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The immunomodulatory effect of oyster mushroom on pre-school children

The immunomodulatory effect of oyster mushroom on pre-school children Šustrová, M., Procházková Ľ, Wimmerová, S. Slovak Medical University in Bratislava

The immunomodulatory effect of beta-glucan – a natural polysaccharide – has been known for centuries. In addition to β-glucan, oyster mushroom contains other beneficial substances as well. These include alpha-glucans, glycoproteins, chitin, etc. Furthermore, oyster mushroom also contains fibre, which has a significant prebiotic effect and boosts the activity of the present probiotic bacteria. Most published studies describe the impact of the isolated β-glucan on the immune system. In our work, we address the effect of the entire oyster mushroom as a complex of mutually potentiating substances on the improvement of the overall condition of pre-school children, and in particular its impact on the immune system of a developing child. This article offers a holistic view of the benefits of the administration of oyster mushroom in the form of a syrup to children.

A double-blind study involved 153 children aged one to seven years, who were overall low thriving and had recurrent respiratory infections, and had not been subject to immunomodulatory treatment. After the threemonth oyster mushroom treatment, the medical condition of the children significantly improved, and various immune functions have been altered or normalized, in particular the concentration of immunoglobulins, phagocyte activity, leukocyte count and some parameters of cell-mediated immunity.

Fungal polysaccharides and their immunomodulatory effect Bioactive polysaccharides from fungi have been particularly known as medicinal mushrooms in Asia as part of the traditional diet and medicine, but in recent years, more scholars and scientists are paying increasing to them in order to better understand the effects, as well as for their use in the pharmaceutical industry. It is a fact that these biopolymers, especially β-glucans or heteropolysaccharides, have taken a place as anticancer or antimicrobial agents, hypocholesterolemics, and prophylactic drugs (Jeseňak 2015, Jeseňak 2017, Vetvička 2013, Spriet et al 2011, Li et al 2014, Pontes et al 2016). Natural immunostimulant β-1.3/1.6 D-glucan isolated from the oyster mushroom (Pleurotus ostreatus) is a modulator of the immune system, particularly by activating certain features of the natural or specific immunity. According to Ferenčík (2004), polysaccharides consisting of a large number of glucose units, which are found in the cell walls of certain bacteria or higher fungi (Basidiomycetes) – can be ranked as non-specific immunostimulants – substances of a microbial origin. The cell walls of fungi contain 5 major components: β-1-3 glucan, β-1-6-D-glucan, and 1-3-D-glucan, chitin and glycoproteins. In addition to the fungal polysaccharides, the oyster mushroom contains lovastatin, ergosterol (provi-

tamin D, which is responsible for the anti-tumour immunity), aegerolysins – ostreolysin, lectin, mushroom fibre, and when it comes to enzymes – laccase and glucooxygenase (Jablonský et al. 2012).

The immune system of children The immune system develops gradually in the prenatal and postnatal periods. Children under one year of age do not have a completely developed immune system; they are, however, protected by biological substances found in breast milk. Even healthy three-month-old infants have a very low concentration of immunoglobulins. The situation of children who are born with a congenital immune deficiency is different – in the insufficient function of their immune system. Primary immunodeficiencies (PID) are not common and are divided into natural immunity (phagocytosis and complement), specific immunodeficiency (manifested as antibody formation disorders, changes in the specific cell-mediated immunity), or as combined immunodeficiencies. Clinical symptoms depend on the extent and type of the insufficient function of the immune system. The more severe the immunodeficiency, the sooner the clinical symptoms appear in children.

In terms of clinical symptoms, those most common in children include chronic and recurrent infections (as many as 8 infections within one year)

associated with repeated antibiotic therapy, as well as chronic diarrhoea, skin disorders (atopic eczema, candidiasis), growth disorders, or symptoms associated with specific immunodeficient conditions (hypoplasia of cartilage, ataxia, etc.). Another situation are secondary immunodeficiencies (SID), for example, due to improper nutrition of children and a lack of vitamins and certain trace elements, or as a result of chronic diseases.

Indication of nonspecific immunomodulation of children The indication of nonspecific immunomodulatory treatment is used in paediatrics to improve the health condition of children as well as prevent diseases, but also in the recovery process after the causal treatment.

It is also important in patients with primary (PID) and secondary immunodeficiencies (SID). Both groups represent a diverse group of diseases caused by a deficiency or disorder of one or more components of the immune system. The most common clinical symptom of a deficiency is increased susceptibility to infections or other immunopathological disorders. Nowadays, approximately 120 disease units are known. The most common is congenital IgA deficiency with an occurrence in 1 to 1,200 live births (Hrubiško, 2016). Some authors also classify human trisomy 21 (Down syndrome) and other

The immunomodulatory effect of oyster mushroom on pre-school children

chromosomal aberrations at PID. In recent years, the incidence of Down syndrome has moved from 1 in 700 to 1 in 1,200 live births (Šustrová, 2016). Secondary immunodeficiency diseases are much more common than primary ones. They may arise throughout life in different age periods, and namely as a result of various activators such as infections, toxic damage, malnutrition, metabolic disorders or other environmental factors.

The correct development of the immune system of children is not only affected by genetic predispositions, but also by their environment. Some of the functions are activated after contact with necessary antigens, e.g. in the past, families with numerous children, contact with pets, etc. were beneficial. Additionally, the food was homemade, simple and mostly fresh, without the addition of preservatives.

