Development of in vitro three-dimensional co-culture system for metabolic syndrome therapeutic agents.

AIMS: There are many obstacles to overcome in the development of new drugs for metabolic diseases, including efficacy and toxicity problems in later stages of drug development. To overcome these problems and predict efficacy and toxicity in early stages, we constructed a new model of insulin resistance in terms of communication between 3T3-L1 adipocytes and RAW264.7 macrophages by three-dimensional (3D) culture. RESULTS: In this study, results focused on the functional resemblance between 3D co-culture of adipocytes and macrophages and adipose tissue in diabetic mice. The 3D mono-culture preadipocytes showed good cell viability and induced cell differentiation to adipocytes, without cell confluence or cell-cell contact and interaction. The 3D co-cultured preadipocytes with RAW264.7 macrophages induced greater insulin resistance than two-dimensional and 3D mono-cultured adipocytes. Additionally, we demonstrated that 3D co-culture model had functional metabolic similarity to adipose tissue in diabetic mice. We utilized this 3D co-culture system to screen PPARγ antagonists that might have potential as therapeutic agents for diabetes as demonstrated by an in vivo assay. CONCLUSION: This in vitro 3D co-culture system could serve as a next-generation platform to accelerate the development of therapeutics for metabolic diseases.

Absolute configuration of antifibrotic (+)-episesamin isolated from Lindera obtusiloba BLUME.

Fractionation of a 70% ethanolic extract from twigs of Lindera obtusiloba BLUME (Japanese spicebush, Tohaku) yielded five fractions of different polarity. The antifibrotic activity within the chloroform phase was best assessed by an in vitro bioassay using rat hepatic stellate cell (HSC) proliferation and their autocrine transforming growth factor beta (TGF-beta) expression as sensitive fibrosis-associated read out. Chromatography of the chloroform extract on Sephadex LH-20 or liquid-liquid extractions yielded a crystalline compound as an active principle, which was identified from NMR and ESI-MS analyses, its melting point, and its optical rotation as (7S,7'R,8R,8'R)-3,4:3',4'-bis(methylenedioxy)-7,9':7',9-diepoxy-lignane [(+)-episesamin]. X-Ray diffraction confirmed the structure and provided, for the first time, directly its absolute configuration. (+)-Episesamin blocked proliferation and the profibrotic autocrine TGF-beta expression HSC without significant cytotoxicity.

Evaluation of maturity parameters and heavy metal contents in composts made from animal manure.

The objective of this study was to evaluate a variety of maturity indices and heavy metal concentrations in the composting process. A full-scale study was performed on animal manures (dairy:beef:swine = 50%:30%:20%, w/w) mixed with sawdust using a self-screwed turner over a time period of 63 days. Several chemical compost characteristics and germination indices were monitored on samples collected weekly at a prototype composting facility. NH(4)-N concentrations decreased during the composting time, on the contrary NO(3)-N concentrations increased sharply after 30 days. As composting proceeded, the humic acids content increased from 1.45% to 2.70% of the organic carbon content, whereas the fulvic acids content decreased gradually. Temperature in the compost pile and concentration of NH(3) averaged 32.4 degrees C and 12.9 ppm at the initial composting phase and 69.5 degrees C and 47.3 ppm at the active bio-oxidation composting phase, respectively. Seed germination and germination index (GI) increased during the composting process, and the compost samples at the end of composting phase had GI values greater than 120. Among the composting processes, there were significant differences (p<0.05) in concentration of Zn, Cu and Pb in the compost. In conclusion, the suggested maturity parameter values and metal concentrations of composted animal manures can provide better understanding of maturation process of animal manure compost.

Effect of spraying biological additives for reduction of dust and bioaerosol in a confinement swine house.

The aim of this on-site experiment is to evaluate and compare efficiencies of currently utilized biological additives to reduce emissions of dust and bioaerosol in a confinement swine house. The mean reduction rate of total dust only after spray ranged was approximately 30% for all the treatments, compared to initial level before spraying additives which was found to reduce the initial level of total dust significantly (p < 0.05). The mean reduction rate of all the treatments at 1 hr after spraying was about 24% which was 6% lower than only after spray. Since 3 hr after spraying, however, total dust level fluctuated inconstantly for all the treatments, besides application of soybean oil. The mean reduction rates of all the treatments only after spraying as compared to initial level before spraying were about 53% for total airborne bacteria (p < 0.01) and 51% for total airborne fungi (p < 0.01), respectively. At 1 hr after spraying, the reduction rate of total airborne fungi averaged to about 35% for all the treatments (p < 0.05), while insignificant reductions of total airborne bacteria were found only in the treatments with salt water, soybean oil, artificial spice, and essential oil (p > 0.05). The fluctuations of total airborne bacteria and fungi, which were similar to total dust, were observed for all the treatments 3 hr after spray.

Monitoring of aerial pollutants emitted from Swine houses in Korea.

This on-site survey study was performed to determine the concentrations and emissions of aerial contaminants in the different types of swine houses in Korea and then to present beneficial information available for Korean pig producers to manage optimal air quality in swine house. The swine houses investigated in this research were selected based on three criteria; manure removal system, ventilation mode and growth stage of swine. Mean concentrations of aerial pollutants in swine houses were 8 ppm for ammonia, 300 ppb for hydrogen sulfide, 2 mg m(-3) for total dust, 0.6 mg m(-3) for respirable dust, 4 log(cfu m(-3)) for total airborne bacteria and 3 log(cfu m(-3)) for total airborne fungi, respectively. Mean emissions based on pig (liveweight; 75 kg) and area (m(2)) were 250 and 340 mg h(-1) for ammonia, 40 and 50 mg h(-1) for hydrogen sulfide, 40 and 50 mg h(-1) for total dust, 10 and 15 mg h(-1) for respirable dust, 1.0 and 1.3 log(cfu) h(-1) for total airborne bacteria and 0.7 and 1.0 log(cfu) h(-1) for total airborne fungi, respectively. In general concentrations and emissions of gases were relatively higher in the swine houses managed with deep-pit manure system with slats and mechanical ventilation mode than the different swine housing types whereas those of particulates and bioaerosol were highest in the naturally ventilated swine houses with deep-litter bed system.

Temporal and spatial distributions of aerial contaminants in an enclosed pig building in winter.

Optimal management of indoor air quality in an enclosed swine house, especially in winter, is indispensable for preventing the transmission of infectious respiratory disease to workers and animals. Studies are needed to elucidate the correlation of aerial contaminants and climate factors. It was observed that indoor air contamination in an enclosed pig building was the highest at 2:00--5:00 PM, followed by 8:00--11:00 PM and 8:00--11:00 AM. It was assumed that this was attributed to the increase of swine activities in the afternoon. In general, the concentration of total dust and total airborne bacteria in an enclosed pig building was found to have a significant correlation with temperature and relative humidity (P<0.05). There were significant correlations between total dust and total airborne bacteria, between total dust and ammonia, and between total dust and odor at the 95% confidence level. In conclusion, temperature and total dust concentration correlated significantly with all parameters except hydrogen sulfide (H(2)S). This could have been due to the drying of swine feces due to the increase in interior temperature and the resuspension of feed deposited on the floor of the pig building by the swines' activity, resulting in a high generation of dust that adsorbed and carried the airborne bacteria and odorous compounds in the enclosed pig building. It was proved that an adsorptive capacity of dust for ammonia was higher than for hydrogen sulfide.