Establishment of a multiplex RT-PCR assay for the rapid detection of fish cytokines

To monitor the expression of cytokine genes in Japanese pufferfish, a novel platform for quantitative multiplexed analysis was developed. This custom-designed multiplex RT-PCR assay was used to analyze the expression profiles of 19 cytokine genes, including pro-inflammatory (IL-1β, IL-6, IL-17A/F3, IL-18, TNF-α, TNF-N), anti-inflammatory (IL-4/13A, IL-4/13B, IL-10), T-cell proliferation/differentiation (IL-2, IL-15, IL-21, TGF-β1), B-cell activation/differentiation (IL-7, IL-6, IL-4/13A, IL-4/13B), NK cell stimulation (IL-12p35 and IL-12p40), induction of anti-viral activity (I-IFN-1 and IFN-γ), and monocyte/macrophage progenitor cell proliferation (M-CSF1b) cytokines in head kidney cells under immune stimulatory conditions. The expression profiles were dissimilar in the unstimulated control and immune-stimulated cells. Moreover, increased expression profile was observed due to different stimulations for IL-1β, IL-6, IL-10, IL-12p35, IL-12p40, IL-21, TNF-α, TNF-N, I-IFN-1 and IFN-γ genes. These results suggest that cytokine genes could be used as biomarkers to know the immune status of fish. The constructed multiplex RT-PCR assay will enhance understanding on immune regulation by cytokines in fish.     

Evaluating the effect of liming on N<Subscript>2</Subscript>O fluxes from denitrification in an Andosol using the acetylene inhibition and <Superscript>15</Superscript>N isotope tracer methods

We assessed the effect of liming on (1) N₂O production by denitrification under aerobic conditions using the ¹⁵N tracer method (experiment 1); and (2) the reduction of N₂O to N₂ under anaerobic conditions using the acetylene inhibition method (experiment 2). A Mollic Andosol with three lime treatments (unlimed soil, 4 and 20 mg CaCO₃ kg^?1) was incubated at 15 and 25 °C for 22 days at 50% and then 80% WFPS with or without 200 mg N kg^?1 added as ¹⁵N enriched KNO₃ in experiment 1. In experiment 2, the limed and unlimed soils were incubated under completely anaerobic conditions for 44 h (with or without 100 mg N kg^?1 as KNO₃). In experiment 1, limed treatments increased N₂O fluxes at 50% WFPS but decreased these fluxes at 80% WFPS. At 25 °C, cumulative N₂O and ¹⁵N₂O emissions in the high lime treatment were the lowest (with at least 30% less ¹⁵N₂O and total N₂O than the unlimed soil). Under anaerobic conditions, the high lime treatment showed at least 50% less N₂O than the unlimed treatment at both temperatures with or without KNO₃ addition but showed enhanced N₂ production. Our results suggest that the positive effect of liming on the mitigation of N₂O evolution from soil was influenced by soil temperature and moisture conditions.     

Intake of Heated Leaf Extract of Coriandrum sativum Contributes to Resistance to Oxidative Stress via Decreases in Heavy Metal Concentrations in the Kidney

Plant Foods for Human Nutrition - Coriandrum sativum (coriander) is an annual herb of the Apiaceae family and has been used as a traditional remedy. Here we examined whether heated leaf extract of...     

Impact of nitrogen cycling associated with production and consumption of food on nitrogen pollution of stream water

We evaluated the impact of nitrogen (N) cycling on N pollution of stream water, with emphasis on N disposed of (hereafter referred to as “disposal N”) from human and livestock excrement and the N surplus in cropland, compared to the total-N concentration of stream water, in seven zones of Asahikawa City characterized by various types of land use. In order to estimate N cycling, we used the Nitrogen Flow Model, composed of the N budgets of human, livestock, and cropland subsystems. The urban area with a population density of over 4,000 persons km^-2 generated a very large amount of disposal N (about 2,700 kg N ha^-1 cropland y^-1). Based on the amount of disposal N and the volume of domestic sewage water used, the N concentration estimated for the urban area was 34 mg N L^-1, which found in the effluent from the sewage treatment facility (24–28 mg N L^-1), regardless of the season. Thus, it was indicated that most of the disposal N in the urban area was discharged directly to streams through the sewage treatment facilities, contributing to a point source of N pollution of stream water. In addition, the disposal N from livestock facilities was larger in pig and poultry farming areas than in other farming areas, contributing to some extent to a potential source of N pollution. As a result, the concentrations increased above 1 mg N L^-1 in the urban and surrounding areas. On the other hand, the N surplus in cropland was practically determined by the N flows associated with chemical fertilizer, livestock excrement as manure, and crop uptake. The N surplus was similar among the seven zones, ranging from 69 to 99 kg N ha^-1 y^-1. The N concentration estimated from the amount of N surplus and 50% of mean annual precipitation as a discharge rate was 13.6–19.5 mg N L^-1. Most of the surplus N was indicated to be leached out. However, the total-N concentration measured in the major streams flowing through Asahikawa City was mostly below 1 mg N L^-1 except for the urban and surrounding areas. The surplus N in cropland may not reach the streams, even if N leaching occurs, probably due to N removal by plant uptake, denitrification, and sedimentation in the riparian zone and stream channels. Thus the effect of agricultural practices on N pollution of stream water was not appreciable.