• BMB Reports
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• 제52권4호
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• pp.289-294
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• 2019

The present study evaluated the role of AHNAK in Bartonella henselae infection. Mice were intraperitoneally inoculated with $2{\times}10^8$ colony-forming units of B. henselae Houston-1 on day 0 and subsequently on day 10. Blood and tissue samples of the mice were collected 8 days after the final B. henselae injection. B. henselae infection in the liver of Ahnak-knockout and wild-type mice was confirmed by performing polymerase chain reaction, with Bartonella adhesion A as a marker. The proportion of B. henselae-infected cells increased in the liver of the Ahnak-knockout mice. Granulomatous lesions, inflammatory cytokine levels, and liver enzyme levels were also higher in the liver of the Ahnak-knockout mice than in the liver of the wild-type mice, indicating that Ahnak deletion accelerated B. henselae infection. The proportion of CD4+interferon-${\gamma}$ ($IFN-{\gamma}^+$) and $CD4^+$ interleukin $(IL)-4^+$ cells was significantly lower in the B. henselae-infected Ahnak-knockout mice than in the B. henselae-infected wild-type mice. In vitro stimulation with B. henselae significantly increased $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected wild-type mice, but did not increase $IFN-{\gamma}$ and IL-4 secretion in the splenocytes obtained from the B. henselae-infected Ahnak-KO mice. In contrast, $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, IL-10, RANTES, and tumor necrosis $factor-{\alpha}$ secretion was significantly elevated in the splenocytes obtained from both B. henselae-infected wild-type and Ahnak-knockout mice. These results indicate that Ahnak deletion promotes B. henselae infection. Impaired $IFN-{\gamma}$ and IL-4 secretion in the Ahnak-knockout mice suggests the impairment of Th1 and Th2 immunity in these mice.

• 한국수정란이식학회지
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• 제16권3호
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• pp.223-231
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• 2001

The success of nuclear transplantation with mammalian oocytes depends critically on the potential of oocytes activation, which mainly caused to prevent the re-accumulation of maturation promoting factor (MPF). This study was conducted to compare the effect of combined treatment of lonomycin with a Hl-histone kinase inhibitor (dimethylaminopurine, DMAP) or cdc2 kinase inhibitor (sodium pyrophosphate, SPP) on activation of bovine oocytes. In vitro matured bovine oocytes with the first polar body (PB) and dense cytoplasm were assigned to 3 experimental groups. For activation treatment, oocytcs were exposed to 5 $\mu$M lonomycin for 5 min (Group 1), and followed by 1.9 mM dimethylaminopurine (DMAP) for 3 h (Group 2) or followed by 2 mM sodium pyrophosphate (SPP) for 3 h (Group 3). The activation effects in the three treatments and the control group (untreated) were judged by the extrusion of the second PB and formation of a pronucleus (PN). Differences among groups were analysed using one-way ANOVA after arc-sine transformation of proportional data. All three treatments led to high activation rates (90% to 95%), with significant difference from the control. However, the extrusion of the second PB and the rate of PN formation differed remarkably among treatments. In Group I and 3, about 95% of the oocytes had extruded the second polar body, but one PN had formed in a higher proportion of oocytes in Group 3 than in Group 1 (90% vs. 5%). In experiment 2, the rates of cleavage and development into blastocysts in Group 1 were significantly lower than those of Group 2 and 3 (8.7% and 0% vs. 50.5% and 11.6%, and 44.6% and 7.2%, respectively, P<0.05). In experiment 3, ~80% of parthenotes in Group 1 were developed with haploid chromosomal sets. However, when ionomycin was followed immediately by DMAP (Group 2). only 20% of parthenotes were haploid. In Group 3, combined treatment with ionomycin and SPP, the appearance of abnormal chromosomal tracts was significantly (P〈0.05) reduced and the proportion of haploid parthenotes was increased to 85% (17/20) than in Group 2. These results demonstrate that SPP acted as a cdc2 kinase inhibitor and formed the haploidy in oocyte activation. Thus, the present study suggests that cdc2 kinase inhibitor, such as sodium pyrophosphate, may have an effective role in oocyte activation for the production of cloned embryos/animals by nuclear transplantation.

• Asian Pacific Journal of Cancer Prevention
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• 제15권12호
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• pp.4969-4976
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• 2014

