• Animal Bioscience
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• v.35 no.3
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• pp.399-409
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• 2022

Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing β-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or half maximal effective concentration (EC₅₀) levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC₅₀ levels (3,910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.

• The Journal of Advanced Prosthodontics
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• v.6 no.2
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• pp.79-87
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• 2014

PURPOSE. The aim of this study was to determine the efficiency of Erbium, Chromium: Yttrium-Scandium-Gallium-Garnet laser in different output powers for removing permanent resin cement residues and therefore its influence on microshear bond strength compared to other cleaning methods. MATERIALS AND METHODS. 90 extracted human molars were sectioned in 1 mm thickness. Resin cement was applied to surface of sliced teeth. After the removal of initial cement, 6 test groups were prepared by various dentin surface treatment methods as follows: no treatment (Group 1), ethylene diamine tetra acetic acid application (Group 2), Endosolv R application (Group 3), 1.25 W Erbium, Chromium:Yttrium-Scandium-Gallium-Garnet laser irradiation (Group 4), 2 W Erbium, Chromium:Yttrium-Scandium-Gallium-Garnet laser irradiation (Group 5) and 3.5 W Erbium, Chromium:Yttrium-Scandium-Gallium-Garnet laser irradiation (Group 6). The topography and morphology of the treated dentin surfaces were investigated by scanning electron microscopy (n=2 for each group). Following the repetitive cementation, microshear bond strength between dentin and cement (n=26 in per group) were measured with universal testing machine and the data were analyzed by Kruskal Wallis H Test with Bonferroni correction (P<.05). Fracture patterns were investigated by light microscope. RESULTS. Mean microshear bond strength ${\pm}$ SD (MPa) for each group was $34.9{\pm}17.7$, $32.1{\pm}15.8$, $37.8{\pm}19.3$, $31.3{\pm}12.7$, $44.4{\pm}13.6$, $40.2{\pm}13.2$ respectively. Group 5 showed significantly difference from Group 1, Group 2 and Group 4. Also, Group 6 was found statistically different from Group 4. CONCLUSION. 2 W and 3.5 W Erbium, Chromium: Yttrium-Scandium-Gallium-Garnet laser application were found efficient in removing resin residues.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.5
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• pp.638-645
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• 2013

Interferon-tau (IFNT) is thought to be the conceptus protein that signals maternal recognition of pregnancy in ruminants. We and others have observed that OCT4 expression persists in the trophectoderm of ruminants; thus, both CDX2 and OCT4 coexist during the early stages of conceptus development. The aim of this study was to examine the effect of CDX2 and OCT4 on IFNT gene transcription when evaluated with other transcription factors. Human choriocarcinoma JEG-3 cells were cotransfected with an ovine IFNT (-654-bp)-luciferase reporter (-654-IFNT-Luc) construct and several transcription factor expression plasmids. Cotransfection of the reporter construct with Cdx2, Ets2 and Jun increased transcription of -654-IFNT-Luc by about 12-fold compared with transfection of the construct alone. When cells were initially transfected with Oct4 (0 h) followed by transfection with Cdx2, Ets2 and/or Jun 24 h later, the expression of -654-IFNT-Luc was reduced to control levels. OCT4 also inhibited the stimulatory activity of CDX2 alone, but not when CDX2 was combined with JUN and/or ETS2. Thus, when combined with the other transcription factors, OCT4 exhibited little inhibitory activity towards CDX2. An inhibitor of the transcriptional coactivator CREB binding protein (CREBBP), 12S E1A, reduced CDX2/ETS2/JUN stimulated -654-IFNT-Luc expression by about 40%, indicating that the formation of an appropriate transcription factor complex is required for maximum expression. In conclusion, the presence of OCT4 may initially minimize IFNT expression; however, as elongation proceeds, the increasing expression of CDX2 and formation of the transcription complex leads to greatly increased IFNT expression, resulting in pregnancy establishment in ruminants.

