• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1708-1713
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• 2002

Endometrial explants obtained from does between days 13 and 21 of pregnancy were cultured in a modified minimum essential medium in the presence of [$^35S$]methionine and [$^3H$]-leucine. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. No marked qualitative changes in patterns of protein production by caprine endometrium between days 13-21 of pregnancy. At least 11 proteins showed consistently a clear spot or a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa). Limited N-terminal sequence analysis of these two isoforms showed that the protein had complete homology with bovine placental and plasma retinol-binding protein (RBP) over the first 20 amino acids. Through use of the antiserum raised against bovine placental RBP, immunoreactive RBP was detected in cultures conditioned by uterine explants prepared at days 13, 15 and 21 of pregnancy. In the present study, proteins synthesized and secreted by caprine endometrium during periattachment period of early pregnancy were characterized. The pregnant endometrium secreted a number of neutral-to-acidic proteins which constituted, in part, the histotroph. A vitamin A-transport protein, RBP, was identified in cultures conditioned by endometrium of days 13-21 of pregnancy. The uterine endometrium is the only source of retinol for embryonic tissues. The uterine RBP appears to transport retinol locally toward embryonic tissues. Secretion of RBP by caprine endometrium of days 13, 15 and 21 of pregnancy suggested that retinol played an important role in conceptus development during periattachment period of early pregnancy.

• Parasites, Hosts and Diseases
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• v.59 no.6
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• pp.557-564
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• 2021

Macrophages play a key role in chronic inflammation, and are the most abundant immune cells in the tumor microenvironment. We investigated whether an interaction between inflamed prostate cancer cells stimulated with Trichomonas vaginalis and macrophages stimulates the proliferation of the cancer cells. Conditioned medium was prepared from T. vaginalis-infected (TCM) and uninfected (CM) mouse prostate cancer (PCa) cell line (TRAMP-C2 cells). Thereafter conditioned medium was prepared from macrophages (J774A.1 cell line) after incubation with CM (MCM) or TCM (MTCM). When TRAMP-C2 cells were stimulated with T. vaginalis, protein and mRNA levels of CXCL1 and CCL2 increased, and migration of macrophages toward TCM was more extensive than towards CM. Macrophages stimulated with TCM produced higher levels of CCL2, IL-6, TNF-α, their mRNAs than macrophages stimulated with CM. MTCM stimulated the proliferation and invasiveness of TRAMP-C2 cells as well as the expression of cytokine receptors (CCR2, GP130, CXCR2). Importantly, blocking of each cytokine receptors with anti-cytokine receptor antibody significantly reduced the proliferation and invasiveness of TRAMP-C2 cells. We conclude that inflammatory mediators released by TRAMP-C2 cells in response to infection by T. vaginalis stimulate the migration and activation of macrophages and the activated macrophages stimulate the proliferation and invasiveness of the TRAMP-C2 cells via cytokine-cytokine receptor binding. Our results therefore suggested that macrophages contribute to the exacerbation of PCa due to inflammation of prostate cancer cells reacted with T. vaginalis.

• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.163-170
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• 1986

Balb/c mouse spleen cells in vitro sensitized against ICR spleen cells were cultured in conditioned media(CM). The CM was produced by ICR spleen cells stimulated with Concanavalin-A(Con-A), and sensitized lymphoid cells were grown in CM. ICR mouse spleen cells were appeared to be a good generator of IL-2. Optimal growth was seen in growth medium containing 20% fetal calf serum. and 25% CM. When cultures were initiated at 1, 5, $10{\times}10^4\;cells/ml$, the cells were increased in numbers by about 20, 13, 5-fold, respectively, every 9 days. Such growth pattern was sustained for about 4-6 weeks and thereafter the cell growth was diminished gradually. Direct immunofluorescence indicated that 93% of the lymphoid cells grown in CM(for 10 days) expressed Thyl surface antigen. And the cells grown in CM were cytotoxic to the sesitizing ICR mouse spleen cells though cytotoxicity level was not high. According to these results, the cells grown in CM were considered to be cytotoxic T lymphocytes. The lymphoid cells grown for 20 days were nearly unresponsive to Con-A and therefore dependent only IL-2 to be used for IL-2 assay.

