Polyunsaturated Fatty Acids and Modulation of Cholesterol Homeostasis in THP-1 Macrophage-Derived Foam Cells

Transformation of macrophages to foam cells is determined by the rates of cholesterol uptake and efflux. This study uses a real time RT-PCR technique to investigate the role of conjugated linoleic acid (CLA), α-linolenic acid (ALA) and eicosapentaenoic acid (EPA) in the regulation of the ATP-binding cassette A1 (ABCA1) and liver X receptor α (LXR) genes, which are involved in cholesterol homeostasis. Accordingly, these fatty acids significantly reduced the total, free and esterified cholesterols within the foam cells. While the expression of the ABCA1 and LXRα genes was increased in the presence of the pharmacological LXRα ligand, T0901317, their mRNA expression was not significantly affected by CLA, ALA and EPA. These results suggest that although polyunsaturated fatty acids have an effect on cholesterol homeostasis, they cannot change the expression of the ABCA1 and LXRα genes. Alternatively, several other genes and proteins may be involved

Preparation and Characterization of Monoclonal Antibody Against Truncated Recombinant Nuclear Matrix Protein (NMP22)

Background: Bladder cancer is a major worldwide health problem. Diagnosis of acute and chronic bladder carcinoma is based on the detection of a number of tumor markers such as nmp22 (nuclear matrix protein 22). Nmp22 is one of the tumor markers used for detecting the recurrence of bladder cancer. Objectives: The aim of this study was to develop hybrid cell producing monoclonal antibodies (MAbs), specifically for nmp22 using hybridoma technology. Materials and Methods: Complete and incomplete adjuvant with recombinant truncated nmp22 antigens were emulsified and injected to BALB/c mice. The spleen was removed and the splenocytes were fused with sp2/0 myeloma cells. Characterization of the produced mAb was carried out. Results: As a result of the fusion, hybridoma cells were produced and exhibited high- titer antibodies. By testing of colons based on the EnzymeLinkedImmunosorbent Assay (ELISA), 135hybridomacolonswere selected, out of whicheight high titer clones including 54D-6F-2E-36-7H-2A-8E-1D-6D were detected and one of the clones named FPR92 was selected, and the produced mAb was further characterized. The produced antibody belongs to the IgG class and its light chain was kappa. With respect to affinity, the mAb was included as high affinity 3�10-7 reacting with NMP22 recombinant protein .The western blot of cancerous bladder tissue showed the presence of 40 and 55 kD proteins as major bands that reacted with this mAb. Conclusions: Based on a double limiting dilution protocol, a type of monoclonal antibody, named FPR92 was produced and characterized. Keywords: Nmp22, Hybridoma, mAb, FPR92, Characterizatio

Preparation and preliminary studies of [64Cu]-antiMUC-1 for breast cancer targeting

PR81 is a monoclonal antibody that binds with high affinity to MUC1 that over expressed on breast tumors. PR81 is considered a suitable targeting molecule that was radiolabeled using Cu-64 for positron imaging studies. The monoclonal antibody was conjugated with DOTA moiety and after purification was evaluated for radiochemical purity, immunoreactivity, cell toxicity and structure integrity as well as biodistribution study in normal rats. The radiolabeled antibody prepared with acceptable radiochemical purity (> 93.2 ± 0.6 %, ITLC; specific activity; 4.6 µCi/µg), protein structure integration, significant cytotoxicity and significant immunoreactivity retention was assessed by radioimmunoassay (RIA). Animal biodistribution of the 64Cu-DOTA-PR81 was consistent with other radiolabeled antibodies. The results showed that 64Cu-DOTA-PR81 may be considered for tumor imaging for ultimate diagnosis and follow-up of MUC1 expression in oncology