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By
of
DOCTOR OF PHILOSOPHY
in
Agricultural Science
(Food Science and Technology)
2016
ANTI-ALLERGY PROPERTIES OF SOME
FUNCTIONAL FOODS
By
LIST OF TABLES……………………………………………… V
LIST OF FIGURES ……………………………………………. VIII
LIST OF ABBREVIATIONS………………………………….. X
1. INTRODUCTION…………………………………………… 1
2. REVIEW OF LITERATURE……………………………..... 3
2.1. Functional foods and Immunodulatory activities…………… 3
2.1.1. Pomegranate ……………………………………………… 8
2.1.2. Echinacea purpurea.............................................................. 13
2.1.3. Spirulina plotensis................................................................ 17
2.1.4. Stevioside............................................................................. 22
2.2 Nutrition, Immunity and allergy…………………………...... 25
2.2.1. Role of Nutrients in Immune Functions............................. 28
2.2.2. Food Allergy....................................................................... 29
2.2.3. Diets and Food Allergy........................................................ 31
2.3. Allergy.................................................................................... 33
2.3.1. Diagnosis of allergy............................................................. 35
2.3.2. Allergy Reactions................................................................ 36
2.3.3. Autoimmune diseases.......................................................... 38
2. 3.4. Types of allergies diseases.................................................. 39
2.3. 4.1 Urticaria…………………………………………………. 39
2.3.4.2. Rhinitis………………………………………………….. 41
2.3.4.3. Asthma…………………………………………………... 42
2.3.4.4. Immunodeficiency………………………………………. 44
2.4. Oral Immunotherapy………………………………………… 46
3. MATRIALS AND METHODS……………………………… 47
3.1. MATERIALS………………………………………………. 47
3.1.1. Raw materials…………………………………………….. 47
3.1.2. Chemicals............................................................................. 47
3.1.3. Preparation of fresh pomegranate juice (PJ)……………... 47
3.1.4. Preparation of echinacea extract (EP)…………………… 48
3.1.5. Preparation of spirulina extract (SP)…………………….. 48
3.1.6. Preparation of the formulated pomegranate beverage
(FPB)……………………………………………………... 48
3.1.7. Preparation of Immune Functional Extract (IFE).............. 48
3.1.8. Preparation (IFE) of extract mixtures addition to the
Vaccine…………………………………………………. 50
3.1.9. Food allergen Extraction (vaccine)..................................... 50
3.2. METHODS ……………………………………………….. 50
3.2.1. Physicochemical analyses………………………………… 50
3.3.2.1.1.Determination of pH, titratable acidity(TA),and total soluble
solids (TSS) in Extract and mixtures.................. 50
3.2.1.2. Determination of phenolic compounds in Extract and
mixtures............................................................................ 51
3.2.1.3. Determination of flavonoids compounds in Extract and
mixtures.............................................................................. 51
3.2.1.4. Determination of total phenols content Extract and mixtures
………………………………………………… 51
3.2.1.5. Determination of total flavonoids Extract and mixtures... 51
3.2.1.6. Determination of vitamin C…………………………….. 52
3.2.1.7. Determination of antioxidants activity............................. 52
3.2.2. Sensory evaluation of formulated pomegranate beverage
(FPB)……………………………………………………. 52
3.2.3. Medical biochemical analysis……………………………. 53
3.2.3.1. Immunotherapy Technique............................................... 56
3.2.3.2. Phagocytosis inhibition assay…………………………… 57
3.2.3.3 Eosinophils....................................................................... 58
3.2.3.4. Skin prick test................................................................... 58
3.2.3.5. Total Immunoglobulin IgE…………………………….. 60
3.2.3.6. Immunoglobulin IgE Specificity ……………………….. 62
3.2.3.7. IgG, IgM, IgA by Immunodiffusion................................. 64
3.2.3.8. Radial Immunodiffusion (RID) C3 and C4…………….. 64
3.2.3.9. Peak expiratory flow rate "PEFR" or Pulmonary function test
monitoring method.................................................... 64
3.2.4. Preparation of culture media for fungal and bacteriological
studies of extract and mixture extracts................................. 65
3.2.4.1. Cultivation of fungi........................................................... 65
3.2.4.2. Cultivation of bacteria………………………………..... 65
3.3. Statistical analyses.................................................................. 66
4. RESULTS AND DISCUSSION…………………………….. 67
4.1. Physico-Chemical characteristics of extracts and extract
mixtures.................................................................................. 67
4.2. Determination of total phenols and flavonoids of extracts and
extract mixtures ………………………………………… 69
4.3. Vitamin C contents of extract mixtures by HPLC………….. 70
o
LIST OF FIGURES
No. Page
4.1 Comparison between the using of mixtures extract IFE as 106
regard skin prick test in urticaria group 2……………
4.2 Comparison between the using of mixtures extract as regard 108
skin prick test in rhinitis group 2 ……………….
