Ok Bong Kim;Na Young Kim;Jae Hoon Jeong;Si Wouk Kim;Hye Gwang Jeong;Seong Myeong Yoon;Jong Kun Park;Jung Sup Lee
Animal cells and systems
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v.3
no.2
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pp.229-236
/
1999
The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.
Oh, Sangnam;Park, Mi Ri;Son, Seok Jun;Kim, Younghoon
Food Science of Animal Resources
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v.35
no.4
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pp.459-465
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2015
Bovine milk provides essential nutrients, including immunologically important molecules, as the primary source of nutrition to newborns. Recent studies showed that RNAs from bovine milk contain immune-related microRNAs (miRNA) that regulate various immune systems. To evaluate the biological and immunological activity of miRNAs from milk products, isolation methods need to be established. Six methods for extracting total RNAs from bovine colostrums were adopted to evaluate the isolating efficiency and expression of miRNAs. Total RNA from milk was presented in formulation of small RNAs, rather than ribosomal RNAs. Column-combined phenol isolating methods showed high recovery of total RNAs, especially the commercial columns for biofluid samples, which demonstrated outstanding efficiency for recovering miRNAs. We also evaluated the quantity of five immune-related miRNAs (miR-93, miR-106a, miR-155, miR-181a, miR-451) in milk processed by temperature treatments including low temperature for long time (LTLT, 63℃ for 30 min)-, high temperature for short time (HTST, 75℃ for 15 s)-, and ultra heat treatment (UHT, 120-130℃ for 0.5-4 s). All targeted miRNAs had significantly reduced levels in processed milks compared to colostrum and raw mature milk. Interestingly, the amount of immune-related miRNAs from HTST milk was more resistant than those of LTLT and UHT milks. Our present study examined defined methods of RNA isolation and quantification of immune-specific miRNAs from small volumes of milk for use in further analysis.
Background: The greatest threats to the lives of cancer patients consist of both complications due to metastasis and the recurrence of original cancer. However, the existing forms of chemotherapy do not prevent these adverse events adequately because they suppress the patients' immune systems. A new methodology is required. Aim: To provide further validation for cancer immunotherapy (involving the use of traditional oriental herbal medicine). Method: Related articles, in both Korean and English were reviewed. Results: Cancer immunotherapy involving the use of traditional oriental herbal medicine can create inhospitable conditions for cancer cells and strengthen patients' immunological functioning. These effects are a demonstration of the principles of "strengthening healthy qi (扶正)" and "eliminating pathogens (祛邪)". As a result, immunotherapy protects against metastasis and original cancer recurrence by preventing the growth of cancer cells. This is very similar to the concept of a cancer dormancy therapy. Conclusion: It is strongly urged by the authors that more advanced studies be carried out on this promising therapy in the future in order to improve patients' quality of life and increase their survival time.
The impact of stress on immune function is known to be associated with the interactions among the central nervous system(CNS), neuroendocrine system, and immune system. The main pathways between stress and immune system are wiring of lymphoid organs and neuroendocrine system. Immune system also produces neuropeptides, which modulate immune system. Mediators of psychosocial influences on immune function are found to be peptides released by the pituitry, hormones, md autonomic nervous system. Hypothalamus integrates endocrine, neural and immune systems. Particularly, paraventricular nucleus appears to play a central role in this integration. On the other hand, endocrine system receives feedback from the immune system. The major regulatory pathways which pituitary modulates include the hypothalamic-pituitary-adrenal-thymic(HPAT) axis, hypothalamic-pituitary-gonadal-thymic(HPGT) axis, pineal-hypothalamic-pituitary(PHP) axis. Bidirectional pathways such as feedforward and feedback pathways are suggested in the interaction between stress and immune system. It suggests that psychosocial inputs affect immune function, but also that immunological inputs affect psychosocial function. Thus, prospective studies for elucidating the relationship between stress and immune function should incorporate measures of immune function as well as measures of endocrine, autonomic, and brain activities at the same time.