• Korean Journal of Environmental Agriculture
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• v.38 no.4
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• pp.254-261
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• 2019

BACKGROUND: Agricultural water is known to be one of the major routes in bacterial contamination of fresh vegetable. However, there is a lack of fundamental data on the microbial safety of agricultural water in Korea. METHODS AND RESULTS: We investigated the density of indicator bacteria in the surface water samples from 31 sites collected in April, July, and October 2018, while the groundwater samples were collected from 20 sites within Jeollabuk-do in April and July 2018. In surface water, the mean density of coliform, fecal coliform, and Escherichia coli was 2.7±0.55, 1.9±0.71, and 1.4±0.58 log CFU/100 mL, respectively, showing the highest bacterial density in July. For groundwater, the mean density of coliform, fecal coliform, and E. coli was 1.9±0.58, 1.4±0.37, and 1.0±0.33 log CFU/ 100mL, respectively, showing no significant difference between sampling time. The survival of E. coli O157:H7 were prolonged in water with higher organic matter contents such as total nitrogen (TN), and nitrate-nitrogen (NO₃-N). The reduction rates of E. coli O157:H7 in the water showed greater in order of 25, 35, 5, and 15℃. CONCLUSION: These results can be utilized as fundamental data for prediction the microbiological contamination of agricultural water and the development of microbial prevention technology.

• Korean Journal of Food Science and Technology
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• v.46 no.5
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• pp.609-615
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• 2014

The cytoprotective effect of Moringa oleifera Lam. (drumstick tree) on neuronal cells was investigated to confirm the physiological benefits associated with this natural food resource. First, the drumstick tree extract was chemically analyzed to determine inherent nutritional constituents. Calcium and potassium were identified as the major mineral constituents, and palmitic acid (C16:0, 16.33%) and gadoleic acid (C20:01, 66.34%) were detected as the major fatty acids. Moreover, drumstick tree extract contained 94.78 mg/100 g vitamin E and 112.61 mg/100 g niacin. PC12 cells were used to study the cytoprotective effects of drumstick tree extract. Intracellular accumulation of reactive oxygen species was significantly reduced when $H_2O_2$ treated-neuronal cells were cultured in a medium containing the methanolic extract of drumstick tree, compared to cells treated with only $H_2O_2$. Cell viability assay using MTT showed that the extract protected cells against $H_2O_2$-induced neurotoxicity and inhibited LDH leakage from the cell membrane. Caspase assay showed that the extract exerted cytoprotective effect against apoptosis. Consequently, these data suggest that drumstick tree is a useful natural resource with positive effects on human health.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.2
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• pp.114-120
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• 2007

The augmentation of soft tissue defects is one of the critical problems in the oral and maxillofacial surgery. Various types of graft materials, both autologous and non-autologous, have been used for the augmentation of soft tissue in the facial region. However, it is not easy to choose an ideal material for soft tissue augmentation because each has its advantages and disadvantages. An ideal graft material should meet the following criteria : it should not leave a scar at the area from which it was taken; should have less likelihood of causing infection; should feel natural after implanted; and should be not absorbed. Among the materials meeting these criteria, human dermis and artificial dermis are commonly used for clinical purposes. The present study was aimed to investigate and compare the resorption rate and the histological change following the use of the autologous dermis, the human homogenous dermis $Alloderm^{(R)}$, and the artificial dermis $Terudermis^{(R)}$ to reconstruct the soft tissue defect. Twenty mature rabbits of either sex, weighing about 2 ㎏, were used. Each rabbit was transplanted with the autologous dermis, $Alloderm^{(R)}$, and $Terudermis^{(R)}$ size $1{\times}1-cm$ at the space between the external abdominal oblique muscle and the external abdominal oblique fascia. They were then divided into 4 groups (n=5 each) according to the time elapsed after the surgery: 1, 2, 4, and 8 weeks. The resorption rate was calculated by measuring the volume change before and after the transplantation, and H-E stain was preformed to observe the histological changes. The resorption rate after 8 weeks was 21.5% for the autologous dermis, 16.0% $Alloderm^{(R)}$, and 36.4% $Terudermis^{(R)}$, suggesting that $Alloderm^{(R)}$ is the most stable while $Terudermis^{(R)}$ is the most unstable. In microscopic examinations, the autologous dermis graft was surrounded by inflammatory cells and showed foreign body reactions. The epidermal inclusion cyst was observed in the autologous dermis graft. $Terudermis^{(R)}$ and $Alloderm^{(R)}$ demonstrated neovascularization and the progressive growth of new fibroblast. The results suggest that $Terudermis^{(R)}$ and $Alloderm^{(R)}$ can be availably for substituting the autologous dermis.

