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 Although individual and experimental variations w Empty Although individual and experimental variations w

Сообщение  wangqian в Чт Май 15, 2014 12:11 pm

In the first session, supplier ABT-888 the mice were placed into the open field cham ber with two identical, non toxic objects, 12 cm from the back and sidewalls of the open field box, and 16 cm apart from each other. A 8 min session, in which the time explor ing each object was recorded, an area 2 cm2 surrounding the object is defined such that nose entries within 2 cm of the object were recorded as time exploring the object. The animal was then returned to its home cage, and either 24 or 4 h later, placed into the open field box again. This time, there were two new objects, one identical to the previous objects, and one novel object. The mice were given another 6 min to explore, and the amount of time exploring each object was recorded. Mice that spent 7 s exploring the objects were omitted from the analysis.<br><br> Results were recorded as an object discrimination ratio, which is calculated by dividing the time the mice spent exploring a novel object, divided by purchaseAfatinib the total amount of time exploring the two objects. Synaptosomes preparations and Western blot analysis For synaptic preparations, isolated hippocampi were homogenized in Hepes sucrose buffer supplemen ted with protease and phosphatase inhibitors. Homoge nates were centrifuged at 800 g for 10 min. The supernatant was separated into supernatant and pellet by spinning at 9,200 g for 15 min. P2 contains the crude synaptosomal fraction. Synaptosomes fractions were analyzed by western blot using the following anti bodies, APP to detect mAPP and imAPP, APPCTF to detect CTF and B CTF, BRI2 to detect mBri2.<br><br> It is well established that both cerebral hypoperfusion stroke and type 2 diabetes are risk factors for Alzhei mers disease. Oxidative and supplier AG-1478 nitrosative stres ses are common denominators for these age related diseases. Oxidative stress is associated with b amy loid peptide accumulation in the brains of AD patients. Ab is generated by sequential proteolytic cleavages of the transmembrane amyloid precursor pro tein by two membrane bound proteases, b secretase and the g secretase complex com posed of presenilin 1, nicastrin, APH 1 and PEN 2. In AD brains, the specific regions affected by Ab deposition correlate with increased BACE1 protein levels and activity. Together with the observation that amyloid pathology was diminished in mice deficient in BACE1, these findings strongly suggest that BACE1 elevation leads to enhanced Ab production and deposition in AD.<br><br> Given the central role of Ab in AD pathogenesis and the fact that BACE1 is the rate limit ing enzyme in APP processing and Ab generation, BACE1 remains one of the most important therapeutic targets for treating AD. Compelling evidence indicates that BACE1 expression is tightly regulated at both the transcriptional and transla tional levels. A number of transcriptional factors have been identified that positively or negatively regulate BACE1 gene expression under both basal and cell stressed conditions, such as inflammation. Some studies suggest that BACE1 is regulated by specific microRNAs, post transcriptionally. In addition to inflammation, other conditions have also been shown capable of causing increased BACE1 expression in the brain, including oxida tive stress, traumatic brain injury, hypoxia and ische mia.

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