Atmospheric metal and phosphorus concentrations, inputs, and their biogeochemical significances in the Japan/East Sea.

Atmospheric metals and phosphorus over the southern Japan/East Sea were investigated in order to evaluate their sources, concentrations and inputs, and to identify their biogeochemical roles in this marginal sea. Aerosols were collected on the east coast of Korea from February 2002 to April 2003 (n=101) as well as at a remote island (Ulleung) and on a ship from February 2002 to June 2003 (n=13). The aerosols were analyzed for Al, Co, Cu, Ni, P, Pb and Zn. Simultaneous collections of aerosols at both coast and offshore were performed, and several high dust aerosols (Al>5 microg m(-3)) were collected at both regions. At the coastal site, both dust mineral and pollutants were transported by westerly winds from the Asian continent, but local emissions were significant (e.g., Cu, Ni, P and Zn) as well during the summer monsoon (May-August). The experimental relationships between the coast and offshore sites were defined. From these relationships, it was possible to obtain the annually averaged atmospheric metal and P concentrations over the southern Japan/East Sea, which has increased by over 2 times for the last decade. Through the estimation of atmospheric metal and phosphorus fluxes and comparisons with inputs from the Tsushima Warm Current, the atmospheric pathway was found to be a significant source for Al, Pb and Zn.

Stabilization of dengue virus polymerase in de novo initiation assay provides advantages for compound screening.

Dengue virus (DENV) NS5 protein comprises an N-terminal methyltransferase domain and a C-terminal RNA-dependent RNA polymerase domain (RdRp). DENV RdRp is responsible for viral RNA synthesis via a de novo initiation mechanism and represents an attractive target for anti-viral therapy. Herein we describe the characterization of its de novo initiation activities by PAGE analyses and the knowledge gained was used to develop a fluorescent-based assay. A highly processive and robust assay was achieved by addition of cysteine in the assay buffer. This stabilized the apo-enzyme, and rendered optimal de novo initiation activity while balancing its intrinsic terminal transferase activity. Steady-state kinetic parameters of the NTP and RNA substrates under these optimal conditions were determined for DENV1-4 FL NS5. Heavy metal ions such as Zn(++) and Co(++) as well as high levels of monovalent salts, suppressed DENV polymerase de novo initiation activities. This assay was validated with nucleotide chain terminators and used to screen two diverse small library sets. The screen data obtained was further compared with concurrent screens performed with a DENV polymerase elongation fluorescent assay utilizing pre-complexed enzyme-RNA. A higher hit-rate was obtained for the de novo initiation assay compared to the elongation assay (∼2% versus ∼0.1%). All the hits from the latter assay are also identified in the de novo initiation assay, indicating that the de novo initiation assay performed with the stabilized apo-enzyme has the advantage of providing additional chemical starting entities for inhibiting this enzyme.

Nationwide monitoring of mercury in wild and farmed fish from fresh and coastal waters of Korea.

Mercury (Hg) concentrations were monitored in wild and cultured fish collected from fresh and coastal waters in the Korean peninsula from April 2006 to August 2008 nationwide. Total Hg concentrations were reported for 5043 fish samples, including 78 species from 133 locations. Significant interspecies variation was noted in the Hg levels. The average Hg concentration in each fish species ranged from 6.31 µg kg(-1) for mullet (Mugil cephalus) to 200 µg kg(-1) for mandarin fish (Siniperca scherzeri). Among the species collected, the maximum concentration of Hg, 1720 µg kg(-1), was measured in an Amur catfish (Silurus asotus). Only wild freshwater fish exceeded the WHO ingestion standard. Wild freshwater piscivorous fish samples from a large artificial upstream lake contained the highest Hg levels. Hg concentrations were compared between fish groups categorized as wild and farmed fish from freshwater and coastal waters. Although the wild freshwater fish had similar size ranges, their Hg concentrations were higher than those of the other groups. Compared to the feed of farmed marine and freshwater fishes, the prey of wild freshwater fish had a higher Hg concentration, and the total Hg concentrations in freshwater and associated sediment samples were higher than those in coastal water and associated sediment samples. In the freshwater environment, piscivorous fish bioaccumulated two times more Hg than carnivorous and omnivorous fish and four times more than planktivorous fish. The difference in Hg concentrations among trophic groups might have been due to differences in the size of fish, in addition to the variations among different trophic groups. These data will be useful for developing the fish consumption advisory as a management measure to reduce Hg exposure.

Atmospheric transport of water-soluble ions (NO3(-), NH4(+) and nss-SO4(2-)) to the southern East Sea (Sea of Japan).

Atmospheric deposition of different types of aerosols over the southern East Sea has received little attention in terms of seawater biogeochemistry. We investigated the concentrations of water-soluble ions (NO3(-), NH4+ and nss-SO4(2-)) in the aerosols associated with air mass transport patterns arriving at the east coast of Korea, adjacent to the southern East Sea, in order to determine source regions affecting chemical composition of aerosols and to assess the atmospheric pathway as a significant controlling mechanism of the biogeochemistry in this marginal sea. Concentrations of certain elements (Al, Na, Ca, V, Zn and Pb) together with the water-soluble ions were measured in the aerosol samples (n=34) collected during the period March 2002-February 2003. The geometric mean concentrations of the water-soluble ions were NO3(-) 2.98 (0.56-16.22), NH4+ 1.42 (0.37-6.73) and nss-SO4(2-) 2.47 (0.17-17.35) microgm(-3). The backward trajectories revealed that air masses passing slowly over eastern China contributed more to increases in the concentrations of water-soluble ions than those associated with fast-moving northwesterly and maritime winds. Therefore, the correlation between the NH4+ and NO3+ concentrations increased, suggesting that gas-phase NH3 and HNO3 was forming fine-mode NH4NO3. The atmospheric N input accounted for approximately 10% of new production over the southern East Sea on an annual scale, while it accounted for over approximately 25% of new production during the water column stratification seasons (summer and early fall).

Antileishmanial high-throughput drug screening reveals drug candidates with new scaffolds.

Drugs currently available for leishmaniasis treatment often show parasite resistance, highly toxic side effects and prohibitive costs commonly incompatible with patients from the tropical endemic countries. In this sense, there is an urgent need for new drugs as a treatment solution for this neglected disease. Here we show the development and implementation of an automated high-throughput viability screening assay for the discovery of new drugs against Leishmania. Assay validation was done with Leishmania promastigote forms, including the screening of 4,000 compounds with known pharmacological properties. In an attempt to find new compounds with leishmanicidal properties, 26,500 structurally diverse chemical compounds were screened. A cut-off of 70% growth inhibition in the primary screening led to the identification of 567 active compounds. Cellular toxicity and selectivity were responsible for the exclusion of 78% of the pre-selected compounds. The activity of the remaining 124 compounds was confirmed against the intramacrophagic amastigote form of the parasite. In vitro microsomal stability and cytochrome P450 (CYP) inhibition of the two most active compounds from this screening effort were assessed to obtain preliminary information on their metabolism in the host. The HTS approach employed here resulted in the discovery of two new antileishmanial compounds, bringing promising candidates to the leishmaniasis drug discovery pipeline.