Anti-Tau monoclonal antibody, clone 3C12

Infection of rat pulmonary microvascular endothelial cells with the bacterium Pseudomonas aeruginosa induces the production and release of cytotoxic oligomeric tau and beta amyloid (A beta). Here, we characterized these cytotoxic amyloids. Cytotoxic behavior and oligomeric tau were partially resistant to digestion with proteinase K, but cytotoxicity was abolished by various denaturants including phenol, diethylpyrocarbonate (DEPC), and 1,1,1,3,3,3-hexafluoro-2-isopropanol (HFIP). Ultracentrifugation for 8 h at 150 000 g was required to remove cytotoxic activity from the supernatant. Ultracentrifugation, DEPC treatment, and immunodepletion using antibodies against A beta also demonstrated that cytoprotective protein(s) are released from endothelial cells during P. aeruginosa infection. Mass spectrometry of endothelial cell culture media following P. aeruginosa infection allowed identification of multiple potential secreted modulators of A beta, including cystatin C, gelsolin, and ApoJ/clusterin. Immunodepletion, co-immunoprecipitation, and ultracentrifugation determined that the cytoprotective factor released during infection of endothelial cells by P. aeruginosa is cystatin C, which appears to be in a complex with A beta. Cytoprotective cystatin C may provide a novel therapeutic avenue for protection against the long-term consequences of infection with P. aeruginosa.

Background: Thymoquinone (TQ), a biologically active ingredient of Nigella sativa, has anti-inflammatory, antioxidative and neuroprotective properties. Therefore, it could be a good candidate in the recovery of Alzheimer's disease (AD) pathology rather than current symptomatic reliefs. Purpose: In the present study, we examined the molecular healing effects of TQ in amyloid beta 1-42 (A beta(1-)(42)) peptide-infused AD rat hippocampus. Study design: A micro-osmotic pump containing aggregated A beta(1-)(42) was cannulated into the hippocampus of adult female rats. After two weeks infusion, the dose of TQ (10 mg/kg or 20 mg/kg) was determined according to the HPLC results of cerebrospinal fluid and TQ was given to rats intragastrically for 15 days. Methods: The memory performance of rats was determined by Morris water maze test. Afterwards, the acetylcholinesterase (AChE) level were measured by ELISA. Histopathological examinations of hippocampal tissue were performed for cell survival by Nissl staining, for detection of amyloid plaque deposits by Congo red staining and for determination of degenerating neurons by Fluoro Jade C staining. MicroRNA/mRNA levels and protein expressions of AD-related genes and proteins were analyzed by Real-Time Polymerase Chain Reaction and Western Blotting, respectively. Results: Administration of TQ enhanced the memory performance of A beta(1-)(42 )infused rats and it also ameliorated the neuronal loss in the cornu ammonis (CA1), but not in the dentate gyrus (DG). In addition, TQ treatment decreased the fibril deposition whose accumulation was significantly higher in the A beta(1-)(42)-infused animals compared to that of the control group. The expression profiles of mir29c and Box which significantly upregulated in the A beta(1-)(42)-infused animals were attenuated by TQ. Furthermore, administration of TQ decreased the expressions of A beta, phosphorylated-tau, and BACE-1 proteins. There was no significant therapeutic effect of TQ on the AKT/GSK3 beta or MAPK signaling pathways which were affected due to A beta(1-)(42) infusion. Conclusion: TQ has the capacity to recover the neuropathology by removing A beta plaques and by restoring neuron viability. All might have established the molecular basement of the consolidation in the memory observed by means of TQ treatment.