• KSBB Journal
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• v.19 no.3
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• pp.174-177
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• 2004

A method of collection and long-term storage of viable lily (Lilium longiflorum) pollen grains were developed for their in vitro growth and transformation in consistency. Petroleum ether, n-heptane, cyclohexane and benzene, as pollen collection medium, were determined less toxic to pollen growth in vitro than others tested. Pollen grains, however, lost their growth activity if stored in these solvents more than a week, So, a serial performance, that is, pollen grain collection in these solvents, air-drying and immediate transfer to low temperature condition was determined desirable for keeping the viability much longer. Pollen grains from this storage showed a successful transformation in vitro with a cDNA encoding tissue plasminogen activator (TPA) protein using Agrobacterium via vacuum infiltration according to western blotting analysis.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.693-698
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• 1996

Haemaphysalis longiscornis is the common cattle tick of great economic importance in Korea. Chemical control using dips or sprays has been the traditional method of attempting to kill these ticks during the infestation period. However, the presence of resistant forms to chemical, the rising costs of acaricides and environmental problems have made it almost impossible to use these chemicals on a regular basis according to the pest problem. For this reason, vaccination against ticks and breeding for host resistance against ticks are being studied. In order to determine the common proteins and antigens according to developmental stages, SDS-PAGE and western blotting were performed. In SDS-PAGE 103.3kD and 98.3kD proteins were observed as common proteins, and these proteins were observed as common antigens in western blotting. Unimmunized rabbits were infestated three times with H longicornis. The weight of the second and the third engorged ticks were 0.153g and 0.104g respectively. This weight is 69% and 47% of the first engorged ticks weight respectively. Immunized rabbits by adult ticks antigen and control were infested with H longicornis. The control taked 3-4 days to fully engorge, but the immunized rabbits taked about 7 days. So adult tick antigen may be effective to render the immunity to host.

• Korean Journal of Animal Reproduction
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• v.22 no.3
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• pp.265-276
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• 1998

In an attempt to evaluate the function of MAP kinase of porcine oocytes and to develop a method of assessment for kinase activity, we used MBP as a substrate to detect the MAP kinase activity of porcine oocytes matured in in vitro. The MAP kinase which had lower activity during the first 20 hours of culture started to show an increased amount of activity at 25 hours at which a collapse in nuclear membrane was induced. Significant (P<0.05) a, pp.ared at 30 hours of being cultured. The gel phosphorylation method, MBP which has been known to be a substrate for kinase such as cdc2 kinase, was phosphorylated at two positions corresponding to ERK 1 (44kDa) and ERK2 (42 kDa) which are known as mammalian MAP kinase. The existence of MARKK and MAP kinase were identified with western blotting at 0 hour culture of immature GV oocytes. The amount of those proteins did not increase during 40 hours of culture, which suggest that the increase of MAP kinase activity was caused by phosphorylaton rather than due to change in protein amount. MAPKK and MAP kinase were shown to be dephosporylated with deactivated at M 1 stage by inhibition of protein synthesis with cycloheximide added at the strat following the cultrue. We have reulsts that indicate the existedence of MAP kinase cascade which was activated simultaneously with start of porcine oocyte maturation (GVBD).

• BMB Reports
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• v.47 no.7
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• pp.388-392
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• 2014

Berberine, a type of isoquinoline alkaloid isolated from Chinese medicinal herbs, has been reported to have various pharmacological activities. Studies have demonstrated that berberine has beneficial effects on vascular remodeling and alleviates restenosis after vascular injury. However, its mechanism of action on vascular smooth muscle cell migration is not fully understood. We therefore investigated the effect of berberine on human aortic smooth muscle cell (HASMC) migration. Boyden chamber assay was performed to show that berberine inhibited HASMC migration dose-dependently. Real-time PCR and Western blotting analyses showed that levels of matrix metalloproteinase (MMP)-2, MMP-9, and urokinase-type plasminogen activator (u-PA) were reduced by berberine at both the mRNA and protein levels. Western blotting assay further confirmed that activities of c-Fos, c-Jun, and NF-${\kappa}B$ were significantly attenuated. These results suggest that berberine effectively inhibited HASMC migration, possibly by down-regulating MMP-2, MMP-9, and u-PA; and interrupting AP-1 and NF-${\kappa}B$ mediated signaling pathways.

• KSBB Journal
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• v.23 no.6
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• pp.554-556
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• 2008

Fresh Oenanthe javanica DC. leaves still attached to stem architecture were immersed in NaOH solution for 3 min before agroinfiltration and co-cultivation. MTT assay revealed that NaOH solution containing up to 0.7% was still safe for the leaf viability. Fluorometric GUS enzyme analysis showed that 0.5% NaOH-treated leaf tissues were efficiently transformed by vacuum infiltration for 20 min with Agrobacterium cells at a density of $OD_{600}=0.5$ to 1.0. These conditions worked well for the expression of HBsAg, which was confirmed by western blotting and ELISA.

• Korean Journal of Veterinary Service
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• v.32 no.1
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• pp.19-25
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• 2009

Porcine reproductive and respiratory syndrome (PRRS) virus is the etiological agent of diseases characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRS virus is a small enveloped virus containing a positive-sense, single-stranded RNA genome. In the present study, ORF6 gene of Korean PRRS virus isolate, CNV, was cloned and expressed in baculovirus expression system. The ORF6 gene and expressed protein in the recombinant virus were confirmed by PCR/indirect fluorescence antibody (IFA) test and Western blotting, respectively. The recombinant protein with a molecular weight of approximately 24KDa was confirmed by Western blotting using His6 and PRRS virus-specific antiserum. Expressed ORF6 protein was applied for IFA to detect antibody against PRRS virus using field porcine sera. However, the sensitivity and specificity of developed IFA using expressed ORF6 protein were considerably low compared to those of commercial ELISA kit. This results suggest that IFA using expressed ORF6 protein could not be used as a diagnostic test for PRRS virus infection without further improvements.

