• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1178-1188
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• 2014

In this study, the yeast Pichia pastoris was genetically modified to assemble minicellulosomes on its cell surface by the heterologous expression of a truncated scaffoldin CipA from Clostridium acetobutylicum. Fluorescence microscopy and western blot analysis confirmed that CipA was targeted to the yeast cell surface and that NtEGD, the Nasutitermes takasagoensis endoglucanase that was fused with dockerin, interacted with CipA on the yeast cell surface, suggesting that the cohesin and dockerin domains and cellulose-binding module of C. acetobutylicum were functional in the yeasts. The enzymatic activities of the cellulases in the minicellulosomes that were displayed on the yeast cell surfaces increased dramatically following interaction with the cohesin-dockerin domains. Additionally, the hydrolysis efficiencies of NtEGD for carboxymethyl cellulose, microcrystal cellulose, and filter paper increased up to 1.4-fold, 2.0-fold, and 3.2-fold, respectively. To the best of our knowledge, this is the first report describing the expression of C. acetobutylicum minicellulosomes in yeast and the incorporation of animal cellulases into cellulosomes. This strategy of heterologous cellulase incorporation lends novel insight into the process of cellulosome assembly. Potentially, the surface display of cellulosomes, such as that reported in this study, may be utilized in the engineering of S. cerevisiae for ethanol production from cellulose and additional future applications.

• 한국수산과학회지
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• 제46권6호
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• pp.785-792
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• 2013

Two studies were conducted to investigate the effects of dietary nucleotides (NT) on the growth performance, hematological parameters, and innate immune responses of red seabream Pagrus major. In Expt I, six experimental diets were formulated: a control, four that contained each NT at a level of 0.15% (inosine monophosphate, IMP; adenosine monophosphate, AMP; guanosine monophosphate, GMP; and uridine monophosphate, UMP), and one with a 1:1:1:1 mixture of NTs (IMP, AMP, GMP, and UMP). In Expt II, five experimental diets were formulated that contained 0, 0.1, 0.2, 0.4, and 1.0% IMP (commercial product). Triplicate groups of juvenile (initial body weight 33.1 g) and growing (initial body weight 120 g) red seabream were fed one of the experimental diets to apparent satiation for 8 weeks in Expt I and 12 weeks in Expt II. In Expt I, fish fed diets with NT had higher growth performance than the control group. The nitroblue tetrazolium and lysozyme activities were higher in fish fed the mixed-NT diet, and lowest in the control group. In Expt II, the final body weight and feed utilization of fish fed the 0.1% IMP diet were significantly higher than those of fish fed the control or 1.0% IMP diets. Diet palatability was improved significantly when 0.1% IMP was added. The lysozyme activity was higher in fish fed diets with 0.4-1.0% than in the control group. These results suggest that supplementation of 0.15% IMP and Mixed-NTs in diet can enhance the growth and immune responses in juvenile red seabream. The optimum IMP level appears to be 0.2% in practical feed formulation for growing red seabream.

• 한국수정란이식학회지
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• 제23권1호
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• pp.31-36
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• 2008

We attempted to control the maturation promoting factors (MPF) activity and nuclear remodeling of somatic cell nuclear transfer (NT) bovine embryos. Bovine ear skin fibroblasts were fused to enucleated oocytes treated with either 5 mM caffeine for 2.5 h or 0.5 mM vanadate for 0.5 h and activated. The nuclear remodeling type of the reconstituted embryos was evaluated 1.5 h after activation. MPF activity was assessed in enucleated and chemical treated oocytes before the injection of a donor cell. Effect of chemicals on the embryonic development was evaluated with parthenogenetic embryos. MPF activity increased significantly by caffeine treatment, but decreased by vanadate treatment (p<0.05). Caffeine or vanadate had no deleterious effect on the parthenogenetic embryo development. In caffeine treated group, premature chromosome condensation (PCC) was occurred in 72.2% of NT embryos (p<0.05). In contrast, vanadate induced the formation of a pronucleus-like structure (PN) in a high frequency (68.9%, p<0.05) without PCC (NPCC). Blastocyst development of NT embryos increased by treating with caffeine (30.3%), whereas decreased by treating with vanadate (11.4%) compared to control (22.1%, p<0.05). The results indicate that caffeine or vanadate can control of MPF activity and remodeling type of NT embryos, resulting in the increased or decreased in vitro development.

• Journal of Electrochemical Science and Technology
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• 제1권1호
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• pp.39-44
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• 2010

1.5 V and 3.0 V-class film-type primary batteries were designed for radio frequency identification (RFID) tag. Efficient fabrication processes such as screen-printings of conducting layer ($25{\mu}m$), active material layer ($40{\mu}m$ for anode and $80{\mu}m$ for cathode), and electrolyte/separator/electrolyte layer ($100{\mu}m$), were adopted to give better performances of the 1.5 V-class film-type Leclanch$\acute{e}$ primary battery for battery-assisted passive (BAP) RFID tag. Lithium (Li) metal is used as an anode material in a 3.0 V-class film-type $MnO_2||$Li primary battery to increase the operating voltage and discharge capacity for application to active sensor tags of a radio frequency identification system. The fabricated 3.0 V-class film-type Li primary battery passes several safety tests and achieves a discharge capacity of more than 9 mAh $cm^{-2}$.