• The Korean Journal of Pharmacology
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• v.17 no.2
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• pp.37-45
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• 1981

The importance of thyroid hormones for the survival of rats in the cold is along-established fact. Hypothyroid animals are unable to survive in a cold environment. It was also reported that acute exposure of rats, guinea pigs and rabbits to cold produced an increased secretion of TSH and thereby thyroid hormone secretion within 10 to 30 min, but this increase of thyroid activity disappeared quite rapidly during warming. However, in human study no significant difference was found in the concentration of $T_4$, TSH and cortisol between summer and winter. But plasma $T_3$ concentration was increased significantly in winter in 56 adult men. On the other hand, it has been also known that catecholamines are important in the maintenance of body temperature of rat exposured to cold. Abundant evidences suggest that the sympathetic nervous system is involved in the activation of nonshivering thermogenesis and that thyroid hormone metabolism and secretion are influenced by catecholamines and consequently by the activity of the sympatheticadrenal system. Many of the metabolic effects of catecholamines are associated with an increase in the level of cAMP mediated through activation of adenylate cyclase which converts ATP to cAMP. Other studies have shown that thyroid hormones affect the amount of adenylate cyclase present in the adipose tissue. On the other hand. it was also reported that a particulate cAMP phosphodiesterase activity in fat cells was modulated by the action of thyroid hormones. The objective of the present study was to determine the interaction between thyroid activity and cyclic nucleotides during acute exposure to cold. Albino rats weighing around 200 g were used as the experimental animal. The room temperature group was kept at $25^{\circ}C$ and the cold-exposured group was kept at $4^{\circ}C$ for 1 week or 2 weeks. Each group was subdivided into three subgroups; control, KI, and MTU group. At the end of experiment the animals were etherized and blood was taken from abdominal aorta for $T_4,\;T_3$ and cyclic nucleotides. The determinations of $T_3,\;T_4$ and cyclic nucleotides were carried out with a radioimmunoassay(RIA) method. The results were summerized as followings. 1) A significant increase of thyroid weight was observed in rats exposured to cold for 2 weeks. Furthermore, in rats administered MTU while to exposure to cold the thyroid weight was also increased significantly. 2) After 2 weeks $T_3$ concentration in the plasma of cold-exposured rats was significantly increased in KI group and MTU group as well as in control group. On the contrary, after 2 weeks of cold exposure $T_4$ level was decreased in control group. 3) In the case of cyclic nucleotides, plasma cAMP was increased in the control group after 1 or 2 weeks of cold exposure. However, cAMP level in plasma was rather significantly decreased in KI group and MTU group as well as in control group.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.947-953
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• 2004

