• BMB Reports
• /
• v.30 no.5
• /
• pp.303-307
• /
• 1997

Grb2, which is composed of a Src homology 2 (SH2) domain and two Src homology 3 (SH3) domains, is known to serve as an adaptor protein in signaling for Ras activation. Thus, a blocker of the Grb2 interactions with other proteins can be a potential candidate for an anticancer drug. In this study, we have developed a high throughput screening method for SH3 domain binding ligands and blockers. Firstly, we made and purified the glutathione S-transferase (GST)-fusion proteins with the Grb2 SH2 and SH3 domains, and the entire Grb2. This method measures the binding of a biotin-labeled oligopeptide, derived from a Grb2/SH3 binding motif in the hSos, to the GST-fusion proteins, which are precoated as glutathione S-transferase fusion protein on a solid phase. When $1\;{\mu}g$ of each fusion protein was used to coat the wells, both N- and C- terminal SH3 the domains as well as the whole of Grb2 were able to interact with the biotin-conjugated ligand peptide, while the SH2 domain and GST alone showed no binding affinity. Although N- and C- terminal SH3 domains showed an increase of binding to the ligand peptide in proportion to the amount of peptide, the GST fusion protein with Grb2 demonstrated much higher binding affinity. GST-Grb2 coating on the solid phase showed a saturation curve; 66 and 84% of the maximal binding was observed at 100 and 300 ng/$100\;{\mu}l$, respectively. This binding assay system was peptide sequence-specific, showing a dose-dependent inhibition with the unlabeled peptide of SH3 binding motif. Several other peptides, such as SH2 domain binding motifs and PTB domain binding motif, were ineffective to inhibit the binding to the biotin-conjugated ligand peptide. These results suggest that our method may be useful to screen for new anticancer drug candidates which can block the signaling pathways mediated by SH3 domain binding.

• Korean Journal of Materials Research
• /
• v.15 no.6
• /
• pp.413-418
• /
• 2005

Perovskite $Pb(Ni_{1/3}Nb_{2/3})O_3-Pb(Zr,Ti)O_3[PNN-PZT]$ ceramics were synthesized by conventional ceramic processing technique. In order to modify piezoelectric properties for sensor application in this system, NiO addition was considered to provide $Ni^{+2}$ as an acceptor, which was known to occupy with B site in the structure. The effect of NiO addition up to $8\;mol\%$ on the following piezoelectric properties as well as sintering properties was investigated. When NiO added more than $1\;mol\%$, average grain size was decreased and second phase was found to form. Moreover, the second phase caused decrease in relative dielectric constant $(\varepsilon_{33}T/\varepsilon0)$, electro-mechanical coupling factor $(k_p)$, and piezoelectric charge constant $(d_{33})$, while increasing mechanical quality factor $(Q_m)$. When $1\;mol\%$ NiO was added, density, dielectric properties and piezoelectric properties were abruptly increased.

• The Korean Journal of Nuclear Medicine
• /
• v.9 no.1
• /
• pp.51-58
• /
• 1975

It is a great value to find an early detection of involvement of ureteric obstruction in the carcinoma of cervix. Little or no knowledge of the condition of the kidneys or the lower urinary tract are able to elucidate by the biochemical studies such as blood nitrogen or urine creatinine in carcinoma of cervix. Findings of urography delineates the condition of urinary tract stasis in the renal pelvis and ureters, however, slight stasis maybe difficult to demonstrate. On the other hand isotope nephrography is accepted as a sensitive method to observe renal function especially in regarding to the excretory function of kidney. It was attempted to analysis the findings of urography conjunction with isotope nephrography in 50 cases of unselected patients with invasive carcinoma of cervix through pre and post irradiation follow up studies. Urography was done as a routine procedure and.analysed emphasising changes of collecting systems and ureter condition. Isotope nephrography was carried out by means of $15{\mu}ci\;I^{131}$-Hippuran injected intravenously and the curves were analysed as follows. Parameter were; time of maximum amplitude ($T_{max}$), half time of maximum amplitude ($T\frac{1}{2}$), Kac and Kex value calculated from these two parameters in Tobe's method. The excretion index by Aurell defines the ratio between the maximum activity and the activity measured on the slope of the third phase ten minites after it has reached its maximum. Results: 1. 28.8% had an abnormal IVP suggestive of ureteric involvement before irradiation therapy and the patient of stage III and IV were the great part. 2. 21.7% had abnormal findings of per-irradiation renogram whom showed normal IVP. The other group showed normal IVP which group also showed normal renogram prior irradiation. 3. The more severe the ureteric involvement, the change of excretion index was greater. 4. Even in stage I and II patient, abnormal renogram was revealed in 12 cases (39.4%) among 31 cases. 5. All cases of TAH showed abnormal findings of IVP and renogram. 6. No. definite change of renogram was obtained just after the irradiation therapy (point $A:8000{\sim}9000rads,\;B:5000{\sim}6000rads,\;Co:11000{\sim}13000rads$). Each 3 month follow up study was performed and comparing with preirradiation study which showed significant changes of excretion index of renogram were 42.8% in $6{\sim}9$ month follow-up and 75% in $9{\sim}12$ month, respectively. 7. It seems to be important to observe the parameter Kex and excretion index of renogram to determine early abnormality of kidney excretory function by means of post-irradiation follow up study.