Background: miR-152 is involved in the genesis and development of several malignancies. However, its role in HCC has not been fully clarified. The aim of this study was to investigate the clinicopathological significance of miR-152 and its effect on the malignant phenotype of HCC cells. Methods: miR-152 expression was detected using real-time quantitative RT-PCR in 89 pairs of HCC formalin-fixed paraffin-embedded and their adjacent tissues. Functionally, in vitro effects and mechanisms of action of miR-152 on proliferation, viability, caspase activity, apoptosis and motility were explored in HepG2, HepB3 and SNU449 cells, as assessed by spectrophotometry, fluorimetry, fluorescence microscopy, wound-healing and Western blotting, respectively. Results: miR-152 expression in HCC was downregulated remarkably compared to that in adjacent hepatic tissues. miR-152 levels in groups of advanced clinical stage, larger tumor size and positive HBV infection, were significantly lower than in other groups. A miR-152 mimic could suppress cell growth, inhibit cell motility and increase caspase activity and apoptosis in HCC cell lines. Furthermore, Western blotting showed that the miR-152 mimic downregulated Wnt-1, DNMT1, ERK1/2, AKT and TNFRS6B signaling. Intriguingly, inverse correlation of TNFRF6B and miR-152 expression was found in HCC and bioinformatics confirmed that TNFRF6B might be a target of miR-152. Conclusions: Underexpression of miR-152 plays a vital role in hepatocarcinogenesis and lack of miR-152 is related to the progression of HCC through deregulation of cell proliferation, motility and apoptosis. miR-152 may act as a tumor suppressor miRNA by also targeting TNFRSF6B and is therefore a potential candidate biomarker for HCC diagnosis, prognosis and molecular therapy.

• 식물병연구
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• 제20권3호
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• pp.227-233
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• 2014

초산(acetic acid)과 젖산(lactic acid)의 혼합액(MX)의 심비디움 배양토에 오염된 소나무재선충(Bursaphelenchus xylophilus)의 방제효과를 조사하였다. 실내실험에서 초산과 혼합액은 5.0-1.0% (pH 2.6-4.2)에서 젖산은 5.0%에서만 100%가까운 살선충율을 나타내었고 이보다 낮은 농도(0.5-0.1%, pH 5.1-6.9)에서는 초산과 혼합액보다 젖산에서 살선충력이 유의적으로 낮아졌다. 대부분의 농도에서 혼합액의 살선충율이 초산과 젖산의 평균보다 유의적으로 높게 나타나 혼합액의 살선충력 상승효과가 있을 것으로 판단된다. 폿트실험에서 유기산 혼합액(0.5%, 0.25%), 살선충제(fosthiazate, 표준량, 배량) 및 길항세균(Paenibacillus polymyxa GBR-1) 배양 희석액($10^7CFU/ml$) 등의 처리에 의해 심비디움 배양토에서의 소나무재선충의 밀도가 처리간에는 유의적인 차이가 없이 무처리에 비해 유의적으로 감소하였다. 모든 처리에서 약해가 나타나지 않았고, 처리 후 시기별 pH와 2년생 심비디움의 생육에 무처리 대조구와 유의적인 차이가 나타나지 않았다. 이러한 결과와 이들 식용 유기산의 안전성과 가격을 종합하여 볼 때, 심비디움의 재배 관리 중 생육단계에 따라 이 유기산 혼합액의 지속적인 사용은 검역해충의 방제에 있어서 친환경적이고 실제적으로 유효한 하나의 방법일 것이다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.3-3
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• 2018

For centuries, Panax ginseng Meyer (Korean ginseng) has been widely used as a medicinal herb in Korea, China, and Japan. Ginsenosides are a class of triterpene saponins and recognized as the bioactive components in Korean ginseng. Ginsenosides, which can be classified broadly as protopanaxadiols (PPD), protopanaxatriols (PPT), and oleanolic acids, have been shown to flaunt a vast array of pharmacological activities such as immune-modulatory, anti-inflammatory, anti-tumor, anti-diabetic, and antioxidant effects. In recent years, a number of ginseng and ginsenoside researches have increasingly gained wide attention owing to its unique pharmacological properties. Although good efficacies of ginsenosides have been reported, lack of target specific delivery into tumor sites, low solubility, and low bioavailability due to modifications in gastro-intestinal environments limit their biomedical application in clinical trials. As a result to this major challenge, nanotechnology and drug delivery techniques play a significant role to solve this problematic issue. Thus, we reported the preparation of poly-ethylene glycol (PEG) and glycol chitosan (GC) functionalized to ginsenoside (Compound K and PPD) conjugates via hydrolysable ester bonds with improved aqueous solubility and pH-dependent drug release. In vitro cytotoxicity assays revealed that PEG-CK, and PPD-CK conjugates exhibited lower cytotoxicity compared to bare CK and PPD in HT29 cells. However, GC-CK conjugates exhibited higher and similar cytotoxicity in HT29 and HepG2 cells. Furthermore, GC-CK-treated RAW264.7 cells did not exhibit significant cell death at higher concentration of treatment which supports the biocompatibility of the polymer conjugates. They also inhibited nitric oxide production in lipopolysaccharide (LPS)-induced RAW64.7 cells. In addition to polymer-ginsenoside conjugates, silver (AgNps) and gold nanoparticles (AuNps) have been successfully synthesized by green chemistry using different m. The biosynthesized nanoparticles demonstrated antimicrobial efficacy, anticancer, anti-inflammatory, antioxidant activity, biofilm inhibition, and anticoagulant effect. Special interest on the effective delivery methods of ginsenoside to treatment sites is the focus of metal nanoparticle research.In short, nano-sizing of ginsenoside results in an increased water solubility and bioavailability. The use of nano-sized ginsenoside and P. ginseng mediated metallic nanoparticles is expected to be effective on medical platform against various diseases in the future.