The antioxidative and hepatoprotective potentials of two anthraquinones, alaternin (2-hydroxy-emodin) and emodin, to scavenge and/or inhibit hydroxyl radicals generated by the Fenton reaction and to protect tacrine-induced cytotoxicity in human liver derived HepG2 cells were evaluated, respectively. The inhibitory activity on hydroxyl radical generated in a cell-free chemical system (FeSO$_4$/$H_2O$$_2$) was investigated by a fluorescence spectrophotometer using a highly fluorescent probe, 2$^1$,7$^1$-dichlorofluorescein. The hydroxyl radical scavenging activity was determined by electron spin resonance spectroscopy using 5,5-dimethy-1-pyrroline-N-oxide as hydroxyl radicals trapping agents. Tacrine-induced HepG2 cell toxicity was determined by a 3-［4,5-dimethylthiazole-2yl］-2,5-diphenyltertrazolium bromide assay. Although the scavenging activity of alaternin on hydroxyl radical was similar to that of emodin in dose-dependent pat-terns, the inhibitory activity exhibited by the former on hydroxyl radical generation was stron-ger than that of the latter, with $IC_{50}$/ values of 3.05$\pm$0.26 $\mu$M and 13.29$\pm$3.20 $\mu$M, respectively. In addition, the two anthraquinones, alaternin and emodin showed their hepatoprotective activ-ities on tacrine-induced cytotoxicity, and the EC$_{50}$ values were 4.02 11M and 2.37 $\mu$M, respec-tively. Silymarin, an antihepatotoxic agent used as a positive control exhibited the EC$_{50}$ value of 2.00 $\mu$M. These results demonstrated that both alaternin and emodin had the simultaneous antioxidant and hepatoprotective activities.ies.

• Journal of Industrial and Engineering Chemistry
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• v.67
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• pp.244-254
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• 2018

Surface modified poly ${\text\tiny{L}}$-lactic acid (PLLA) samples with hydroxyapatite (HA), heparin and bone morphogenetic protein-2 (BMP-2) mediated by polydopamine (pDA) coating (PLLA/pDA/HA/Hep/BMP-2) were prepared, and their effects on the enhancements of bone formation and osseointegration were evaluated in vitro and in vivo as compared to PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. The changes in surface chemical compositions, morphologies and wettabilities were observed by X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM), atomic force microscopy (AFM) and water contact angle measurements. Pre-coating of HA particles with pDA provided uniform and homogeneous anchoring of particles to PLLA surface. In addition, the strong ionic interaction between heparin and pDA led PLLA surface readily heparinized for loading of BMP-2. In vitro experiments revealed that the levels of alkaline phosphatase (ALP) activity, calcium deposition, and osteocalcin (OCN) gene expression were higher in MG-63 human osteosarcoma cell lines grown on PLLA/pDA/HA/Hep/BMP-2 than on control PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. In vivo studies using micro-computed tomography (micro-CT) also showed that PLLA/pDA/HA/Hep/BMP-2 screw exhibited greatest value of bone volume (BV) and bone volume/tissue volume (BV/TV) among samples. Histological evaluations with H&E and Von Kossa staining demonstrated that a combination of HA and BMP-2 contributed to the strong osseointegration.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.1
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• pp.29-33
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• 2005

A DNA vaccine methodology using eukaryote expression vectors to produce immunizing proteins in the vaccinated hosts is a novel approach to the development of vaccine and immuno-therapeutics, and it has achieved considerable success over several infectious diseases and various cancers. To further enhance its efficiency, attempts were made to develop novel plasmid vectors containing multiple immunostimulatory CpG motifs, for rapid and strong immune response. First, a 2.9 kb compact plasmid vector (pVAC), containing CMV promoter, polycloning site, BGH poly(A) terminator, ampicillin resistance gene and pBR322 origin was constructed. A pVAC-hEPO was also constructed, which contained a human erythropoietin gene, for evaluating the transfection efficiency of naked plasmid DNA both in vitro and in vivo. To examine the adjuvant effect of multi-CpG motifs on naked plasmid DNA, 22 and 44 enriched and unmethylated CpG motifs were introduced into pVAC to generate pVAC-ISS1 and pVAC-ISS2, respectively. $100{\mu}g$ of pSecTagB, pVAC, pVAC-ISS1 or pVAC-ISS2 were each injected intramuscularly into the tibilias anterior muscle of Balb/c mice. The level of interleukin-6 induced in the mice injected with pVAC-ISS1 and pVAC-ISS2 were significantly elevated after 12 hours, which were almost 2 and 2.5 times higher than that in the mice injected with pSecTagB, respectively. These results suggest that DNA vaccine plasmids with enriched CpG motifs can induce rapid secretion of interleukin-6 by lymphocytes. In conclusion, these vectors can contribute to the development of adjuvant-free DNA vaccinations against infectious diseases and various cancers.