Introduction: Ranibizumab is a mouse monoclonal antibody fragment antigen-binding (Fab) against human vascular endothelial growth factor-A (VEGF-A), inhibiting angiogenesis. This antibody is commercially produced in Escherichia coli host and used to treat wet agerelated macular degeneration (AMD). Methods: In this study, the heavy and light chains of ranibizumab were expressed in Pichia pastoris. The expressed chains were incubated overnight at 4° C for interaction. The formation of an active structure was evaluated based on the interaction with substrate VEGF-A using an indirect ELISA, and an electrochemical setup. Furthermore, reconstruction of Split enhanced green fluorescent protein (eGFP) reporter, chimerized at the C-terminus of the heavy and light chains, was used to characterize chains’ interaction. Results: P. pastoris efficiently expressed designed constructs and secreted them into the culture medium. The anti-Fab antibody detected the constructed Fab structure in western blot analysis. Reconstruction of the Split reporter confirmed the interaction between heavy and light chains. The designed ELISA and electrochemical setup results verified the binding activity of the recombinant Fab structure against VEGF-A. Conclusion: In this work, we indicated that the heavy and light chains of ranibizumab Fab fragments (with or without linkage to Split parts of eGFP protein) were produced in P. pastoris. The fluorescence of reconstructed eGFP was detected after incubating the equal ratio of chimeric-heavy and light chains. Immunoassay and electrochemical tests verified the bioactivity of constructed Fab. The data suggested that P. pastoris could be considered a potential efficient eukaryotic host for ranibizumab production.

Background: Sugarcane is a monocotyledonous crop that is cultivated in the tropical and subtropical regions of the world. One of the most important criteria, influencing the efficiency of the sugarcane transformation is known to be related to physical and biological factors during the transformation procedure. Objectives: The objective of this research was to study the response of callus induction and embryogenic callus production and to identify the major parameters controlling DNA delivery by particle bombardment into sugarcane (Saccharum officinarum L.) cv. NCo310. Materials and Methods: For callus induction and embryogenic callus production, leaf base segments were subjected to in vitro culture medium supplemented with two plant growth regulators (2,4-D and Dicamba). Results showed that 1 mg.L -1 2, 4-D was significantly influential in callus induction and embryogenic callus production. Considering both physical and biological factors, the efficiency of DNA (uidA gene) delivery was assessed by scoring transient GUS (gene (b-glucuronidase)) expression in bombarded tissues. Results: The highest transient GUS expression was obtained when callus was bombarded with the construct harboring rice Act1 promoter, and having 9 cm target distance, 25 inHg vacuum pressure, 1 mm gold particles, 12.5 mg DNA per bombardment and one day pre-culture prior to the bombardment. Conclusions: A bombardment procedure suitable for elite sugarcane varieties was developed, which allowed high-efficiency DNA delivery combined with reduced damage to target tissues.

The paper summarizes the current status of the reporter gene technology and their basics. reporter gene technology is widely used to monitor cellular events associated with gene expression and signal transduction. Based upon the splicing of transcriptional control elements to a variety of reporter genes, it “reports” the effects of a cascade of signaling events on gene expression inside cells. The principal advantage of these assays is their high sensitivity, reliability, convenience and adaptability to large scale measurements. The reporter gene systems b-galactosidase (LacZ), luciferases (LUC), b-glucoronidase (GUS), green fluorescent protein (GFP) and GFP-like proteins, widely used presently, are compared. The potentialities of a new reporter system, based on thermostable lichenase (b-1, 3-1, 4-gluconases) of Clostridium thermocellum are described in detail. Advantages and disadvantages of the used reporter systems, and also the opportunities for development and application of bifunctional reporter systems are considered. Perspectives of reporter genes for studying gene expression regulation in prokaryotic and eukaryotic organisms are elucidated.

We report a case of an 87-year-old man where coronary CT angiography incidentally demonstrated a “Split” right coronary artery (RCA) featuring a “Split” posterior descending artery. This case focusses on the morphological description of this variant as well as its differentiation from a “dual” or “duplicated” RCA.

The Shiite Infallible Imams hadiths reported by the Sunni reporters and their authenticity are of the most important issues which are discussed by hadith scholars in the field of diraya (hadith scholarship) and fiqh (jurisprudence). Generally speaking, we can divide the Shilte hadith scholars into two groups: those who are against the reliability of authenticated hadith and those who are for. Ismai Bn Abi Ziyyad Sukuni is one of the Sunni reporters who has transmitted many reports from Imam Sadiq, peace be upon him, especially in the field of fiqh (jurisprudence).the more we have information about this reporter, the better we can verify reliability of his reports which are found in the Shiite hadith sources. In light of Sukuni's reliability which has been confirmed by the Shiite thinkers and his reports which are practiced by jurisprudents, it is wise to conclude that the reports transmitted by him are reliable and trustworthy.