The glycoprotein hormone family consists of follicle-stimulating hormone (FSH; GTH1), luteinizing hormone (LH; GTH2), and thyroid-stimulating hormone (TSH), which are secreted by the pituitary gland in all mammalian species, and chorionic gonadotropin, which is secreted by placental trophoblast cells in primates and equids. These hormones consist of non-covalently associated α-, β- subunits. Within a species, the amino acid sequence of α-subunit is identical across all glycoprotein hormones and is encoded by a single gene. The αβ dimer is the active form of the hormone, and biological specificity is conferred by the β-subunit. Both of α and β subunit of eel FSH has two N-glycosylation sites (α-subunit: Asn56 and Asn79; β -subunit: Asn5 and Asn22, respectively). In the present study, we constructed deglycosylated mutants at single and double sites in each subunits of eel FSH for identification of Asn linked oligosaccharides' biological role. Mutant cDANs were cloned into pcDNA3 expression vector and transiently transfected into CHO suspension cells. The quantity of rec-eelFSHs were quantified by sandwich ELISA system, using monoclonal antibodies produced in our lab. The wild type rec-FSH protein was detected at the predicted molecular weight of 34 kDa by western blot. The molecular weight of deglycosylated mutants at single site decreased with about 4 kDa and of mutants at double sites decreased with 8 kDa. After PNGase treatment in the rec-eel FSH proteins, molecular weight also decreased to 7-8 kDa. We generated stably parental cell lines, engineered to express a β-arrestin 2EA fusion protein, expressing eel FSHR and C-terminal deleted mutant. 2 out of 5 receptor cells each were selected by G-418 and we tested these cell lines in a receptor functionality using PathHunter β arrestin assay (DiscoverX). Follicle stimulating hormone acts through binding to its specific receptor. Binding of ligand to the receptor activates the adenosine 3',5'-cyclic monophosphate (cAMP) pathway (McFarland et al., 1989; Ji and Ji, 1991a; Rose, 1998) and the inositol 1phosphate (IP1) the second messenger systems. After stimulation of eelFSH receptor stably transfected Parental CHO cells with FSH wild type and mutant hormones as a ligand, production of cAMP and IP-1 were evaluated (Cisbio). cAMP IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79; and βΔ5 were 33.1; 1154.7; 22; 410 and 311.9 ng/ml, respectively. IP-1 IC-50 values by eelFSHwt; αΔ56; αΔ79; αΔ56_79 and βΔ5 were 6.8; 7.1; 4.4; 3.8 and 10.2 ng/ml, respectively too. The cAMP activation was greatly decreased in the αΔ56αmutant. Thus, the site of α56 oligosaccharide in the eel plays an pivotal role for the cAMP stimulation using eel FSH receptor cell lines. In the IP-one assay, the activity in the αΔ56 and βΔ5 mutants was a little decreased than the wt. The biological roles of N-linked oligosaccharides in GPCR internalization are going to be estimated by measuring β arrestin recruitment system.

In this study, we developed and validated microanalysis methods for the determination of linear alkylbenzenesulfonate (LAS), sodium lauryl sulfate (SLS), and alpha olefin sulfonate (AOS). The conditions for the analysis of the surfactants using HPLC with FLD, RID, and ELSD detectors were investigated. The methods were validated by determining the linearity, limits of detection (LODs), limits of quantification (LOQs), recovery, precision, and accuracy. LAS analysis by FLD revealed calibration curves that were linear in the range of 10-200 mg/L for an LAS mixture. The calibration curves for C10-C13 had correlation coefficients of 0.995, 0.997, 0.996, and 0.997, respectively. SLS analysis using RID generated a linear calibration curve in the range of 10-300 mg/L. The calibration curve for SLS C12 had a correlation coefficient of 0.9994. AOS analysis using ELSD resulted in a correlation coefficient of 0.9940. For LAS, the LODs and LOQs were 0.09-0.56 and 0.30-1.87 mg/L, respectively. For SLS C12, the LOD and LOQ were 0.07 and 2.33 mg/L, respectively. For AOS C14, the LOD and LOQ were 16.55 and 21.83 mg/L, respectively. The recoveries were 97.17-98.84% for LAS C10-C14, 97.94% for SLS C12, and 96.11% for AOS C14. The established methods provide acceptable precision and accuracy. Our methods could be useful for the detection of anionic surfactants in dishwashing detergents.

Floor damping materials used in floating floor system to diminish the floor noise have been made with low density and dynamic stiffness. Owing to this low density and dynamic stiffness, the deflection in these materials under long-term loading and cracking of the floor finishing mortar in the floating floor system may occur. This paper presents the results of long-term loading effects on the deflection of different types of floor damping materials. The experimental program involved the long-term loading tests for 490 days loading period on sixteen specimens. Specimens were classified as DM1(Damping Materials) to DM8, depending upon the four main parameters; types, bottom shapes and densities of floor damping materials and amount of loading. Results indicated that the long-term deflection of all specimens of damping materials remained unchanged after 200 days at all loading amounts, except the specimens made up of Polystrene, in which long-term deflection remained unchanged after 160 days at 250 N load and 100 days 500 N load. In this paper, two types of correlation expressions were shown in the deflection range prior to the range where deflection remained constant; two analyses by ISO 20392 and linear regression. In comparison of two analyses and experimental results on the difference of deflection of 16 specimens, the difference of deflection was below 0.4 mm in those analyses in case of that total deflection was below 10 mm. Restrictively, it was judged that the analysis for the deflection of specimens made up of Polystrene is more appropriate using ISO 20392.

This study was carried out to investigate and assess the cold tolerance of 24 species broad-leaved evergreen trees in southern region, South Korea, and propose the selection for urban greening responsive to the climate change. The cold stressed impact of each species was measured and calculated by the electrolyte leakage (EL) method, and then the lethal temperature was predicted by the non-linear regression analysis. The scattered plots and fitted curves of most species tended to show sigmoidal response curve. On assessing the EL values and sigmoidal response curve pattern with different temperature, the differences were obviously showed among all the species. Also, among the species within the same family, the differences were obviously showed. The maximum temperature difference among the species was over 10℃. Between Ilex rutunda and Ilex integra within the same family, Aquifoliaceae, it was over 10℃. The results indicate that there are significant differences in cold tolerance among different species in the same region, which are not affected by any environmental factors but affected by any genetical factors. Thus it is valuable to assess the cold tolerance on most broad-leaved evergreen trees in southern region, South Korea. As a result, Euonymus japonicus, Trachelospermum asiaticum, Dendropanax morbiferus, Ilex integra, Machilus thunbergii, Ilex x wandoensis, Cinnamomum japonicum, Distylium racemosum, and Castanopsis sieboldii may have better cold tolerance and survive the region closer to middle region, South Korea compared to the others.