhythmicity were not present (24). The levels of

albumin and IgA in the saliva were determined by immunonephelometry using the Behring

and the Dade Behring BN II antisera. The obtained results were grouped according to the

following levels: Albumin at 50rng/L of saliva being the norm; 100mg/L being a sign of

irritation; 500 mg/L a sign of infection; and amounts greater than 500 mg/L signiffing an

infection in progress. As for the IgA, normal amounts range between 20 and 150 mgll of

saliva.

The ratio of IgA to albumin was then calculated at all levelsin relation to the

individual groups observed. The preference of this index being a reduction in the amount of

lgA transferred into the saliva as a result of infection.

Data analysis:

Differences in findings for the individual groups observed were compared for each of the

groups before and after the application of Alveo and later also in interrelation to the placebo

groups and the control files. The average values of the findings were calculated as well as the

absolute values and later also the logarithmic values of the mean-root-square errors at95% Cl

by means of statistical software such as Microsoft Excel,Instat 3 and Prism 4 (USA). The

differences were then evaluated using the Smirnov-Kolmogorov test, the t-test, the A}'{OVA

test, the Tukey-Kramer test.

Ethics:

This case study was conducted in accordance to the Declaration of Helsinki (Fourth revision

1996) and the guidelines for clinical testing in the Czech Republic. Permission was obtained

prior to the start of the study from all the parents of the children tested with the formula.

Results:

Table I contains the findings before and after the application of the formula and the placebo

over the course of 4 weeks (equal to the consumption of one bottle of product). The

comparison in the amounts of IgA and albumin in the saliva of the placebo controlled group

did not show any significant changes before or after application. On the contrary, the children

who were given the ALVEO formulation and who suffered from chronic upper respiratory

tract infections showed statistically a highly significant decrease in the amount of IgA and

albumin alike.

In studying the occumences in clinical evidence, we discovered that the clinical

symptoms in children who consumed the ALVEO formula decreased by almost two thirds.

Discussion:

Even in the more advanced countries there are both historical and cultural reasons for the use

and application of alternative medicine rather than conventional treatments. It is important to

acknowledge that for half the world's population this type of treatment is actually the norm.

The use of the term "supplementary"- alternative medicine (complementary - altemative -

medicine - CAM) stems form the close relationship between the classical and alternative

approaches to treatment. This can include altemative physical procedures such as acupuncture

and acupressure as well as the application of herbal remedies as practiced by Indian, Japanese

and Chinese cultures and in procedures such as phytotherapy and so on (19,29).

An example would be in the treatment of allergies where a blend of various botanicals

was used. The formulation Pollen contains a mixture of 12 different pollens. The reported

effect of this product was a decrease in the frequency of allergy symptoms and a lower intake

of antibiotics (2). The use of various herbal homeopathic formulas was observed in a series of

clinical trials in individuals suffering from allergies, asthma, bronchitis as well as other health

conditions (7, 8, 10, 13, 15, lg,zg,and 30). Althoughthe discussion about the positive effecs

of homeopathic products on the market remains intense, it is necessary to take a truly

qualifred stance on the matter; the evidence for which can only be provided by scientific and

complex studies (1, 2, and l2). Presently there are no trial results that completely negate the

effects of phytotherapeutic products; nor are there any results that entirely support them (2).

We discovered that from the total 26 ingredients contained in the tested formula, at least 16

have an effect on the modulation of immunity response while others seem to function via

modulation of CNS (10, 30). In present day USA, herbal extracts and the formulas containing

herbal extmcts are considered to be medicines, food and even dietary supplements. The latter

are commonly refened to as DSHEA (4). Some herbal products are considered to be

adaptogens - products which have a normalizing (restorative) influence on the body and help

defend it against stressors (4, 10). We still require more research about the antioxidant

tendencies of herbal extracts, their effects on the regulation of enzymatic functions and their

precise influence on the entire immune system (1, 7, and 8). In our observations at the Health

Institute we have noticed that herbal extracts have positive effects on the health of children

who are already successfully responding to treatment. But even with some of our patients, we

realize that short-term application of these herbs (over the course of one month) is not enough

to positively influence the observed immunity parameters.