Ligand-receptor clustering event is the most important step in leukocyte adhesion and spreading on endothelial cells. Intercellular adhesion molecule-1 (ICAM-1) has been shown to enhance leukocyte adhesion, but the molecular event during the process of adhesion is unclear. To visualize the dynamics of ICAM-1 movement during adhesion, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 fused to a green fluorescent protein (IC1_GFP/CHO) and examined them under the fluorescence microscopy. The transfection of IC1_GFP alone in these cells was sufficient to support the adhesion of K562 cells that express $\alpha$L$\beta$2 (LFA-1) integrin stimulated by CBR LFA-1/2 mAb. This phenomenon was mediated by ICAM-1-LFA-1 interactions, as an mAb that specifically inhibits ICAM-1-LFA-1 interaction (RRl/l) completely abolished the adhesion of LFA-1* cells to IC1_ GFP/CHO cells. We found that the characteristic of adhesion was followed almost immediately (~10 min) by the rapid accumulation of ICAM-1 on CHO cells at a tight interface between the two cells. Interestingly, ICI_GFP/CHO cells projected plasma membrane and encircled approximately half surface of LFA-1+ cells, as defined by confocal microscopy. This unusual phenomenon was also confirmed on HUVEC after adhesion of LFA-1* cells. Neither cytochalasin D nor 2,3-butanedione 2-monoxime an inhibitor of myosin light chain kinase blocked LFA-1-mediated ICAM-1 clustering, indicating that actin cytoskeleton and myosin-dependent contractility are not necessary for ICAM-1 clustering. Taken together, we suggest that leukocyte adhesion to endothelial cells induces specialized form of ICAM-1 clustering that is distinct from immunological synapse mediated by T cell interaction with antigen presenting cells.
Park Jong-Sun;Cho Jae-Youl;Kim Sung-Soo;Bae Hyun-Jin;Han Jeung-Whan;Lee Hyang-Woo;Hong Sung-Youl
Archives of Pharmacal Research
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v.29
no.5
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pp.384-393
/
2006
Protein carboxylmethylation methylates the free carboxyl groups in various substrate proteins by protein carboxyl O-methyltransferase (PCMT) and is one of the post-translational modifications. There have been many studies on protein carboxylmethylation. However, the precise functional role in mammalian systems is unclear. In this study, immunoglobulin, a specific form of $\gamma-globulin$, which is a well-known substrate for PCMT, was chosen to investigate the regulatory roles of protein carboxylmethylation in the immune system. It was found that the anti-BSA antibody could be carboxylmethylated via spleen PCMT to a level similar to $\gamma-globulin$. This carboxylmethylation increased the hydrophobicity of the anti-BSA antibody up to 11.4%, and enhanced the antigen-binding activity of this antibody up to 24.6%. In particular, the Fc region showed a higher methyl accepting capacity with 80% of the whole structure level. According to the amino acid sequence alignment, indeed, 7 aspartic acids and 5 glutamic acids, as potential carboxylmethylation sites, were found to be conserved in the Fc portion in the human, mouse and rabbit. The carboxylmethylation of the anti-BSA antibody was reversibly demethylated under a higher pH and long incubation time. Therefore, these results suggest that protein carboxylmethylation may reversibly regulate the antibody-mediated immunological events via the Fc region.
Objective: The aim of this study was to evaluate the effects of three feeding systems, i.e., indoor feeding (CON), indoor feeding with 4-h daily access to grazing artificial pasture (ITGP), and indoor feeding with 8-h daily access to grazing artificial pasture (IEGP), on the plasma antioxidant and immunological capacity, slaughter characteristics, meat quality and economic efficiency of Huang-huai lambs. Methods: Thirty-three healthy Huang-huai rams with similar body weight (approximately 5 mo of age, 28.96±1.01 kg) were assigned equally to three experimental groups. When finished fattening, six lambs from each group were collect blood samples for plasma analyses and then slaughtered to determine slaughter characteristics and obtain biceps brachii muscle for further analysis of meat quality and fatty acid profile. Results: Compared to CON group, animals submitted to ITGP and IEGP groups resulted in greater contents of serum glutathione peroxidase, immunoglobulins (IgA, IgG, and IgM), polyunsaturated fatty acids (PUFA), n-6 PUFA, and PUFA/saturated fatty acid (FA) ratio and lower palmitic /oleic acid ratio (p<0.05). Moreover, animals in ITGP group exhibited a higher (p<0.05) loin eye area, content of meat crude protein (CP), and eicosetrienoic acid compared to CON group, while slaughter performance was superior (p<0.05) to that of the IEGP group. The economic efficiency of ITGP group was 70.12% higher than that of CON group, while the IEGP group exhibited a decrease of 92.54% in economic efficiency compared to the CON group. Conclusion: Restricted grazing time combined with indoor feeding was more effective in conferring superior body health, carcass traits and economic efficiency in Huang-huai lambs, as well as higher CP content and healthier FA composition in the resulting meat.
