Apo-9′-Fucoxanthinone,Isolated from Sargassum muticum,Inhibits CpG-Induced Inflammatory Response by Attenuating the Mitogen-Activated Protein Kinase Pathway

Sargassum muticum (S. muticum) is a brown edible alga and widely distributed in Korea. This report was designed to evaluate the anti-inflammatory properties of apo-9′-fucoxanthinone (APO-9′) isolated from S. muticum on pro-inflammatory cytokine production. S. muticum extract (SME) exhibited significant inhibitory effects on pro-inflammatory cytokine production in bone marrow-derived macrophages (BMDMs) and dendritic cells (BMDCs). APO-9′ pre-treatment in the CpG DNA-stimulated BMDMs and BMDCs showed a strong dose-dependent inhibitory effect on interleukin (IL)-12 p40, IL-6 and tumor necrosis factor (TNF)-α production with IC₅₀ values ranging from 5.31 to 13.79. It exhibited a strong inhibitory effect on the phosphorylation of ERK1/2 and on activator protein (AP)-1 reporter activity. APO-9′ pre-treatment exhibited significant inhibition of CpG DNA-induced production of inducible nitric oxide synthase. Taken together, these data suggest that SME and APO-9′ have a significant anti-inflammatory property and warrant further studies concerning the potentials of SME and APO-9′ for medicinal use.     

A preliminary study of dietary protein requirement of juvenile marbled flounder (Pseudopleuronectes yokohamae)

An 8-week feeding trial was conducted to determine the optimal dietary protein level for juvenile marbled flounder. Five semi-purified test diets were formulated to contain different protein levels (CP) including 42.7%, 47.4%, 53.3%, 58.8%, and 64.5% (dry matter), named as CP42.7, CP47.4, CP53.3, CP58.8, and CP64.5, respectively. Five hundred and twenty-five juveniles (6.0 ± 0.1 g) were randomly distributed into 15 tanks (300 L tanks), resulting in 35 fish per tank (n = 3 tanks). Fish were fed the test diets 5 times per day until satiation. The CP58.8 resulted in the highest gain in weight and the best efficiency in feed utilization among the tested protein levels (P < 0.05). Fish fed the CP58.8 diet showed significantly higher whole-body protein and lipid contents than the fish that were fed the other diets (P < 0.05). Fish fed the CP53.3, CP58.8, and CP64.5 diets showed a significantly higher dorsal-muscle lipid content than the fish that were fed the CP42.7 and CP47.4 diets (P < 0.05). The one-slope straight broken-line regression analysis on the results of the thermal growth coefficient and feed conversion ratio indicated that the estimated optimum dietary protein level was 58.8%. Taken together, it is suggested that the dietary protein level of 58.8% is optimal for better growth and high efficiency in feed utilization for the juvenile marbled flounder.     

Comparison of arylalkylamine N-acetyltransferase and melatonin receptor type 1B immunoreactivity between young adult and aged canine spinal cord

Melatonin affects diverse physiological functions through its receptor and plays an important role in the central nervous system. In the present study, we compared immunoreactivity patterns of arylalkylamine N-acetyltransferase (AANAT), an enzyme essential for melatonin synthesis, and melatonin receptor type 1B (MT2) in the spinal cord of young adult (2~3 years) and aged (10~12 years) beagle dogs using immunohistochemistry and Western blotting. AANAT-specific immunoreactivity was observed in the nuclei of spinal neurons, and was significantly increased in aged dog spinal neurons compared to young adult spinal neurons. MT2-specific immunoreactivity was found in the cytoplasm of spinal neurons, and was predominantly increased in the margin of the neuron cytoplasm in aged spinal cord compared to that in the young adult dogs. These increased levels of AANAT and MT2 immunoreactivity in aged spinal cord might be a feature of normal aging and associated with a feedback mechanism that compensates for decreased production of melatonin during aging.     

