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Factors Affecting the Survival of Frozen Thawed Bovine In Vitro Produced Blastocysts

  • Gustafsson, H.;Larsson, B.;Shamsuddin, M.;Jaakma, U.;Emanuelson, U.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.1
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    • pp.7-12
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    • 2001
  • Factors Affecting the Survival of Frozen Thawed Bovine In Vitro Produced Blastocysts. The effect of some factors on the post-thaw survival of a total of 240 in vitro produced bovine blastocysts was investigated using logistic regression analysis. The explanatory variables tested were: type of culture medium before freezing (TCM 199 supplemented with BSA, BSAITS (BSA+insulin+transferrin+selenium), ECS (estrous cow serum) with or without BOEC (bovine oviductal epithelial cells), age of the blastocyst (Day 7, Day 8+9), morphological appearance before freezing (distinct=Q1 or indistinct=Q2 inner cell mass) and type of cryoprotectant (glycerol, 1.0 M or ethylene glycol, 1.6 M). The survival after thawing based on the post-thaw quality and the development after co-culture with BOEC for 24 and 48 hours. Day 7 blastocysts had an almost three times better chance of survival than Day 8+9 blastocysts. Q1, Day 8+9 blastocysts had higher odds to survive after 48 hours in culture than Q2 blastocysts (p<0.05). Blastocysts produced in BSAITS medium had the best chances of survival; however, the odds were not always significant. Blastocysts frozen in glycerol had a better post-thaw quality rating than those frozen in ethylene glycol; however, the difference in post-thaw development at culture was not significant. The relationship between post-thaw quality and post-thaw development at culture was significant (p<0.05). The developmental stage and/or age of the embryo and culture medium where development up to blastocyst takes place affect the post-thaw survival of the bovine embryos.

Purification and Characterization of a Cytochrome P-450 from Pravastatin-Producing Streptomyces sp. Y-110.

  • Park, Joo-Woong;Lee, Joo-Kyung;Kwon, Tae-Jong;Yi, Dong-Hee;Park, Yong-Il;Kang, Sang-Mo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1011-1017
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    • 2001
  • Streptomyces sp. Y-110 cytochrome P-450, induced by the addition of compactin -Na into the culture medium, was purified from the cell extract to apparent homogeniety, mainly by DEAE-Sepharose, hydroxyapatite, and Mono Q column chromatyography. The sepcific activity of purified enzyme on its substrate, compactin-Na, was determined to be 15 nmol of pravastatin per mg protein. The molecular mass of this enzyme on SDS-PAGE was $37{\pm}0.5$ kDa, pI was 4.5, and its CO difference spectrum showed maximum absorption peaks at 452 and 550nm, respectively. The N-terminal amino acid sequence was determined to be Met>Thr>Cys>Thr>Pro>Val>Thr>Val>The>Gly>Ala>Ala>Gly>Gln>Ile>Gly>Tyr>Ala>Leu. Its apparent $K_m$ on compactin-Na was $1.294{\mu}M{\cdot}min^-1,\;and\;V_{max}\;was\;1.028{\mu}M{\cdot}min^-1$. The maximum substrate concentration ($K_s$) for reaction was $270 {\mu}M$and thus $1/[K_s]$ was $3.7{\mu}M$. These physicochemical characteristics and kinetic behavior of this enzyme were compared and shown to be different from those of Streptomyces cytochrome P-450 enzymes reported, suggesting that this enzyme may be an additional member of the Streptomyces cytochrome P-450 family.

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Gene Cloning, Expression, and Characterization of a $\beta$-Agarase, AgaB34, from Agarivorans albus YKW-34

  • Fu, Xiao Ting;Pan, Cheol-Ho;Lin, Hong;Kim, Sang-Moo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.257-264
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    • 2009
  • A $\beta$-agarase gene, agaB34, was functionally cloned from the genomic DNA of a marine bacterium, Agarivorans albus YKW-34. The open reading frame of agaB34 consisted of 1,362 bp encoding 453 amino acids. The deduced amino acid sequence, consisting of a typical N-terminal signal peptide followed by a catalytic domain of glycoside hydrolase family 16 (GH-16) and a carbohydrate-binding module (CBM), showed 37-86% identity to those of agarases belonging to family GH-16. The recombinant enzyme (rAgaB34) with a molecular mass of 49 kDa was produced extracellularly using Escherichia coli $DH5{\alpha}$ as a host. The purified rAgaB34 was a $\beta$-agarase yielding neoagarotetraose (NA4) as the main product. It acted on neoagarohexaose to produce NA4 and neoagarobiose, but it could not further degrade NA4. The maximal activity of rAgaB34 was observed at $30^{\circ}C$ and pH 7.0. It was stable over pH 5.0-9.0 and at temperatures up to $50^{\circ}C$. Its specific activity and $k_{cat}/K_m$ value for agarose were 242 U/mg and $1.7{\times}10^6/sM$, respectively. The activity of rAgaB34 was not affected by metal ions commonly existing in seawater. It was resistant to chelating reagents (EDTA, EGTA), reducing reagents (DTT, $\beta$-mercaptoethanol), and denaturing reagents (SDS and urea). The E. coli cell harboring the pUC18-derived agarase expression vector was able to efficiently excrete agarase into the culture medium. Hence, this expression system might be used to express secretory proteins.