• Food Science and Biotechnology
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• v.15 no.4
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• pp.534-542
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• 2006

The effects of ethanol fractions of three different rice grain extracts, Jakwangchalbyeo, Hwasunchalbyeo, and Ilpumbyeo, on apoptotic cell death in the rat hepatoma H4IIE cell line were investigated using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell viability assay. One hundred mg/mL Jakwangchalbyeo extract significantly reduced cell viability to 69.5, 57.2, and 46.1% within 24, 48, and 72 hr, respectively. Fluorescence-activated cell sorting (FACS) analyses were also performed to characterize the cell death pattern caused by treatment with the rice grain extracts. Apoptotic cell death was clearly observed with time after treatment with the Jakwangchalbyeo extract. In Western blotting analysis, degradation of the 116 kDa poly-ADP-ribose polymerase (PARP) molecule was observed with concomitant formation of an 89 kDa product 24, 48, and 72 hr after treating cells with the Jakwangchalbyeo extract. This indicates that an apoptotic process caused cell death in these cells. In conclusion, red-pericarp Jakwangchalbyeo extract induced apoptotic cell death in H4IIE cells to a larger extent than the other rice extracts.

• The Korean Journal of Oral and Maxillofacial Pathology
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• v.42 no.6
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• pp.167-174
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• 2018

Natural products are vastly utilized as a source of chemotherapeutic agents for human cancers. Kochia scopraia is traditionally used for the cure of urological and dermatological diseases. Recently, methanol extract of Kochia scoparia (MEKS) has been shown to have anti-cancer activity to various human cancers. However, there is no report demonstrating the anti-cancer activity of MEKS in human mucoepidermoid carcinoma (MEC) cells. In this study, the authors studied the effects of MEKS on the cell proliferation and underlying mechanism in YD15 human MEC cells. MEKS decreased YD15 cell proliferation proven by trypan blue exclusion assay and induced apoptosis, evidenced by cell cycle analysis and western blotting. Autophagy induction by MEKS was verified by western blotting. In addition, MEKS regulated the expression of phosphorylated Akt, phosphorylated p38 and Nrf2 protein. This results can imply that MEKS might be a potential candidate for the treatment of human MEC cells.

• Biomolecules & Therapeutics
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• v.27 no.4
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• pp.373-380
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• 2019

Sphingosine kinase 1 and its product, sphingosine 1-phosphate (S1P), as well as their receptors, have been implicated in inflammatory responses. The functions of receptors $S1P_1$ and $S1P_2$ on cell motility have been investigated. However, the function of $S1P_3$ has been poorly investigated. In this study, the roles of $S1P_3$ on inflammatory response were investigated in primary peritoneal macrophages. $S1P_3$ receptor was induced along with sphingosine kinase 1 by stimulation of lipopolysaccharide (LPS). LPS treatment induced inflammatory genes, such iNOS, COX-2, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$. TY52156, an antagonist of $S1P_3$ suppressed the induction of inflammatory genes in a concentration dependent manner. Suppression of iNOS and COX-2 induction was further confirmed by western blotting and NO measurement. Suppression of $IL-1{\beta}$ induction was also confirmed by western blotting and ELISA. Caspase 1, which is responsible for $IL-1{\beta}$ production, was similarly induced by LPS and suppressed by TY52156. Therefore, we have shown $S1P_3$ induction in the inflammatory conditions and its pro-inflammatory roles. Targeting $S1P_3$ might be a strategy for regulating inflammatory diseases.

• The Korean Journal of Physiology and Pharmacology
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• v.27 no.2
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• pp.131-141
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• 2023

Compelling evidence has demonstrated the critical role of circular RNAs (circRNAs) during lung adenocarcinoma (LUAD) progression. Herein, we explored a novel circRNA, circ_0129047, and detailed its mechanism of action. The expression of circ 0129047, microRNA-665 (miR-665), and protein tyrosine phosphatase receptor type B (PTPRB) in LUAD tissues and cells was determined using reverse transcription quantitative polymerase chain reaction and Western blotting. Cell Counting Kit8 and colony formation assays were conducted to detect LUAD cell proliferation, and western blotting was performed to quantify apoptosis-related proteins (Bcl2 and Bax). Luciferase reporter and RNA immunoprecipitation assays were used to validate the predicted interaction between miR-665 and circ_0129047 or PTPRB. A xenograft assay was used for the in vivo experiments. Circ_0129047 and PTPRB were downregulated in LUAD tissues and cells, whereas miR-665 expression was upregulated. Overexpression of circ_0129047 suppresses LUAD growth in vivo and in vitro. Circ_0129047 is the target of miR-665, and the miR-665 mimic ablated the antiproliferative and pro-apoptotic phenotypes of LUAD cells by circ_0129047 augmentation. MiR-665 targets the 3'UTR of PTPRB and downregulates PTPRB expression. PTPRB overexpression offsets the pro-proliferative potential of miR-665 in LUAD cells. Circ_0129047 sequestered miR-665 and upregulated PTPRB expression, thereby reducing LUAD progression, suggesting a promising approach for preventing LUAD.