• Title/Summary/Keyword: animal cell

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Effects of Probiotic and Prebiotic on Average Daily Gain, Fecal Shedding of Escherichia Coli, and Immune System Status in Newborn Female Calves

  • Roodposhti, Pezhman Mohamadi;Dabiri, Najafgholi
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.9
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    • pp.1255-1261
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    • 2012
  • Thirty two Holstein female calves (initial body weight = $40{\pm}3.0$ kg) were used to investigate the effects of probiotic and prebiotic on average daily gain (ADG), fecal E. coli count, white blood cell count, plasma IgG1 level and cell-mediated immune response to injection of phytohemagglutinin in suckling female calves. Calves were assigned randomly to one of the four treatments, including whole milk without additives (control), whole milk containing probiotic, whole milk containing prebiotic and whole milk containing probiotic and prebiotic (synbiotic). Average daily gain was greater in calves fed probiotic, prebiotic and synbiotic at weeks 6, 7 and 8 (p<0.05). E. coli count was significantly lower in calves fed probiotic, prebiotic and synbiotic on d 56 (p<0.05). There was no significant difference between treatments in blood samples and cell-mediated response. This study showed that addition of probiotic, prebiotic and combination of these additives to milk enhanced ADG and reduced fecal E. coli count in preruminant calves.

Anti-Helicobacter pylori, Cytotoxic, and Anti-inflammatory Activities of White Ginseng Extract

  • Jee, Hee-Sook;Chang, Kyung-Hoon;Moon, Sun-Hee;Park, Se-Ho;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.1106-1109
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    • 2008
  • The anti-Helicobacter pylori activity, cytotoxicity, and anti-inflammatory activity of white ginseng extract (WGE) were investigated in vitro in this study. The antimicrobial effects of WGE toward H. pylori strains 52 J99, SSI, and 51 were tested using the disk diffusion method. Among these H. pylori strains, H. pylori 52 was the most sensitive, having the largest inhibition zone (19 mm), followed by J99, SSI, and 51. The zone of inhibition due to WGE increased significantly with increasing dosage. The cytotoxicity of WGE toward the human cancer cell lines A-549 (human lung carcinoma), HEC-1-B (human endometrial adenocarcinoma), HeLa (human uterin adenocarcinoma), and SW-156 (human kidney carcinoma) was measured using the 3-(4,5-dimethylthizol-2-yl)-2,5-diphenylate-tetrazolium bromide (MTT) assay. WGE exhibited an inhibitory effect on cell growth at 2.0 mg/mL for all tumor cell lines. An analysis of anti-inflammatory activity using the RAW 264.7 cell line showed that the inhibition of nitric oxide (NO) production increased as the WGE content increased. These results demonstrate the potential of WGE to be used as a health-promoting substance.

Adiponectin Gene Cloning and Its Expression in Insect Cell Expression System

  • Yuh, In Suh;Sheffield, Lewis G.
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.193-198
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    • 2012
  • This study was to examine expression of the recombinant full-length adiponectin (recombinant adiponectin) in insect ovarian cell culture system and to characterize structural properties of the recombinant adiponectin secreted in medium. Gene construct encoding the recombinant adiponectin contained N-terminal collagen-like domain (110 Amino Acids, AAs), C-terminal globular domain (137 AAs) and C-terminal peptides for detection with V5 antibody (26 AAs included adaptor peptide) and purification using the 6xHis tag (6 AAs). The approximate molecular weight of the product (monomer) was 35 kDa. Molecular mass species of the expressed recombinant adiponectin were monomer (~35 kDa), dimer (~70 kDa), trimer (~105 kDa) and hexamer (~210 kDa). The major secreted species were the LMW forms, such as monomer, dimer, and trimer. There was MMW of hexamer as minor form. HMW multimers (~300 kDa) were shown as a tracer or not detected on the SDS-PAGE in several experiments (data not shown). The multimer forms in this study were not compatible to those in animal or human serum and adipose tissue by other researcher's study in which the major multimer forms were HMW. By protein denaturing experiments with reducing reagent (${\beta}$-MeOH), anionic detergent (SDS) and heat ($95^{\circ}C$) on the SDS-PAGE, not all adiponectin multimers seemed to have disulfide bond linked structure to form multimers. The recombinant adiponectin which expressed in insect ovarian cell culture system seemed to have the limitation as full physiological regulator for the application to animal and human study.

