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Profiling Bartonella infection and its associated risk factors in shelter cats in Malaysia

  • Nurul Najwa Ainaa Alias;Sharina Omar;Nur Indah Ahmad;Malaika Watanabe;Sun Tee Tay;Nor Azlina Aziz;Farina Mustaffa-Kamal
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.38.1-38.12
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    • 2023
  • Background: Poor disease management and irregular vector control could predispose sheltered animals to disease such as feline Bartonella infection, a vector-borne zoonotic disease primarily caused by Bartonella henselae. Objectives: This study investigated the status of Bartonella infection in cats from eight (n = 8) shelters by molecular and serological approaches, profiling the CD4:CD8 ratio and the risk factors associated with Bartonella infection in shelter cats. Methods: Bartonella deoxyribonucleic acid (DNA) was detected through polymerase chain reaction (PCR) targeting 16S-23S rRNA internal transcribed spacer gene, followed by DNA sequencing. Bartonella IgM and IgG antibody titre, CD4 and CD8 profiles were detected using indirect immunofluorescence assay and flow cytometric analysis, respectively. Results: B. henselae was detected through PCR and sequencing in 1.0% (1/101) oral swab and 2.0% (1/50) cat fleas, while another 3/50 cat fleas carried B. clarridgeiae. Only 18/101 cats were seronegative against B. henselae, whereas 30.7% (31/101) cats were positive for both IgM and IgG, 8% (18/101) cats had IgM, and 33.7% (34/101) cats had IgG antibody only. None of the eight shelters sampled had Bartonella antibody-free cats. Although abnormal CD4:CD8 ratio was observed in 48/83 seropositive cats, flea infestation was the only significant risk factor observed in this study. Conclusions: The present study provides the first comparison on the Bartonella spp. antigen, antibody status and CD4:CD8 ratio among shelter cats. The high B. henselae seropositivity among shelter cats presumably due to significant flea infestation triggers an alarm of whether the infection could go undetectable and its potential transmission to humans.

Digestibility of amino acids in fish meal and blood-derived protein sources fed to pigs

  • Park, Chan Sol;Adeola, Olayiwola
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1418-1425
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    • 2022
  • Objective: An experiment was conducted to determine the standardized ileal digestibility (SID) of amino acids (AA) in fish meal (FM) and blood-derived protein sources including spray-dried porcine plasma (SDPP), porcine red blood cells (PRBC), and blood meal (BM) fed to growing pigs. Methods: Ten barrows (mean initial body weight of 22.1±1.54 kg) surgically fitted with T-cannulas at the distal ileum were allotted to a duplicated 5×4 incomplete Latin square design with 5 experimental diets and 4 periods. Four experimental diets were prepared to contain FM, SDPP, PRBC, or BM as the sole source of nitrogen. A nitrogen-free diet was prepared and included to estimate the basal ileal endogenous losses of AA. For the 7-day experimental period, pigs were fed for 5 days as adaptation, and ileal digesta samples were collected for 9 hours on days 6 and 7. Results: The SID of crude protein in BM (48.0%) was less (p<0.05) than in FM, SDPP, and PRBC (83.4%, 83.9%, and 87.3%, respectively). Pigs fed the diet containing BM had less (p<0.05) SID of AA, except isoleucine and proline, than those fed the diet containing FM, SDPP, or PRBC. Among FM, SDPP, and PRBC, there was no difference in the SID of crude protein and all AA, except isoleucine. The SID of isoleucine in PRBC and BM (62.7% and 48.3%, respectively) was less (p<0.05) than in FM and SDPP (88.0% and 84.9%, respectively). The SID of lysine in FM, SDPP, PRBC, and BM was 85.4%, 84.9%, 89.7%, and 51.9%, respectively. Conclusion: The SID of most AA was not different among FM, SDPP, and PRBC, but BM had lower SID of most AA than FM, SDPP, and PRBC.

