effect of acrylamide on stomach, cerebellum and testis of the albino rat (ppt) 3.12.2008

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Acrylamide has several effects toxic and carcinogenic. 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. The stomach, testis and cerebellum specimens were collected form fifty adult male albino rats. The animals were divided into two main groups. Group (I) was subdivided into 3 subgroups, control, Ia that received acrylamide in a dose of 25mg/kg/10days (P.O.) and Ib that received acrylamide in a dose of 25mg/kg/10days (I.P.). Group (II) was subdivided into 3 subgroups, control, IIa that received acrylamide in a dose of 50mg/kg/10days (P.O.) and IIb that received acrylamide in a dose of 50mg/kg/10days (I.P.). At the end of the experiment, all rats were anesthetized using ether inhalation. Specimens from testis, cerebellum and stomach were extracted and processed for light and electron microscopic examination. For the light microscope serial sections 5-6µm thick obtained, stained with H&E (Testis, Cerebellum and Stomach); Feulgen stain (Testis) and silver stain (Modified Glees) (Cerebellum). Testicular specimens were studied with the Image analysis technique so as to assess cellular apoptosis. Moreover, DNA cytometry of the basal compartment of the testis was performed.In the present work, it was found that the testis of the group treated with a dose of 25mg/kg/10days showed mild affection whether acrylamide was administered orally or intraperitoneally. On the other hand, the testis of the group treated with a dose of 50mg/kg/10days showed damage especially with the intraperitoneal administration in comparison to the oral treatment. This was in the form of degeneration of germ cells, numerous multinucleated giant cells with sloughed seminiferous epithelium, and vacuolations in-between the germ cells. In addition, 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.Image analysis of the feulgen stained sections was performed to evaluate the carcinogenicity of acrylamide on testis. It was found that acrylamide at a dose of 50mg/kg/10days showed increased abnormal DNA content in cells, which was considered a reliable marker for malignancy. As regards effect of acrylamide on the cerebellum, it was concluded that the Purkinje cells were target cells to the acrylamide. Rats treated at a dose of 25mg/kg/10days showed that Purkinje cells somata appeared unaffected, while degeneration of Purkinje cells dendrites and corresponding axons were evident in the molecular and white matter respectively. Furthermore, rats treated with 50mg/kg/10days showed argyrophilic Purkinje cells somata and their dendrites were clearly evident in the Purkinje cell and molecular layers respectively. Also, Purkinje cell axon degeneration was observed in the white matter in the form of fragmented, linearly arrayed debris. The intraperitoneal treatment showed more damage as compared to the oral one.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.The present study pointed out the hazards of acrylamide and its possible effect on human health. Recommendations are necessary to decrease acrylamide level in different foods and ways to decrease acrylamide formation during preparation of the different foods should be advertised.

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    Effect of

    Acrylamide

    on

    Stomach, Cerebellum

    and Testis of the Albino

    Rat

    By

    Hesham

    Noaman

    Abdel

    Raheem

    M.D. THESIS

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    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

    (Professorof Anatomy).

    Doctor:

    Shahira

    Samir

    Zaki

    (Assistant

    Professor of Anatomy).

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    Introduction

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    Acrylamide

    Chemistry

    Uses

    CH2=CHCONH2

    water treatment,

    enhanced oil recovery,

    papermaking aids,

    soil conditioning agents,

    sewage and waste treatment,

    ore processing,

    permanent

    -

    press fabrics.

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    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:

    0.3

    -

    0.8

    g/kg

    BW/day.

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    0

    5

    10

    1520

    25

    30

    ug/day

    Biscuit / Cookies

    All Other Foods

    Exposure to

    Acrylamide

    in Food

    French Fries &

    Potatoes

    Breads

    Potato Chips

    Cereal

    Coffee

    Cakes

    Dried Foods

    Pop Corn

    Salty Snacks

    Chocolate

    Nuts/Seeds/

    Butters

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    Effect of temperature

    Acrylamide levels in potato chips friedfor 4 minutes increased with frying

    oil temperature. (ppb=parts per billion)

    160 C

    27ppb

    170 C

    70ppb

    180 C

    326ppb

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    Effect of time

    Acrylamide levels in potato chips fried at 180oCincreased with frying time.

    3.5 m

    12ppb

    4 m

    46ppb

    4.5 m

    227ppb

    5 m

    973ppb

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    Aim of thework

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    The current project aimed at exploring the harmfuleffects 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.

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    Material and

    Methods

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    The stomach, testis and cerebellum specimens

    were collected form fifty adult male albino rats.

    Animals were divided to two main groups: I, IIeach group was subdivided into 3 subgroups.

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    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

    cytometrywas performed.

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    Results

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    I- Effect of Acrylamide on testis

    Animals that received 25mg/kg BW/10daysshowed mild affection.

    Animals that received 50mg/kg BW/10days

    showed evident damage especially with theintraperitoneal administration. This was in the

    form of degeneration of germ cells, numerous

    multinucleated giant cells with sloughed

    seminiferous epithelium, and vacuolations in-between the germ cells.

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

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    Control

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

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

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

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    Control

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

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

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    ApoptosisFeulgen 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

    intraperitoneally6/10

    50mg/kg/10days/orally 9/10

    50mg/kg/10days/intraperitoneally 15/10

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    St d f DNA C t t

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    Control 25mg/kg/10days whether orally or intraperitoneally

    50mg/kg/10days/orally 50mg/kg/10days/ intraperitoneally

    Study of DNA Cytometry

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    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, whiledegeneration of their dendrites and axons were evidentin the molecular and white matter respectively.

    In addition, Rats treated with 50mg/kg/10days

    showed argyrophilic degenerated Purkinje cells somata.

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    Control

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    25mg/kg/10days/Orally

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

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

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    Control

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

    Control

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

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    III- Effect of Acrylamide on

    StomachThe 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.

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    50mg/kg/10days/IP

    Control

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    Control

    50mg/kg/10days/.P.

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

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    Conclusion

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    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, 0.4-5 g/kg body

    weight/day,yet the cumulative effects of suchtoxicant on human health are still waiting to be fully

    identified.

    Further studies focusing on the influence of

    acrylamide on different organs in smaller doses forprolonged periods could aid in the full

    understanding of hazards implicated by acrylamide.

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