Effects of L-proline on cellular responses of hen erythrocytes subjected to thermal stress.

Little is known on the protective effects of L-proline on hen erythrocytes. The aim of the study was to determine the protective effects of this amino acid at concentrations of 50 µg/mL, 100 µg/mL, 200 µg/mL in hen erythrocytes subjected to temperatures 41 °C, 43 °C and 45 °C for 1 h and 4 h. The following cellular parameters were determined: viability, morphological alterations, caspase 3/7 activity, heat shock protein HSP70 1A activity and glutathione level. The results showed that exposure to 43 °C and 45 °C resulted in a decrease of viability and increased morphological alterations of the non-treated erythrocytes. Caspase 3/7 activity was increased only at 45 °C, however HSP70 1A activity and glutathione level were increased in the temperature-dependent manner. On the other hand, erythrocytes additionally exposed to L-proline showed alterations of the parameters when compared to the non-treated cells. L-proline at 50 µg/mL and 100 µg/mL increased caspase 3/7 activity at both 41 °C and 43 °C, however it was less augmented at all the concentrations at 45 °C. Glutathione level was decreased in heat-stressed (at 43 °C and 45 °C) hen erythrocytes treated with L-proline (at 50 µg/mL and 100 µg/mL) but it was increased at 200 µg/mL. HSP70 1A activity was augmented in a concentration- and temperature-dependent manner. The results indicate that proapoptotic or antiapoptotic effects of L-proline depend on its concentration and temperature of heat stress and thermoprotective effects induced by the amino acid on some parameters in hen erythrocytes may be a result of stimulation of antioxidative defense and stimulation of HSP70 1A activity.

Activity and Safety of Inhaled Itraconazole Nanosuspension in a Model Pulmonary Aspergillus fumigatus Infection in Inoculated Young Quails.

Pulmonary aspergillosis is frequently reported in parrots, falcons, and other birds held in captivity. Inhalation is the main route of infection for Aspergillus fumigatus, resulting in both acute and chronic disease conditions. Itraconazole (ITRA) is an antifungal commonly used in birds, but its administration requires repeated oral dosing, and the safety margin is narrow. To investigate the efficacy of inhaled ITRA, six groups of ten young quails (Coturnix japonica) were inoculated intratracheally with 5 × 10(6) spores (3 groups) or 5 × 10(7) spores (3 groups). Animals were exposed to nebulized ITRA nanosuspension as 10 % suspension or 4 % suspension, once daily for 30 min, starting 2 h after inoculation for 6 days. Control groups were exposed to nebulized saline for the same period of time. Survival and clinical scores were evaluated, and animals were subjected to gross pathology. In control animals, aspergillosis resulted in systemic disease without pulmonary or air sac granulomas. Animals died from multiple organ failure. Inhalation of 10 % ITRA nanosuspension blocked lethality and prevented disease-related symptoms in the quails exposed to the low dose of spores, while the disease course in quails inoculated with the high-spore dose was retarded. Inhalation of 4 % ITRA nanosuspension was less effective. Both inhalations were well tolerated, and gross pathology did not reveal signs of local toxicity. The data indicate that inhaled administration of 10 % ITRA nanosuspension is capable of alleviating an acute A. fumigatus infection in quails. A lower ITRA concentration may be only active in chronic pulmonary aspergillosis.