• IMMUNE NETWORK
• /
• 제10권2호
• /
• pp.35-45
• /
• 2010

Background: Expression of recombinant antibodies and their derivatives fused with other functional molecules such as alkaline phosphatase in Escherichia coli is important in the development of molecular diagnostic reagents for biomedical research. Methods: We investigated the possibility of applying a well-known Fos-Jun zipper to dimerize $V_H$ and $V_L$ fragments originated from the Fab clone (SP 112) that recognizes pyruvate dehydrogenase complex-E2 (PDC-E2), and demonstrated that the functional zipFv-112 and its alkaline phosphatase fusion molecules (zipFv-AP) can be produced in the cytoplasm of Origami(DE3) trxB gor mutant E. coli strain. Results: The zipFv-AP fusion molecules exhibited higher antigen-binding signals than the zipFv up to a 10-fold under the same experimental conditions. However, conformation of the zipFv-AP seemed to be influenced by the location of an AP domain at the C-terminus of $V_H$ or $V_L$ domain [zipFv-112(H-AP) or zipFv-112(L-AP)], and inclusion of an AraC DNA binding domain at the C-terminus of VH of the zipFv-112(L-AP), termed zipFv-112(H-AD/L-AP), was also beneficial. Cytoplasmic co-expression of disulfide-binding isomerase C (DsbC) helped proper folding of the zipFv-112(H-AD/L-AP) but not significantly. Conclusion: We believe that our zipFv constructs may serve as an excellent antibody format bi-functional antibody fragments that can be produced stably in the cytoplasm of E. coli.

• Reproductive and Developmental Biology
• /
• 제36권2호
• /
• pp.121-131
• /
• 2012

After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q-ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.

• 한국식품위생안전성학회지
• /
• 제18권3호
• /
• pp.146-152
• /
• 2003

당 대사에 관여하는 Pyruvate dehydrogenase phosphatase (PDP)는 해당과정에서의 대사 산물인 pyruvate 를 acetyl CoA로 만들어 구연산 회로로 진입시켜주는 효소인 Pyruvate dehydrogenase complex(PDP)의 활성을 조절하는 중요한 효소이다. PDP의 catalytic subunit는 PDP의 dihydrolipoamide acetyltransferase(E2), PDP regulatory subunit (PDPr), 그리고 칼슘 결합 도메인 등으로 구성되어 있는 것으로 추측되어지고 있다. 본 연구에서는 PDP 단백질을 분리정제하고 결정화 하고자하였다. PDP는 catalytic subunit(PDPc, Mr 52,600 Da)과, regulatory subunit (PDPr, 95,600 Da)으로 구성되어 있으며 칼슘 존재하에 PDPc는 dihydrolipoamide acetyltransferase(E2) component와 결합하여 기질인 인산 E1 component의 탈인산화율을 증가시킨다. PDPc는 intrinsic 칼슘 결합부위를 가지며 두 번째 칼슘 부위는 E2 존재 하에 형성된다. 이러한 특이한 상호반응을 이용한 GSH-Sepharose-GST-L2 matrix를 이용하여 약 1000 U/mg의 specific activity를 갖는 순수 PDPc를 약 80%의 yield로 얻어 결정화에 사용하였다.

• 한국약용작물학회지
• /
• 제18권6호
• /
• pp.416-420
• /
• 2010

Root yield and quality of ginseng cultured in paddy soil was low relatively compared with that of upland soil because of moisture injury in root during rainy season. Drainage class in soils generally divided into 6 classes, and it is possible to cultivate ginseng practically in imperfectly drainage class (IDC). This study carried out to select the varieties that is suitable for paddy soil, which is easy to be generated rusty-colored root and physiological-discolored leaf. Experiment plot arranged with the condition of soil humidity contents such as poorly drainage class (PDC) and imperfectly drainage class (IDC), and upland soil. Growth characteristics and root yield were investigated in four-year-old ginseng of varieties, Cheonpoong (CP), Yeonpoong (YP), Hwangsookjong (HS), and Jakyeongjong (JK). CP among four varieties showed the highest yield in IDC and CP was the lowest ratio in leaf discoloration and rusty-colored root. HS was followed by CP in the order of root yield, but it had the weakness that the ratio of rusty-colored root was high respectively.