Characterization of Streptomyces sp. AMLK-135 Producing Anti- MRSA Antibiotics

  • Lee, Min-Jeong;Lim, Dae-Seog;Lee, Myung-Sub;Yoon, Won-Ho;Kim, Chang-Han
    • Journal of Microbiology and Biotechnology
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    • v.7 no.6
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    • pp.397-401
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    • 1997
  • The present research program was conducted to characterize a strain of actinomycetes producing an anti methicillin-resistant Staphylococcus aureus (MRSA) antibiotic. Soil samples were collected from various sites in Korea and a number of actinomycetes were isolated from the soil samples by applying selective agar for actinomycetes. Among over 400 isolates, a strain (AMLK-135) producing anti-MRSA antibiotic against S. aureus TK 784 was selected. According to the morphological and physiological characteristics, the strain AMLK-135 was confirmed to belong to the genus Streptomyces. From the results of species identification with the TAXON program, the strain AMLK-135 was shown to belong to major cluster 5 (Streptomyces exfoliatus), but it had a low simple matching coefficient ($S_{SM}$ SM/) value to member organisms of major cluster 5. Percentage ($\%$) of strain further away of the strain AMLK-135 was low (1.9400) and it was placed further away than the outer-most members in major cluster 5. Therefore, the strain AMLK-135 was identified as a new species of the genus Streptomyces.

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A Case of Nasal Diffuse Large B-Cell Lymphoma in a Cat

  • Kim, Seong-Jun;Kim, Jun-Young;Oh, Dong-Keun;Cho, Jun-Ho;Park, Hee-Myung;Kang, Min-Hee
    • Journal of Veterinary Clinics
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    • v.35 no.4
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    • pp.141-143
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    • 2018
  • A 6-year-old spayed female Turkish Angora cat presented with sneezing, nasal discharge, and decreased appetite lasting for 21 days. Skull radiography revealed slightly increased density of soft tissue in the left nasal cavity. Computed tomography (CT) scan revealed an extensive mass with nasal septum destruction and moderate contrast enhancement in the left nasal cavity. After surgical biopsy, histopathological examination confirmed that the mass was an infiltrative round cell neoplasm, composed of sheets of large neoplastic cells. Immunohistochemical analysis revealed that most of the neoplastic cells were strongly positive for CD79a and weakly positive for PAX5. Additionally, numerous mature lymphocytes were found to be positive for CD3. This is the first reported case of nasal diffuse large B-cell lymphoma (DLBCL) in a Turkish Angora cat in Korea.

Molecular Characterization of Porcine DNA Methyltransferase I

  • Lee, Yu-Youn;Kang, Hye-Young;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.34 no.4
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    • pp.283-288
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    • 2010
  • During normal early embryonic development in mammals, the global pattern of genomic DNA methylation undergoes marked. changes. The level of methylation is high in male and female gametes. Thus, we cloned the cDNA of the porcine DNA methyltransferase 1 (Dnmt1) gene to promote the efficiency of the generation of porcine clones. In this study, porcine Dnmt1 cDNA was sequenced, and Dnmt1 mRNA expression was detected by reverse transcription-polymerase reaction (RT-PCR) in porcine tissues during embryonic development. The porcine Dnmt1 cDNA sequence showed more homology with that of bovine than human, mouse, and rat. The complete sequence of porcine Dnmt1 cDNA was 4,774-bp long and consisted of an open reading frame encoding a protein of 1611 amino acids. The amino acid sequence of porcine DNMT1 showed significant homology with those of bovine (91%), human (88%), rat (76%), and mouse (75%) Dnmt1. The expression of porcine Dnmt1 mRNA was detected during porcine embryogenesis. The mRNA was detected at stages of porcine preimplantation development (1-cell, 2-cell, 4-cell, 8-cell, morula, and blastocyst stages). It was also abundantly expressed in tissues (lung, ovary, kidney and somatic cells). Further investigations are necessary to understand the complex links between methyltransferase 1 and the transcriptional activity in cloned porcine tissues.

Traf4 is required for tight junction complex during mouse blastocyst formation

  • Lee, Jian;Choi, Inchul
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.4
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    • pp.307-313
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    • 2021
  • Traf4 (Tumor necrosis factor Receptor Associated Factor 4) is a member of the tumor necrosis factor receptor (TNFR) - associated factors (TRAFs) family. TRAF4 is overexpressed in tumor cells such as breast cancer and associated with cytoskeleton and membrane fraction. Interestingly, TRAF4 was localized with tight junctions (TJs) proteins including OCLN and TJP1 in mammary epithelial cells. However, the expression patterns and biological function of Traf4 were not examined in preimplantation mouse embryos although Traf4-deficient mouse showed embryonic lethality or various dramatic malformation. In this study, we examined the temporal and spatial expression patterns of mouse Traf4 during preimplantation development by qRT-PCR and immunostaining, and its biological function by using siRNA injection. We found upregulation of Traf4 from the 8-cell stage onwards and apical region of cell - cell contact sites at morula and blastocyst embryos. Moreover, Traf4 knockdown led to defective TJs without alteration of genes associated with TJ assembly but elevated p21 expression at the KD morula. Taken together, Traf4 is required for TJs assembly and cell proliferation during morula to blastocyst transition.

