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Isolation, Purification, and Characterization of a Thermostable Xylanase from a Novel Strain, Paenibacillus campinasensis G1-1

  • Zheng, Hongchen;liu, Yihan;Liu, Xiaoguang;Wang, Jianling;Han, Ying;Lu, Fuping
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.930-938
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    • 2012
  • High levels of xylanase activity (143.98 IU/ml) produced by the newly isolated Paenibacillus campinasensis G1-1 were detected when it was cultivated in a synthetic medium. A thermostable xylanase, designated XynG1-1, from P. campinasensis G1-1 was purified to homogeneity by Octyl-Sepharose hydrophobic-interaction chromatography, Sephadex G75 gel-filter chromatography, and Q-Sepharose ion-exchange chromatography, consecutively. By multistep purification, the specific activity of XynG1-1 was up to 1,865.5 IU/mg with a 9.1-fold purification. The molecular mass of purified XynG1-1 was about 41.3 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Sequence analysis revealed that XynG1-1 containing 377 amino acids encoded by 1,134 bp genomic sequences of P. campinasensis G1-1 shared 96% homology with XylX from Paenibacillus campinasensis BL11 and 77%~78% homology with xylanases from Bacillus sp. YA-335 and Bacillus sp. 41M-1, respectively. The activity of XynG1-1 was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Ni^{2+}$, $Fe^{2+}$, $Fe^{3+}$, $Zn^{2+}$, SDS, and EDTA. The purified XynG1-1 displayed a greater affinity for birchwood xylan, with an optimal temperature of $60^{\circ}C$ and an optimal pH of 7.5. The fact that XynG1-1 is cellulose-free, thermostable (stability at high temperature of $70^{\circ}C{\sim}80^{\circ}C$), and active over a wide pH range (pH 5.0~9.0) suggests that the enzyme is potentially valuable for various industrial applications, especially for pulp bleaching pretreatment.

A Study on the Effect of Acupuncture on Anesthesia and the Mode of Action (The Second Report) (자침(刺鍼)이 마취(痲醉)에 미치는 작용기전(作用機轉) 연구(硏究) (제(第) 2 보(報)))

  • Park, Hee-soo;Park, Kyoung-sik
    • Journal of Acupuncture Research
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    • v.19 no.4
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    • pp.140-151
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    • 2002
  • This study was carried to identify whether acupuncture at several meridian points can affect the human anesthesia or not through the analysis of serum intermediates, such as, melatonin, serotonin, TSH, cortisol in human serum. We investigated the effect of accupuncture on the change of serum intermediates which have frequently been made a subject of discussion owing to its hyponic and sedative properties in practice. The result obtained were as follow ; 1. Serum melatonin levels was hardly affected at 40 minutes after general acupuncture on Hab-Gog, Nae-Gwan, and the other meridian points as compared with that($5.64{\pm}1.02pg/ml$) of resting control group. But on day 1 after acupuncture, these tend to increase significantly ($7.95{\pm}2.05pg/ml$ to $8.21{\pm}1.57pg/ml$, p <0.05). On the other hand, control group under sleep showed the serum level of $7.39{\pm}1.03pg/ml$. Additional acupuncture at In-Dang induced the increased melatonin level, especially at 40minutes after acupuncture(p <0.05). 2. Serotonin level in resting control serum recorded $51.14{\pm}8.17{\mu}g/ml$. Acupuncture at determined meridian point intend to increase average level of serotonin, but not significant. However observation on day 1 after acupuncture at Hab-Gwan meridian point and under sleeping showed the significantly increased serotonin level, $74.05{\pm}35.83{\mu}g/ml$, $70.44{\pm}13.08{\mu}g/ml$, respectively. 3. TSH level in intact human serum recorded $0.85{\pm}0.24{\mu}IU/ml$. and the serum exposed to acupuncture showed the irregular pattern of TSH level in a mean range of $0.76{\pm}0.44{\mu}IU/ml$ to $1.06{\pm}0.38{\mu}IU/ml$, regardless of meridian point or time after acupuncture. 4. The values of serum cortisol in control group were $11.76{\pm}2.06{\mu}g/dl$ in resting, $7.51{\pm}2.85{\mu}/dl$ under sleep, respectively. The level of serum cortisol in 40minutes after acupuncture was markedly (p <0.05) reduced to the serum levels of $5.65{\pm}2.58{\mu}g/dl$ (Hab-Gog, Nae-Gwan), $7.58{\pm}3.21{\mu}g/dl$ (Gyo-Gam, Sin-Mun besides Hab-Gog, Nae-Gwan), $6.43{\pm}3.54{\mu}g/dl$ (In-Dang besides to Hab-Gog, Nae-Gwan), respectively, as compared with control, intending to increase a little on day 1 after acupuncture. From the above results, the analysis of serum intermediates suggest that acupunctuation at meridian points applied to at this study act upon the phase of light anaesthesia or hypnosis, at the same time affect pituitary-adrenal axis rather than hypothalamus-pituitary axis in the secretary system of hormone and also don't affect dorsal-raphe nucleus according to the observation of change transition in serum intermediates, such as TSH, serotonin, and cortisol.