Today‘s children are growing up in an isolated home environment, mostly in single-child families, with no access to the natural way of life. Their diet lacks essential nutrients, particularly vitamins and trace elements.

There are currently many immunomodulatory preparations, however, relevant evidence of their effectiveness is lacking for most of them. The only exceptions are immunomodulators of biological origin, including transfer factors – leukocyte lysates.

Human transfer factor Immodin is an original Czechoslovak product that has found wide application in the immuno-allergenic practice in the treatment of certain primary and secondary immunodeficiencies. However, due to its painful application, it is not suitable for the treatment of pre-school children. The second preparation is xenogeneic transfer factor Imunor, produced from swine leukocytes, and as opposed to the subcutaneously administered Immodin, it is applied orally. Orally applied preparations also include bacterial lysates and some anti-virals, such as Isoprinosine, intramuscular or intravenous immunoglobulins, and interleukins (biological treatment). The latter preparations are expensive and are only suitable for severe immunity disorders. Non-specific immunomodulatory drugs include some of the vitamins (vitamin D) and trace elements (selenium, zinc).

Nowadays, in lighter forms of low thriving and immunity disorders, various alternative forms of natural preparations are very popular, as they

are very well tolerated without any significant adverse symptoms. Recently, preparations containing beta-glucan from different manufacturers (Beta-Glucan, ImunoglukanP4H, and others) have become popular. In our practice, the preparation of the whole oyster mushroom in the form of a syrup from Terezia Company has proven to be very successful.

Patient groups and methodology The study was performed from March 2016 to May 2017. It was approved by the Ethics Committee of the Slovak Medical University on October 1, 2016. The children in the study were selected only after parental consent and the signing of an informed consent. We have created two groups of children: the first group consisted of children who were given oyster mushroom in a fruit syrup for a period of three months, and the other group consisted of children who were given the same dosage of pure fruit syrup without the oyster mushroom (control group) for three months.

Both groups were divided into two other subgroups according to their diagnoses. The first subgroup consisted of children with a primary immunodeficiency – mostly children with Down syndrome (DS), and the second subgroup consisted of children with recurrent respiratory infections (nonDS). Before the application of the oyster mushroom in the syrup, each child had undergone a complete immunological examination. After the three-month application, we repeated the control immunological examination. The control group in both groups consisted mostly of siblings.

3. Thereof children with a primary immunodeficiency DS control: 24 4. Children with respiratory infections nonDS control: 28 Seven children did not finish their participation in the study for a variety of reasons. Three children were intolerant of the fruit syrup, and in case of four children the treatment was interrupted due to the use of antibiotic treatment.

Initial and final immunological examinations: 1. Blood count + sedimentation 2. Biochemical examination including acute phase proteins (CRP, A2M) 3. Antibody-mediated immunity: IgG, IgA, IgM, IgE, basic specific and food allergens – atopy panel 4. Cell-mediated immunity by flow cytometry: expression of CD3, CD4, CD8, CD16+56, CD19 and a CD4/CD8 ratio 5. Complement: C3 and C4 complement components 6. Nonspecific immunity by flow cytometry: phagocyte activity and phagocyte index

Oyster mushroom preparation in a syrup (Baby Imun) – a daily dose of 5 mL contains 250 mg of oyster mushroom (118.2 mg of β-glucan ± 3.75 mg). 100 mL of the preparation contains 1.3 kg of fruit.

Pear flavour contains 100% natural apple concentrate, fructose syrup, 100% natural pear concentrate, 100% pure oyster mushroom powder.

Black cherry flavour contains 100% natural apple concentrate, fructose syrup, 100% natural black cherry concentrate, 100% pure oyster mushroom Conditions for the selection of powder. the children: For a minimum of one month they had not taken any anti- Administration of the oyster biotics or other immunomodulatory mushroom – children aged 1 to 3 one therapy in the course of the previous teaspoon on an empty stomach befoyear. At the time of the first sampling, re breakfast, children over 3 years of they did not have any acute infections age one teaspoon twice a day on an (CRP under 5.0 µm/l). empty stomach before breakfast and before dinner.

Total number of subjects in the study: 160 Results Final number of subjects who finished The oyster mushroom was administhe oyster mushroom application and tered in children with recurrent resbecame part of the evaluation: 153 piratory infections, low thriving, aged one to seven. The selection criteria Total number of subjects using the were strict to avoid interference with oyster mushroom: 101 any other treatment. For the study, 1. Thereof children with a primary im- we selected 67 children with Down munodeficiency: DS: 43 syndrome and a disorder of some fun2. Children with recurrent respiratory ctions of the immune system (IS) from infections: nonDS: 58 all over Slovakia, aged one to seven The total number of children who only (43.79%). Delayed maturation of the IS received the fruit syrup without the is clinically manifested by recurrent inoyster mushroom: 52 fections of the respiratory tract. Other 2.

The immunomodulatory effect of oyster mushroom on pre-school children

prevalent diagnoses were recurrent infections 37.5% (n=58), inhalant allergies 10% (n=15), children with a transitional IgA deficiency 5.2% (n=8) and 3% of other diagnoses (n=5).

