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Animal Experiment: Male Wistar Rats Essay

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MATERIALS AND METHODS
Animals
4-6 weeks old male Wistar rats weighing 80-120 g at study initiation, were obtained/purchased from the Animal House of Ahvaz Jondishapour University of Medical Sciences (AJUMS), Ahvaz, Iran. The rats were housed in an animal room maintained at 23 ± 3 °C and a relative humidity of 30-70 % with the altering light/dark cycle of 12h during the experiment and for at least, one week prior to sensitization period (for acclimatization purposes). The animals were housed in stainless-steel wire cages in groups of 5 and had free access to PN-free standard laboratory rodent chow and water.
All protocols involving the examined animals, were orchestrated in conformity with the instructions for Laboratory Animal Experiments in AJUMS Animal Research & Care Center.

Reagents
The reagents used in this research, were purchased from the sources enclosed in parentheses:
Alum [Al(OH)3] (Alhydrogel 2.0%, Serva Chemical Co., U.S.A), Cholera toxin (C-3012, Sigma Chemical Co., St. Louis, Mo, U.S.A), DMSO (Merck Chemical Co., Germany), Evan's blue dye (Merck Chemical Co., Germany), Rat histamine kit (LDN Chemical Co., Germany), K3-EDTA (Sigma Chemical Co., St. Louis, Mo, U.S.A), PBS (Merck Chemical Co., Germany), Rat Total IgE kit (ICL Chemical Co, U.S.A).
Unshelled/incrusted crude PNs were supplied from Safee-Abad Tree-plantar Research Station in the town of Dezfool, Khuzestan Province, Iran.

Antigen/Allergen Preparation
In the present study, PN proteins were used as allergens which were extracted from fresh crude PNs as follows, briefly:
PN bodies were pulverized by a mill and the resulted paste was defatted by n-Hexane (1:3 v/v, 3 times). Subsequent to separation process, residues were dried out and deodorized via gentle he...


... middle of paper ...


...t response was defined as a wheal reaction showing up as a blue area/circle measuring greater than 5 mm in diameter when read at 20 min post id PN-challenge.

Detection of Vascular Leakage
Five min before intra-peritoneal PN challenge-dose adminstration, 7 rats from each group received 200 μL of Evan's blue dye (5 mg/mL PBS) by tail vein injection. Footpads (& paws) of mice were examined for signs of vascular leakage (visible blue color) 40-45 min after ip administering of 200 μg of the filter-sterilized CPE.

STATISTICAL ANALYSIS
Data were analyzed using the SigmaStat statistical software package (SPSS, Chicago, Ill, Version 15). Statistical differences between two groups were analyzed by 1-way ANOVA. A probability value of less than 0.05 was considered statistically, significant.



Works Cited

Adjuvant, Anaphylactic Parameters, Peanut Allergy, Wistar-strain Model


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