• Title/Summary/Keyword: GAL4

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Chlorphenesin Galactoside Production using Immobilized β-galactosidase-producing Escherichia coli (고정화된 β-galactosidase 생산 대장균을 이용한 chlorphenesin galactoside 생산)

  • Jung, Kyung-Hwan
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1164-1168
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    • 2015
  • Previous research showed that chlorphenesin galactoside (CPN-Gal), a preservative in cosmetics, was safer than CPN against human skin cells [9]. To establish a stable and long-term process for CPN-Gal production, we investigated the repeated-batch process. In this process, β-gal-producing recombinant Escherichia coli cells were immobilized in calcium alginate beads, and CPN was converted to CPN-Gal by the transgalactosylation reaction. The process was conducted in a 300 ml flask, which contained E. coli cell-immobilized alginate beads, 33.8 mM of CPN, and 400 g/l of lactose. The pH and temperature were 7.0 and 40℃, respectively. During the repeated-batch operation, four consecutive batch operations were conducted successfully until 192 hr. The conversion yield of CPN to CPN-Gal was 64% during 192 hr, which was higher than the values in previous reports [3, 13]. Thereafter, however, the conversion yield gradually decreased until the operation was finished at 336 hr. Western blotting of immobilized E. coli cells revealed that β-gal gradually decreased after 192 hr. In addition, alginate beads were cracked when the operation was finished. It is probable that, including this loss of E. coli cells by cracks, deactivation, and product inhibition of E. coli β-gal might lead to a gradual decrease in the production of CPN-Gal after 192 hr. However, as the purification of β-gal is not necessary with β-gal-producing recombinant E. coli cells, β-gal-producing E. coli cells might be a practical and cost-effective approach for enzymatically synthesizing CPN-Gal. It is expected that this process will be extended to long-term production process of CPN-Gal for commercialization.

Natural and synthetic pathogen associated molecular patterns modulate galectin expression in cow blood

  • Asiamah, Emmanuel Kwaku;Ekwemalor, Kingsley;Adjei-Fremah, Sarah;Osei, Bertha;Newman, Robert;Worku, Mulumebet
    • Journal of Animal Science and Technology
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    • v.61 no.5
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    • pp.245-253
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    • 2019
  • Pathogen-associated Molecular Patterns (PAMPs) are highly conserved structural motifs that are recognized by Pathogen Recognition receptors (PRRs) to initiate immune responses. Infection by these pathogens and the immune response to PAMPS such as lipopolysaccharide (LPS), Peptidoglycan (PGN), bacterial oligodeoxynucleotides [CpG oligodeoxynucleotides 2006 (CpG ODN2006) and CpG oligodeoxynucleotides 2216 (CpG ODN2216)], and viral RNA Polyinosinic-Polycytidylic Acid (Poly I:C), are associated with infectious and metabolic diseases in animals impacting health and production. It is established that PAMPs mediate the production of cytokines by binding to PRRs such as Toll-like receptors (TLR) on immune cells. Galectins (Gal) are carbohydrate-binding proteins that when expressed play essential roles in the resolution of infectious and metabolic diseases. Thus it is important to determine if the expression of galectin gene (LGALS) and Gal secretion in blood are affected by exposure to LPS and PGN, PolyI:C and bacterial CpG ODNs. LPS increased transcription of LGALS4 and 12 (2.5 and 2.02 folds respectively) and decreased secretion of Gal 4 (p < 0.05). PGN increased transcription of LGALS-1, -2, -3, -4, -7, and -12 (3.0, 2.3, 2.0, 4.1, 3.3, and 2.4 folds respectively) and secretion of Gal-8 and Gal-9 (p < 0.05). Poly I:C tended to increase the transcription of LGALS1, LGALS4, and LGALS8 (1.78, 1.88, and 1.73 folds respectively). Secretion of Gal-1, -3, -8 and nine were significantly increased in treated samples compared to control (p < 0.05). CpG ODN2006 did not cause any significant fold changes in LGALS transcription (FC < 2) but increased secretion of Gal-1, and-3 (p < 0.05) in plasma compared to control. Gal-4 was however reduced in plasma (p < 0.05). CpG ODN2216 increased transcription of LGALS1 and LGALS3 (3.8 and 1.6 folds respectively), but reduced LGALS2, LGALS4, LGALS7, and LGALS12 (-1.9, -2.0, -2.0 and; -2.7 folds respectively). Secretion of Gal-2 and -3 in plasma was increased compared to control (p < 0.05). Gal-4 secretion was reduced in plasma (p < 0.05). The results demonstrate that PAMPs differentially modulate galectin transcription and translation of galectins in cow blood.

