Simultaneous zinc and vitamin A deficiency are common health problems in

Simultaneous zinc and vitamin A deficiency are common health problems in developing countries. level was significantly higher in both control and deficient groups than the serum IgA level. Results indicated that zinc and vitamin A deficiency is usually associated with a lower production of IgA. Micronutrient intervention strategies addressing IgA-related gastrointestinal infections are needed. = 11 common excess weight 27-29 g) and aged 6 weeks were assigned into two control and deficient groups. The control group (= 6) Cd151 were fed a normal diet containing sufficient retinyl acetate as vitamin A and zinc (5000 IU vitamin A/kg 60.2 mg zinc/kg) and the deficient group (= 5) maintained on a low vitamin A and zinc (550 IU vitamin A/kg supplemented in the form of retinyl acetate 5.2 mg zinc/kg) diet as the experimental group for a period of five months. There was no difference in lipid carbohydrate and protein content of the diets of groups. Also mineral and vitamin content of diets was kept constant except for vitamin A and zinc as mentioned above. The diet contained per 100 gram: Casein: 20 Sugar: 22.4 α-Corn starch: 44.6 Soybean oil: 5 Cellulose: 2 Mineral mixture: 5 and Vitamin mixture: 1. Composition of the mineral mixture was as follows: CaHPO4: 14.56 NaCl: 4.66 Fe-citrate: 3.18 MgSO4: 7.17 MnSO4: 0.12 CuSO4: 0.03 KI: 0.01 g/100 g of mixture. The vitamin mixture contained vitamin D3: 0.25 mg vitamin E acetate: 500 mg vitamin K3: 520 mg vitamin B complex: 2.2 gram vitamin C: 3 g and inositol: 600 mg per 100 g. The mineral and vitamin combination were free of zinc and vitamin A respectively. Zinc contamination was avoided by using deionized water and wearing polyethylene gloves for the preparation of animals’ diet. Blood collection At the end of the experiment mice were killed under deep anesthesia with Nembutal (Dainippon Pharmaceutical Co. Ltd. Japan). The blood collected via cardiac puncture and sera Moexipril hydrochloride was immediately collected after centrifugation at 5000 g and stored in -20°C until used. Preparation of intestinal mucosa extracts Segments of jejunum intestine were precisely removed from Moexipril hydrochloride the nearly 2 cm after stomach to the ileocecal valve. Mucosal piles were immediately weighed and maintained in 1 ml of cold phosphate buffer saline (PBS) and homogenized with a digital Teflon homogenizer Moexipril hydrochloride (AS ONE model: AN Japan) for 1 minute at 4000 g. After centrifugation for 20 minutes at 4°C with the speed of 13 500 g supernatants were used for IgA assessments. Determination of serum and intestinal mucosa total immunoglobulin A Serum and intestinal mucosa total IgA levels were determined by enzyme-linked immunosorbent assay (ELISA) method using a mouse IgA ELISA quantitation kit (Bethyl Laboratories Inc TX U.S.A) according to the manufacturer’s instructions. In brief goat anti-mouse IgA-affinity purified antibody was dispensed in each well and incubated for an hour at room temperature. They were then washed with a wash solution containing 50 mM Tris 0.14 M NaCl 0.05% Tween 20 for three times. After blocking non specific binding sites and incubating for 30 minutes at room temperature standards and samples were added and incubated for 60 minutes at room temperature. Horseradish peroxidase-conjugated Goat anti-mouse IgA was added to each well and incubated for 60 min at room temperature. Finally after addition of an appropriate substrate absorbance was read with ELISA reader (Corona Electric MTP-32 Japan) at 450 nm. Determination of serum zinc and retinol The concentration of zinc in serum was determined using an inductively coupled plasma mass spectrometer (ICP-MS; model 8500 Shimadzu Tokyo Japan) as previously described [23]. Vitamin A measurement in serum was carried out by high performance liquid Moexipril hydrochloride chromatography (HPLC) according to a method described previously [24] using a HPLC system (Shiseido Co. Ltd Tokyo Japan). Statistics Data were first checked for distribution using Kolmogorov-Smirnov test. Statistical analysis was done using independent sample < 0.05. Results Animal growth and food intake As shown in Fig. 1 there were no significant differences in final Moexipril hydrochloride body weights among two groups. Animals kept on a low vitamin A and zinc diet (deficient group) had significantly more (< 0.05) food intake as compared to the controls. Fig. 1 Mean ± SD of food intake and body weight.