• Archives of Pharmacal Research
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• v.20 no.3
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• pp.225-233
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• 1997

A murine leukemia x LM fibroblast hybrid cell line with immune augmenting properties stimulated resistance to Mycobacterium avium complex (MAC) in mouse peritoneal macrophages, and in immune deficient beige mice (C57BL/6/bgj/bgj). The proliferation of MAC in mouse peritoneal macrophages was inhibited by medium conditioned by the growth of the hybrid cells (hybrid cell-CM). Under similar circumstances, media conditioned by the growth of LM cells (LM cell-CM), a mouse fibroblast cell line used as one parent in forming the hybrid cell, was exhibited no inhibitory effect. Treatment of mouse peritoneal macrophages with hybrid cell-CM, but not with LM cell-CM, stimulated the expression of each of four previously described macrophage activation antigens, suggesting that the hybrid cells formed immunomodulators in addition to those formed by LM cells. Furthermore, the morphology of the macrophages following treatment with hybrid cell-CM was clearly distinguishable from that following exposure of the cells to LM cell-CM. The therapeutic effects of hybrid cells on the progression of MAC-infection were indicated by the prolonged survival of MAC-infected immune-deficient beige mice. One hundred percent of treated animals survived more than 60 days, while untreated animals died in approximately 22 days.

• Bulletin of the Korean Chemical Society
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• v.30 no.12
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• pp.2943-2948
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• 2009

Copper-dimethylglyoxime complex (CuDMG) modified Copper electrode (Cu/CuDMG) showed a catalytic activity towards cyclohexanol oxidation in NaOH solution. The modified electrode prepared by the dimethylglyoxime anodic deposition on Cu electrode in the solution contained 0.20 M $NH_4Cl\;+\;NH_4OH\;(pH\;9.50)\;and\;1\;{\times}\;10^{-4}$ M dimethylglyoxime. The modified electrode conditioned by potential recycling in a potential range of -900${\sim}$900 mV vs. Ag/AgCl by cyclic voltammetry in alkaline medium (1 M NaOH). The results show that the CuDMG film on the electrode behaves as an efficient catalyst for the electro-oxidation of cyclohexanol in alkaline medium via Cu (III) species formed on the electrode.

• Tuberculosis and Respiratory Diseases
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• v.51 no.4
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• pp.315-324
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• 2001

Background : IL-8 is a potent chemotactic cytokine that plays an important role in the host defense mechanism against M. tuberculosis by recruiting inflammatory cells to the site of the infection. Lung epithelial cells, as well as alveolar macrophages are known to produce IL-8 in response to M. tuberculosis. IL-8 gene expression is mainly regulated on the level of transcription by NF-${\kappa}B$. This study investigated whether or not A549 cells produce IL-8 in NF-${\kappa}B$ dependent mechanism in response to macrophages phagocytosing M. tuberculosis. Methods : Peripheral blood monocytes that were obtained from healthy donors were cultured for 24 h with M. tuberculosis and a conditioned medium(CoMTB) was obtained. As a negative control, the conditioned medium without M. tuberculosis (CoMCont) was used. A549 cells were stimulated with M. tuberculosis, CoMCont and CoMTB and the IL-8 concentration in the culture media was measured by ELISA. The CoMTB induced IL-8 mRNA expression in the A549 cells was evaluated using RT-PCR, and CoMTB induced $I{\kappa}B{\alpha}$ degradation was measured using western blot analysis. CoMTB induced nuclear translocation and DNA binding of NF-${\kappa}B$ was also examined using an electrophoretic mobility shift assay(EMSA), and the CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity was measured using a luciferase reporter gene assay. Results : CoMTB induced IL-8 production by A549 cells($46.8{\pm}4.8\;ng/ml$) was higher than with direct stimulation with M. tuberculosis ($6.8{\pm}2.9\;ng/ml$). CoMTB induced IL-8 mRNA expression increased after 2 h of stimulation and was sustained for 24 h. $I{\kappa}B{\alpha}$ was degraded after 10 min of CoMTB stimulation and reappeared by 60 min. CoMTB stimulated the nuclear translocation and DNA binding of NF-${\kappa}B$. The CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity($13.6{\pm}4.3$ times control) was higher than either CoMCont($2.0{\pm}0.6$ times control) or M. tuberculosis ($1.4{\pm}0.6$ times control). Conclusion : A conditioned medium of peripheral blood monocytes phagocytosing M. tuberculosis stimulates NF-${\kappa}B$ dependent IL-8 production by the lung epithelial cells.