A significant result of the oyster mushroom application – Baby Imun – was a significant clinical improvement in children. The parents have evaluated the success rate of the treatment as follows: 1. No infections during the treatment and for 3 months after the treatment: 98 children 2. A reduction in the number of respiratory infections by at least 50%: 25 children 3. A reduction in the frequency of infections by 25%: 15 children 4. The parents did not notice any changes: 15 children 5. No antibiotic treatment: 133 children 6. Antibiotic treatment: once in the case of 15 children 7. Repeated antibiotic treatment: 6 children out of 153 The laboratory results of the blood count, cellular and humoral immunity and phagocytosis had been determined before the administration of the oyster mushroom and after the end of the treatment. The most remarkable positive change we have seen was an increase in the concentration of IgA in both groups, a modification (normalization) of the leukocyte count in children with Down syndrome, nonspecific immunity – in phagocyte activity and phagocyte index. Some significant differences were only in one group – the children with recurrent respiratory infections (nonDS) or children with Down syndrome. We have also evaluated the changes between the two groups. We have compared: 1.

Differences in individual parameters for the entire group (DS and nonDS) compared with the review before and after the oyster mushroom treatment.

2. Differences in the DS group (Down syndrome) in individual parameters compared with the DS review before and after the application of the treatment.

3. Differences in the nonDS file (recurrent respiratory infections) in individual parameters versus the review before and after the administration of the treatment, and compared with the nonDS review before and after the administration of the treatment.

4. Differences between the DS and nonDS compared before and after the administration of the treatment.

5. Differences between the groups,

DS and nonDS review before and after values of the results of all the children before and after the treatment, howethe administration of the treatment. ver, ranged from 70% to 90%.

In the evaluation of all parameters in the five categories, we have only se- Number of leukocytes. In children lected the most prominent. with Down syndrome, the number of leukocytes increased, although the Increase in the concentration of differences were not significant, and IgA. The most significant effect was in the nonDS children, the number an increase in the concentration of of leukocytes insignificantly decreaIgA in both groups. The IgA levels sed. The differences in the number of significantly increased in children with leukocytes between the DS and the Down syndrome (p=0.001) compared nonDS groups before the treatment to the review (p=0.02), but also in the were significant p=0.001. nonDS group p=0.001, while no differences were demonstrated in the con- Changes in cell-mediated immunitrol group. In the whole group, there ty. Similarly to the number of leukocywere differences in both groups (DS tes, the results of the two groups di+ nonDS) before and after treatment ffered. We did not see any significant p<0.001 versus the review. In the fi- differences in the number of T-lymnal chart we present the difference phocytes (CD3) and subpopulations between the DS and nonDS groups before and after the oyster mushroom before the oyster mushroom tre- treatment as shown in (CD3) and atment (p=0.021) and after the oyster (the CD4/CD8 ratio). The differences in the number of β-lymphocytes mushroom treatment (p=0.02).

(CD19) in the entire group of DS and Differences in the concentration nonDS children before and after the of IgG and IgM. The differences in oyster fungus treatment were signifithe concentration of IgG were less cant p=0.001, as well as in the DS and significant. The concentration of IgG nonDS control group p<0,001. in children with Down syndrome increased after the oyster mushroom Discussion treatment p=0.082, and in the control In our study, we could not rely on the group p=0.003. In nonDS children, relevant results of other studies, becauthere were significant differences se so far no works observing the effect p=0.045, but in the control group they of the whole oyster mushroom on the were not significant. The differences improvement of immune system funbetween the DS and the nonDS files ctions in small children, and therefore were significant even after the oyster also the improvement of the overall mushroom treatment p=0.004, as well health condition, have been published. as in the control group p=0.002. In Our results clearly show that there has the concentration of IgM, there were been a normalization of several immusignificant differences between the nological parameters. This study is also DS and nonDS groups before the tre- unique in the fact that we have been atment (p<0001) as well as after the able to compare two different groups, treatment (p=0.001). namely children with recurrent respiratory infections and children with Down Changes in non-specific immunity. syndrome. We have gained the trust At first, the phagocyte activity during of the parents, which is also apparent the oyster mushroom application in- from the fact that out of 160 children, creased, however, one month after up to 153 children have completed the end of the treatment, it decreased. the study. Repeated sampling of the Significant changes were only in the biological material of young children nonDS group p=0.004, but not in the is a significant challenge for both the control group. In contrast, the num- children and their parents. ber of phagocyte particles decreased after the administration (phagocyte For us, it was important to learn that index PhI) in children with Down syn- with the improvement of their health drome p<0.001 versus the insignifi- and a decline in the frequency – and cant differences in the control group. for most children even the absence – In nonDS children, there were diffe- of infections, the total number of leurences before and after the treatment kocytes in the children with recurrent p=0.025; however, there were insigni- infections decreased. It was otherwise ficant changes in the control group. pleasing to learn that in children with There were also significant differen- Down syndrome the number of leuces in the comparison of the DS and kocytes increased – one could say that nonDS groups prior to the treatment the number of leukocytes normalized. p=0,051, but not after the oyster Children with Down syndrome are chamushroom treatment. The reference racterized by leukopenia (lymphope

The immunomodulatory effect of oyster mushroom on pre-school children

nia or neutropenia). In our practice, the oyster mushroom is the first product that was able to normalize the number of leukocytes. The change in leukocytes in the children of both groups is associated with a change in phagocyte activity and phagocyte index.