Anti-tumor Effect of 4-1BBL Modified Tumor Cells as Preventive and Therapeutic Vaccine

  • Hong Sung Kim
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.312-316
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    • 2022
  • We have previously reported that genetically modified tumor cells with 4-1BBL have anti-cancer effects in a CT26 mouse colorectal tumor model. In this study, genetically modified tumor cells with 4-1BBL were evaluated for their potential as candidates for preventive and therapeutic cancer vaccine. To identify the effect of preventive and therapeutic vaccine of genetically modified tumor cells with 4-1BBL, tumor growth pattern of CT26-4-1BBL as a cancer vaccine was examined compared to CT26-beta-gal. In therapeutic vaccination, CT26-WT was inoculated into mice and then vaccinated mice with doxorubicin (Dox)-treated CT26-beta-gal and CT26-4-1BBL (single or three times). Triple vaccination with Dox-treated tumor cell inhibited tumor growth compared to single vaccination. Vaccination with CT26-4-1BBL showed an efficient tumor growth inhibition compared to vaccination with CT26-beta-gal. For preventive vaccination, Dox-treated CT26-beta-gal and CT26-4-1BBL was vaccinated into mice with three times and then administered mice with CT26-WT. Preventive vaccination with CT26-4-1BBL showed no tumor growth. Preventive vaccination with CT26-beta-gal also led to tumor-free mice. These results suggest that genetically modified tumor cells with 4-1BBL can be used as therapeutic or preventive cancer vaccine.

Studies on the Effect of 'Gal Geun Tang' upon Stress Resistance of Rats (갈근탕(葛根湯) 투여가 흰 쥐의 스트레스 저항 반응에 미치는 영향에 관한 연구)

  • Kwon, Chang-Ho
    • Korean Journal of Pharmacognosy
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    • v.5 no.4
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    • pp.217-222
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    • 1974
  • Oxygen consumption rate has been determined to study the effect of 'Gal Geun Tang' upon stress resistance of rats. The oxygen consumption rate of liver tissue increased slightly when normal rats were given the Gal Geun Tang infusion only. When rats were given the infusion after an injection of typhoid vaccine, the elevated boby temperature and oxygen consumption rate due to typhoid vaccine fell to normal level. From this result, it is concluded that 'Gal Geun Tang' has a function that may normalize abnormally strained liver metabolism. Oxygen consumption rate of the rats significantly increased when rats were exposed to cold condition. However, under the same condition it rather maintained the normal consumption rate when the rats were given previously "Gal Geun Tang". It is suggested that 'Gal Geun Tang' has an effects that impairs the immune response from cold stress. The rats that were given Cinnamomi Ramulus('Ge Ji Tang') showed less effect than the group of 'Gal Geun Tang'.

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A study on the development of Korean Indigenous beverages in a research on GAL-Soo (한국 고유음료류 개발에 관한 연구중 갈수에 관한 연구)

  • 오승희
    • Journal of the Korean Professional Engineers Association
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    • v.15 no.1
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    • pp.14-23
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    • 1982
  • 1) The characteristic of the recipe for GAL-SOO is to make odorous, pharmaceutical cereal into plaster by honey-mixed boiling so as to be drunken whenever it needed. But the cereal which included sugar was boiled without adding sugar. 2) The part of cereal which was used mostly in making GAL-SOO was seeds of cereals. 3) The history of GAL-SOO was derived from GU GA PIL YONG, a document of WON dynasty of GHINA, but it was developped according to our taste in this county. 4) The pharmaceutical action of GAL-SOO was mainly to strengthen stomach, digestion and appeasing thirst. 5) GAL-SOO tastes so sour and sweet that it was splendid to drink especially in summer as a beverage. 6) The Value of developping GAL-SOO as a beverage is highly approved because of its expediency of recipe and tough endurance storing under ordinary temperature.