• Journal of the Korean Applied Science and Technology
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• v.38 no.1
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• pp.186-195
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• 2021

In this study, when stem cell culture solution is used as a cosmetic ingredient, one of the most prominent problems is that the ingredients generally have low thermal stability. Therefore, in this study, in order to find out how the stem cell culture medium is heated or preserved at high temperature, the effect of various effects of stem cells on the various effects of the stem cells was investigated. Investigated. As a result of the experiment, the wound healing assay confirmed that the cell migration increased after 6 hours, and after 24 hours, it was confirmed that the cell mobility was increased and cell division was promoted, thereby being concentrated. As a result of investigating the amount of transdermal water loss by preparing a cosmetic product containing stem cell culture solution, it was confirmed that the culture solution addition group showed an improvement rate of 31% compared to the non-added group, thereby helping in skin wound recovery. As a result of this, it is considered that this point should be considered when the stem cell culture medium is used as an active ingredient in cosmetics in the future.

• BMB Reports
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• v.43 no.8
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• pp.541-546
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• 2010

We utilized a mammalian expression system to purify and characterize autotaxin (ATX)/lysophospholipase D, an enzyme present in the blood responsible for biosynthesis of lysophosphatidic acid. The human ATX cDNA encoding amino acids 29-915 was cloned downstream of a secretion signal of CD5. At the carboxyl terminus was a thrombin cleavage site followed by the constant domain (Fc) of IgG to facilitate protein purification. The ATX-Fc fusion protein was expressed in HEK293 cells and isolated from conditioned medium of a stable clone by affinity chromatography with Protein A sepharose followed by cleavage with thrombin. The untagged ATX protein was further purified to essential homogeneity by gel filtration chromatography with a yield of approximately 5 mg/liter medium. The purified ATX protein was enzymatically active and biologically functional, offering a useful tool for further biological and structural studies of this important enzyme.

• Environmental Engineering Research
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• v.23 no.2
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• pp.216-222
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• 2018

The growth characteristics of Spirulina platensis were investigated using four photo-bioreactors with $CO_2$-mixed air flows. Each reactor was operated under a specific condition: 3% $CO_2$ at 50 mL/min, 3% $CO_2$ at 150 mL/min, 6% $CO_2$ at 50 mL/min, and 6% CO2 at 150 mL/min. The 3% $CO_2$ at 150 mL/min condition produced the highest algal growth rate, while the 6% $CO_2$ at 150 mL/min conditioned produced the lowest. The algal growth performance was suitably assessed by the linear growth curve rather than the exponential growth. The medium pH decreased from 9.5 to 8.7-8.8 (3% $CO_2$) and 8.4-8.5 (6% $CO_2$), of which trends were predicted only by the pH-carbonate equilibrium and the reaction kinetics between dissolved $CO_2$ and $HCO_3{^-}$. Based on the stoichiometry between the nutrient amounts and cell elements, it was predicted that depleted nitrogen (N) at the early stage of the cultivation would reduce the algal growth rates due to nutrient starvation. In this study, use of the photobioreactors capable of good light energy distribution, proper ranges of $CO_2$ in bubbles and medium pH facilitated production of high amounts of algal biomass despite N limitation.

• The Korean Journal of Zoology
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• v.39 no.2
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• pp.215-222
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• 1996

In the previous studies, we have demonstrated that prorenin converting enzyme (PRECE) was identical to the epidermal grouch factor-binding protein (EGF-BP) type B, which was a member of the mouse glandular kallikrein family, To examine whether or not EGF-BP type A and C are involved in the processing of prorenin, we have cloned the CDNAS of the EGF-BP type h and C from a library of male ICR mouse submandibular gland (SMGI. And then CHO cells were transfected with the EGF-BP expression plasmids. and stable cell lines expressing a high level of the EGF-BPS precursor were obtained. The conditioned medium was then treated with trypsin, which has been knotvn to effectively convert the EGF-BP type A and C precursor to the active forms. 수ubsequentlv, the prorenin converting activity of the trypsin-treated or untreated medium was examined. PRECE converted exactly prorenin to renin, but the prorenin converting activities of EGF-BP type A and C were not detected. From these results, it seems that only type B of these EGF-BPs is involved in processing Ren 2 prorenin in mouse SMG.