Evaluations of Spectral Analysis of in vitro 2D-COSY and 2D-NOESY on Human Brain Metabolites (인체 뇌 대사물질에서의 In vitro 2D-COSY와 2D-NOESY 스펙트럼 분석 평가)

  • Choe, Bo-Young;Woo, Dong-Cheol;Kim, Sang-Young;Choi, Chi-Bong;Lee, Sung-Im;Kim, Eun-Hee;Hong, Kwan-Soo;Jeon, Young-Ho;Cheong, Chae-Joon;Kim, Sang-Soo;Lim, Hyang-Sook
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.1
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    • pp.8-19
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    • 2008
  • Purpose : To investigate the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar nuclear Overhauser effect/enhancement (NOE) interaction through 2D- correlation spectroscopy (COSY) and 2D- NOE spectroscopy (NOESY) techniques. Materials and Methods : All 2D experiments were performed on Bruker Avance 500 (11.8 T) with the zshield gradient triple resonance cryoprobe at 298 K. Human brain metabolites were prepared with 10% $D_2O$. Two-dimensional spectra with 2048 data points contains 320 free induction decay (FID) averaging. Repetition delay was 2 sec. The Top Spin 2.0 software was used for post-processing. Total 7 metabolites such as N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), lutamine (Gln), glutamate (Glu), myo-inositol (Ins), and lactate (Lac) were included for major target metabolites. Results : Symmetrical 2D-COSY and 2D-NOESY pectra were successfully acquired: COSY cross peaks were observed in the only 1.0-4.5 ppm, however, NOESY cross peaks were observed in the 1.0-4.5 ppm and 7.9 ppm. From the result of the 2-D COSY data, cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methylene protons (CH2(3,$H{\alpha}$)) at 2.50ppm and methylene protons ($CH_2$,(3,$H_B$)) at 2.70 ppm were observed in NAA. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. From the result of 2-D NOESY data, cross peaks between the NH proton at 8.00 ppm and methyl protons ($CH_3$) were observed in NAA. Cross peaks between the methyl protons ($CH_3$(3)) at 1.33 ppm and methine proton (CH(2)) at 4.11 ppm were observed in Lac. Cross peaks between the methyl protons (CH3) at 3.03 ppm and methylene protons (CH2) at 3.93 ppm were observed in Cr. Cross peaks between the methylene protons ($CH_2$(3)) at 2.11 ppm and methylene protons ($CH_2$(4)) at 2.35 ppm, and between the methylene protons($CH_2$ (3)) at 2.11 ppm and methine proton (CH(2)) at 3.76 ppm were observed in Glu. Cross peaks between the methylene protons (CH2 (3)) at 2.14 ppm and methine proton (CH(2)) at 3.79 ppm were observed in Gln. Cross peaks between the methine proton (CH(5)) at 3.27 ppm and the methine proton (CH(4,6)) at 3.59 ppm, and between the methine proton (CH(1,3)) at 3.53 ppm and methine proton (CH(2)) at 4.05 ppm were observed in Ins. Conclusion : The present study demonstrated that in vitro 2D-COSY and NOESY represented the 3-bond and spatial connectivity of human brain metabolites by scalar coupling and dipolar NOE interaction. This study could aid in better understanding the interactions between human brain metabolites in vivo 2DCOSY study.

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Advanced Treatment of Sewage and Wastewater Using an Integrated Membrane Separation by Porous Electrode-typed Electrolysis (분리막/다공 전극형 전기분해 조합공정을 이용한 하.폐수의 고도처리)

  • Choi, Yong-Jin;Lee, Kwang-Hyun
    • Membrane Journal
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    • v.22 no.2
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    • pp.95-103
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    • 2012
  • To treat nitrate and non-biodegradable organics effectively in sewage, industrial wastewater and livestock wastewater, the activated sludge process integrated by a membrane separation and a porous electrode- electrolysis was proposed and its efficiency was investigated. The proposed system was consisted of 3 processes; activated sludge, membrane filtration and electrolysis. In the study, the membrane filtration played a role in reducing the load of the electrolysis to operate the proposed process stably. The electrolysis consisted of a porous electrode to increase the efficiency due to the extension of the specific surface area. Additionally, redox reaction in the electrolysis was induced by decomposing influent water as current was applied. As a result, hydrogen free radicals and oxygen radicals as intermediates were produced and they acted as oxidants to play a role in decomposing non-degradable organics. It was environmentally-friendly process because intermediates produced by porous electrode were used to treat waste matters without supplying external reagent. Experimental data showed that the proposed process was more excellent than activated sludge process. SS removal efficiencies of the proposed process, membrane filtration and activated sludge process were about 100%, about 100% and about 90%, respectively. COD removal efficiencies of the proposed system, membrane filtration and activated sludge process were about 92%, about 84% and about 78%, respectively. T-N removal efficiencies of the proposed system, membrane filtration and activated sludge process were about 88%, about 67%, and about 58%, respectively. The SS data showed that SS was efficiently removed in the single of the membrane filtration. The COD/T-N data showed that COD/T-N of membrane hybrid process was treated by removing a little soluble organics and SS, and that COD/T-N of electrolysis hybrid process was treated by oxidize organics with high removal rate.

