Inflammatory Stress Processes Research Articles

An adaptation for exhaled breath condensate collection in rabbits

ABSTRACTAim: To set up and test the feasibility of a handmade apparatus adapted for exhaled breath condensate (EBC) collection in medium-sized animals. Materials and Methods: The apparatus was produced using an 18-mm thick u-shaped borosilicate glass. The u-shaped tube body is 25 cm in diameter, and the horizontal portions are 12 cm in diameter. The base consists of a tube joint 14/20 or 14 mm thick by 20 cm in diameter, and has a length of 5 cm. This has a hole that is plugged for condensate flow to a 1.5 mL polypropylene microtube that stores the condensate during collection. Was placed inside a styrofoam box and immersed in crushed ice and salt to ensure cooling. The temperature was monitored and maintained throughout the collection at −10°C. One of the outputs of the u-shaped tube was connected to the expiratory limb of the ventilator. Results: An experimental model of ALI, induced by oleic acid (OA) was adopted to determine the concentration of biomarkers of oxidative stress: malondialdehyde (MDA), glutathione (GSH), and nitrite/nitrate (NOx). The proposed model allows measurement of NOx, MDA, and GSH. However, the NOx and MDA levels in the EBC were not significant. It was only possible to observe an upward trend, which suggests a temporal evolution of the presence of these markers in the EBC. Conclusion: The EBC collection method adapted is effective to generate sufficient content that allows to determine the levels of different biomarkers, such as NOx, MDA, and GSH, that are involved in oxidative and inflammatory stress processes during respiratory diseases.

Effect of transient cerebral ischemia on the expression of receptor for advanced glycation end products (RAGE) in the gerbil hippocampus proper.

The receptor for advanced glycation end products (RAGE) is a multi-ligand receptor of the immunoglobulin superfamily that has been implicated in multiple neuronal and inflammatory stress processes. In this study, we examined changes in RAGE immunoreactivity and its protein levels in the gerbil hippocampus (CA1-3 regions) after 5min of transient global cerebral ischemia. The ischemic hippocampus was stained with cresyl violet, neuronal nuclei (a neuron-specific soluble nuclear antigen) antibody and Fluoro-Jade B (a marker for neuronal degeneration). 5days after ischemia-reperfusion, delayed neuronal death occurred in the stratum pyramidale of the CA1 region. RAGE immunoreactivity was not detected in any regions of the CA1-3 regions of the sham-group; the immunoreactivity was markedly increased only in the CA1 region from 3days after ischemia-reperfusion. On the other hand, RAGE immunoreactivity was newly expressed in astrocytes, not in microglia. Western blot analysis showed that RAGE protein level was highest at 5days post-ischemia. In brief, both the RAGE immunoreactivity and protein level were distinctively increased in astrocytes in the ischemic CA1 region from 3days after transient cerebral ischemia. These results indicate that the increase of RAGE expression in astrocytes after ischemia-reperfusion may be related to the ischemia-caused activation of astrocytes in the ischemic CA1 region.

Effect of transient cerebral ischemia on the expression of receptor for advanced glycation end products in the gerbil hippocampus proper

The receptor for advanced glycation end products (RAGE) is a multiligand receptor of the immunoglobulin superfamily that has been implicated in multiple neuronal and inflammatory stress processes. In the present study, we investigated changes in RAGE immunoreactivity and its protein levels in the gerbil hippocampus (CA1 to 3 regions) after 5 minutes of transient global cerebral ischemia.

Neuronal RAGE expression modulates severity of injury following transient focal cerebral ischemia

Inflammation has a significant role in the neurological injury that follows stroke. The receptor for advanced-glycation end products (RAGE) is a multiligand member of the immunoglobulin superfamily that has been implicated in multiple neuronal and inflammatory stress processes. To directly test the role of neuronal RAGE in stroke, we employed two cohorts of transgenic mice, one over-expressing full-length functional human RAGE in neurons, and the other a human RAGE transgene in which deletion of the cytoplasmic domain of the receptor in neurons suppresses signal transduction stimulated by ligands (referred to as dominant negative or DN-RAGE). We found a statistically significant increase in stroke volume in the RAGE over-expressing cohort compared to normal controls, and a trend towards decreased stroke volume in the DN RAGE cohort. These results indicate that RAGE signaling directly contributes to pathology in cerebral ischemia.