The safety of herbal formulations depends on their.orrrportdon; though the fact

rcmains that the effects of many herbal extracts are still not known. For example, there are

more than 100 different components in tea tree oil alone (29).

Preference should be given to those products, which have undergone a series of

laboratory and clinical tests, and most importantly, which have been approved by the

appropriate bodies in the respective country. Many widely available folk remedies may

actually turn out to be potentially harmful, Therefore, it is important to always notify the

public about the possible side-effects and risks involved with taking any particular formula

(29). Chamomile can for example trigger an allergic reaction in individuals who have been

sensibilized by the allergens of ragweed or chrysanthemum (29). Other herbs such as Garden

angelica and rue can highly increase one's photosensitivity.

Herbs and herbal products demonstrate significant antimicrobial activity. This activity

is mainly influenced by the phenolic and polyphenolic substances, which also act as strong

antiviral, antibacteiial and antifungalagents (7, l3); Terperroids also dernonstrde

antibacterial, antifungal, antiviral and protozoaic activity. Similarly it is with alkaloids,

lectins, polypeptides and other substances (7, 13, and l8). Some ingredients in ALVEO such

as licorice are important besause they induce interferon activity, increase the activity of

natural killer (or NK) cells and display antiviral activity including against HIV. Glyconhizin

has anti-inflanrmatory and anti-allergic properties (7, 8, 15, andlS). Some ingredients

contained in the formulation not only demonstrate significant antimicrobial activity but they

also have the ability to modulate the immune system. These traits have been confirmed in a

series of clinical trials, mainly in individuals inflicted with respiratory illnesses (2' 20).

Additional benefits of the formula are many. There is evidence of favorable impact on skin

infections, immune dysfunctions, and fatigue syndrome (29). The use of the formula as a

preventative stems from, not only its effects on the modulation of the immune system, but

also from its effects against stress and fatigue as well as many its other benefits (21,22).

In addition to observing the clinical behaviors or reactions of an individual to an

ALVEO regime, we also decided to measure the parameters of salivary immunity and IgA,

from which we further calculated the ration of Albumin to IgA. The reason for this was to

check if our results were corrdct. We evaluated the diagnoses of the children prior to the study

and after its completion. The collection of saliva was carried out id a standard manner and

always at the same time of day (23,25). There is little possibility of comparing the

immunological findings after the phytotherapy with those of other studies because of the lack

of literature available on this subject. Only isolated studies study the effects of ginseng on the

levels of IgA (9, 11). The measuring of IgA in saliva alone is fairly inaccurate because it is

way too easily affected by the presence of inflammation or damage to the mucous lining of

the oral cavity. This is why we decided to also measure the amounts of albumin in the saliva.

The presence of which signifies the presence of an inflammation, irritation or infection (3, 16,

17,20,23,25,28). We also cannot omit the effects of tonsillectomy on the amounts of both

observed components. as in our study on body fat (5, l2). Circadian and seasonal rhythmicity

can also significantly influence the findings of certain proteins in saliva (25). This is why it is

so important to strictly adhere to a collection schedule. The influence of physical strain on

salivary immunity is also worth a mention (5).

In closing, we can state that we observed the benefits of regular application of the

ALVEO formulation in children. The conclusions we reached suggest a limitation caused by

tbe short application p€riod We believe tbat a oourse of snrdy over the period of three montbs

L

t_I

would be optimal. Furthermore, new studies show that the inclusion of new indicators in

saliva would be appropriate for the purpose of nutritional immunology (l l).

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Appendix:

Albumin in saliva of children before (1) and after (2)application of Alveo and placebo

140

o 130

E tzo@

5 110

E 100E90cE80Ero

-\

i SlgA in saliva of children befor€ (1) and afier (2) iI application of Alveo and placebo

25t)o

fr zooob rsoCD

E roo

8so0

i SlgAl I SlgA2 , SlgAl SlgA2

Controlgrdp Case grouP

it

Josef Richter, MD, PhD, \Dpt of Immunology, Regional Public Helth Instute, Usti nad

Labem, Czech republic.

Josef.richter@ zuusti.cz

(W fi'"'e-L{'u'