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 .

Algae from the inland ice of Greenland. [Translation from: Kongl.Vetenskaps-Akademiens Forhandlingar 1871(2) 293-296, 1871.]

On the large inland ice cover of Greenland, where the temperature even during the short summer, falls below freezing some time during the daily 24 hours, it is still possible to find organic life. The author found in melted ice water several species of algae on his ”walk” in the latter part of July 1870. The article describes the finds and tries to identify the algae to family level.

The content of organic matter in some waters in the Moscow region [Translation from: C.R. Acad. Soc. URSS, 61, 293-296, 1948]

There is at the moment no direct method of determining the organic matter content of natural waters. In 1940/41 8 different water bodies in central Russia were studied and their organic matter identified. The author concludes that there is currently no easy method to determine organic matter in water. A number methods need to be applied.

The food of yellowfin and skipjack tunas in the Eastern Tropical Pacific Ocean

ENGLISH: Knowledge of the kinds of organisms eaten by tunas, and the relative importance of different kinds of organisms in different situations, is of importance to our understanding of regional and local aggregations and of the behavior of the tropical tunas. Determination of the relative importance of benthic forms in the stomachs of tunas captured in the vicinity of islands and offshore banks, compared with the forms found in the stomachs of those from oceanic areas, distant from these shoal areas, is useful to the understanding of the mechanisms by which the islands and banks attract tuna. A thorough knowledge of the diet may also reveal more about the seasonal and annual variations in the distribution of these fish along the coasts of Mexico, Central America and elsewhere. A knowledge of the diet of the tunas is basic to quantitative studies of the zooplankton and nekton in the Eastern Pacific Ocean with reference to the production of tunas. Studies of the diet of the tropical tunas by examination of stomach contents was, therefore, commenced in late 1957. SPANISH: El conocimiento de las clases de organismos de que se alimentan los atunes, y la importancia relativa de las diferentes clases de organismos en situaciones diferentes, es de valor para nuestra comprensión de las agrupaciones regionales y locales, y de los hábitos de los atunes tropicales. La determinación de la importancia relativa de las formas bénticas en los estómagos de los atunes capturados en la vecindad de las islas y de los bancos apartados de la costa, comparadas con las formas encontradas en los estómagos de los atunes procedentes de áreas oceánicas, distantes de estas áreas poco profundas, es útil para comprender el mecanismo por el cual las islas y los bancos atraen al atún. Un conocimiento completo de la dieta puede revelar también algo más acerca de las variaciones estacionales y anuales en la distribución de estos peces a lo largo de las costas de México, América Central y otras partes. El conocimiento de la dieta del atún es básico para los estudios cuantitativos del zooplancton y del necton en el Océano Pacífico Oriental, con referencia a la producción de los atunes. El estudio de la dieta de los atunes tropicales mediante el examen del contenido estomacal, fué comenzado, por lo tanto, a fines de 1957.

Anuario Estadistico de Pesca 1984

This document is in Spanish. Catch statistics for Mexican waters 1983. (PDF has 293 pages.)

Anuario Estadistico de Pesca 1985

This document is in Spanish. Los cuatro capítulos que integran este Anuario constituyen la cobertura básica de las distintas fases de la actividad pesquera. Con ello se integra la informaci6n cuantitativa referente a cada una de las fases, desde la identificaci6n de sus prop6sitos, hasta los medios y técnicas con que se realizan y los resultados finales alcanzados en el año de referencia. En el Capitulo 1, relativo a la fase primaria o extractiva, se han considerado no s610 las cifras correspondientes a la captura, se incluyen tambien los correspondientes a los bienes de capital -embarcaciones, artes y equipos de pesca e infraestructura portuaria- y a la poblaci6n dedicada a la extracci6n. Para la etapa industrial, Capítulo 11, se presenta la informaci6n de los activos fijos dedicados a la actividad pesquera, así como el resultado de su operaci6n por principales procesos: congelado, enlatado, reducci6n y otros como el seco-salado y el ahumado. Para la fase comercial, capítulo 111, se incluye la info~ maci6n referente a los volúmenes disponibles de productos pesqueros, su valor y las cifras relativas al comercio ex terior. Se consigna, igualmente, la estadística concerniente al consumo de estos productos. Por último, en el Capítulo IV se integran las informaciones provenientes de diferentes unidades administrativas del sector, como las relativas a la capacitaci6n, al empleo y al financiamiento de la actividad. Catch statistics for Mexican waters 1985. (PDF has 293 pages.)

Potentials for marine fish hatchery production in Nigeria

The development of fish hatcheries and fish fingerling production in Nigeria has all along concentrated largely on the freshwater fish species without paying enough attention to the more numerous fish species that abound in our coastal/marine environment. This paper, therefore, tries to highlight some basic technologies (in term of design and management) of marine fish hatchery based on the author's experiences in Southeast Asia. Appropriate adaptable technologies for the production of our indigenous species such as the Snappers, Groupers etc. are also discussed. General recommendations are made for marine fish hatchery development in Nigeria

Ecosystem study Altenwoerth: impacts of a hydroelectric power-station on the River Danube in Austria

The aim of this article is to briefly describe the effects of the Altenwoerth Barrage, on the River Danube, on some physical variables and their consequent effects on water chemistry and the biota of the river. The methods used for biological sampling are summarised, especially those used in the limnological part of the study, and the macroinvertebrate and fish fauna listed. Comparisons are then made between the impounded section of river immediately above the dam and two unimpounded free-flowing sections of the river. Further developments on the Danube are considered.