Compositional Variation of Arsenopyrites in Arsenic and Polymetallic Ores from the Ulsan Mine, Republic of Korea, and their Application to a Geothermometer (울산광산산(蔚山鑛山産) 유비철석(硫砒鐵石)의 조성변화(組成變化) 및 지질온도계(地質溫度計)에 대(對)한 적용(適用))

  • Choi, Seon-Gyu;Chung, Jae-Ill;Imai, Naoya
    • Economic and Environmental Geology
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    • v.19 no.3
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    • pp.199-218
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    • 1986
  • Arsenopyrite in arsenic and polymetallic ores from calcic Fe-W skarn deposit of the Ulsan mine, Republic of Korea, has been investigated by means of electron microprobe analysis and X-ray diffractometry. As a result, it is revealed that the Ulsan arsenopyrite may be classified into the following three species with different generation on the basis of its mode of occurrence, chronological order during polymetallic mineralization and chemical composition; arsenopyrites I, II and III. 1) Arsenopyrite I-(Ni, Co)-bearing species belonging to the oldest generation, which has crystallized together with (Ni, Co)-arsenides and -sulpharsenides in the early stage of polymetallic mineralization. In rare cases, it contains a negligible amount of antimony. It occurs usually as discrete grains with irregular outline, showing rarely subhedral form, and is diffused in skarn zone. The maximum contents of nickel and cobalt are 10.04 Ni and 2.45 Co (in weight percent). Occasionally, it shows compositional zoning with narrow rim of lower (Ni+Co) content. 2) Arsenopyrite II-arsenian species, in which (Ni+Co) content is almost negligible, may occur widely in arsenic ores, and its crystallization has followed that of arsenopyrite I. It usually shows subhedral to euhedral form and is closely associated with $l{\ddot{o}}llingite$, bismuth, bismuthinite, chalcopyrite, sphalerite, bismuthian tennantite, etc. It is worthy of note that arsenopyrite II occasionally contains particles consisting of both bismuth and bismuthinite. 3) Arsenopyrite III-(Ni, Co)-free, S-excess and As-deficient species is close to the stoichiometric composition, FeAsS. It occurs in late hydrothermal veins, which cut clearly the Fe-W ore pipe and the surrounding skarn zone. It shows euhedral to subhedral form, being extremely coarse-grained, and is closely associated with pyrite, "primary" monoclinic pyrrhotite, galena, sphalerite, etc. Among three species of the Ulsan arsenopyrite, arsenopyrite I does not serve as a geothermometer, because (Ni+Co) content always exceeds 1 weight percent. In spite of the absence of Fe-S minerals as sulphur-buffer assemblage, the presence of $Bi(l)-Bi_2S_3$ sulphur-buffer enables arsenopyrite II to apply successfully to the estimation of either temperature and sulphur fugacity, the results are, $T=460{\sim}470^{\circ}C$, and log $f(S_2)=-7.4{\sim}7.0$. With reference to arsenopyrite III, only arsenopyrite coexisting with pyrite and "primary" monoclinic pyrrhotite may serve to restrict the range of both temperature and sulphur fugacity, $T=320{\sim}440^{\circ}C$, log $f(S_2)=-9.0{\sim}7.0$. These temperature data are consistent with those obtained by fluid inclusion geothermometry on late grandite garnet somewhat earlier than arsenopyrite II. At the beginning of this paper, the geological environments of the ore formation at Ulsan are considered from regional and local geologic settings, and physicochemical conditions are suspected, in particular the formation pressure (lithostatic pressure) is assumed to be 0.5kb (50MPa). The present study on arsenopyrite geothermometry, however, does not bring about any contradictions against the above premises. Thus, the following genetical view on the Ulsan ore deposit previously advocated by two of the present authors (Choi and Imai) becomes more evident; the ore deposit was formed at shallow depth and relatively high-temperature with steep geothermal gradient-xenothermal conditions.