• International Journal of Oral Biology
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• 제40권2호
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• pp.71-77
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• 2015

The activation of glial cells in the spinal cord has been contribute to the initiation and maintenance of pain facilitation induced by peripheral inflammation and nerve injury. The present study investigated effects of botulinum toxin type A (BoNT-A), injected subcutaneously or intracisternally, on the expression of microglia and astrocytes in rats. Complete Freund's Adjuvant (CFA)-induced inflammation was employed as an orofacial chronic inflammatory pain model. A subcutaneous injection of $40{\mu}L$ CFA into the vibrissa pad was performed under 3% isoflurane anesthesia in SD rats. Immunohistochemical analysis for changes in Iba1 (a microglia marker) and GFAP (an astrocyte marker), were performed 5 days after CFA injection. Subcutaneous injection of CFA produced increases in Iba1 and GFAP expression, in the ipsilateral superficial lamia I and II in the medullary dorsal horn of rats. Subcutaneous treatment with BoNT-A attenuated the up-regulation of Iba1 and GFAP expressions induced by CFA injection. Moreover, intracisternal injection of BoNT-A also attenuated the up-regulated Iba1 and GFAP expressions. These results suggest that the anti-nociceptive action of BoNT-A is mediated by modulation activation of glial cells, including microglia and astrocyte.

• Journal of Veterinary Science
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• 제22권6호
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• pp.80.1-80.13
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• 2021

Background: Hyperbaric gaseous cryotherapy (HGC) is a type of cryotherapy used in human medicine for rehabilitation after orthopedic surgeries. Because HGC is known to reduce acute or chronic pain, research is needed to prove its effectiveness in veterinary medicine. Objectives: To compare the effects of HGC between the HGC treatment group and the nontreatment (NT) group on postoperative swelling, range of motion, lameness score, postoperative pain, and kinetic measurements after stifle joint surgery in dogs. Methods: Dogs were randomized in an HGC group or NT groups. In the HGC group, HGC was applied once a day for a total of 2 days after surgery. All parameters were measured postoperatively and at 1, 2, 10, and 28 days after surgery. Results: Twenty dogs were enrolled: 10 in the HGC group and 10 in the NT group. Soft tissue swelling was not significantly different between groups at any time point. In the HGC group, pain scores decreased significantly 24 h after surgery and 48 h after surgery. Dogs in the HGC group showed a significantly decreased lameness and improvement for all kinetic measurements beginning 48 h after surgery. In addition, the HGC group indicated a significant increase in range of motion as compared with the NT group at 28 days after surgery. Conclusions: HGC plays a powerful role in decreasing initial postoperative pain. Furthermore, the improvement in pain affects the use of the operated limb, and the continued use of the limb eventually assists in the quick recovery of normal function.

• Journal of Animal Science and Technology
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• 제65권1호
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• pp.57-68
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• 2023

Flavor is an important sensory trait of chicken meat. The free amino acid (FAA) and nucleotide (NT) components of meat are major factors affecting meat flavor during the cooking process. As a genetic approach to improve meat flavor, we performed a genome-wide association study (GWAS) to identify the potential candidate genes related to the FAA and NT components of chicken breast meat. Measurements of FAA and NT components were recorded at the age of 10 weeks from 764 and 767 birds, respectively, using a White leghorn and Yeonsan ogye crossbred F2 chicken population. For genotyping, we used 60K Illumina single-nucleotide polymorphism (SNP) chips. We found a total of nine significant SNPs for five FAA traits (arginine, glycine, lysine, threonine content, and the essential FAAs and one NT trait (inosine content), and six significant genomic regions were identified, including three regions shared among the essential FAAs, arginine, and inosine content traits. A list of potential candidate genes in significant genomic regions was detected, including the KCNRG, KCNIP4, HOXA3, THSD7B, and MMUT genes. The essential FAAs had significant gene regions the same as arginine. The genes related to arginine content were involved in nitric oxide metabolism, while the inosine content was possibly affected by insulin activity. Moreover, the threonine content could be related to methylmalonyl-CoA mutase. The genes and SNPs identified in this study might be useful markers in chicken selection and breeding for chicken meat flavor.