• Journal of Microbiology and Biotechnology
• /
• 제23권2호
• /
• pp.251-259
• /
• 2013

An endoxylanase gene (PoxynA) that belongs to the glycoside hydrolase (GH) family 11 was cloned from a xylanolytic strain, Penicillium oxalicum B3-11(2). PoxynA was overexpressed in Trichoderma reesei QM9414 by using a constitutive strong promoter of the encoding pyruvate decarboxylase (pdc). The high extracellular xylanase activities in the fermentation liquid of the transformants were maintained 29~35-fold higher compared with the wild strain. The recombinant POXYNA was purified to homogeneity, and its characters were analyzed. Its optimal temperature and pH value were $50^{\circ}C$ and 5.0, respectively. The enzyme was stable at a pH range of 2.0 to 7.0. Using beechwood as the substrate, POXYNA had a high specific activity of $1,856{\pm}53.5$ IU/mg. In the presence of metal ions, such as $Cu^{2+}$, and $Mg^{2+}$, the activity of the enzyme increased. However, strong inhibition of the enzyme activity was observed in the presence of $Mn^{2+}$ and $Fe^{2+}$. The recombinant POXYNA hydrolyzed birchwood xylan, beechwood xylan, and oat spelt xylan to produce short-chain xylooligosaccharides, xylopentaose, xylotriose, and xylobiose as the main products. This is the first report on the expression properties of a recombinant endoxylanase gene from Penicillium oxalicum. The properties of this endoxylanase make it promising for applications in the food and feed industries.

• Molecules and Cells
• /
• 제45권7호
• /
• pp.454-464
• /
• 2022

DJ-1 is one of the causative genes of early-onset familial Parkinson's disease (PD). As a result, DJ-1 influences the pathogenesis of sporadic PD. DJ-1 has various physiological functions that converge to control the levels of intracellular reactive oxygen species (ROS). Based on genetic analyses that sought to investigate novel antioxidant DJ-1 downstream genes, pyruvate dehydrogenase (PDH) kinase (PDK) was demonstrated to increase survival rates and decrease dopaminergic (DA) neuron loss in DJ-1 mutant flies under oxidative stress. PDK phosphorylates and inhibits the PDH complex (PDC), subsequently downregulating glucose metabolism in the mitochondria, which is a major source of intracellular ROS. A loss-of-function mutation in PDK was not found to have a significant effect on fly development and reproduction, but severely ameliorated oxidative stress resistance. Thus, PDK plays a critical role in the protection against oxidative stress. Loss of PDH phosphatase (PDP), which dephosphorylates and activates PDH, was also shown to protect DJ-1 mutants from oxidative stress, ultimately supporting our findings. Further genetic analyses suggested that DJ-1 controls PDK expression through hypoxia-inducible factor 1 (HIF-1), a transcriptional regulator of the adaptive response to hypoxia and oxidative stress. Furthermore, CPI-613, an inhibitor of PDH, protected DJ-1 null flies from oxidative stress, suggesting that the genetic and pharmacological inhibition of PDH may be a novel treatment strategy for PD associated with DJ-1 dysfunction.

• 한국의류산업학회지
• /
• 제24권2호
• /
• pp.260-266
• /
• 2022

Cellulose fiber is a material used in various fields. It is the most used type of fiber because of its excellent hygroscopicity and dyeability. Recently, as natural fiber materials have been highlighted due to the influence of eco-friendliness and well-being, bamboo fiber has become a commonly used eco-friendly fiber. Cellulose fibers are part of the -OH hydroxyl group, which means they are more chemically reactive than synthetic fibers. In this study, the cationization properties of bamboo-cotton blended fabrics cationized using CHPTAC (3-chloro-2-hydroxypropyl trimethyl ammonium chloride) in the PDC (padding-drying-curing) method were investigated. Various characteristics according to cationization were studied through elemental analysis, FT-IR (fourier-transform infrared spectroscopy) analysis, X-ray diffraction analysis, TGA (thermogravimetric) analysis, and SEM (scanning electron microscope) analysis. The nitrogen content of the cationized bamboo-cotton blended fabric increased with an increase in the concentration of the cationizing agent CHPTAC, and it was seen to be highly bound to cellulose molecules. As a result of the FT-IR analysis, both 100% pure cotton fabrics and CHPTAC-0 and CHPTAC-150 fabrics were seen to be typical cellulose. As a result of the X-ray diffraction analysis, both 100% pure cotton fabrics and CHPTAC-0 and CHPTAC-150 fabrics showed typical cellulose I structures. As a result of the X-ray diffraction analysis, both 100% pure cotton fabrics and CHPTAC-0 and CHPTAC-150 fabrics showed typical cellulose I structures. As the cationization progressed, micropores appeared on the surface of the blended fabric.