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Construction of the Detection System of Endocrine Disrupters using Yeast Two-Hybrid System with Human Estrogen Receptor ligand Binding Domain and Co-activators (Human Estrogen Receptor Ligand Binding Domain (hER LBD)과 Co-activator로 구성된 효모 Two-Hybrid System을 이용한 내분비계장애물질 검출계의 구축)

  • 이행석;조은민;류재천
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.175-182
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    • 2002
  • Endocrine disruptors (EDs) are the chemicals that affect endocrine systems through activation or inhibition of steroid hormone response. It is necessary to have a good system to evaluate rapidly and accurately endocrine-disrupting activities of suspected chemicals and their degradation products. The key targets of EDs are nuclear hormone receptors, which bind to steroid hormones and regulate their gene transcription. We constructed a co-expression system of Gal4p DNA binding domain (DBD)- ligand binding domain of human estrogen receptor $\alpha$ or $\beta$, and Gal4p transactivation domain (TAD)-co-activator AIB-1, SRC-1 or TIF-2 in Saccharomyces cerevisiae with a chromosome-integrated lacZ reporter gene under the control of CYC1 promoter and Gal4p binding site (GAL4 upstream activating sequence, GAL4$_{UAS}$). Expression of this reporter gene was dependent on the presence of estrogen or EDs in the culture medium. We found that the two-hybrid system with combination of the hER$\beta$ LBD and co-activator SRC-1 was most effective in the xenoestrogen-dependent induction of reporter activity. The extent of transcriptional activation by those chemicals correlated with their estrogenic activities measured by other assay systems, indicating that this assay system is efficient and reliable for measuring estrogenic activity. The data in this research demonstrated that the yeast detection system using steroid hormone receptor and co-activator is a useful tool for identifying chemicals that interact with steroid receptors.s.

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Hepatoprotecive Effects of Alnus japonica Extract on Experimental Liver Injury Models (오리나무 추출물(AI-1367)의 간질환 동물모델에서의 간 보호효과)

  • Zhao, Yu-Zhe;Lee, Sung-Hee;Huh, Jae-Wook;Ra, Jeong-Chan;Sohn, Dong-Hwan
    • YAKHAK HOEJI
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    • v.56 no.2
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    • pp.99-107
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    • 2012
  • The protective effect of AI-1367 (Alnus japonica extract) on liver injury was investigated. Primary rat hepatocyte intoxication was induced by tert-butyl hydroperoxide (tBH), carbon tetrachloride ($CCl_4$), or D-glactosamine (D-GalN). Liver injury was induced by $CCl_4$, D-GalN or MCD (methionine choline deficient)-diet in mouse. The cellular leakage of lactate dehyrogenase and cell viability followed by the treatment of hepatotoxicants were significantly improved by AI-1367 treatment at a concentration range of 5~50 ${\mu}g/ml$ for tBH, 5~50 ${\mu}g/ml$ for D-GalN, and 5~100 ${\mu}g/ml$ for $CCl_4$, respectively. Treatment with AI-1367 (20, 10, 5 mg/kg, p.o.) on liver injury induced by subcutaneous injection of $CCl_4$ or D-GalN reduced significantly the levels of aspartate transaminase and alanine transaminase in serum. Histological observations revealed that fatty acid changes, hepatocyte necrosis and inflammatory cell infiltration in $CCl_4$ (D-GalN)-induced liver injury was improved by administration of AI-1367. AI-1367 treatment (10, 5, 2.5 mg/kg, p.o.) also significantly recovered the body weight change and serum levels of aspartate transaminase, alanine transaminase and triglyceride in liver injury induced by MCD diet. From these results, AI-1367 shows protective effects against tBH, $CCl_4$, D-GalN, or MCD diet-induced hepatotoxicity in vitro or in vivo.

Reproductive Characteristic of Transgenic Massachusetts General Hospital Miniature Pigs for Xenotransplantation (Massachusetts General Hospital 미니돼지 유래 이종이식용 형질전환 돼지의 번식 특성 분석)

