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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Restriction of Metabolizable Energy in Broiler Growers and Its Impact on Grower and Breeder Performance

  • Sunder, G. Skyam;Kumar, Ch. Vijaya;Panda, A.K.;Raju, M.V.L.N.;Rao, S.V. Rama;Gopinath, N.C.S.;Reddy, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.8
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    • pp.1258-1265
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    • 2007
  • Metabolizable energy (ME) required for basal metabolism, activity and growth was considered as the criterion for targeting specific increases in body weight (100 g/week) of broiler chicks during the grower phase (5-20 weeks) and its impact was evaluated on breeder performance. Broiler female chicks (460) from a synthetic dam line were randomly distributed to 4 test groups with 23 replicates of 5 birds each and housed in cages. The first group (ME-100) was offered a calculated amount of ME by providing a measured quantity of grower diet (160 g protein and 2,600 kcal ME/kg) which increased with age and weight gain (133-294 kcal/bird/day). The other three groups were offered 10 or 20% less ME (ME-90 and ME-80, respectively) and 10% excess ME (ME-110) over the control group (ME-100). From 21 weeks of age, a single breeder diet (170 g protein and 2,600 kcal ME/kg) was uniformly fed to all groups and the impact of grower ME restriction on breeder performance evaluated up to 58 weeks. The targeted body weight gain of 1,600 g in a 16-week period was achieved by pullets of the ME-100 group almost one week earlier by gaining 8.7 g more weight per week. However, pullets in the ME-90 group gained 1,571 g during the same period, which was closer to the targeted weight. At 20 weeks of age, the conversion efficiency of feed (5.21-5.37), ME (13.9-14.1 kcal/g weight gain) and protein (0.847-0.871 g/g weight gain), eviscerated meat yield, giblet and tibia weights were not influenced by ME restriction, but the weights of abdominal fat and liver were higher with increased ME intake. Reduction of ME by 10% in the grower period significantly delayed sexual maturity (169.3 d), but increased egg production (152.5 /bird) with better persistency. Improved conversion efficiency of feed, ME and protein per g egg content were also observed in this group up to 56 weeks. The fertility and hatchability at 58 weeks of age were higher in the ME-90 group compared to the control and 10% excess ME feeding. In conclusion, the present study revealed the possibility of achieving targeted weight gain in broiler growers by feeding measured quantities of ME during the rearing period with consequential benefits in breeder performance.

Development of Chemiluminescence Immunoassay(CIA) & ELISA for the Detection of Anti-sperm Antibodies in Male Serum (항정자 항체 검출을 위한 CIA 및 ELISA 개발을 위한 기초 연구)

  • Kim, S.C.;Lee, K.S.;Kim, Y.K.;Kim, C.K.;Choi, K.H.;Kwon, O.J.;Kim, J.B.
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.1
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    • pp.71-80
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    • 1990
  • New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

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The Crystallographic and Magnetic Properties of $Fe_{1-x}Co_x$System ($Fe_{1-x}Co_x$계의 결정구조와 자기적인 성질)

  • 김정기;한경훈;서정철
    • Journal of the Korean Magnetics Society
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    • v.9 no.4
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    • pp.190-195
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    • 1999
  • The crystallographic and magnetic properties of the system of $Fe_{1-x}Co_x$(x=0.2 and 0.4) prepared by microwave arc-melting with the maximum power of 3.5 kW and a iron-foil with thickness of 25 ${\mu}{\textrm}{m}$ have been studied by the methods of X-ray diffraction and the measurement of the magnetic hysteresis using the vibrating sample magnetometer at room temperature. The samples were prepared in three different ways: First, pellet form pressed under the pressure of 9,000 N/$\textrm{cm}^2$. Second, the sheet cold rolled. Third, thin sheet treated with the temperature of 90$0^{\circ}C$. The X-ray diffraction pattern of the sample prepared by the first method shows that the crystal structure of the sample is bcc as same as that of Fe with a good uniformity. The iron-foil has the coercivity of 43 Oe and the initial slope of magnetization of 0.328 emu/gOe. The coervicity and magnetization of the sample prepared by the second method increased as the Co content increased. But the initial slop of the magnetization decreased as the Co content increased. This means that the displacement of domain wall is suppressed by the increases of coercivity as the Co content increased. The saturation magnetization of the samples made by the third method increased. On the other hand, the coercivity of these samples decreased. The increase of saturation magnetization of the samples seems to be related to the changes in X-ray intensity after heat treatment. Also some magnetic parameters of the samples were calculated by using a simple model and compared with other values.