After the activation of leukocytes and increased ingestion after the oyster mushroom treatment, the phagocyte activity and phagocyte index temporarily increased, but within two months both parameters normalized. Multiple authors (Jeseňak et al. 2015, Vetvicka et al. 2014) describe an increase in the phagocyte activity after the β-glucan treatment. However, sampling was done immediately after the treatment, when β-glucan was directly affecting the ingestion. On the other hand, the laboratory tests were performed in various places in Slovakia and the Czech Republic.

Levels of immunoglobulins IgA and IgG significantly increased in both groups, and the increase was especially apparent in the concentration of IgA, where we noticed significant differences in comparison with the control group. The concentration of IgA is zero after childbirth and then it gradually increases, which is associated with the antigenic stimuli. As we have mentioned earlier, many children who have been brought up in a home with no siblings or without any natural contact with other children have a transient IgA hypogammaglobulinaemia.

After entering the children collective facilities, they are not prepared to bear the increased antigenic burden. The oyster mushroom has proven to be an excellent stimulator for increasing the concentration of IgA in transient hypogammaglobulinaemia.

For us and the satisfied parents, the most important knowledge was the improvement of the health condition of the children. A similarly significant benefit was the simple administration and availability of the oyster mushroom syrup.

Conclusion The administration of the oyster mushroom syrup was effective in both groups of children. Several functions of the immune system improved or were modified, and there was a significant improvement in the health of the children. In contrast to declared immunopreparations, for the first time we have seen a significant improvement in antibody-mediated immunity (a significant increase in the concentration of IgA in the entire group) and the normalization of the leukocyte count in children with Down syndrome. We did not notice any side effects or intolerances to the oyster mushroom syrup.

The results of the laboratory tests, our experience and the satisfaction of the parents have convinced us that the oyster mushroom syrup is a suitable alternative for the treatment of hypogammaglobulinaemia, for leukopenia treatment of children with Down syndrome, and for the treatment of children with recurrent respiratory infections. We recommend a three-month treatment for children prior to joining collective facilities.

References: 1. Ferenčík, M., Rovenský, J., Shoenfeld, Y., Maťha, V.: Imunitný systém – dobrý obranca, ale aj možný diverzant. Slovac Acadenic Press, s.r.o., Bratislava, ISBN 80-89104-45-2, s. 273 2. Hrubiško, M. a spol.: Zvyčajné a menej časté indikácie humánneho transfer faktoru Immodin. Neintervenčná multicentrická observačná retrospektívna štúdia hodnotiaca indikácie dávkovanie Immodinn-u v rutinnej klinickej praxi. 2016. Klinická imunológia a alergológia, ISSN 1335-0013, vol. 26, č. 1, 2016 s. 20-29.

3. Hrubiško, M.: Miesto biologicky aktívnych polysacharidov v imunomodulačnej liečbe. 2016. Pediatrics, ISSN 1336-863X, No1, č. 11, s 4-6.

4. Jablonský, V.A., Šašek, V., Vančuříková, Z.: Huby ako liek, 2012. Ottovo nakladatelství ISBN 978-80-7451-258-2, s.200.

5. Jeseňak, M.: Imunogukán P4H – klinická účinnosť podporená medicínou dôkazov. 2016. Klinická imunológia a alergológia, ISSN 1335-0013, vol. 26, č. 1, s.39.

6. Jeseňak, M., Urbančíková, I., Banovcin, P.: Respiratory Tract Infections and the Role of Biologically Active Polysaccharides in Their Management and Prevention. 2017. Nutrients ISSN 2072-6643. Vol. 779, No 9, s. 2-12.

7. Li, F., Kin X., Liu, B., Zhuang, W., Scalabrin, D.: Follow-up formula consuption in 3-to 4 year olds and respiratory infections. 2014. Pediatrics, ISSN 0022-3476, No 133, s. 1533-1540.

8. Pontes, M.V. et al: Cow´s milk based beverage consuption in 1-to 4 year-olds and allergic manifestations: an RCT. 2016. Nutrition Journal, ISSN 2937-016-0138-0, No 10, s. 1-10.

9. Spriet, I., Desmet, S., Hansen, R., et al: No interference of the 1-3- No 1 β-D-glucan containing supplement imuni X with the 1-3- β-D-glucan serum test. 2011, Mycoses, ISSN 14390507, Vol 54, No 5, s. 352-353.

10. Šustrová, M.: Medicínske aspekty ranej starostlivosti o deti s Downovým syndrómom. História starostlivosti na Slovensku. 2016. In: Aktuální trendy sociální práce (Česko-slovenské perspektivy rozvoje) Sborník z medzinárodní vědecké konference. Příbram: Ústav sv. Jana Nepomuka Neumanna, ISBN 978-80-906146-8-0, s. 108-115.

11. Šustrová, M.: Zvýšená prevalencia autoimunitných chorôb u jedincov s Downovým syndrómom s parciálnou a translokačnou formou. 2016.

In: Alergie. Časopis pro kontinuální vzdělávání v alergologii a klinické imunologii, ISSN 1212-3536, ročník 18, s. 59.