  • Ji, Soo-Jeong;Lee, Gunsup;Park, Sang Hyoun;Kim, Kyung Woon;Byun, Sung-June;Ock, Sun A;Hwang, Seongsoo;Woo, Jae-Seok;Oh, Keon Bong
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.165-170
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    • 2017
  • Pigs have been extensively used as mediators of xenotransplantation research. Specifically, the Massachusetts General Hospital (MGH) miniature pig was developed to fix major histocompatibility antigens for use in xenotransplantation studies. We generated transgenic pigs for xenotransplantation using MGH pigs. However, it has not been studied yet whether these pigs show similarity of reproductive physiological characteristics to wild types of MGH miniature pig. In this study we analyzed the estrous cycles and pregnancy characteristics of wild type (WT) and transgenic MGH miniature pigs, which were ${\alpha}1,3$-galactosyltransferase (GalT) heterozygous and homozygous knock-out, and membrane cofactor protein (MCP) inserted in its locus, $GalT^{-MCP/+}$ and $GalT^{-MCP/-MCP}$ pigs. Estrous cycles of WT, $GalT^{-MCP/+}$ and $GalT^{-MCP/-MCP}$ pigs were $20.9{\pm}0.74$, $20.1{\pm}1.26$, and $17.3{\pm}0.87days$, respectively, and periods of estrous were $3.2{\pm}0.10$, $3.1{\pm}0.12$, and $3.1{\pm}0.11days$. The periods of gestation of WT, $GalT^{-MCP/+}$ and $GalT^{-MCP/-MCP}$ pigs were $114.2{\pm}0.37$, $113.3{\pm}0.67$, and $115.4{\pm}0.51days$, respectively. Litter sizes of WT, $GalT^{-MCP/+}$ and $GalT^{-MCP/-MCP}$ pigs were $4.8{\pm}0.35$, $4.8{\pm}1.11$ and $3.0{\pm}0.32$ respectively. There were no significant differences on estrous cycle, periods of estrous and gestation, and litter size among WT, $GalT^{-MCP/+}$ and $GalT^{-MCP/-MCP}$ pigs, meaning that GalT knock-out and additional expression MCP of the MGH miniature pig did not effect on reproduction traits. These results provide relevant information to establish breeding system for MGH transgenic pig, and for propagation of $GalT^{-MCP/-MCP}$ pig to supply for xenotransplantation research.

Alpha 1,3-Galactosyltransferase Deficiency in Miniature Pigs Increases Non-Gal Xenoantigens

  • Min, Gye-Sik;Park, Jong-Yi
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.511-518
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    • 2011
  • To avoid hyperacute rejection of xenografts, ${\alpha}1,3$-galactosyltransferase knock-out (GalT KO) pigs have been produced. In this study, we examined whether Sia-containing glycoconjugates are important as an immunogenic non-Gal epitope in the pig liver with disruption of ${\alpha}1,3$-galactosyltransferase gene. The target cells were then used as donor cells for somatic cell nuclear transfer (scNT). A total of 1,800 scNT embryos were transferred to 10 recipients. One recipient developed to term and naturally delivered two piglets. Real-time RT-PCR and glycosyltransferase activity showed that ${\alpha}2,3$-sialyltransferase (${\alpha}2,3ST$) and ${\alpha}2,6$-sialyltransferase (${\alpha}2,6ST$) in the heterozygote GalT KO liver have higher expression levels and activities compared to controls, respectively. According to lectin blotting, sialic acidcontaining glycoconjugate epitopes were also increased due to the decreasing of ${\alpha}$-Gal in heterozygote GalT KO liver, whereas GalNAc-containing glycoconjugate epitopes were decreased in heterozygote GalT KO liver compare to the control. Furthermore, the heterozygote GalT KO liver showed a higher Neu5Gc content than control. Taken together, these finding suggested that the deficiency of GalT gene in pigs resulted in increased production of Neu5Gc-bounded epitopes (H-D antigen) due to increase of ${\alpha}2,6$-sialyltransferase. Thus, this finding suggested that the deletion of CMAH gene to the GalT KO background is expected to further prolong xenograft survival.

Study of oriental medical science documentory records of "So-gal" (소갈(消渴)에 관한 문헌적(文獻的) 고찰(考察))

  • Gong, Sang-Eun;Yoon, Il-Ji;Heo, Dong-Seok;Oh, Min-Seok
    • Journal of Haehwa Medicine
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    • v.16 no.1
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    • pp.107-114
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    • 2007
  • 1. So-gal IS also called as so-jung, So-dan, Pe-so, Jung-so. Gyuk-so. 2. Cause of So-gal is impairment of Jin-Aek. induced by mal intake. stress, overstrain, intoxication, aging. The process is done by Cho-yeol. 3. Symptoms of So-gal is classified m to three categories Sang-So stands for polydipsia, Jung-So stands for polyphagia and weight loss, and Ha-So stands for polyuria. 4. In treating of So-gal, Chung-Hwa Bo-eum Bo-yang is the main principle. The fundamental basis of treating So-gal is Chi-shin. 5. It is not easy to perfectly classify symptoms of So-gal into three categories Therefore further inquiry is required on classification methods of So-gal.

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