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2022년도 추계학술대회
• /
• pp.132-132
• /
• 2022

The application of direct seeding cultivation reduces time, labor, and cost. However, this application often has poor seedling establishment and leads to lower yield as compared to transplanting system. The tolerance to anaerobic and low temperature germination is important to improve seedling establishment and the wide-spread application of direct seeding method. This study was carried out to evaluate the responses of three japonica cultivars to different temperatures (15℃, 18℃, 21℃, 24℃, and 27℃) and different flooding levels (1 cm, 4 cm, 7 cm, 10 cm) during germination and seedling development. The mean survival percentage significantly increased (P<0.05) when the flooding level decreased and when temperature increased. There were significant effects of the interaction between temperature and water depth on survival percentage and seedling height. When temperature decreased from 27℃ to 15℃, the germination duration significantly increased from 6.4 days to 16.3 days while the germination speed, survival percentage, and seedling height decreased from 5.3 seeds day^-1, 61.9% and 190.6 cm to 2.2 seeds day^-1, 33.2%, and 47 cm, respectively. The increase in temperature under submergence condition was associated with the increased expression of Amy3D and ALDH2a but the decreased expression of ADH1 and PDC1. The results of this research would be used for further studies and breeding programs to improve rice seedling establishment and the application of direct seeding cultivation.

• Investigative Magnetic Resonance Imaging
• /
• 제18권3호
• /
• pp.232-243
• /
• 2014

목적: 축추이하 경추 손상시 발생하는 연속적, 비연속적 연골하골 압박손상의 빈도와 원발부위 손상 패턴에 대해 알아보고자 하였다. 대상과 방법: 축추이하 경추 손상이 있는 환자 47명의 전산화 단층촬영, 자기공명영상, 의무기록을 후향적으로 검토하여 연속적, 비연속적 연골하골 압박손상의 발생유무와 수, 레벨, 손상 부위를 기록하였다. 원발부위 손상 패턴을 알아보기 위하여 손상된 원발부위의 레벨, 수, 손상 형태, 전/후방 추간판인대복합체의 손상유무, 후방인대복합체의 손상유무, 척수 손상유무를 분석하고 손상 기전을 분석하였다. 분석된 원발손상 패턴과 손상기전은 Mann-Whitney U test, Pearson's chi square test, Fisher's exact test의 통계적 기법으로 연골하 압박손상의 발생유무와 연관성을 조사하였다. 결과: 총 18명에게서 (18/47, 38.29%) 연골하 압박손상이 발생했으며 그 중 9명은 원발부위와 인접하여 연속적으로, 다른 9명은 원발부위와 떨어져 비연속적으로 발생하였다. 3번 흉추에 가장 흔하게 발생하였고 (15/47, 31.91%), 세 개의 레벨에 걸쳐있는 경우가 가장 많았다 (6/18, 33.33%). 모든 연골하 압박손상은 척추체의 전상방 부위나 상종판 주변에 발생하였으며 강한 외력의 외상과 연관되었다. 연골하 압박손상의 발생은 원발부위의 손상형태와 후방인대복합체의 손상과 통계학적으로 유의한 연관성을 보였다. 비연속적 연골하 압박손상은 연속적 연골하 압박손상에 비해 비교적 상부 경추에 원발 손상부위가 있고 후방추간판인대 복합체의 손상을 동반하는 경우가 통계학적으로 유의하게 많았다. 그외 분석한 다른 인자들은 통계학적으로 유의한 결과를 보이지 않았다. 결론: 이차성 연골하 압박손상은 흔하며 강한 외력에 의한 굴곡압박성 경추 외상과 관련 있을 것이다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제25권8호
• /
• pp.1124-1131
• /
• 2012

Four Thai - rumen fistulated male swamp buffaloes (Bubalus bubalis), about four years old with $400{\pm}20kg$ liveweight, were randomly assigned according to a $4{\times}4$ Latin square design to receive dietary treatments. The treatments were: ground corn cob (GCC) replacement for cassava chip (CC) in concentrate at 0% (T1); GCC replacement at 33% (T2); GCC replacement at 67% (T3); and GCC replacement at 100% (T4), respectively. During the experiment, concentrate was offered at 0.5% BW while 5% urea-treated rice straw was given at ad libitum. The result revealed that there was no effect of GCC replacement on DMI among treatments. In addition, digestibilities of DM, OM and CP were not different while aNDF linearly increased with an increasing level of GCC replacement. However, GCC replacement did not affect rumen fermentation such as ruminal pH, $NH_3$-N and VFA concentration; except C3 proportion which was the highest at 33% replacement while the lowest was at 100% replacement. All replacements of GCC resulted in similar protozoal and bacterial populations and microbial protein synthesis (MPS). Purine derivatives (PD) concentration in urine and PD to creatinine (PDC) index were varied with time of urination and among treatments at 0 to 8 and 8 to 16 h post feeding and higher values were shown among the GCC replacement groups. However at 16 to 24 h-post feeding, it was untraceable. In addition, creatinine concentration was similar among all treatments at every sampling time. Based on the above results, GCC can be used as an energy source for swamp buffalo fed with rice straw. Spot sampling of urine can be used for purine derivatives determination.