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Effect on Quality of Pan-Fired Green Tea by 1st Pan-Firing Time (덖음차 제조공정 중 첫 덖음시간이 품질에 미치는 영향)

  • Park, J.H.;Han, J.S.;Choi, H.K.
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.2
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    • pp.101-106
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    • 1999
  • These studies were carried out to investigate the quality of green tea processed by different 1st-pan-firing time and to select the best conditions of green tea processing. The results were as follows. At harvesting tea plant leaf, tea plants had 4 leaves, each leaf size was $6.4cm\;{\times}\;2.4cm$, and weight of 100 buds was 40.2g. The content of moisture of green tea was decreased with the prolongation of 1st-pan-firing time, and the yield of green tea at 10 min. pan-firing time was the highest value of 20.12%. After processing green tea, 1st 10 min. pan-fired one had much more contents of total nitrogen(TN 5.41%) and total amino acid(T.A.A; 3,282mg/100g) and less content of tannin(13.22%) and caffeine(1.64%) than the other treatments. The content of chlorophyll and vitamin C went down with the extension of 1st pan-firing period. Sixteen kinds of free amino acid including theanine as a good taste component of green tea were isolated and measured by HPLC. T.A.A contents were in the range of $2,447{\sim}3,192mg/100g$ and among various pan-firing methods, 10 min. 1st pan-fired one had the highest content of T.A.A. The taste-determinants such as theanine, aspartic acid, serine, glutamic acid and arginine in green tea liquor, occupied as much as $89.4{\sim}90.0%$ of T.A.A.s. Among these, theanine was more than 50%, Lightness and yellowness of green tea were increased with delay of pan-firing time, while green color appeared to be more distinct at 10 min. 1st pan -firing time than other treatments. Besides, it gained the highest score of sensory evaluation among them. These above results suggested that 10 min. pan-fired green tea was considered to be the best level as pan-firing time of green tea processing.

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Study of Quality Control of Traditional Wine Using IT Sensing Technology (IT 센싱 기술을 이용한 전통주 발효의 품질관리 연구)

  • Song, Hyeji;Choi, Jihee;Park, Chan-Won;Shin, Dong-Beom;Kang, Sung-Soo;Oh, Sung Hoon;Hwang, Kwontack
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.6
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    • pp.904-911
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    • 2015
  • The objective of this study was to investigate the quality characteristics of traditional wine using an radio-frequency identification (RFID) system annexed to a fermenter. In this study, we proposed an RFID-based data transmission scheme for monitoring fermentation of traditional alcoholic beverages. The pH, total acidity, total sugar, soluble sugar, free sugar, alcohol content, and organic acids of were investigated and subjected to fermentation of traditional alcoholic beverages three times. The pH ranged from 7.98, 7.95, and 7.68 at day 0, decreased drastically to 3.31~2.96 at day 2, and then slowly increased to the end point, finally reaching 3.34 at day 20. Acidity tended to increase quickly with time, especially for all samples after day 2. The fermentation environment induced a sudden increase acidity in reactants and indicated a low pH. The total sugars during fermentation quickly decreased to the range of 20.3, 22.43, and 19.2% at day 2, and the slope of reduction steadily decreased to 5.1, 6.1, and 4.8% at day 10. On the other hand, the alcohol content showed the reverse trend as total sugars. The alcohol content also showed the same pattern as total acids, showing the highest alcohol content of 17.3% (v/v) on day 20. In this study on traditional wine fermentation using an RFID system, we showed that pH, soluble sugar, and alcohol content can be adopted as key indicators for quality control and standardization of traditional wine manufacturing.