Antagonistic Activity against Dirty Panicle Rice Fungal Pathogens and Plant Growth-Promoting Activity of Bacillus amyloliquefaciens BAS23

  • Saechow, Sukanya;Thammasittirong, Anon;Kittakoop, Prasat;Prachya, Surasak;Thammasittirong, Sutticha Na-Ranong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1527-1535
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    • 2018
  • Bacterial strain BAS23 was isolated from rice field soil and identified as Bacillus amyloliquefaciens. Based on dual culture method results, the bacterium BAS23 exhibited potent in vitro inhibitory activity on mycelial growth against a broad range of dirty panicle fungal pathogens of rice (Curvularia lunata, Fusarium semitectum and Helminthosporium oryzae). Cell-free culture of BAS23 displayed a significant effect on germ tube elongation and mycelial growth. The highest dry weight reduction (%) values of C. lunata, H. oryzae and F. semitectum were 92.7%, 75.7%, and 68.9%, respectively. Analysis of electrospray ionization-mass spectrometry (ESI-MS) and $^1H$ nuclear magnetic resonance (NMR) spectroscopy revealed that the lipopeptides were iturin A with a C14 side chain (C14 iturinic acid), and a C15 side chain (C15 iturinic acid), which were produced by BAS23 when it was cultured in nutrient broth (NB) for 72 h at $30^{\circ}C$. BAS23, the efficient antagonistic bacterium, also possessed in vitro multiple traits for plant growth promotion and improved rice seedling growth. The results indicated that BAS23 represents a useful option either for biocontrol or as a plant growth-promoting agent.

Cloned Placenta of Korean Native Calves Died Suddenly at Two Months after Birth Displays Differential Protein Expression

  • Kim Hong Rye;Kang Jae Ku;Lee Hye Ran;Yoon Jong Taek;Seong Hwan Hoo;Jung Jin Kwan;Park Chang Sik;Jin Dong Il
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.63-68
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    • 2005
  • Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, $H^+-transporting$ ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.

Ultrasonographic Diagnosis of the Symptomatic Ganglion around the Bicipital Groove - Case Report - (증상을 유발한 이두구 주변의 결절종에 대한 초음파 진단 - 증례보고 -)

  • Han, Suk Ku;Kim, Seok Hyun;Song, Ha Jung;Kim, Ryul;Park, Hyun Woo;Song, Hyun Seok
    • The Journal of Korean Orthopaedic Ultrasound Society
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    • v.8 no.1
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    • pp.21-25
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    • 2015
  • Diagnosis of the long head tendon of biceps is not easy. and an ultrasonography is helpful at the office. We report a case who visited with the right shoulder pain and was diagnosed as a mass around the bicipital groove using the ultrasonography, finally undertook an arthroscopic resection. On the histologic examination, synovial lining cell were not found, the wall was thickened. He diagnosed as ganglion.

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Association between Hair Mineral Content and Nutritional Status in Vegetarians and Non-Vegetarians (채식과 비채식인의 모발 내 무기질 함량과 영양상태의 관련성)

  • Cho, Jung-Hee;Kim, Mi-Kyoung;Kim, So-Hyeon;Cho, Sang-Woon;Park, Yoo-Kyoung
    • Journal of Nutrition and Health
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    • v.44 no.3
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    • pp.203-211
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    • 2011
  • Previous studies have shown that plant-based vegetarian diets, which typically contain a variety of antioxidants and dietary fiber, help reduce the risk of heart disease, diabetes, obesity, and cancer. However, some studies have reported that vegetarian diets can lead to deficiencies in protein and trace minerals compared to non-vegetarian diets. This study was conducted to compare anthropometric measurements, blood parameters, dietary intake, and hair mineral status in long-term vegetarians (MV; moderate vegan, LV; lacto-ovo vegetarian) and non-vegetarians (NV). Thirty MV (12 males, 18 females; mean age, $50.58{\pm}5.05$ years), 15 LV (11 males, four females; mean age, $49.45{\pm}4.97$ years), and 30 NV (15 males, 15 females; mean age, $48.90{\pm}3.62$ years) participated. No significant differences were observed for age, height, weight, or body mass index, but body fat was significant lower in MV and LV males than that in NV males. White blood cell counts of MV, LV, and NV male subjects were significantly different. Dietary intake data showed that plant protein and plant iron intake were significantly higher in MV and LV than those in NV. Animal protein, animal fat, and animal iron intake were significantly higher in NV than those in MV and LV. A hair mineral analysis showed that calcium and iron were higher in the hair of MV and LV than those in NV. Zinc concentration in hair was not significantly different among the groups. The results suggest that vegetarian diets are adequate to sustain mineral status to at least the same degree as that of non-vegetarian diets.