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Crystal Structure and Magnetic Properties of Iron Doped La-Sr-Mn-O (철을 미량 치환한 La-Sr-Mn-O의 결정학적 및 자기적 특성 연구)

  • Ahn, Geun-Young;Shim, In-Bo;Kim, Sam-Jin;Kim, Chul-Sung
    • Journal of the Korean Magnetics Society
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    • v.12 no.1
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    • pp.14-19
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    • 2002
  • The iron-doped perovskite La$_{0.67}$Sr$_{0.33}$Mn$_{0.99}$$^{57}$Fe$_{0.01}$O$_3$compound has been studied by x-ray diffraction, Mossbauer spectroscopy, and vibrating sample magnetometry. The single phase of the polycrystalline La$_{0.67}$Sr$_{0.33}$Mn$_{0.99}$$^{57}$Fe$_{0.01}$O$_3$powder has been prepared by a waterbased solgel method. Crystalline La$_{0.67}$Sr$_{0.33}$Mn$_{0.99}$$^{57}$Fe$_{0.01}$O$_3$was a rombohedral structure with lattice parameters a$_{0}$=5.480 $AA$, $alpha$=60.259$^{circ}$. Mossbauer spectra of La$_{0.67}$Sr$_{0.3}$/Mn$_{0.99}$$^{57}$Fe$_{0.01}$O$_3$have been taken at various temperatures ranging from 20 to 400 K. As the temperature increases toward the Curie temperature, T$_{c}$=375 K, the Mossbauer spectra show line broadening and the difference between the 1,6 and 3,4 linewidths is caused by the anisotropic hyperfine field fluctuation. The anisotropic field fluctuation of +H (P$_{+}$=0.80) is greater than -H (P$_{-}$=0.20). We calculated that the anisotropy energy was 124.01 erg/cm$^3$for T=150 K which is associated with the large line broadening.

Effects of $Dy_2$$O_3$ composition for the photoluminescence and long-phosphorescent characteristics of stuffed tridymite $SrAl_2$$O_4$ : $Eu^{2+}$ phosphors (Stuffed tridymite계 $SrAl_2$$O_4$ : $Eu^{2+}$ 형광체의 발광 및 장잔광특성에 미치는 $Dy_2$$O_3$의 영향)

  • 이영기;김병규
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.11 no.2
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    • pp.71-77
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    • 2001
  • We investigated photoluminescence, long-phosphorescent and crystalline properties with various $Dy_2$$O_3$ compositions (0.0~9.5mol%) in $SrAl_2$$O_4$ : $Eu^{2+}$,$Dy^{3+}$ phosphor powders prepared by the solid state reaction. The highest emission wavelength (520nm) of photoluminescence spectra was not affected by the Dy doping concentrations. The$SrAl_2$$O_4$single phase which was determined by X-ray diffraction and photoluminescence was appeared for the concentrations of $Dy_2$$O_3$$\leq$1.0 mol%. After removal of the pulsed Xe-lamp excitation (360nm), also, excellent long phosphorescent properties of the phosphors were obtained with the concentrations of $Dy_2$$O_3$$\leq$1.0mol%, although the decay time for all phosphors decrease exponentially.

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Helicopter-borne and ground-towed radar surveys of the Fourcade Glacier on King George Island, Antarctica (남극 킹조지섬 포케이드 빙하의 헬리콥터 및 지상 레이다 탐사)

  • Kim, K.Y.;Lee, J.;Hong, M.H.;Hong, J.K.;Shon, H.
    • Geophysics and Geophysical Exploration
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    • v.13 no.1
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    • pp.51-60
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    • 2010
  • To determine subglacial topography and internal features of the Fourcade Glacier on King George Island in Antarctica, helicopter-borne and ground-towed ground-penetrating radar (GPR) data were recorded along four profiles in November 2006. Signature deconvolution, f-k migration velocity analysis, and finite-difference depth migration applied to the mixed-phase, single-channel, ground-towed data, were effective in increasing vertical resolution, obtaining the velocity function, and yielding clear depth images, respectively. For the helicopter-borne GPR, migration velocities were obtained as root-mean-squared velocities in a two-layer model of air and ice. The radar sections show rugged subglacial topography, englacial sliding surfaces, and localised scattering noise. The maximum depth to the basement is over 79m in the subglacial valley adjacent to the south-eastern slope of the divide ridge between Fourcade and Moczydlowski Glaciers. In the ground-towed profile, we interpret a complicated conduit above possible basal water and other isolated cavities, which are a few metres wide. Near the terminus, the GPR profiles image sliding surfaces, fractures, and faults that will contribute to the tidewater calving mechanism forming icebergs in Potter Cove.