4.3 Comparison between the using of mixtures extract IFE as 109
regard skin prick test in asthma group 2……………..
4.4 Comparison between the using of mixtures extract (IFE) and 110
addition to vaccine (MIX) as regard skin prick test in urticaria
group 2……………………………………..
4.5 Comparison between the using of mixtures extract addition to 111
vaccine MIX as regard skin prick test in rhinitis group
2……………………………………….....
4.6 Comparison between the using of mixtures extract addition to 113
vaccine as regard skin prick test in asthma…
4.7 Comparison between mixtures extracted (IFE) AD and adds 114
to vaccine as regard season in urticaria group 2 …
4.8 Comparison between mixtures extracted (IFE) BC and added 116
to vaccine (MIX) as regard season in rhinitis group
2…………………………………………………..
4.9 Comparison between mixtures extracted (IFE) BC and added 117
to vaccine (MIX) as regard season in asthma group
2…………………………………………………..
4.10 Comparison between mixtures extracted (IFE) BC and added 119
to vaccine (MIX) as regard family history in urticaria group
2………………………………………...
4.11 Comparison between mixtures extracted (IFE) BC and added 120
to vaccine (MIX) as regard family history in rhinitis group
2………………………………………….
4.12 Comparison between mixtures extracted (IFE) BC and added 121
to vaccine (MIX) as regard family history in asthma group
2…………………………………….........
4.13 Comparison of Sp.IgE level AD, BC and AB extract mixtures 124
(IFE) added to vaccine as regard (MIX) in patient
group…………………………………………..
4.14 Comparison of T.IgE level AD, BC and AB extract mixtures 126
(IFE) added to vaccine as regard (MIX) in patient group 2
and 3 ………………………………….
4.15 Comparison of IgG mg/dl level AD, BC and AB extract 128
mixtures (IFE) added to vaccine as regard (MIX) in patient
group...………….................................................
4.16 Comparison of IgA mg/dl level AD, BC and AB extract 132
mixtures (IFE) added to vaccine as regard (MIX) in patient
group 2………………………………………….
4.17 Comparison of IgM mg/dl level AD, BC and AB extract 134
mixtures (IFE) added to vaccine as regard (MIX) in patient
group 2…………………………………………..
4.18 Comparison of EoE% AD, BC and AB extract mixtures (IFE) 136
added to vaccine as regard (MIX) in patient
group2……………………………………………………
4.19 Comparison of phagocytosis % AD, BC and AB extract 138
mixtures (IFE) added to vaccine as regard (MIX) in patient
group2…………………………………………..
4.20 Comparison of Peak expiratory flow rate% AD, BC and AB 141
extract mixtures (IFE) added to vaccine as regard (MIX) in
rhinitis and asthma ............................................
4.21 Immunodeficiency level mg/dl before using the mixtures 143
extract and after immunotherapy extracts mixture IFE in
AD in comparison to the control………………………...
4.22 Bacterial and fungal growth reviews for food material 177
extracts (1) Pomegranate (2) Echinacea (3) Spirulina (4)
Stevioside and extract mixtures prepared for patients with
(5) Rhinitis BC(6) Urticaria and Immunodeficiency AD (7)
Asthma AB allergies compared to control (CB: Control
Bacterial , CF: Control Fungal ) after 24 hr and 48 hr
incubation time……………………………………
5. SUMMARY AND CONCLUSION
The results showed that reductive power, which means the antioxidant
activity in pomegranate juice PJ was high 92.95% compared with extracts
SP, EP and ST which are as follows (71.09, 69.02 and 67.14%), respectively.
the results also showed a significant increase in antioxidant activity between
the mixtures used in the improvement of the functions of the immune system
and reduce the symptoms of food allergies for urticaria, asthma, rhinitis and
immunodeficiency. It was noted the existence of a significant increase in both
the total soluble solids and pH levels in both extracts and mixtures used. And
no significant differences in total solids between mixtures used in the
sensitivity of the rhinitis and asthma.
The results obtained from phenols in the beverage product using high-
performance liquid chromatography HPLC showed the increase of phenolic
acid content of pryogaiol, tyrosol, chlorogenic, epicatechin, vanillic,
aminobenzoic then catechol.
The most important medical results obtained extracts and mixtures used
in cases of food allergies and immunodeficiency