The conceptus which are resulted by mating between two genetically non-identical partners can be considered to be an allograft to the mother science which is not rejected by the mother's immunological attack. The present studies have been, therefore, attempted in order to elucidate the mechanism by which protection of the fete-placental allograft, between the C3H/HeJ female mouse and DBA/2 male mouse occurred. For this purpose, firstly systemic immunity was investigated by measuring T and B lymphocytes subsets. Natural killer cell activity in maternal splenic tissue and by observing the effects of pregnancy serums, progesterone and hCG on immune systems. Secondly, local immunity also investigated by measuring T lymphocytes subsets, natural killer cell activity in lymph nodes draining the uterus. The subsets of Thy-1.2$^+$ cells and L 3T4$^+$ cells decreased slightly while the subsets of Ly2$^+$ cell increased significantly compared with those of the control group beyond the mid-gestational stage. The subsets of B cell gradually in-creased from the mid-gestational stage untill delivery. The natural killer cell activity in the maternal splenic tissue significantly increased during the period of 5th to 8th day of gestation. The natural killer cell activity was significantly suppressed by the pregnancy serums and non-pregnant serums compared with those of serum-free group. The treatment of hCG significantly suppressed natural killer cell activity in the dose dependent manner (1 unit/ml-1000 unit/ml) while pro-gesterone increased the natural killer cell activity at phamarcological dose only. In the lymph nodes draining the uterus, the subsets of Thy-1.2$^+$ cells significantly increased during the period of implantation and L3T4$^+$ cell subsets slightly increased during the mid-gestational stage. The subsets of Ly2$^+$ cell increased significantly during the mid-gestational stage, but decreasing slightly be-fore delivery. The natural killer cell activity was significantly elevated after the implantation period in the lymph nodes draining the uterus. The natural killer cell activity of the lymph nodes draining the uterus was higher than those of splenic tissue during the same periods of gestation. It is therefore, concluded that during the pregnancy, the phenomena which the fete-placental allograft has not been rejected and rather protected from the maternal immunological attack might be due to local immune suppression in fete-maternal interface tissues rather than systemic immune suppression. And the subsets of Thy-1.2$^+$ cells and L3T4$^+$ cells mainly contribute to accepting allograft in early stage of pregnancy, while the subsets of Ly2$^+$ cell and the subsets of B cell increased significantly compared with those of the control group beyond the mid-gestational stage, so their role in systemic immunity and local immunity gradually increased from the mid-gestational stage until delivery.
Insulin-like growth factor-I (IGF-I) and IGF-II have structure like insulin. In contrast to insulin, however, the bioavaility of IGFs is modulated by the IGF-binding protein (IGFBPs). Each of IGFBPs was different with molecular masses, biological characteristics, and immunological properties.. Human fibroblasts secrete IGFBPs that can modify IGF-I action. In diabetes mellitus, the most study of IGF systems have been investigated in insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, and streptozotocin-in-duced animals in vivo. Recently, a little research regarding the IGFs system has been proposed in por-tion of cell in vitro. In this study, effects of low or high glucose condition on IGFBP-5 in GM10 was investigated. By western blotting analysis, IGFBP-5 level decreased in cells cultured at high glucose, but IGFBP-5 level of mRNA didn't change. IGFBP-5 protease that cleaves IGFBP-5 in conditioned me-dium had was inhibited by EDTA and heparin, like serine protease and metalloprotease. Furthermore, the protease activity was increased in high glucose cultivated condition. In results of gelatin zymog-raphy, molecular weight of proteolytic metalloenzymes was indentified 69-kDa and protease activity was increased in time-dependent manner. Although the mechanism has yet to be determined, IGFBP-5 proteolysis in GM10 cells cultured with high glucose may increase effects of IGFs to decrease the glu-cose level through dissociation of IGFs from IGFBPs. Therefore, we suggest that IGF- I and IGFBPs could be potential models in study of pathophysiology such as diabetes mellitus.
Kim, Jin-Sook;Kim, Seung-Cheol;Jeon, Bo-Young;Park, Seung-Kyu
Korean Journal of Microbiology
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v.45
no.3
/
pp.281-285
/
2009
In this study, we compared the BacT/Alert liquid culture system with Ogawa and $L\ddot{o}wenstein$-Jensen (L-J) media for sputum culture and drug susceptibility test (DST) of Mycobacterium tuberculosis. Rapid liquid culture systems have been widely employed both for primary cultures of M. tuberculosis from clinical specimens and for drug susceptibility test because of its greater sensitivity and faster turn-around time than the conventional egg-based culture methods (Ogawa, $L\ddot{o}wenstein$-Jensen media). Sputum specimens were decontaminated with N-acetyl-L-cysteine (NALC)-4% NaOH and inoculated into the BacT/Alert culture bottles and Ogawa media. 95 from among 135 sputa were smear-positive, 97 (71.9%) were culture-positive by the BacT/Alert culture system, while 89 (65.9%) were positive by Ogawa media. The mean time to culture-positive by the BacT/Alert process system was about 11.3 days, which was significantly shorter than that by Ogawa media (22.4 days). Of 32 M. tuberculosis cultures examined for drug sensitivity, the concordant rate between the two methods (BacT/ Alert liquid culture system, $L\ddot{o}wenstein$-Jensen media) ranged from 87.5% for isoniazid and 90.6% for rifampicin.
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