Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .

Genome-wide computational identification of microRNAs and their targets in the deep-branching eukaryote Giardia lamblia

Using a combined computational program. we identified 50 potential microRNAs (miRNAs) in Giardia lamblia. one of the most primitive unicellular eukaryotes. These miRNAs are unique to G. lamblia and no homologues have been found in other organisms; miRNAs.

Performance prediction of graphene nanoribbon and carbon nanotube transistors

Carbon nanotubes (CNTs) and graphene nanoribbons (GNRs) field-effect transistor (FET) can be the basis for a quasi-one- dimensional (Q1D) transistor technology. Recent experiments show that the on-off ratio for GNR devices can be improved to level exploration of transistor action is justified. Here we use the tight-binding energy dipersion approximation, to assess the performance of semiconducting CNT and GNR is qualitatively in terms of drain current drive strength, bandgap and density of states for a specified device. By reducing the maximum conductance 4e2/h by half, we observed that our model has a particularly good fit with 50 nm channel single walled carbon nanotube (SWCNT) experimental data. Given the same bandgap, CNTs outperform GNRs due to valley degeneracy. Nevertheless, the variation of the device contacts will decide which transistor will exhibit better conductivity and thus higher ON currents. © 2011 American Institute of Physics.

Explosion pressure prediction via polynomial mathematical correlation based on advanced CFD Modelling

Computational Fluid Dynamics CFD can be used as a powerful tool supporting engineers throughout the steps of the design. The combination of CFD with response surface methodology can play an important role in such cases. During the conceptual engineering design phase, a quick response is always a matter of urgency. During this phase even a sketch of the geometrical model is rare. Therefore, the utilisation of typical response surface developed for congested and confined environment rather than CFD can be an important tool to help the decision making process, when the geometrical model is not available, provided that similarities can be considered when taking into account the characteristic of the geometry in which the response surface was developed. The present work investigates how three different types of response surfaces behave when predicting overpressure in accidental scenarios based on CFD input. First order, partial second order and complete second order polynomial expressions are investigated. The predicted results are compared with CFD findings for a classical offshore experiment conducted by British Gas on behalf of Mobil and good agreement is observed for higher order response surfaces. The higher order response surface calculations are also compared with CFD calculations for a typical offshore module and good agreement is also observed. © 2011 Elsevier Ltd.

Stability enhancement by casing grooves: The importance of stall inception mechanism and solidity

This paper concerns the optimisation of casing grooves and the important influence of stall inception mechanism on groove performance. Installing casing grooves is a well known technique for improving the stable operating range of a compressor, but the wide-spread use of grooves is restricted by the loss of efficiency and flow capacity. In this paper, laboratory tests are used to examine the conditions under which casing treatment can be used to greatest effect. The use of a single casing groove was investigated in a recently published companion paper. The current work extends this to multiple-groove treatments and considers their performance in relation to stall inception mechanisms. Here it is shown that the stall margin gain from multiple grooves is less than the sum of the gains if the grooves were used individually. By contrast, the loss of efficiency is additive as the number of grooves increases. It is then shown that casing grooves give the greatest stall margin improvement when used in a compressor which exhibits spike-type stall inception, while modal activity before stall can dramatically reduce the effectiveness of the grooves. This finding highlights the importance of being able to predict the stall inception mechanism which might occur in a given compressor before and after grooves are added. Some published prediction techniques are therefore examined, but found wanting. Lastly, it is shown that casing grooves can, in some cases, be used to remove rotor blades and produce a more efficient, stable and light-weight rotor. © 2010 by ASME.