Comparison of echogram measurements against data expectations and assumptions for distinguishing seafloor substrates

Defining types of seafloor substrate and relating them to the distribution of fish and invertebrates is an important but difficult goal. An examination of the processing steps of a commercial acoustics analyzing software program, as well as the data values produced by the proprietary first echo measurements, revealed potential benef its and drawbacks for distinguishing acoustically distinct seafloor substrates. The positive aspects were convenient processing steps such as gain adjustment, accurate bottom picking, ease of bad data exclusion, and the ability to average across successive pings in order to increase the signal-to-noise ratio. A noteworthy drawback with the processing was the potential for accidental inclusion of a second echo as if it were part of the first echo. Detailed examination of the echogram measurements quantified the amount of collinearity, revealed the lack of standardization (subtraction of mean, division by standard deviation) before principal components analysis (PCA), and showed correlations of individual echogram measurements with depth and seafloor slope. Despite the facility of the software, these previously unknown processing pitfalls and echogram measurement characteristics may have created data artifacts that generated user-derived substrate classifications, rather than actual seafloor substrate types.

Factors influencing cannibalism and abundance of walleye pollock (Theragra chalcogramma) on the eastern Bering Sea shelf, 1982–20

Cannibalism is thought to be an inf luential top-down process affecting walleye pollock (Theragra chalcogramma) recruitment in the eastern Bering Sea (EBS). In summer, many age-1 pollock occupy the same depths as those of adult walleye pollock, making them vulnerable to cannibalism. We examine factors that inf luence the occurrence and amount of cannibalism, as well as the abundance and co-occurrence of predator and prey walleye pollock. Large walleye pollock were generally found in deeper waters and avoided cold temperatures; whereas, age-1 walleye pollock were found in broader bottom depth and temperature ranges. The occurrence of cannibalism was highest in the area where predator and prey walleye pollock co-occurred and the amount of cannibalism was highest on the middle and outer EBS shelf. Both the occurrence and amount of cannibalism were influenced by location, bottom temperature and bottom depth, and the abundance of prey walleye pollock. The abundance of both large and small walleye pollock decreased during the 1982–2006 survey period in the EBS and, hence, the occurrence and amount of cannibalism also decreased. The occurrence and amount of cannibalism observed in the diet samples from the summer survey were good indicators of year class strength, as estimated by the stock assessment model. There was more cannibalism of age-1 walleye pollock when predicted recruit abundance was highest, indicating that summer cannibalism on age-1 walleye pollock, a top-down process, does not control walleye pollock recruitment in the EBS.

Potential causes of mortality for horseshoe crabs (Limulus polyphemus) during the biomedical bleeding process

Biomedical companies catch and bleed horseshoe crabs for the production of Limulus amebocyte lysate (LAL), a product used for protecting public health (Berkson and Shuster, 1999). LAL is a clotting agent, derived solely from horseshoe crab blood cells, which is used to detect the presence of pathogenic gramnegative bacteria in injectable drugs and implantable medical and dental devices (Mikkelsen, 1988; Novitsky, 1991). In addition, LAL is used in many diagnostic tests for such illnesses as gram-negative bacterial meningitis and typhoid fever (Ding and Ho, 2001). Because the LAL test allows one to detect femtogram levels of endotoxin (Ding and Ho, 2001), it is the most effective test for detecting endotoxin contamination, and its increasing use in medical and pharmaceutical laboratories makes it a highly valued product.

Life history and production of Capitella capitata (Polychaeta: Capitellidae) in Río de la Plata Estuary (Argentina)

A benthic survey was carried out from November 1998 to December 1999 in the tidal flats of Bahía Samborombón (Río de la Plata estuary, Argentina), in order to study the population structure, reproductive aspects, growth and secondary production of Capitella capitata (Fabricius, 1780). Growth was analyzed using ELEFAN routine, and the secondary production was estimated by Hynes and Coleman's method (1968). C. capitata did not present periods of very important recruitments throughout the year; however, the abundance of smallest size classes was higher during summer and autumn. The summer cohort showed a growth rate (K) of 2.05 and a seasonal growth oscillation (C) of 0.6, pointing out that worms grew very slowly during winter months. The life span of this cohort was 13 months. The autumn cohort showed a lower growth rate (K= 1.5) and its growth was lowest during winter. The life span was 15 months for this cohort. C. capitata in Punta Rasa presented an extended reproductive period, with absence of activity during winter months. The type of eggs and larvae suggest that C. capitata has benthic larval development in the study area, destining its reproductive effort to the production of a low number of eggs, and assuring larvae survival through incubation in brooding tubes. The annual mean biomass in Punta Rasa was 0.117 g m-2 (AFDW), with a mean secondary production of 0.23 g m-2 y-1 and a P/B ratio of 1.96 y-1. The relatively low density, biomass production and P/B ratio of C. capitata in Punta Rasa can be considered as reference values for this species inhabiting undisturbed or moderately disturbed areas.