One Case of Insulinoma (인슐린 분비성 췌장세포암 1례)

  • Ahn, Jae-Hee;Seo, Hye-Sang;Yoon, Sung-Chul;Yoon, Kyung-Woo;Choi, Soo-Bong;Lee, Hyun-Woo
    • Journal of Yeungnam Medical Science
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    • v.5 no.2
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    • pp.205-211
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    • 1988
  • Insulinoma is a rare tumor, occurring more often in women and in the older age range. Eighty percent of patients have a single benign tumor, usually 2cm in diameter, located with about equal frequency in body, head or tail of pancreas and amenable to surgical cure. About 10% have multiple tumors. The remaining 10% of patients have metastatic malignant insulinoma. The symptom of insulinoma is characterized by the periodic attack of hypoglycemia of blood sugar level below 50mg%, by fasting or exertion, and rapid relief of symptom by oral or intravenous administration of glucose. Symptom often lead to misdiagnosis as a neurologic or psychiatric disorder. A case described by authors was 44-year old female with the chief complaints of the loss of consciousness, epileptic seizure although she has been treated by anticonvulsants. Serum blood sugar and insulin level during fasting sugested insulinoma but abdominal computed tomography shows no definitive mass in pancreas. Celiac angiography revealed insulinoma. She transfered to the defartment of General Surgery and was performed enucleation. Microscopic findings shows the islet cell tumor of pancreas. A brief review of the literature was made.

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Stage-specific Expression of Lanosterol 14${\alpha}$-Demethylase in Mouse Oocytes in Relation to Fertilization and Embryo Development Competence

  • Song, Xiaoming;Ouyang, Hong;Tai, Ping;Chen, Xiufen;Xu, Baoshan;Yan, Jun;Xia, Guoliang;Zhang, Meijia
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.3
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    • pp.319-327
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    • 2009
  • Follicular fluid meiosis-activating sterol (FF-MAS) has been suggested as a positive factor which could improve the oocyte quality and subsequent embryo development after in vitro fertilization. However, FF-MAS is a highly lipophilic substance and is hard to detect in studying the relationship between MAS and quality of oocyte maturation. The present study focused on the expression of lanosterol 14${\alpha}$-demethylase (LDM), a key enzyme that converts lanosterol to FF-MAS, on mouse oocyte maturation and its potency on development. LDM expression was strong in gonadotropin-primed germinal vesicle stage oocytes, weak after germinal vesicle breakdown (GVBD), and then strong in MII stage oocytes. The LDM-specific inhibitor azalanstat significantly inhibited oocyte fertilization (from 79.4% to 68.3%, p<0.05). Also, azalanstat (5 to 50 ${\mu}M$) decreased the percentage of blastocyst development dosedependently (from 78.7% to 23.4%, p<0.05). The specific inhibition of sterol ${\Delta}14$-reductase and ${\Delta}7$-reductase by AY9944 accumulates FF-MAS and could increase blastocyst development rates. Additionally, in the AY9944 group, the rate of inner cell mass (ICM)/ total cells was similar to that of in vivo development, but the rate was significantly decreased in azalanstat treatment. In conclusion, LDM, the key enzyme of FF-MAS production, may play an important role in fertilization and early development of the mouse embryo, especially in vitro.

Association of Genetic Polymorphisms at 1q22 but not 10q23 with Gastric Cancer in a Southern Chinese Population

  • Yang, Xue-Xi;Li, Fen-Xia;Zhou, Cui-Ping;Hu, Ni-Ya;Wu, Ying-Shong;Li, Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2519-2522
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    • 2012
  • Objective: Data from a recent genome-wide association studiesy of gastric cancer (GC) and oesophageal squamous cell carcinoma in Chinese living in the Taihang Mountains of north-central China suggest that 1q22 and 10q23 are susceptibility-associated regions for GC. However, this has not been confirmed in southern Chinese populations. The aim of this study was to investigate whether these polymorphisms at 1q22 and 10q23 are associated with the risk of GC in a southern Chinese population. Methods: We selected seven top significant associated single nucleotide polymorphisms (SNPs) at 1q22 and 10q23 and conducted a population-based case-control study in a southern Chinese population. Genotypes were determined using MassARRAYTM system (Sequenome, San Diego, CA). Results: Two SNPs at 1q22, rs4072037 and rs4460629, were significantly associated with a reduced risk of GC, best fitting the dominant genetic model. Logistic regression models adjusted for age and sex showed that rs4072037 AG and GG (OR=0.64, P=0.017, compared with AA) and rs4460629 CT and TT (OR=0.54, P=0.0016, compared with TT) significantly reduced the risk of GC. However, no significant results for the five SNPs at 10q23 were obtained in this study. Conclusion: These outcomes indicate that 1q22 is associated with GC susceptibility in this southern Chinese population, while an association for the locus at 10q23 was not confirmed.