• Journal of Technologic Dentistry
• /
• v.20 no.1
• /
• pp.37-49
• /
• 1998

The specimens used were Ag-25 Pd-15 Cu ternary alloy and Au addition alloy. These alloys were melted and casted by induction electric furnace and centrifugal casting machine in Ar atmosphere. These specimens were solution treated for 2hr at $800^{\circ}C$ and were then quenched into iced water, and aged at $350{\sim}550^{\circ}C$ Age- hardening characteristics of the small Au-containing Ag-Pd-Cu dental alloys were investigated by means of hardness testing. X-ray diffraction and electron microscope observations, electrical resistance, ergy dispersed spectra and electron probe microanalysis. Principal results are as follows : Hardening occured in two stages, i.e., stage I in low temperature and stage II in high temperature regions, during continuous aging. The case of hardening in stage I was due to the formation of the $L1_0$ type face-centered tetragonal PdCu-ordered phase in the grain interior and hardening in stage I was affected by the Cu concentration. In stage II, decomposition of the ${\alpha}$ solid solution to a PdCu ordered phase($L1_0$ type) and an Ag-rich ${\alpha}2$ phase occurred and a discontinuous precipitation occurred at the grain boundary. From the electron microscope study, it was conclued that the cause of age-hardening in this alloy is the precipitation of the PdCu ordered phase, which has AuCu I type face-centered tetragonal structure. Precipetation procedure was ${\alpha}{\to}{\alpha}+{\alpha}_2+PdCu {\to}{\alpha}_1+{\alpha}_2+PdCu$ at Pd/Cu = 1.7 Ag-Pd-Cu alloy is more effective dental alloy as ageing treatment and is suitable to isothermal ageing at $450^{\circ}C$.

• Proceedings of the KSAR Conference
• /
• 2000.10a
• /
• pp.10-12
• /
• 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.

• Korean Journal of Animal Reproduction
• /
• v.24 no.4
• /
• pp.357-364
• /
• 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.

• Korean Journal of Environmental Agriculture
• /
• v.34 no.2
• /
• pp.139-148
• /
• 2015

BACKGROUND: The aim of this study was to isolate and identify algicidal bacterium that tends to kill the toxic dinoflagellate Alexandrium catenella, and to determine the algicidal activity and algicidal range of algicide. METHODS AND RESULTS: Among of algicidal bacteria isolated in this study, NH-3 isolate was the strongest algicidal activity against A. catenella. NH-3 isolate was identified on the basis of biochemical characteristics and analysis of 16S rRNA gene sequences. The NH-3 isolate showed over 99% homology with Arthrobacter oxydans, and was designated as Arthrobacter sp. NH-3. The optimal culture conditions were $25^{\circ}C$, initial pH 7.0, and 2.0% (w/v) NaCl concentration. The algicidal activity of Arthrobacter sp. NH-3 was significantly increased to maximum value in the late of logarithmic phase. Arthrobacter sp. NH-3 showed algicidal activity through indirect attack, which excreted active substance into the culture filtrate. When 10% culture filtrate of NH-3 was applied to A. catenella, 100% of algal cells were destroyed within 30 h. In addition, the algicidal activities were increased in dose and time dependent manners. The pure algicide was isolated from the ethyl acetate extract of the culture filtrate of NH-3 by using silica gel column chromatography and high performance liquid chromatography (HPLC). We investigated the algicidal activity of this algicide on the growth of harmful algal bloom (HAB) species, including A. catenella. As a result, it showed algicidal activity against several HAB species at a concentration of $100{\mu}g/mL$ and had a relatively wide host range. CONCLUSION: Taken together, our results suggest that Arthrobacter sp. NH-3 and its algicide could be a candidate for controlling of toxic and harmful algal blooms.

