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 .

The changes in various iron compounds in culture media for algae. [Translation from: Archiv fur Hydrobiologie Supplemente Band 38(1/2) 151-169, 1970.]

The purpose of this work is a contribution to the quantitative record of the use of iron by planktonic algae. Preliminary experiments with Chlorella to determine the rate of iron intake in the presence of inorganic sources of iron did not produce the desired result. The crucial point of this work is the investigation of the influence of various external factors on the stability of FeEDTA (FeEDTA = Ferric(III)-compound of ethylene-diamine tetra-acetic acid), since this compound appears to be particularly well-suited as a source of iron for planktonic algae (e.g. TAMIYA et al. 1953). Cultures of Chlorella fusca in a light thermostat were used in experimental research. Methods and results are discussed.

For the determination of iron (II) and total iron with 2,2'-dipyridyl in mineral waters. Reduction of iron (III) with ascorbic acid [Translation from: Zeitschrift fur Analytische Chemie 209 340-341, 1965]

The determination of bi- and trivalent iron in proximity, in mineral waters has gained in significance, on biological and technical grounds. This short paper describes the procedure of the determination of bivalent iron and total iron in a water sample.

PICES Press, Vol. 12, No. 2, July 2004

The future of PICES [pdf, 1.7 MB] Paris by day - Symposium on "Quantative ecosystem indicators in fisheries management" [pdf, 0.2 MB] The Bering Sea: Current status and recent events [pdf, 0.4 MB] The state of the western North Pacific in the second half of 2003 [pdf, 0.7 MB] The state of the eastern North Pacific entering spring 2004 [pdf, 0.4 MB] PICES-IFEP Workshop on "In-situ iron enrichment experiments in the eastern and western subarctic Pacific" [pdf, 1.4 MB] Canadian SOLAS/PICES-IFEP session on "Response of the upper ocean to meso-scale iron enrichment" [pdf, 0.3 MB] Fisheries and ecosystem responses to recent regime shifts [pdf, 0.8 MB] PICES Interns [pdf, 0.8 MB] Did a regime shift occur in 1998 around Japan?- Highlights from a symposium addressing this question [pdf, 0.8 MB] The Global Ocean Carbon Observing System - Connecting national programs and regional networks [pdf, 1.7 MB] The North Pacific Ecosystem Metadatabase [pdf, 1.2 MB] International GLOBEC Symposium on "Climate variability and Sub-Arctic marine ecosystems" [pdf, 0.2 MB] PICES Calendar [pdf, 0.2 MB] PICES/GLOBEC Symposium on "Climate variability and ecosystem impacts on the North pacific: A basin-scale synthesis" [pdf, 0.2 MB]

Report of the 2004 Workshop on In Situ Iron Enrichment Experiments in the Eastern and Western Subarctic Pacific

Foreword 1. BACKGROUND AND OBJECTIVES (pdf, 0.1 Mb) 2. 2004 WORKSHOP SUMMARY (pdf, < 0.1 Mb) 2.1. What have we learned from the enrichment experiments? 2.2 What are the outstanding questions? 2.3 Recommendations for SEEDS-II 3. EXTENDED ABSTRACTS OF THE 2004 WORKSHOP 3.1 Synthesis of the Iron Enrichment Experiments: SEEDS and SERIES (pdf, 0.5 Mb) Iron fertilization experiment in the western subarctic Pacific (SEEDS) by Atsushi Tsuda The response of N and Si to iron enrichment in the Northeast Pacific Ocean: Results from SERIES by David Timothy, C.S. Wong, Yukihiro Nojiri, Frank A. Whitney, W. Keith Johnson and Janet Barwell-Clarke 3.2 Biological and Physiological Responses (pdf, 0.2 Mb) Zooplankton responses during SEEDS by Hiroaki Saito Phytoplankton community response to iron and temperature gradient in the NW and NE subarctic Pacific Ocean by Isao Kudo, Yoshifumi Noiri, Jun Nishioka, Hiroshi Kiyosawa and Atsushi Tsuda SERIES: Copepod grazing on diatoms by Frank A. Whitney, Moira Galbraith, Janet Barwell-Clarke and Akash Sastri The Southern Ocean Iron Enrichment Experiment: The nitrogen uptake response by William P. Cochlan and Raphael M. Kudela 3.3 Biogeochemical Responses (pdf, 0.5 Mb) What have we learned regarding iron biogeochemistry from iron enrichment experiments? by Jun Nishioka, Shigenobu Takeda and W. Keith Johnson Iron dynamics and temporal changes of iron speciation in SERIES by W. Keith Johnson, C.S. Wong, Nes Sutherland and Jun Nishioka Dissolved organic matter dynamics during SEEDS and SERIES experiments by Takeshi Yoshimura and Hiroshi Ogawa Formation of transparent exopolymer particles during the in-situ iron enrichment experiment in the western subarctic Pacific (SEEDS) by Shigenobu Takeda, Neelam Ramaiah, Ken Furuya and Takeshi Yoshimura Atmospheric measurement by Mitsuo Uematsu 3.4 Prediction from Models (pdf, 0.3 Mb) Modelling iron limitation in the North Pacific by Kenneth L. Denman and M. Angelica Peña A proposed model of the SERIES iron fertilization patch by Debby Ianson, Christoph Voelker and Kenneth L. Denman 4. LIST OF PARTICIPANTS FOR THE 2004 WORKSHOP (pdf, < 0.1 Mb) APPENDIX 1 Report of the 2000 Planning Workshop on Designing the Iron Fertilization Experiment in the Subarctic Pacific (pdf, 1 Mb) APPENDIX 2 Terms of Reference for the Advisory Panel on Iron fertilization experiment in the subarctic Pacific Ocean (pdf, < 0.1 Mb) APPENDIX 3 Historical List of Advisory Panel Members on Iron fertilization experiment in the subarctic Pacific Ocean (pdf, < 0.1 Mb) APPENDIX 4 IFEP-AP Annual Reports (pdf, 0.1 Mb) APPENDIX 5 PICES Press Articles (pdf, 0.6 Mb) (194 page document)

Ni-resist iron for propellers

Ni-Resist is the name applied to a group of alloy irons to which a sufficient amount of alloying element (mainly nickel) has been added to produce an austenitic matrix. Because of the alloy content and the austenitic matrix, Ni-Resist irons exhibit much better corrosion resistance and toughness than do ordinary cast irons.

Studies on arsenical creosote as a wood preservative for marine structures. Part 2. Observation on leaching, corrosion and resistance to borer attack

A detailed study on arsenical creosote with reference to leaching, corrosion and anti-borer properties was carried out. Results showed that aging had very little effect on the preservative which suggested better fixation of the preservative into the wood. Corrosion of mild steel, galvanised iron, aluminium-magnesium alloy (M57S) and copper panels in the preservative was found to be negligible. Normal creosote and low temperature creosote of Regional Research Laboratory, Hyderabad, both fortified with arsenic trioxide resisted borer damage on wooden panels for a period of over five months in the port of Cochin. The performance of low temperature creosote fortified with arsenic was found to be equally satisfactory when compared to normal creosote fortified in the same manner. A loading of 208.6 Kgs/ml³ for Haldu (Adina cordifolia) and 138 Kgs/m³ for Mango (Mangifera indica) in the case of normal creosote and 177 Kgs/m³ for Mango the case of RRL creosote were found to be sufficient for treating the wood.