Results of Preoperative Concurrent Chemoradiotherapy for the Treatment of Rectal Cancer (직장암의 수술 전 동시적 항암화학방사선치료 결과)

  • Yoon, Mee-Sun;Nam, Taek-Keun;Kim, Hyeong-Rok;Nah, Byung-Sik;Chung, Woong-Ki;Kim, Young-Jin;Ahn, Sung-Ja;Song, Ju-Young;Jeong, Jae-Uk
    • Radiation Oncology Journal
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    • v.26 no.4
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    • pp.247-256
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    • 2008
  • Purpose: The purpose of this study is to evaluate anal sphincter preservation rates, survival rates, and prognostic factors in patients with rectal cancer treated with preoperative chemoradiotherapy. Materials and Methods: One hundred fifty patients with pathologic confirmed rectal cancer and treated by preoperative chemoradiotherapy between January 1999 and June 2007. Of the 150 patients, the 82 who completed the scheduled chemoradiotherapy, received definitive surgery at our hospital, and did not have distant metastasis upon initial diagnosis were enrolled in this study. The radiation dose delivered to the whole pelvis ranged from 41.4 to 46.0 Gy (median 44.0 Gy) using daily fractions of $1.8{\sim}2.0\;Gy$ at 5 days per week and a boost dose to the primary tumor and high risk area up to a total of $43.2{\sim}54\;Gy$ (median 50.4 Gy). Sixty patients (80.5%) received 5-fluorouracil, leucovorin, and cisplatin, while 16 patients (19.5%) were administered 5-fluorouracil and leucovorin every 4 weeks concurrently during radiotherapy. Surgery was performed for 3 to 45 weeks (median 7 weeks) after completion of chemoradiotherapy. Results: The sphincter preservation rates for all patients were 73.2% (60/82). Of the 48 patients whose tumor was located at less than 5 cm away from the anal verge, 31 (64.6%) underwent sphincter-saving surgery. Moreover, of the 34 patients whose tumor was located at greater than or equal to 5 cm away from the anal verge, 29 (85.3%) were able to preserve their anal sphincter. A pathologic complete response was achieved in 14.6% (12/82) of all patients. The downstaging rates were 42.7% (35/82) for the T stage, 75.5% (37/49) for the N stage, and 67.1% (55/82) for the overall stages. The median follow-up period was 38 months (range $11{\sim}107$ months). The overall 5-year survival, disease-free survival, and locoregional control rates were 67.4%, 58.9% and 84.4%, respectively. The 5-year overall survival rates based on the pathologic stage were 100% for stage 0 (n=12), 59.1% for stage I (n=16), 78.6% for stage II (n=30), 36.9% for stage III (n=23), and one patient with pathologic stage IV was alive for 43 months (p=0.02). The 5-year disease-free survival rates were 77.8% for stage 0, 63.6% for stage I, 58.9% for stage II, 51.1% for stage III, and 0% for stage IV (p<0.001). The 5-year locoregional control rates were 88.9% for stage 0, 93.8% for stage I, 91.1% for stage II, 68.2% for stage III, and one patient with pathologic stage IV was alive without local recurrence (p=0.01). The results of a multivariate analysis with age (${\leq}55$ vs. >55), clinical stage (I+II vs. III), radiotherapy to surgery interval (${\leq}6$ weeks vs. >6 weeks), operation type (sphincter preservation vs. no preservation), pathologic T stage, pathologic N stage, pathologic overall stage (0 vs. I+II vs. III+IV), and pathologic response (complete vs. non-CR), only age and pathologic N stage were significant predictors of overall survival, pathologic overall stage for disease-free survival, and pathologic N stage for locoregional control rates, respectively. Recurrence was observed in 25 patients (local recurrence in 10 patients, distant metastasis in 13 patients, and both in 2 patients). Acute hematologic toxicity ($\geq$grade 3) during chemoradiotherapy was observed in 2 patients, while skin toxicity was observed in 1 patient. Complications developing within 60 days after surgery and required admission or surgical intervention, were observed in 11 patients: anastomotic leakage in 5 patients, pelvic abscess in 2 patients, and others in 4 patients. Conclusion: Preoperative chemoradiotherapy was an effective modality to achieve downstaging and sphincter preservation in rectal cancer cases with a relatively low toxicity. Pathologic N stage was a statistically significant prognostic factor for survival and locoregional control and so, more intensified postoperative adjuvant chemotherapy should be considered in these patients.