12. Vetvicka, V., Richter, J., Svozil, V., Rajnohová Dobiasová, K, Kral, V.: Placebo-driven clinical trials of transfer point glucan #300 in children with chronic respiratory problems. 2013.

Am J Immunol, ISSN 1600-0897, No 9, s. 43-47.

Clinical trial with BABY IMUN

a syrup with oyster mushroom to support children‘s immune system The study was approved by the Ethics Committee of the Slovak Medical University on October 1, 2016.

We only selected children for the study after their parents gave their consent and signed an informed consent.

Clinical observation guarantor: Prof. MUDr. Mária Šustrová, CSc. Clinical Immunologist of the Slovak Medical University, Bratislava

Objective of the study To boost the immune system of children with recurrent respiratory infections

Duration of the study March 2016 - May 2017

Group of patients

A total of 153 children aged 1 - 7 The total number of children using BABY IMUN: 101 1. Number of children with primary immunodeficiency: DS: 43 2. Children with recurrent respiratory infections: nonDS: 58 Total number of children using only fruit syrup without oyster mushroom: 52 3. Number of children with primary immunodeficiency DS examination: 24 4. Number of children with respiratory infections nonDS examination: 28

Conditions for the selection of children At least one month without any antibiotic treatment and one year without any immunomodulatory treatment.

No acute infections at the time of the first sampling (CRP below 5.0 μm/l, alfa 2 macroglobulin normal).

Process

 Monitoring of 19 immunological parameters  Blood sampling prior to the use of BABY IMUN of nonspecific and specific immunity and 2-3 months after the use of BABY IMUN

Usage

 Children aged 1-3: Once a day one teaspoon on an empty stomach before breakfast  Children from 3 years of age: Twice a day one teaspoon on an empty stomach before breakfast and dinner

BABY IMUN 1 teaspoon (5 ml) of syrup contains 250 mg of 100% pure oyster mushroom powder with a declared beta-glucan content of 118.2 mg ± 3.75 mg.

The syrup is based on a fruit concentrate made from fresh fruit. One bottle (100 ml) contains more than 1 kg of fresh fruit concentrate.  NO preservatives  NO sweeteners

 NO color  NO color additives

Food supplement

Selected initial and final immunological examination 1.

2.

3.

4.

5.

Complete blood count + sedimentation Biochemical examination including acute-phase proteins (CRP, A2M) Antibody-mediated immunity (immunoglobulins IgG, IgA, IgM, and IgE) Nonspecific immunity: phagocytic activity (FA) and phagocytic index (FI) Cell-mediated immunity

Legend CRP - acute-phase protein that is formed in the liver, the level in healthy humans is CRP < 6.0 mg/l IgA - Immunoglobulin A: an antibody that is extensively released on the surface of mucous membranes and plays an important role in protecting the body against microorganisms DS - Down syndrome nonDS - without Down syndrome

Conclusions - the most important changes in the immunity profile (graphs) Only some of the most significant results are selected, although we have noticed significant changes in other monitored parameters as well.

Increased IgA concentration - in each group, significant differences in comparison with the control group p < 0,001

3,0

4,0 p=0,02

*

2,5 p=0,001

2,0

2,0

IgA g/L

IgA g/L

3,0

1,5 1,0

1,0 ,5 ,0

,0 Before use

After use

Before use

After use

IgA values in the DS group

IgA values in the nonDS group

 with BABY IMUN  control group

 with BABY IMUN  control group

Increased IgA concentration in both groups, significant differences in comparison with the control group 20,0

4,0

p=0,001

p = 0,014

p=0,018 p < 0,001

15,0

IgG g/L

3,0

IgA g/L

Increased IgG concentration in both groups

2,0

10,0

1,0

5,0

,0

,0 Before use

After use

Before use

After use

IgA levels in the DS + nonDS group

IgG levels in the DS + nonDS group

 with BABY IMUN  control group

 with BABY IMUN  control group

Differences in the leukocyte count

Phagocytic activity (nonDS) p = 0,004

95,0 15,0 90,0 12,5

10,0

FA %

LKC x 103 uL

85,0

7,5

75,0

5,0

2,5

80,0

70,0 p = 0,001

65,0 Before use

After use

Before use

After use

Leukocyte counts in the group of children taking BABY IMUN

Phagocytic activity in the nonDS group of children

 DS group  nonDS group

 with BABY IMUN  control group

Overall The administration of oyster mushroom contained in BABY IMUN was effective. It improved or fixed multiple functions of the immune system and significantly improved the health of the children. In comparison with the proclaimed immunomodulators, we noticed a significant improvement in antibody-mediated immunity (significant increase in IgA concentration in the entire group) and a normalization of leukocyte counts in children with Down syndrome. We did not notice any adverse effects or intolerance to the oyster mushroom in the syrup.

Results in the group of children (DS + nonDS) taking BABY IMUN (n=101)  During the administration and three months after the administration, there were no respiratory

infections in 75 children (74.25 %).  During the administration and three months after the administration, only 15 children (14.85 %)

experienced an insignificant infection (cold) - without ATB treatment.

Overall, the

89 %

of children had

excellent results 11

Number of children using BABY IMUN (n=101)  Children without an infection 75

15 75

 Children with one insignificant infection

(cold) not treated with 15

 Children with infections 11

89 % Summary  It was proven that oyster mushroom acts as an immunomodulator = a substance capable of

changing the extent of the immune response, either by altering the antigenic properties of the antigens, or by affecting the immune mechanisms of the individual.