Effects of Ultrasound and High-Voltage Pulsed Current on Adjuvant-Induced Arthritis in Rats (Adjuvant로 유발된 관절염의 흰쥐에서 초음파와 고전압 맥동 전류 치료의 효과)

  • Lee, Dong-Jin;Lee, Dong-Yeop;Hwang, Don-Young
    • Physical Therapy Korea
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    • v.13 no.3
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    • pp.33-40
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    • 2006
  • The purpose of this study was to assess the effects of High-Voltage Pulsed Current (HVPC) and ultrasound on adjuvant-induced arthritis in rats. Adjuvant arthritis was induced in female Sprauge-Dawley rats by the subcutaneous injection of a single dose of $.1m{\ell}$ of Complete Freund's Adjuvant (CFA) (1 mg of Mycobacterium Butyricum suspended in $.1m{\ell}$ paraffin oil) into the right hind paw. A randomized, parallel-groups design of 24 subjects was used. All rats were randomly assigned to control (n=8), ultrasound (n=8), and HVPC (n=8) were compared with those of injured rats. The rats in the pulsed ultrasound group were treated at 1 MHz frequency with $.5W/cm^2$ intensity in 1:4 mode for 5 minutes per day. The rats in the HVPC group were treated at 120 pulses per second and $50{\mu}s$ phase duration, 20 mA intensity for 30 min per day. Treatment was done in the left and right hind limb for 2 weeks. We evaluated clinical, radiographic, hematologic and histopathologic findings before and after treatment and obtained the following results. 1. Edema of the right hind paw was more significantly reduced in the ultrasound and HVPC groups than the control group on days 9, 12, and 14 (p<.05). Edema of the left hind paw was more significantly reduced in ultrasound and HVPC groups than the control group on days 12, 14 (p<.05). 2. WBC counts of the ultrasound and HVPC groups as compared with the control group were becoming remarkably decreased after the treatment. 3. In radiologic findings, arthritis formation was seen according to the score of arthritis, which was the highest in the control group, upon the observation of radiographs of the left and right hind paws. However, no statistically significant difference was present in the score within three groups. 4. In the histopathologic findings, ultrasound and HVPC groups had effectively suppressed erosions of articular cartilage and inflammatory cell infiltrations. Therefore, the results of the study show that rats that were treated with the ultrasound and HVPC effectively suppressed adjuvant arthritis. However, no statistically significant difference was present between the ultrasound group and the HVPC group.

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The molten KOH/NaOH wet chemical etching of HVPE-grown GaN (HVPE로 성장된 GaN의 용융 KOH/NaOH 습식화학에칭)

  • Park, Jae Hwa;Hong, Yoon Pyo;Park, Cheol Woo;Kim, Hyun Mi;Oh, Dong Keun;Choi, Bong Geun;Lee, Seong Kuk;Shim, Kwang Bo
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.24 no.4
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    • pp.135-139
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    • 2014
  • The hydride vapor phase epitaxy (HVPE) grown GaN samples to precisely measure the surface characteristics was applied to a molten KOH/NaOH wet chemical etching. The etching rate by molten KOH/NaOH wet chemical etching method was slower than that by conventional etching methods, such as phosphoric and sulfuric acid etching, which may be due to the formation of insoluble coating layer. Therefore, the molten KOH/NaOH wet chemical etching is a better efficient method for the evaluation of etch pits density. The grown GaN single crystals were characterized by using X-ray diffraction (XRD) and X-ray rocking curve (XRC). The etching characteristics and surface morphologies were studied by scanning electron microscopy (SEM). From etching results, the optimum etching condition that the etch pits were well independently separated in space and clearly showed their shape, was $410^{\circ}C$ and 25 min. The etch pits density obtained by molten KOH/NaOH wet chemical etching under optimum etching condition was around $2.45{\times}10^6cm^{-2}$, which is commercially an available materials.