Effect of Acrylamide on Stomach, Cerebellum and Testis of the Albino Rat. By Hesham Noaman Abdel Raheem M.D. THESIS

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1 Effect of Acrylamide on Stomach, Cerebellum and Testis of the Albino Rat By Hesham Noaman Abdel Raheem M.D. THESIS

2 Acknowledgement should go to all persons who have assisted, in a way or another, to allow this study to come to light; Doctor: Kariman Mohammed El-Gohari (Professor of Anatomy and Head of Anatomy Department). Doctor: Hassan Mostafa Serry (Professor of Anatomy). Doctor: Shahira Samir Zaki (Assistant Professor of Anatomy).

3 Introduction

4 Acrylamide Chemistry Uses water treatment, enhanced oil recovery, papermaking aids, soil conditioning agents, sewage and waste treatment, ore processing, permanent-press fabrics. CH 2 =CHCONH 2

5 Human Exposure to Acrylamide Acrylamide is a chemical compound formed de novo when certain carbohydrate-rich foods fried, baked, or roasted at high temperatures. It was not known to be in food until April 24, 2002, where Swedish scientists reported high concentrations of in fried, baked foods. Maillard reaction is reaction of reducing sugar (as glucose) with free asparagine, an amino acid found in many foods, especially at high temperature. WHO/FAO: Dietary intake: µg/kg BW/day.

6 Exposure to Acrylamide in Food French Fries & Potatoes Breads Potato Chips Cereal Biscuit / Cookies All Other Foods ug/day Coffee Cakes Dried Foods Pop Corn Salty Snacks Chocolate Nuts/Seeds/ Butters

7 Effect of temperature 160 C 27 ppb 170 C 70 ppb 180 C 326 ppb Acrylamide levels in potato chips fried for 4 minutes increased with frying oil temperature. (ppb=parts per billion)

8 Effect of time 3.5 m 12 ppb 4 m 46 ppb 4.5 m 227 ppb 5 m 973 ppb Acrylamide levels in potato chips fried at 180 o C increased with frying time.

9 Aim of the work

10 The current project aimed at exploring the harmful effects of acrylamide on the structure of the stomach, cerebellum and testis in the albino rat, in an attempt to clarify its potential risk on the human health.

11 Material and Methods

12 The stomach, testis and cerebellum specimens were collected form fifty adult male albino rats. Animals were divided to two main groups: I, II each group was subdivided into 3 subgroups.

13 Specimens from testis, cerebellum and stomach were extracted and processed for light and electron microscopic examination. For the light microscope serial sections obtained, stained with H&E (Testis, Cerebellum and Stomach); Feulgen stain (Testis) and silver stain (Modified Glees) (Cerebellum). An image analyzer was used to assess cellular apoptosis, and in addition, DNA cytometry was performed.

14 Results

15 I- Effect of Acrylamide on testis Animals that received 25mg/kg BW/10days showed mild affection. Animals that received 50mg/kg BW/10days showed evident damage especially with the intraperitoneal administration. This was in the form of degeneration of germ cells, numerous multinucleated giant cells with sloughed seminiferous epithelium, and vacuolations inbetween the germ cells.

16 Control 50mg/kg/10days/ I.P.

17 Control 50mg/kg/10days/ I.P.

18 Control 50mg/kg/10days/ I.P.

19 Control 50mg/kg/10days/ I.P.

20 Control 50mg/kg/10days/ I.P.

21 50mg/kg/10days/ I.P. 50mg/kg/10days/ P.O.

22 Apoptosis Feulgen stain was performed to assess the effect of acrylamide on cellular apoptosis in the testis. It was clearly proved that acrylamide at a dose of 50mg/kg/10days showed increased apoptosis. Control group 5/10 25mg/kg/10days whether orally or intraperitoneally 6/10 50mg/kg/10days/orally 9/10 50mg/kg/10days/intraperitoneally 15/10

23

24 Study of DNA Cytometry Control 25mg/kg/10days whether orally or intraperitoneally 50mg/kg/10days/orally 50mg/kg/10days/ intraperitoneally

25 II- Effect of Acrylamide on Cerebellum The Purkinje cells were target cells to acrylamide. Rats treated at a dose of 25mg/kg/10days showed that Purkinje cells somata appeared unaffected, while degeneration of their dendrites and axons were evident in the molecular and white matter respectively. In addition, Rats treated with 50mg/kg/10days showed argyrophilic degenerated Purkinje cells somata.

26 Control

27 25mg/kg/10days/Orally 25mg/kg/10days/I.P.

28 50mg/kg/10days/Orally 50mg/kg/10days/ I.P.

29 Control Control 25mg/kg/10days/I.P. 50mg/kg/10days/P.O.

30 III- Effect of Acrylamide on Stomach The stomach of animals treated with acrylamide in a dose of 25mg/kg/10days showed no gastric affection, while there were mild degenerative changes and an apparent increase in mucous secreting cells in the group that received 50mg/kg/10days.

31 Control 50mg/kg/10days/IP

32 Control 50mg/kg/10days/Ì.P.

33 Control 50mg/kg/10days/I.P.

34 Conclusion

35 The present study expands the available information concerning the hazards carried by the consumption of acrylamide on testis, cerebellum and stomach. Although the doses of acrylamide utilized in the present investigation were higher than the average dietary daily intake in humans, µg/kg body weight/day, yet the cumulative effects of such toxicant on human health are still waiting to be fully identified. Further studies focusing on the influence of acrylamide on different organs in smaller doses for prolonged periods could aid in the full understanding of hazards implicated by acrylamide.

36

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