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Reference Interval of Serum Thyroid Hormones in Healthy Korean Adults (건강한 한국인 성인의 혈청 갑상선호르몬 참고치)

  • Jang, Yoon-Young;Kim, Chang-Yoon;Hwang, Tae-Yoon;Kim, Kyung-Dong;Lee, Chae-Hoon
    • Journal of Preventive Medicine and Public Health
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    • v.41 no.2
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    • pp.128-134
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    • 2008
  • Objectives : This study was conducted to determine the reference interval of serum thyroid hormones(TSH, $FT_3$, $FT_4$) in healthy Korean adults. Methods : Health examination data from 1,591 healthy Korean adults who visited an university hospital were analyzed. Patients with specific health conditions capable of altering laboratory results were excluded from the study. Serum thyroid hormones were measured using IMMULITE 2000 (DPC, USA, 2002). Subjects were 18-65 years old; 911 were male, and 690 were female. Results : The arithmetic means of TSH, $FT_3$, and $FT_4$ values for male subjects were $1.28{\pm}1.84\;{\mu}IU/ml$, $3.23{\pm}0.57\;pg/ml$, and $1.42{\pm}0.22\;ng/dl$, respectively. In female subjects, the arithmetic means of TSH, FT3, and FT4 values were $1.49{\pm}2.08\;{\mu}IU/ml$, $3.08{\pm}0.54\;pg/ml$, and $1.29{\pm}0.24\;ng/dl$, respectively. The arithmetic mean $FT_4$ value for males decreased with age (p<0.01). The arithmetic mean $FT_3$ value for females increased with age (p<0.01). The arithmetic mean thyroid hormone values of all study subjects differed significantly based on season. The arithmetic mean of male $FT_4$ decreased with increasing BMI (p<0.01). The arithmetic mean of female $FT_3$ increased with increasing BMI (p<0.01). The reference intervals recommended by the IMMULITE 2000 manufacturer are $0.40-4.00\;{\mu}IU/ml$ for TSH, 1.80-4.20 pg/ml for $FT_3$, and 0.80-1.90 ng/dl for $FT_4$ (same values for both genders). Conclusions : There was a significant difference in the interval of thyroid hormones between males and females, but the reference interval of IMMULITE 2000 was not established by gender. There is a need to reestablish the reference interval for thyroid hormones in Korean healthy adults.

Effects of Selenium and Vitamin E on Incidence of Retained Placenta in Holstein Dairy Cows (Selenium 과 Vitamin E 投與가 젖소의 後産停滯에 미치는 영향)

  • Jeong, Gi-Hwa;Park, Sang-Mun;Jeon, Jae-Won;Gwak, Dae-O;Park, Chung-Saeng
    • Korean Journal of Animal Reproduction
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    • v.15 no.1
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    • pp.15-22
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    • 1991
  • To determine the preventive effects of selenium and/or vitamin E on placenta retention in Holstein dairy cows. The cows were divided six groups ; control, Se, vit.E 1x, vit.E 3x, Se + vit./E 1x and Se + vit.E 3x in Cooong Nam province. 50mg of Se as sodium selenite and 700 IU of vit. E as dl-${\alpha}$ tocopherol acetate were injected 21 day perior to expected calving date, and 700 IU vit.E was injected 3 times at 21, 14 and 7 days perior to expected calving date in vit.E 3x groups. The cows which did not expelled their placenta until 12 hours post-parturient were termed as cow of placenta retention. The i.m. injection of a single dose of selenium and vit.E has significantly(P<0.05) affected the incidence of retained placenta by 7.0% versus 25.6% of controls, but a single injection of selenium or vit.E and a combination of selenium 1x and vit.E 3x has not significantly affected. But they affected neiher subsequent reproductive performances nor milk yields.