• Corrosion Science and Technology
• /
• v.16 no.5
• /
• pp.257-264
• /
• 2017

Film of new Ti-15Zr-5Nb alloy formed during galvanic anodizing in orthophosphoric acid solution was characterized by optical microscope, scanning electron microscope (SEM), energy dispersive spectroscopy (EDS), and Raman micro-spectroscopy. Its anticorrosive properties were determined by electrochemical techniques. The film had a layer with nanotube-like porosity with diameters in 500-1000 nm range. The nano layer contained significant amounts of P and O as well as alloying element. Additionally, Raman micro-spectroscopy identified oxygen as oxygen ion in $TiO_2$ anatase and phosphorous as $P_2O_7{^{4-}}$ ion in phosphotitanate compound. All potentiodynamic polarization curves in artificial Carter-Brugirard saliva with pH values (pH= 3.96, 7.84, and 9.11) depending on the addition of 0.05M NaF revealed nobler behavior of anodized alloy and higher polarization resistance indicating the film is thicker and more compact nanolayer. Lower corrosion rates of the anodized alloy reduced toxicity due to less released ions into saliva. Bigger curvature radii in Nyquist plot and higher phase angle in Bode plot for the anodized alloy ascertain a thicker, more protective, insulating nanolayer existing on the anodized alloy. Additionally, ESI results indicate anodized film consists of an inner, compact, barrier, layer and an outer, less protective, porous layer.

• Progress in Superconductivity
• /
• v.11 no.2
• /
• pp.83-86
• /
• 2010

There are theoretical and experimental evidences for the pseudo-gap in electron doped cuprates being due to interaction between electrons and anti-ferromagnetism(AFM). A remaining issue is on how AFM correlates with pseudo-gap, and eventually with superconductivity. To elucidate the issue, we have performed temperature dependent angle-resolved photoemission studies of an e-doped cuprate superconductor $Sm_{2-x}Ce_xCuO_4$(SCCO) x=0.18 at 20K and 150K. In the case of $Nd_{2-x}Ce_xCuO_4$, the most well known e-doped cuprate, pseudo-gap disappears at around 100 K for x=0.17. Our experimental result reveals that the pseudo-gap of SCCO exists even at 150K for x=0.18. This result implies that the AFM of SCCO survives even in x=0.18, which agrees with previously reported phase diagram of SCCO. Yet, the superconductivity disappears around x=0.18 for both NCCO and SCCO in spite of the difference in the magnetic order. This result sheds a light on the disappearance of superconductivity on the over-doped side.

• Journal of Technologic Dentistry
• /
• v.22 no.1
• /
• pp.145-152
• /
• 2000

In order to select an effective antibiotic substance against oral resident bacteria, we were isolated from soil and texonomically analyzed. Seven hundred and eighteen strains were isolated on humic acid- vitamin agar(HV agar) and 220 strains were on methanol medium from three each paddy forest, field and riverside soil samples. So, during the screening of antibiotics from soil, we isolated microorganisms showing powerful antagonistic activity against oral resident bacteria. Microorganism was tested against 25 strains of bacteria, yeast and fungi. Among them, No. 248 strain exhibited the most strongly growth inhibition. So, the taxonomical analysis the isolated strain was found to be unknown Actinomyces sp. and was named No 248. A production of the antibiotics from No. 248 begins at the early exponential phase developed at the 72th hour under the optinum conditions. The property of No. 248 antimicrobial compound was very stable under acid(pH 3.0) and alkali(pH 10.0) treatment, but it was instable in heat treatment at $120^{\circ}C$. For the improvement of antibiotic activity, two mutants were isolated from strain No. 248 by the treatment of mutagenic agents, NTG and hydroxylamine. As a result, the mutant strains excreted the potent antibiotics to inhibit the growth of Candida albicans.