Injection molding of vertebral fixed cage implant

A vertebral cage is a hollow medical device which is used in spine surgery. By implanting the cage into the spine column, it is possible to restore disc and relieve pressure on the nerve roots. Most cages have been made of titanium alloys but they detract the biocompatibility. Currently PEEK (polyether ether ketone) is applied to various implants because it has good properties like heat resistance, chemical resistance, strength, and especially biocompatibility. A new shape of vertebral cage is designed and injection molding of PEEK is considered for production. Before injection molding of the cage, it is needed to evaluate process conditions and properties of the final product. Variables affecting the shrinkage of the cage are considered, e.g., injection time, packing pressure, mold temperature, and melt temperature. By using the numerical simulation program, MOLDFLOW, several cases are studied. Data files obtained by MOLDFLOW analysis are used for stress analysis with ABAQUS, and shrinkage and residual stress fields are predicted. With these results, optimum process conditions are determined.     

Evaluation of a competitive ELISA for antibody detection against avian influenza virus

Active serologic surveillance is necessary to control the spread of the avian influenza virus (AIV). In this study, we evaluated a commercially-available cELISA in terms of its ability to detect AIV antibodies in the sera of 3,358 animals from twelve species. cELISA detected antibodies against reference H1- through H15-subtype AIV strains without cross reactivity. Furthermore, the cELISA was able to detect antibodies produced following a challenge of the AIV H9N2 subtype in chickens, or following vaccination of the AIV H9 or H5 subtypes in chickens, ducks and geese. Next, we tested the sensitivity and specificity of the cELISA with sera from twelve different animal species, and compared these results with those obtained by the hemagglutination-inhibition (HI) test, the "gold standard" in AIV sera surveillance, a second commercially-available cELISA (IZS ELISA), or the agar gel precipitation (AGP) test. Compared with the HI test, the sensitivities and specificities of cELISA were 95% and 96% in chicken, 86% and 88% in duck, 97% and 100% in turkey, 100% and 87% in goose, and 91% and 97% in swine, respectively. The sensitivities and specificities of the cELISA in this study were higher than those of IZS ELISA for the duck, turkey, goose, and grey partridge sera samples. The results of AGP test against duck and turkey sera also showed significant correlation with the results of cELISA (R-value >0.9). In terms of flock sensitivity, the cELISA correlated better with the HI test than with commercially-available indirect ELISAs, with 100% flock sensitivity.     

Comparative study on calretinin immunoreactivity in gerbil and rat retina

Expression of calretinin in retina has been ascribed to multiple biological and functional aspects in the visual system. In this study, we examined the distribution patterns of calretinin immunoreactivity in gerbil and rat retina. In the gerbil, calretinin immunoreactivity was present in bipolar and amacrine cells of the inner nuclear layer and in neurones of the ganglion cell layer. In the rat, amacrine and ganglion cells showed calretinin immunoreactivity, but bipolar cells did not contain calretinin immunoreactivity. In both species, calretinin immunoreactivity was absent in cones, cone bipolars, and horizontal cells. In conclusion, gerbil as well as rat has a rod-dominant retina. The differences in calretinin expression between rat and gerbil require further investigations under various functional and developmental conditions.     

Manipulated Mouse Embryos as Bioassay System for Water Quality Control

Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K⁺ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system.     

Fabrication of electropsun PLGA and small intestine submucosa-blended nanofibrous membranes and their biocompatibility for wound healing

In recent decades, tremendous research has focused on the production of nanoscale fibers using synthetic polymers, with the goal of fabricating nanofibrous scaffolds for wound healing. However, the hydrophobicity of such polymers typically hinders attachment and proliferation of the cells. In this study, we combined poly-d,l-lactide-co-glycolide (PLGA) and small intestine submucosa (SIS) to fabricate blended nanofibers for wound healing by electrospinning. PLGA and SIS were dissolved in 1,1,1,3,3,3-hexafluoro isopropanol to produce different weight ratios of PLGA/SIS-blended nanofibrous membranes (NFM). Physicochemical characterization of the electrospun NFM was performed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, water contact angle analysis, degradation test and tensile testing. The PLGA/SIS-blended NFM showed improved hydrophilicity and tensile strength. Better infiltration, attachment and proliferation of rat granulation fibroblasts of PLGA/SIS-blended NFMs compared to PLGA NFMs were identified by morphological differences determined by SEM and a water-soluble tetrazolium salt assay kit. Based on our results, the PLGA/SIS blended NFMs were found to be suitable for use as a potential material for wound dressing.