 The results of the study demonstrated the effectiveness of the oyster mushroom syrup in the form of

reducing the number of infections in children with recurrent respiratory infections.

 The syrup is suitable for children from 1 year of age.  During the study, the monitored children did not exhibit any side effects = excellent tolerance.

Ingredients: 100% pear concentrate (58,3%), 100% apple concentrate, 100% oyster mushroom (Pleurotus ostreatus) powder, dried sea buckthorn (Hippophae rhamnoides) juice powder.

Warning:

► Food supplement. ► Suitable for children from 1 year of age, pregnant and lactating women, gluten-free diet. ► Keep out of reach of children. ► Do not exceed the recommended daily dosage. ► This is not a substitute for a balanced and healthy diet and lifestyle. ► Store below 25 °C. ► Shake well before use. ► After opening, store in the refrigerator and use within 20 days. ► Do not use in patients with a known hypersensitivity to any of the product‘s components.

Recommended dosage: Children aged 1 to 3: 1 tea spoon once daily before a meal.

Children aged 3 to above: 1 tea spoon twice a day before a meal.

Suitable for long-term use.

Manufacturer: TEREZIA COMPANY s.r.o. | www.terezia.eu

The use of food supplement GLUREGUL in the treatment of patients with type 2 diabetes Study guarantor: MUDr. Denisa Spodniaková – diabetologist Place of study: Banská Bystrica, Slovakia Number of patients: 60 (28 men, 32 women) Age of patients: 33–74 years (average age 58.3 years) Indication: type 2 diabetes mellitus (average length of illness 5.7 years) Treatment of patients: oral antidiabetics + diabetes diet Objective of the observation: To verify the effect of the product on glycaemia, on the compensation of diabetes at 3 months and on the tolerability of the product.

Dosage: 2x 1 capsule for 3 months

Evaluation: comparison of laboratory results before and after administration of the product tolerability of the product side effects evaluation of the subjective condition of patients

Monitored parameters: glycated hemoglobin (HbA1C) glycaemia (blood sugar level)

Results: Administration of the product in type 2 diabetes patients with oral antidiabetic treatment had a statistically significant effect: it reduces glycated hemoglobin levels it stabilizes glycaemia levels 100% customer satisfaction no side effects excellent tolerability of the product

Food supplement

GLUREGUL: contains the rare shaggy ink cap mushroom with organically bound vanadium ginger, cinnamon and heart-leaved moonseed contribute to normal blood sugar levels made without additives and preservatives from a Czech manufacturer with a 30-year tradition

HbA1c The glycated hemoglobin level in each patient before GLUREGUL administration (orange) and after GLUREGUL administration (blue) 14 14

HbA1c (%)

12 12 10 10 8 8 6 6 4 4 2 2 0 0

1 112 22 3 3 34 44 5 55 6 6 67 77 8 889 9 9101112131415161718192021222324252627282930313233343536373839404142434445464748495051525354555657585960 101112131415161718192021222324252627282930313233343536373839404142434445464748495051525354555657585960 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 HbA1c po th After po th HbA1c

HbA1c pred th Before pred th HbA1c

HbA1C before and after administration Result A statistically significant decline in glycated hemoglobin after 3 months of GLUREGUL use. average value before administration: 8% average value after administration (3 months): 6.85 % Before administration

After administration

Normal HbA1c levels Excellent compensation 4.75 %

Acceptable compensation 6.5 %

Poor compensation 7.5 %

Glycaemia The average glycaemia level in each patient before GLUREGUL administration (orange) and after GLUREGUL administration (blue) 20

Glycaemia (mmol/l)

18 16 14 12 10 8 6 4 2 0

10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 1 1 22 3 344 55 6 6 77 8 8 9 9101112131415161718192021222324252627282930313233343536373839404142434445464748495051525354555657585960

Gly Afterpo th

Gly pred th Before

Glycaemia before and after administration Result A statistically significant decline in average glycaemia values after 3 months of administration of GLUREGUL. average value before administration: 9.5 mmol/l average value after administration (3 months): 7.5 mmol/l Before administration

After administration

Normal glycaemia levels Hypoglycaemia

Normal 3.5 mmol/l

Hyperglycaemia 5.5 mmol/l

GLUREGUL Blood sugar level regulation Characteristics: GLUREGUL is a combination of mushroom and plants, which is beneficial for blood sugar level. It is suitable for people dealing with glucose tolerance and overweight. Heart-leaved moonseed helps regulate blood sugar levels. It supports the cleansing effect of the urinary tract. Shaggy ink cap is primarily characterized by its high nutritional value. Ginger and cinnamon help maintain normal blood sugar levels.

Ingredients: 1 capsule contains 320 mg of the GLUREGUL mixture [powder from the dried lyophilised mushroom of shaggy mane (Coprinus comatus) 170 mg, extract from the heart-leaved moonseed/guduchi (Tinospora cordifolia) 80 mg, dried ginger powder (Zingiber officinale) 50 mg, dried cinnamon (Cinnamomum zeylanicum) 20mg].

Capsule shell: gelatine.