Shear bond strength of the three different kinds of resin cement on CAD/CAM ceramic inlay (CAD/CAM 세라믹 인레이에 대한 3종의 레진 시멘트의 전단결합강도에 관한 연구)

  • Baek, Chul-Woo;Park, Cheol-Woo;Park, Jun-Sub;Ryu, Jae-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.51 no.1
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    • pp.20-26
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    • 2013
  • Purpose: The purpose of this study was to evaluate the bond strengths between the latest CAD/CAM ceramic inlay and various resin cements which are used primarily for esthetic restoration. Materials and methods: Cylindrical ceramic blocks(Height: 5 mm, diameter: 3 mm) were fabricated by using Cerec3 and bonded on the dentin of the ninety extracted caries-free molars using three different kinds of resin cement(Unicem$^{(R)}$, Biscem$^{(R)}$, and Variolink II$^{(R)}$) according to the manufacturer's instructions. Ninety specimens were divided into 3 groups according to three different kinds of resin cement. Half of each group were conducted thermocycling under the conditions of the $5-55^{\circ}C$, 5,000 cycle but the other half of them weren't. All specimens were kept in normal saline $37^{\circ}C$, for 24 hours before measuring the bond strength. The shear bond strength was measured by Universal testing machine with a cross head speed of 0.5 mm/min. The results were analyzed statistically by t-test and one-way ANOVA. Results: Unicem$^{(R)}$ group showed the highest shear bond strength despite a slight decline by thermocycling. The shear bond strength of Unicem$^{(R)}$ group and ValiolinkII$^{(R)}$ group were significantly influenced by thermocycling, whereas Biscem$^{(R)}$ group was not influenced (P<.05). There were no significant differences in the bond strength between the three groups without thermocycling, but there was significant differences between Unicem$^{(R)}$ group and Valiolink II$^{(R)}$ group with thermocycling(P<.05). Conclusion: It has been shown to be clinically effective when the self-adhesive resin cements Unicem$^{(R)}$ and Biscem$^{(R)}$ were used instead of the etch-and-rinse resin cement Valiolink II$^{(R)}$ during the bonding of CAD/CAM ceramic inlay restorations with teeth.

eRF1aMC and $Mg^{2+}$ Dependent Structure Switch of GTP Binding to eRF3 in Euplotes octocarinatus

  • Song, Li;Jia, Yu-Xin;Zhu, Wen-Si;Chai, Bao-Feng;Liang, Ai-Hua
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.176-183
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    • 2012
  • Eukaryotic translation termination is governed by eRF1 and eRF3. eRF1 recognizes the stop codons and then hydrolyzes peptidyl-tRNA. eRF3, which facilitates the termination process, belongs to the GTPase superfamily. In this study, the effect of the MC domain of eRF1a (eRF1aMC) on the GTPase activity of eRF3 was analyzed using fluorescence spectra and high-performance liquid chromatography. The results indicated eRF1aMC promotes the GTPase activity of eRF3, which is similar to the role of eRF1a. Furthermore, the increased affinity of eRF3 for GTP induced by eRF1aMC was dependent on the concentration of $Mg^{2+}$. Changes in the secondary structure of eRF3C after binding GTP/GDP were detected by CD spectroscopy. The results revealed changes of conformation during formation of the eRF3C GTP complex that were detected in the presence of eRF1a or eRF1aMC. The conformations of the eRF3C eRF1a GTP and eRF3C eRF1aMC GTP complexes were further altered upon the addition of $Mg^{2+}$. By contrast, there was no change in the conformation of GTP bound to free eRF3C or the eRF3C eRF1aN complex. These results suggest that alterations in the conformation of GTP bound to eRF3 is dependent on eRF1a and $Mg^{2+}$, whereas the MC domain of eRF1a is responsible for the change in the conformation of GTP bound to eRF3 in Euplotes octocarinatus.