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In the present study, we determined the significance of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in Alzheimer's disease (AD). We characterized the expression of TRAIL protein in the cerebrospinal fluid (CSF) and serum with ELISA and TRAIL mRNA in the peripheral blood mononuclear cells (PBMCs) with real-time PCR in 22 patients with AD and 20 control cases. We could not find TRAIL protein in the CSF samples. The concentration of TRAIL protein in sera from patients with AD was not different from controls. However, there was an inverse correlation between serum TRAIL levels and Mini-Mental State Examination scores in AD patients. Also we did not find significant difference in TRAIL mRNA in the PBMCs of patients with AD when compared with control group. Our data indicate that TRAIL serum level decreases in the late stage of disease. [ABSTRACT FROM AUTHOR]

The main pathophysiology in cerebral ischemia is the structural alteration in the neurovascular unit, coinciding with neurovascular matrix degradation. Among the human matrix metalloproteinases (MMPs), MMP-2 and -9, known as gelatinases, are the key enzymes for degrading type IV collagen, which is the major component of the basal membrane that surrounds the cerebral blood vessel. In the present study, we investigated the effects of resveratrol on cytotoxicity, reactive oxygen species (ROS), and gelatinases (MMP-2 and -9) in human cerebral microvascular endothelial cells exposed to 6 hours of oxygen-glucose deprivation and a subsequent 24 hours of reoxygenation with glucose (OGD/R), to mimic ischemia/reperfusion in vivo. Lactate dehydrogenase increased significantly, in comparison to that in the normoxia group. ROS was markedly increased in the OGD/R group, compared to normoxia. Correspondingly, ROS was significantly reduced with 50 μM of resveratrol. The proMMP-2 activity in the OGD/R group showed a statistically significant increase from the control cells. Resveratrol preconditioning decreased significantly the proMMP-2 in the cells exposed to OGD/R in comparison to that in the OGD/R group. Our results indicate that resveratrol regulates MMP-2 activity induced by OGD/R via its antioxidant effect, implying a possible mechanism related to the neuroprotective effect of resveratrol.

We describe the organization of wet-lab special-study modules (SSMs) in the Central Research Laboratory of Dokuz Eylül Medical School, Izmir, Turkey with the aim of discussing the scientific, laboratory, and pedagogical aspects of this educational activity. A general introduction to the planning and functioning of these SSMs is given, along with specific examples. The wet-lab SSMs incorporate several innovative pedagogies: problem-based learning, research-based learning, practical laboratory education, team-based learning, and project-based learning. Oral and written evaluations show that the students find this activity rewarding. The wet-lab SSM model applied in the Research-Lab of Dokuz Eylül School of Medicine represents a format which is effective in training the students in research methodology, practical laboratory work, and independent learning.

Erythropoietin (Epo) exerts neuroprotective, glioprotective, and vascular protective effects in the nervous system. However, the mechanisms of the cytoprotective effect of Epo have not been fully clarified. Here, we investigated whether Epo affects the transcription and activation of nuclear factor erythroid 2-related factor 2 (Nrf2), which is a key transcription factor of the cellular anti-oxidant defense system, and mRNA expression of its target genes including heme oxygenase-1 (HO-1). Epo was added to SH-SY5Y cells at 1 U mL(-1) and cultures were incubated for 24 h and then mRNA expression of Nrf2 target genes were analyzed with real-time PCR. SH-SY5Y cells were incubated with Epo at different time points and the nuclear and cytoplasmic levels of Nrf2 protein expression were examined by Western blotting and immunohistochemistry. Specific inhibitors of mitogen-activated protein kinases (MAPKs) and phosphatidylinositol-3 kinase (PI3K) were used to find out the possible signaling pathways that mediate the activating effect of Epo on Nrf2 activation. In cultured human SH-SY5Y neuroblastoma cells, Western blotting, immunohistochemistry, and real-time PCR analysis demonstrated that Epo-induced nuclear translocation of Nrf2 and upregulates HO-1 expression. Inhibitors of MAPKs and PI3K decreased Epo-induced nuclear translocation of Nrf2 and HO-1 mRNA expression. These results suggest that Epo induces neural HO-1 expression through the activation of PI3K, MAPK, and Nrf2 pathways, and this may unveil a novel mechanism which mediates the cytoprotective responses elicited by Epo., (Copyright (c) 2010 John Wiley & Sons, Ltd.)

Endothelial cells lining the inner blood vessel walls play a key role in the response to hypoxia, which is frequently encountered in clinical conditions such as myocardial infarction, renal ischemia and cerebral ischemia. In the present study we investigated the effects of hypoxia and hypoxia/reoxygenation on gelatinases (matrix metalloproteinase-2 and -9), their inhibitor (TIMP-2) and activator (MT1-MMP), in human umbilical vein endothelial (HUVE) cells. HUVE cells were subjected to 4 h of hypoxia or hypoxia followed by 4 and 24 h of reoxygenation. The pro- and active forms of MMP-2 and MMP-9 were analyzed by gelatin zymography; TIMP-2 protein level was assayed using ELISA, while MT1-MMP activity was measured using an activity assay. The secretion of MMP-2 proform increased significantly in cells subjected to 4 h of hypoxia followed by 4 or 24 h of reoxygenation, compared with the normoxic group. TIMP-2 protein level also increased significantly in the hypoxia/reoxygenation groups, compared with the normoxic group. There were no statistically significant differences in the levels of active MT1-MMP in all groups. This study indicates that MMP-2 and TIMP-2 could be regarded as important components of a mechanism in the pathophysiology of ischemic injury following reperfusion.

The definite diagnosis of Alzheimer's disease (AD) is based on post mortem pathological examination. To date, there is no laboratory test that can discriminate AD patients from healthy individuals. In the perspective of recent knowledge, there are three cerebrospinal fluid (CSF) markers which have the highest sensitivity and specificity: A beta(1-40), A beta(1-42), and p-tau. In the present study, 15 'Probable Alzheimer's Disease' (PAD) patients and 15 control subjects were included. PAD patients were selected from the patients of Dokuz Eylül University Neurology Department Dementia outpatient clinic and control subjects were selected from the patients who were undergone epidural anesthesia because of any surgical operation. The concentrations of Ab1-40, Ab1-42, and p-tau in CSF were quantified by using ELISA. Also, the effects of 'PAD patients' CSF on the survival of PC12 cell line were assessed. There was a significant decrease of Ab1-40 and increase of p-tau in patients with AD when compared with controls. Ab1-42 concentration was not significantly different between groups. There was a positive correlation between duration of the disease and CSF of p-tau concentration in patients with AD. There was no significant difference in cell line viability values between groups., ((c) 2009 John Wiley & Sons, Ltd.)