20 mg

dried cinnamon

6 % 16 %

25 %

50 mg

dried ginger powder

80 mg

extract from the heart-leaved moonseed

53 %

170 mg

powder from the dried lyophilised mushroom of shaggy mane

Recommended dosage: 1 capsule twice a day before meals. Swallow the whole capsule with a drink of water, or open it and sprinkle its content over a meal or into water. Suitable for long-term use.

Warning: not suitable for children up to the age of 3, pregnant and breastfeeding women keep out of children‘s reach do not exceed the recommended daily dose not to be used as a substitute for a varied and balanced diet and a healthy lifestyle store in a dry place with a temperature of up to 25 °C consume the capsule immediately after removal from the blister suitable for a gluten free diet do not use if you are hypersensitive to any of the product‘s ingredients

Food supplement

Treating iron deficiency anemia with food supplement HEMO

Treating iron deficiency anemia

MUDr. Daniela Babincová, Department of Hematology, Malacky Hospital,

Chief Physician MUDr. Henrich Gašparík, PhD. Department of Gynecology, Hospital and Polyclinic Myjava

with food supplement HEMO PLUS Iron deficiency anemia is a common health problem. For diagnosis and proper treatment, the first step identifying the cause of iron deficiency (Fe) and eliminate it by replacing the missing iron. Today, there is a number of iron supplements available, but some patients experience intolerances when taking them, preventing the desired effect of treatment from being achieved. The authors used the HEMOPLUS food supplement for iron substitution, and they hereby present the results of the monitoring of two patient groups.

Introduction Iron is part of all living cells in the body. It is involved in many enzymatic processes and is an essential component for the synthesis of hemoglobin, which ensures the transfer of oxygen in the body.

The body of an adult contains 3.5 - 5 g of iron. Most of it (about 2/3) is found in erythrocytes, the rest is in muscle pigment - myoglobin and other cells. Iron stores - ferritin and hemosiderin are found in mononuclear cells in bone marrow, liver and the spleen.

Iron enters the body through food. It is ionized in the stomach in the presence of HCl (hydrochloric acid) and is absorbed into the blood in the duodenum and in the upper small intestine as bivalent iron. As early as 2-4 hours after a meal, it reaches the blood plasma where it binds to a transport protein - transferrin, which transfers it to the target organs.

Causes of iron deficiency anemia: 1. Insufficient intake in food 2. Increased need for Fe in certain periods of life - adolescence (start of menstruation in girls, pregnancy, breastfeeding , muscle building in boys) 3. Increased iron loss - acute blood loss caused by injuries and surgery, bleeding caused by blood clotting disorders, chronic blood loss in digestive tract disorders (gastric ulcer, hiatus hernia, bowel diverticulosis, Crohn‘s disease, chronic diarrhea, tumors, haemorrhoids), gynecological diseases (hypermenorrhea, metrorrhagia due to myomatous uterus), urological pathologies associated with bleeding in the urine, extensive burns, desquamation processes on the skin and others 4. Impaired iron absorption (lack of HCI in the stomach, after gastrectomy) Symptoms of iron deficiency anemia Manifestations of anemia depend on its severity and rate of onset. Sudden massive blood loss (trauma, surgery) is initially accompanied by a reduction in oxygen carriers - erythrocytes - and a reduction in the circulating blood volume. Dramatic clinical manifestations are a reflection of circulatory failure, not of iron deficiency, and the condition is resolved by replenishment with solutions and the administration of erythrocyte transfusions. Iron deficiency is only manifested after this. Typical iron deficiency anemia usually develops over a long period of time with a long-lasting cause, either insufficient Fe intake or increased blood loss. The patient adapts to the condition, the symptoms of anemia come gradually depending on the degree of severity of the anemia. Subjective indications include increased fatigue, malaise, decreased physical performance, dyspnoea, palpitations, headaches. The basis for a proper diagnosis and treatment is a detailed medical history and physical examination of the patient, followed by laboratory tests. Information about eating habits, digestive problems, gynecologi-

cal problems, blood loss and changes in the patient‘s weight is important. During an objective examination we look for pale skin and examine visible mucous membranes, cracked mouth corners, increased nail and hair brittleness, dry skin.

According to anamnestic data, it is often necessary to perform gastroenterological, gynecological and other examinations.

iron deficiency using food supplement HEMOPLUS produced by Terezia Company.

Patient group and methodology We monitored two groups of patients.

The first group consisted of female patients with iron deficiency anemia with increased menstrual blood loss, bleeding due to myomatous uterus, hiatus hernia, bowel diverticulitis, hemorrhoids, and insufficient iron intake Necessary laboratory tests Blood count (Hb, Ery, HTK, Le, MCV, MCH, through food. Anaemia with a haemoglobin value of 80-100 g/l and a low level of serum MCHC, Tr, blood smear).

Serum iron, transferrin, ferritin, transferrin sa- and reserve iron were an indication to admituration. nister the preparation.

Based on the hemoglobin levels we determine the degree of severity of the anemia: mild (Hb The other group included pregnant women above 100g/l), moderate (Hb 80-100 g/l), severe with iron deficiency anaemia.

(Hb below 80 g/l). The number of erythrocytes in iron deficiency anemia may not always be The first group consisted of 55 women reduced, changes in erythrocyte morphology Entry laboratory examination: Hb, HTK, Ery, are typical. Hypochromia and microcytosis are MCV, MVH, serum iron, ferritin, transferrin. present. Small, pale erythrocytes with marked The patients took the HEMOPLUS preparation bright centers (anulocytes, planocytes) can be 2x2 cps. daily for two months. seen under the microscope.