Effects of Mutagenesis for Glycosylation Sites of Recombinant Human EPO During Production from Cultured CHO Cell

  • Lee, Hyun-Gi;Seong, Hwan-Hoo;Im, Seok-Ki;Chung, Hee-Kyoung;Lee, Poongyeon;Lee, Yeun-Kun;Min, Kwan-Sik;Chang, Won-Kyoung;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.97-97
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    • 2002
  • Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.

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Cell Growth and Antioxidant Activity on Onion Juice Fermentation by Using Lactobacillus plantarum as Animal Probiotics (가축용 생균제 Lactobacillus plantarum을 이용한 양파즙 발효의 균체성장과 항산화 활성)

  • Chang, Woo-Kyung;Cho, Sang-Buem;Kim, Dong-Woon;Lee, Sang-Suk;Kim, Soo-Ki
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1729-1737
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    • 2010
  • This study was conducted to establish the optimal medium condition for the animal probiotic Lactobacillus plantarum by using onion juice. Cell yield and antioxidant activity increased in proportion to high additive levels of onion juice in medium. Onion juice, sucrose and yeast extract were selected as media ingredient factors and the effects of their mixed ratio in medium were evaluated. The full factorial design consisted of 24 experimental runs and was employed to estimate the main effects of the factors and their interactions. Significant positive effects on cell yield and antioxidant activity was shown with yeast extract and onion juice, respectively. Significant interaction was found only between sucrose and yeast extract in antioxidant activity. Finally, we selected an optimal medium that was composed of (g/l) onion juice, 600; sucrose, 15; yeast extract, 5. The efficiency of this optimum medium was estimated by using a 5 l jar fermenter. As a result, the maximum cell yield was $9.7\;{\log}_{10}$ (CFU/ml) at 12 hr. Cell yield at the end of incubation (20 hr) was $8.9\;{\log}_{10}$ (CFU/ml) and it was very similar with the predicted value, $9.0\;{\log}_{10}$ (CFU/ml). Antioxidant activity of culture was maintained at about 60~65% during all incubation time, resulting in a higher-than-predicted activity of 47.1%.

Preventive Effect of Natural Pigments Against Ultraviolet B-induced Cell Death in HaCat Cells

  • Lim, Jae-Chung;Bae, Chun-Sik;Jeong, Soo-Young;Boo, Hee-Ock;Hwang, Seong-Jin;Lim, Seul-Ki;Park, Min-Jung;Kim, Jong-Chun;Kang, Seong-Soo;Han, Ho-Jae;Park, Soo-Hyun
    • Biomedical Science Letters
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    • v.17 no.1
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    • pp.55-60
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    • 2011
  • Skin is a physical barrier against diverse injury and damages. Exposure to ultraviolet (UV) radiation causes detrimental skin injuries such as inflammation and cell death. The value of natural pigments could be applied to many usages including cosmetics. This study was conducted to examine the protective effect of natural pigments extracted from mulberry, balsam pear, purple-colored sweet potato, pehmannia root, gardenia fruit, and black rice against UV-induced cell death in HaCaT cells, human keratinocyte cell lines. In the present study, the exposure of 50 mJ/$cm^2$ UV-B for 24 hr induced cell death in HaCaT cells, which was prevented by the pretreatment of extracts of mulberry, balsam pear, purple-colored sweet potato, rehmannia root, gardenia fruit, and black rice. In addition, the exposure of 50 mJ/$cm^2$ UV-B for 24 hr also increased lipid peroxide (LPO) formation, compared to control in HaCaT cells, which was prevented by the pretreatment of extracts of mulberry, balsam pear, purple-colored sweet potato, rehmannia root, gardenia fruit, and black rice. In conclusion, the extracts of mulberry, balsam pear, purple-colored sweet potato, rehmannia root, gardenia fruit, and black rice prevented the UV-B-induced cell apoptosis via the inhibition of oxidative stress in HaCaT cells.