• Journal of Chest Surgery
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• 제40권9호
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• pp.593-599
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• 2007

배경: 폐절제술이 우심실 기능에 미치는 영향과 BNP (brain natriuretic peptide)의 관계에 대해서는 아직 자세히 알려져 있지 않다. 본 연구에서는 폐절제술 후 변화하는 혈역학적 수치와 우심부전의 보상기전으로서 증가하는 BNP의 관계에 대해서 알아보고 BNP가 폐절제술 후의 우심부전의 지표가 될 수 있는지를 확인하고자 하였다. 대상 및 방법: 폐암으로 폐엽절제술과 전폐절제술을 시행한 12명의 환자에서 면역화학적 방법(Elecsys $1010^{(R)}$, Roche, Germany)을 이용하여 수술 전후의 NT-proBNP 수준을 측정하고, Swan-Ganz 카테터로 수술 전후에 혈역학적 지표를 측정하여 비교하였으며 수술 전후에 심초음파를 시행하여 우심실 및 좌심실압 등의 변화를 비교하였다. 통계처리는 SPSSWIN(version 11.5)를 이용한 Wilcoxon rank sum test와 linear regression을 이용하였다. 결과: NT-proBNP 수준은 수술 전과 비교하여 수술 후 6시간, 수술 후 1일과 2일, 3일, 7일째에 유의하게 증가하였다(p=0.003, 0.002, 0.002, 0.006, 0.004). Swan-Ganz 카테터를 통한 혈역학적 변수 중에서는 평균 폐동맥압이 수술 전과 비교하여 수술 직후와 수술 후 6시간, 수술 후 1일, 2일, 3일째에 유의하게 증가하였으며(p=0.002, 0.002, 0.006, 0.007, 0.008), 평균 우심실압력은 수술 직후와 수술 후 6시간, 수술 후 1일과 3일에서 유의하게 증가하였다(p=0.006, 0.009, 0.044, 0.032). 폐혈관저항지수[폐혈관저항지수=(평균폐동맥압-평균폐동맥쐐기압)/심박출계수]는 수술 후 6시간, 수술 후 2일에서 유의한 증가가 있었다(p=0.008, 0.028). 평균폐동맥압의 수술 후 변화와 NT-proBNP 변화를 회기분석하였을 때 수술 후 6시간에서 유의성이 있었으며(r=0.602, p=0.038) 이후에는 유의성이 없었다. 심초음파 결과는 수술 전후를 비교하여 유의성이 없었다. 결론: 폐절제술 후 6시간의 폐동맥압의 변화와 NT-proBNP의 변화가 유의성이 있었다. 따라서 폐절제술 후 NT-proBNP의 변화는 폐절제술 후 우심실의 조기 혈역학적 변화를 반영하는 척도가 될 수 있으리라 판단된다.

• Pediatric Infection and Vaccine
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• 제7권1호
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• pp.83-93
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• 2000

목 적 : Urabe AM-9 볼거리 백신주는 무균성 뇌막염의 발생 빈도가 높은 것으로 알려져 있다. 백신 접종 후 무균성 뇌막염을 일으키는 경우 Urabe AM-9 백신주의 hemagglutinin-neuraminidase(HN) 유전자의 염기서열 1081번의 G가 A로 치환되어 335번째 아미노산이 glutamic acid에서 lysine으로 바뀌게 됨에 따라 야생형의 mumps 바이러스와 동일한 작용을 나타내는 것으로 알려져 있다. 이에 국내에서 사용중인 Urabe AM-9 백신주의 안전성 여부를 조사하기 위해 일부 백신주와 야생 분리주의 HN 유전자의 염기서열 분석을 실시하였다. 방 법 : 국내에서 사용되고있는 Urabe AM-9주 백신 2종류와 1998~1999년에 볼거리 환자로부터 분리된 야생 분리주를 이용하여 RT-PCR 방법에 의해 HN 유전자를 증폭시킨 후 염기 및 아미노산 서열 분석을 실시하였다. 결 과 : 백신주와 야생 분리주 모두 HN 343, 1476, 1570번 위치에서의 염기 변이는 없었으나 1081번 위치 염기는 모두 야생주와 같은 Lysine/AAA form으로 나타났으며 변이주인 Glutamic acid/GAA와의 혼합 양상은 관찰되지 않았다. HN 1470번 위치의 염기는 백신주 중 계대 배양시 C에서 A로 치환되었으며, HN 1727번 위치 염기는 모두 A에서 G로 치환된 것으로 나타났다. 결 론 : 국내에서 사용중인 2종류의 Urabe AM-9 백신주와 야생 분리주 모두 1081번 위치의 염기가 야생주와 같은 무균성 뇌막염을 일으킬 생물학적 가능성이 있는 것으로 나타났으며, 국내에서도 상기 백신주를 사용한 후 무균성 뇌막염이 발생하고 있을 개연성이 매우 크다고 사료되며 직접적인 연관성에 대한 보완 연구를 위하여 정확하고 믿을만한 자료에 의한 실제 부작용 실태를 파악하여야 한다.

• Molecules and Cells
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• 제41권5호
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• pp.413-422
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• 2018

Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.