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A Prospective Comparison of Fertilizability of in vitro Matured Human Oocytes Obtained from Stimulated Cycle: Conventional Versus ICSI (과배란유도 주기에서 얻어진 체외성숙 난자의 수정능: 고식적 체외수정시술과 세포질내정자주입법의 비교)

  • Jee, Byung-Chul;An, So-Jung;Moon, Jeong-Hee;Hwang, Eun-Ju;Suh, Chang-Suk;Kim, Seok-Hyun;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.4
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    • pp.249-254
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    • 2009
  • Objective: The aim of this study was to compare the fertilization and cleavage rates of human in vitro matured oocytes after fertilized by conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Methods: A total of 135 GV stage oocytes were obtained from 59 women who received ovarian stimulation and IVF during Jan 2007 to Oct 2008. Ovarian hyperstimulation was performed using hMG or recombinant FSH with GnRH antagonist and then ovulation triggered by recombinant hCG. The immature oocytes obtained from stimulation cycles were cultured in IVM medium up to 48 hrs; commercial medium supplemented with rFSH 75 mIU/mL, rhCG 0.5 IU/mL and rEGF 10 ng/mL. The in vitro matured oocytes were fertilized by conventional IVF (41 GV oocytes) or ICSI method (94 GV oocytes). Results: Maturation rate were 51.2% and 59.6% in conventional IVF group and ICSI group, respectively. There was no significant difference in fertilization rates between two groups; 71.4% and 80.4%, respectively. The cleavage rate was also similar in two groups. Conclusion: The presented data suggest that conventional IVF has comparable fertilization and cleavage potential compared with ICSI as the insemination method of immature human oocytes obtained from stimulated cycle.

Nausea and Vomiting after Transcatheter Arterial Chemoembolization for Hepatocellular Carcinoma: Incidence and Risk Factor Analysis

  • Wang, Shi-Ying;Zhu, Wen-Hao;Vargulick, Sonya;Lin, Sam Bill;Meng, Zhi-Qiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5995-6000
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    • 2013
  • Background: Nausea and vomiting after transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) are common in clinical practice, but few studies have reported the incidence and risk factors of such events. Objective: The purpose of this study was to analyze the incidence and risk factors of nausea and vomiting after TACE for HCC. Methods: This study was a single-center retrospective analysis of a prospectively maintained database. Between May 2010 and October 2012, 150 patients with HCC were analyzed for incidence and preprocedural risk factors. Results: The incidence of postembolization nausea and vomiting was 38.8% and 20.9%, respectively, in patients with HCC. Patients who developed nausea had lower levels (<100 IU/L) of serum alkaline phosphatase (ALP) compared to those without nausea ($123.04{\pm}69.38$ vs. $167.41{\pm}138.95$, respectively, p=0.044). Female gender correlated to a higher incidence of nausea as well (p=0.024). Patients who developed vomiting, compared to those who did not, also had lower levels (<100 IU/L) of serum ALP ($112.52{\pm}62.63$ vs. $160.10{\pm}127.80$, respectively, p=0.010), and serum alanine transferase (ALT) ($35.61{\pm}22.87$ vs. $4.97{\pm}29.62$, respectively, p=0.045). There were no statistical significances in the incidences of nausea and vomiting between male patients over 50 years old and female patients who have entered menopause (p=0.051 and p=0.409, respectively). Multivariate analysis by logistic regression analysis demonstrated that female gender and ALP>100 IU/L were the most independent predictive factors of postembolization nausea (odds ratio (OR): 3.271, 95% CI: 1.176-9.103, p=0.023 and OR: 0.447, 95% CI: 0.216-0.927, p=0.030, respectively). ALP>100 IU/L was also the most independent predictive risk factor of postembolization vomiting (OR: 0.389, 95% CI: 0.159-0.952, p=0.039). Conclusions: Postembolizaiton nausea and vomiting are common in patients with HCC. Recognition of the risk factors presented above before TACE is important for early detection and proper management of postembolization nausea and vomiting. Nevertheless, future studies are required.

Clomiphene Citrate Challenge Test and Its Clinical Correlation with Prognostic Factors of Ovarian Response (Clomiphene Citrate 부하검사와 난소 반응 예측 인자와의 연관성에 관한 연구)

