Seroprevalence of human T lymphtropic virus (HTLV) among tissue donors in Iranian tissue bank

Iranian Tissue Bank prepares a wide range of human tissue homografts such as; heart valve, bone, skin, amniotic membrane and other tissues for different clinical applications. The purpose of this study was to determine the seroprevalence of HTLV in tissue donors. About 1,548 tissue donors were studied during a 5-years period by ELISA assays. HTLV_1,2—antibodies were tested for all of donors with other tests upon American Association of Tissue Banks (AATB) standards. About 25 (1.61%) out of 1,548 tissue donors were HTLV positive that 17 donors were male and 8 were female (female/male ratio was approximately 47%). Regarding to the prevalence of HTLV among tissue donors and importance of cell and tissue safety and quality assurance, we recommend that all blood, cell and tissue banks should be involved both routine serological methods and other complementary tests such as polymerase chain reaction (PCR) for diagnosis of HTLV.     

Cyclooxygenase-2 inhibition by novel Bisaryl imidazolyl imidazole derivatives increases Bax/Bcl-2 ratio and upregulates Caspase-3 gene expression in Caco-2 colorectal cancer cell line

Cyclooxygenase-2 (COX-2) inhibitors including celecoxib inhibit cell growth and induce apoptosis in cancer cells. In this study, the relation of Bax (an apoptosis promoter) to Bcl-2 (an apoptosis inhibitor) ratio with the apoptosis co-ordination enzyme, caspase-3 was investigated in correlation with the treatment of 4,5-bisaryl imidazolyl imidazoles as novel selective COX-2 inhibitors in Caco-2 colorectal cancer cells. Recently, the organic reactions under microwave irradiation attracted attention of scientists due to their high reaction rate, mild reaction conditions and the formation of clean products. Therefore, a microwave-assisted method was used to synthesize our compounds. The effects of these COX-2 inhibitors on the proliferation of Caco-2 cells were evaluated by MTT assay. cDNA microarray and clustering analysis were used to evaluate effects of our synthetic compounds on gene expression pattern of 112 genes involved in apoptosis pathways. Bax, Bcl-2 and caspase-3 mRNA expression and their relationship were analyzed by quantitative real-time PCR. Results indicated that proliferation of Caco-2 cells after treatment with 4,5-bisaryl imidazolyl imidazoles on Caco-2 cells were time and dose dependent. We conclude that increase in Bax/Bcl-2 ratio leads to an up-regulation in caspase-3 mRNA expression.     

Recombinant λ-phage nanobioparticles for tumor therapy in mice models

Lambda phages have considerable potential as gene delivery vehicles due to their genetic tractability, low cost, safety and physical characteristics in comparison to other nanocarriers and gene porters. Little is known concerning lambda phage-mediated gene transfer and expression in mammalian hosts. We therefore performed experiments to evaluate lambda-ZAP bacteriophage-mediated gene transfer and expression in vitro. For this purpose, we constructed recombinant λ-phage nanobioparticles containing a mammalian expression cassette encoding enhanced green fluorescent protein (EGFP) and E7 gene of human papillomavirus type 16 (λ-HPV-16 E7) using Lambda ZAP- CMV XR vector. Four cell lines (COS-7, CHO, TC-1 and HEK-239) were transduced with the nanobioparticles. We also characterized the therapeutic anti-tumor effects of the recombinant λ-HPV-16 E7 phage in C57BL/6 tumor mice model as a cancer vaccine. Obtained results showed that delivery and expression of these genes in fibroblastic cells (COS-7 and CHO) are more efficient than epithelial cells (TC-1 and HEK-239) using these nanobioparticles. Despite the same phage M.O.I entry, the internalizing titers of COS-7 and CHO cells were more than TC-1 and HEK-293 cells, respectively. Mice vaccinated with λ-HPV-16 E7 are able to generate potent therapeutic antitumor effects against challenge with E7- expressing tumor cell line, TC-1 compared to group treated with the wild phage. The results demonstrated that the recombinant λ-phages, due to their capabilities in transducing mammalian cells, can also be considered in design and construction of novel and safe phage-based nanomedicines.     

Hydrogels Composed of Cyclodextrin Inclusion Complexes with PLGA-PEG-PLGA Triblock Copolymers as Drug Delivery Systems

Although conventional pharmaceuticals have many drug dosage forms on the market, the development of new therapeutic molecules and the low efficacy of instant release formulations for the treatment of some chronic diseases and specific conditions encourage scientists to invent different delivery systems. To this purpose, a supramolecular hydrogel consisting of the tri-block copolymer PLGA-PEG-PLGA and α-cyclodextrin was fabricated for the first time and characterised in terms of rheological, morphological, and structural properties. Naltrexone hydrochloride and vitamin B12 were loaded, and their release profiles were determined.     

Switchgrass in California: where,and at what price?

This article assesses the potential of California agriculture to supply biofuel feedstock in the form of switchgrass. We construct a fully calibrated, multiregion, multi‐input and multioutput model of agricultural supply for California's Central Valley based on the principles of positive mathematical programming. We exploit agronomic information obtained from a biophysical model to estimate regional production functions for switchgrass. The model predicts the extent and location of potential feedstock production in the Central Valley. Our results suggest that switchgrass adoption rates differ widely among agricultural regions, and that switchgrass is not likely to displace specialty crops by much statewide.     

Optimization of transfection methods for Huh-7 and Vero cells: A comparative study

Availability of an efficient transfection protocol is the first determinant in success of gene transferring studies in mammalian cells which is accomplished experimentally for every single cell type. Herein, we provide data of a comparative study on optimization of transfection condition by electroporation and chemical methods for Huh-7 and Vero cells. Different cell confluencies, DNA/reagent ratios and total transfection volumes were optimized for two chemical reagents including jetPEI? and Lipofectamine? 2000. Besides, the effects of electric field strength and pulse length were investigated to improve electroporation efficiency. Transfection of cells by pEGFP-N1 vector and tracking the expression of GFP by FACS and Fluorescence Microscopy analysis were the employed methods to evaluate transfection efficiencies. Optimized electroporation protocols yielded 63.73 ± 2.36 and 73.9 ± 1.6% of transfection in Huh-7 and Vero cells respectively, while maximum achieved level of transfection by jetPEI? was 14.2 ± 0.69 and 28 ± 1.11% Huh-7 and Vero cells, respectively. Post transfectional chilling of the cells did not improve electrotransfection efficiency of Huh-7 cells. Compared to chemical based reagents, electroporation showed superior levels of transfection in both cell lines. The presented protocols should satisfy most of the experimental applications requiring high transfection efficiencies of these two cell lines.