At least one week after they stopped taking the preparation, the above-mentioned laboraCurrent possibilities of treating iron defi- tory parameters were examined again. ciency anemia 1. When we find the cause of the iron deficien- The other group consisted of 50 pregnant wocy, we remove it (change eating habits, elimi- men with iron deficiency anaemia.

Entry laboratory examination: Hb, Ery, HRK, nate the source of blood loss).

2. Administration of medications and iron pre- MCH, MCV, serum iron, ferritin, transferrin.

The patients took the HEMOPLUS preparation parations. for pregnant women 2x2 cps. for two months.

Today there are many available iron prepara- The same laboratory parameters were examitions: for intravenous and intramuscular admi- ned after one month and after two months of nistration when a rapid increase in blood count is talking the preparation. needed, oral preparations in the form of tablets, capsules, drops. An extreme solution to iron defi- Results In the first group, 44 female patients of 55 ciency anemia is erythrocyte transfusion.

Although most patients tolerate iron prepara- (80 %) achieved full normalization of all the motions well, some report an intolerance causing nitored laboratory parameters. constipation, stomach upset, a metallic taste 8 female patients (14.5 %) achieved partial imin the mouth, urticaria, diarrhea. provement, and in 3 female patients (5.45 %) This has led the authors to the decision to treat the laboratory parameters were not improved

Treating iron deficiency anemia with food supplement HEMO

In all monitored female patients, the MCH value / haemoglobin content in erythrocytes rose, which is essential for oxygen transfer.

No female patient experienced bad side effects, the female patients provided information about subjective improvement of their condition during the treatment.

The three female patients in which we did not notice an improvement of laboratory parameters had a diagnosis of uterus myomatosus, and they were bleeding heavily and repeatedly

during the therapy. Their condition had to be resolved by surgery.

In the other group, we performed the laboratory tests after one month of taking the HEMOPLUS preparation, and we found a partial improvement in the monitored laboratory parameters. After two months of taking the preparation, we noticed a statistically signifi-

cant increase in Hb and Fe. noids from grapes and red beet provide antiTolerance of the preparation was very good, no oxidant and detoxifying effects, sea buckthorn unwanted side effects occurred. is extremely rich in vitamin C that is needed for the resorption of iron. Oyster mushroom with Discussion its content of dietary fibre and probiotics reguPhysicians in many medical disciplines encoun- lates the processes in the intestinal tract, and it ter the need to treat iron deficiency anaemia. If also supports non-specific immunity. the cause of the iron deficiency can be remo- The composition of the preparation is well-baved successfully, its replenishment is a matter lanced, and the effect of particular components of several weeks. However, there are many pa- has a synergic effect on the body. tients who have to rely on long-term treatment with iron. We have many available iron medi- Conclusion cations that are tolerated well by the majority Administration of the HEMOPLUS preparation of patients. However, an intolerance to these in two groups of female patients resulted in a preparations is not a rarity, and in the case of a surprisingly good result, which can be compatient with the need of long-term iron substi- pared to the use of per-oral iron preparations. tution, a serious problem can appear.

We did not notice any manifestations of intoWe encounter persistent bowl obstruction, sto- lerance, and primarily in the group of pregnant mach pains, abdominal discomfort, diarrhoea, women the response to the therapy was very a persisting metallic taste in the mouth, rash. good.

The patients often discontinue the use of the Our experience convinced us that the admimedication, and the expected effect does not nistration of the HEMOPLUS preparation could arrive. be an alternative to the treatment of iron deFor the aforementioned reasons, we tried to use ficiency anaemia to a large extent, not only in the HEMOPLUS natural preparation to resolve the case of patients with an intolerance to iron iron deficiency anaemia. Another motivation preparations. was to help pregnant women with anaemia whose pregnancy sickness escalated when they took iron preparations.

The results of both groups´ monitoring were surprisingly positive. The normalization of the parameters of complete blood count and iron corresponded to the effect of iron preparations, with the bonus of excellent tolerance of the preparation.

Characteristics of the HEMOPLUS preparation In addition to iron, the preparation also contains natural components, which enrich it with B-group vitamins that are important as factors for the maturation of blood cells in bone marrow. FlavoB) Ery –red blood cell count (erythrocytes) mil./mm3

A) Hb – haemoglobin level (red blood pigment) (g/l) 120

5,0

110

4,5

100

4,0

Hb_0 before application

90

80

Ery_0 before application

3,5

Hb_1 1 month after application

Ery_1 1 month after application

Hb_2 2 months after application 3,0

Ery_2 2 months after application Ery_0 Ery_1 Ery_2

Hb_0 Hb_1 Hb_2

C) FeS – level of reserve iron (μmol/l)

D) Quantity of reserve iron in the form of ferritin (μg/l)

50

60 50

40

40

30

30 20 Fe_0 before application

10 0 Fe_0 Fe_1 Fe_2

20

Fe_1 1 month after application

10

Fe_2 2 months after application

0

Ferritin_0 before application Ferritin_1 1 month after application Ferritin_2 2 months after application Ferritin_0 Ferritin_1 Ferritin_2

2.