  • Moon, Shin-Yong;Chae, Hee-Dong;Kim, Kwang-Rye;Suh, Chang-Suk;Kim, Seok-Hyun;Choi, Young-Min;Shin, Chang-Jae;Kim, Jung-Gu;Lee, Jin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.283-292
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    • 1996
  • Objective: To determine the cutoff value of clomiphene citrate challenge test(CCCT) that can predict the normal and abnormal(diminished) ovarian response group and to assess the usefulness of CCCT as a predictor of ovarian reserve. Materials and Methods: From March 1994 to Februry 1996, CCCT was performed to 129 infertile patients and among them, 20 patients whose basal FSH on the third day of menstrual cycle was more than 20 mIU/ml were excluded. At the same time, the same CCCT was performed to the fifteen healthy volunteers with proven fertility to determine the cutoff value of CCCT. Results; 1) A FSH value higher than 23.4 mIU/ml, measured on the 10th day of menstrual cycle, was defined as a abnormal ovarian response. The cutoff value of 23.4 mIU/ml is more than 2 standard deviations(SD) above the mean value of 15 healthy women after CCCT. 2) The abnormal CCCT group, the subpopulation with a FSH value of 23.4 mIU/ml or more, was 7.3%(8/109) and their mean age was higher than the normal CCCT group($36.5{\pm}4.5$ vs. $32.9{\pm}4.8$, P = 0.059). And the percentage of the patients older than 35 years of the abnormal CCCT group was significantly higher than that of the normal CCCT group(62.5% vs. 38.6%, p <0.05). 3) There was no correlation between the hormone values of the third day and the 10th day of menstrual cycle before and after CCCT except between FSH of the third day and the 10th day. Conclusion: The CCCT is a good method to predict the individual ovarian response to COH for ART, especially in the patients who has no other abnormal findings that predict poor prognosis. And it is neccessary to determine the cutoff value of CCCT by the large numbers of randomized study, and combining the previously proven prognostic factors, it can be applicated in many individual centers for evaluate the ovarian response before ART program.

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Serum Steroid Hormone Level and Hematological Characteristics of One-year Cultured eels, Anguilla japonica Based on Total Length and Sex (양식 1년산 뱀장어, Anguilla japonica의 전장 및 성별에 따른 혈중 성호르몬 수준 및 혈액학적 성상)

  • 전민지;한경민;배준영;유진형;이계안;배승철
    • Journal of Aquaculture
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    • v.16 no.4
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    • pp.267-272
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    • 2003
  • This study investigated sex determination through serum steroid hormone level and compared hematological characteristics of one-year old cultured eels, Anguilla japonica. Four different groups were divided with total length at 40∼45, 45∼50, 50∼55, and 55∼60 cm. Males dominated eels of 40∼50 cm in total length group, while females dominated eels of more than 50 cm. All females and males were immature. Hematocrit levels of males (total length: 40∼50 cm) were higher than those of females (total length: 50∼60 cm). Hemoglobin content also differed between males and females. Glutamate oxaloacetate transaminase (GOT) levels of 40∼45 cm males tended to be higher (157$\pm$3.46 IU/L) than that of other sized males, but there were no significant differences between groups (P<0.05). The highest GOT levels were found in of 55∼60 cm females (148$\pm$3.46 IU/L) compared with that of other female groups (P<0.05). Glutamate pyruvate transaminase (GPT) levels of males and females in the 55∼60 cm size range was significantly higher thanothers (P<0.05). The result of this study indicated steroid hormone content of males and females were very low, but testosterone (T) and estradiol-17$\beta$, (E2) contents of females were higher than those of males. For further research of sex determination in cultured eels, it needs to investigate in more defined body length.

Farm Survey on Eggshell Quality and Egg Vitamin D3 Contents in Laying Hens Fed Vitamin D3-enriched Diets (산란계 사료 내 비타민 D3 첨가에 따른 난각품질과 계란내 비타민D3 함량에 관한 농장조사)

  • Dong-Hae Joh;Byung-Yeon Kwon;Da-Hye Kim;Kyung-Woo Lee
    • Korean Journal of Poultry Science
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    • v.50 no.2
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    • pp.73-80
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    • 2023
  • Vitamin D3 is an essential nutrient which plays an important role in calcium metabolism for eggshell formation, in calcium and phosphorus metabolism for bone mineralization, and in maintaining host immunity. Although there have been a great deal of studies investigating the role of vitamin D3 in eggshell quality and vitamin D3 contents in eggs, no attempts have been made to monitor the eggshell quality and vitamin D3 contents in eggs at farm level. Thus, this survey was conducted to measure eggshell quality and vitamin D3 contents in eggs laid from laying hens fed diets containing different levels of vitamin D3. Eggs from four commercial laying hen farms were sampled before and 1, 2, 3 and 5 weeks after provision of the vitamin D3-enriched diets added with the level of 16,500 IU and 29,000 IU. Dietary vitamin D3 did not affect the eggshell color and breaking strength, but increase the eggshell thickness. In addition, vitamin D3 contents in eggs were elevated as vitamin D3 in diets was increased. It is concluded that addition of dietary vitamin D3 into the diets of laying hens at the commercial laying hen farms could improve eggshell quality and vitamin D3 contents in eggs. It is expected that the prediction equation for egg vitamin D3 contents might be produced if more data on vitamin D3 contents in diets and eggs at the farms are to be analyzed.

Production of Porcine Embryos in Different Culture Medium

  • Lee, S. Y.;Park, Y. H.;Park, C. K.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.67-67
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    • 2003
  • Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

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