4.6.2. Anti-Inflammatory Activity

The anti-inflammatory activity of the polysaccharides and their PECs was evaluated on three mouse hind paw edema tests induced by histamine (exudative inflammation), concanavalin A (B-cells dependent inflammation) or delayed type hypersensitivity reaction (T-cells dependent inflammation) [65]. Control and experimental groups consisted of 8-10 animals each. In the histamine and concanavalin A tests of inflammation the substances were injected intraperitoneally (i.p.) in water solutions at doses 10.0 mg/kg (PECs) and 9.0 mg/kg (original κ-CGN or CH). In the test of delayed type hypersensitivity reaction (DTH) the substances were administered intraperitoneally at doses 5.0 mg/kg (PECs) and 4.5 mg/kg (κ-CGN or CH). The last dose values were equal to the highest quota of each polysaccharide in appropriate PEC. The dose of reference substances (indomethacin or diclofenac, "Fluka BioChemica" (Sigma-Aldrich, Saint Louis, MO, USA) was 50 mg/kg [66]. The control group of animals received water. At the end of experiment, the animals were sacrificed by cervical dislocation, the mouse hind paws were cut o ff at the ankle joint and weighed. The ratio of the di fference in weight between the treated and untreated hind paws to the weight of the untreated hind paw was used as an inflammatory edema index (IEI). The anti-inflammatory activity (AIA) was presented as a di fference between 100% and percentage of inflammation index versus to the control group.

#### 4.6.3. Histamine-Induced Mouse Paw Edema Test

The outbred male albino mice were injected with 0.05 mL 0.01% histamine in saline solution into the plantar of the hind paw. The collateral paw was injected with saline solution. The tested compounds (κ-CGN, CH, and their PECs) were administered i.p. one hour before the histamine injection. The reference agen<sup>t</sup> indomethacin was administered by the same way at the dose 50 mg/kg. The animals were sacrificed 5 h after the histamine injection.

#### 4.6.4. Concavaline A-Induced Mouse Paw Edema Test

The male mice C57Bl/6 were injected with 0.02 mL 0.5% concanavaline A (Sigma-Aldrich, Saint Louis, MO, USA) in saline solution into the plantar of the hind paw. The collateral paw was injected with saline solution. The tested compounds were administered i.p. one hour before the concanavaline A injection. The reference agen<sup>t</sup> diclofenac was administered by the same way at the 50 mg/kg dose. The animals were sacrificed one hour after the concanavaline A injection.

#### 4.6.5. Delayed Type Hypersensitivity Reaction (DTH) Test

The tested compounds were administered intraperitoneally to the male mice C57Bl/6. One day later the sheep erythrocytes suspension (10<sup>8</sup> cells) was injected i.p. to control and experimental groups. At the fifth day after immunization the animals were subplantar injected with the sheep erythrocytes suspension (10<sup>8</sup> cells) into the hind paw. The collateral paw was injected with saline solution. The animals were sacrificed by cervical dislocation 24 h after the last antigen injection