• 한국발생생물학회지:발생과생식
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• 제25권1호
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• pp.15-24
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• 2021

Benzo[a]pyrene (B[a]P) is a potent carcinogen and is classified as an endocrine-disrupting chemical. In mammalian testes, Sertoli cells support spermatogenesis. Therefore, if these cells are negatively affected by exposure to xenotoxic chemicals, spermatogenesis can be seriously disrupted. In this context, we evaluated whether mouse testicular TM4 Sertoli cells are susceptible to the induction of cytotoxicity-mediated cell death after exposure to B[a] P in vitro. In the present study, while B[a]P and B[a]P-7,8-diol were not able to induce cell death, exposure to BPDE resulted in cell death. BPDE-induced cell death is accompanied by the activation of caspase-3 and caspase-7. Depolarization of the mitochondrial membrane and cytochrome c release from mitochondria were observed in benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE)-treated cells. These results indicate that TM4 cells are susceptible to apoptosis in a caspase-dependent manner. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that aryl hydrocarbon receptor (AhR) expression was almost undetectable in TM4 cells and that its expression was not altered after B[a]P treatment. This indicates that TM4 cells are nearly AhR-deficient. In TM4 cells, the CYP1A1 protein and its activity were not present. From these results, it is clear that AhR may be a prerequisite for CYP1A1 expression in TM4 cells. Therefore, TM4 cells can be referred to as CYP1A1-deficient cells. Thus, TM4 Sertoli cells are believed to have a rigid and protective cellular machinery against genotoxic agents. In conclusion, it is suggested that tolerance to B[a]P cytotoxicity is associated with insufficient AhR and CYP1A1 expression in testicular Sertoli cells.

• 한국발생생물학회지:발생과생식
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• 제17권3호
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• pp.207-213
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• 2013

This study was performed to investigate the effect of peroxisome proliferators activated receptor-${\gamma}$ (PPAR-${\gamma}$) ligand, pioglitazone, on production of regulated upon activation normal T-cell expressed and secreted (RANTES) and in vitro fertilization (IVF) outcome in infertile patients with endometriosis. Sixty-four infertile patients with stage III or IV endometriosis undergoing IVF were randomly allocated to the study or the control group. The long protocol of GnRH agonist (GnRH-a) was used for controlled ovarian stimulation (COS) in all patients. Patients in the study group were treated with pioglitazone at a dose of 15 mg/day orally from the starting day of GnRH-a treatment to the day of hCG injection. Blood samples were drawn for serologic assay of RANTES on the first day of GnRH-a treatment and the day of hCG injection. There were no differences between the study and control groups in patient characteristics. There were also no differences between the two groups in COS duration, and the numbers of retrieved oocytes, fertilized oocytes and embryos transferred. The clinical pregnancy rate per cycle was higher in the study group, but this difference was not statistically significant. However, embryo implantation rate was significantly higher in the study group of 12.5% compared with 8.6% in the control group (P<0.05). The serum RANTES levels after pioglitazone treatment were significantly lower than those before pioglitazone treatmen in the study group (P<0.05). Our data suggest that pioglitazone treatment can suppress RANTES production and improve the embryo implantation rate in patients with endometriosis undergoing IVF.

• 대한수의학회지
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• 제29권3호
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• pp.283-289
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• 1989

In order to measure in vitro cell mediated immunity in the guinea pigs sensitized with the killed bacilli of Mycobacterium bovis ($AN_5$), M avium (serotype 2), M tuberculosis and M intracellulare (serotype 8), leukocyte adherence inhibition (LAI) test was established using the antigens of purified protein derivatives (PPD) tuberculin. By using LAI test, specificity of cell-mediated immune responses of the guinea pigs inoculated with various Mycobacterium spp was investigated, and comparison between values of LAI and skin test was also made to evaluate the specificity of the newly designed test. The results obtained throughout the experiments were summarized as follows; 1. The optimal concentration of PPD antigens for LAI test was 1 to 2mg per ml of medium. 2. When the leukocytes of guinea pigs sensitized with both M bovis($AN_5$) and M avium (serotype 2) for 2 to 8 weeks were incubated with homologous or heterologous PPD antigens, mean values of LAI test were $61.2{\pm}11.2$ and $65.6{\pm}5.1%$ in homologous PPD antigens respectively, while $30.0{\pm}3.7$ and $32.8{\pm}5.7%$ in heteNlogous PPD antigens, showing the prominently high value of LAI in the homologous syst,em (p<0.01). 3. When the leukocytes of guinea pigs sensitized with both M tuberculosis and M intracellulare (serotype 8) for 2 to 8 weeks were incubated with homologous and heterologous PPD antigens, mean values of LAI test were $67.9{\pm}2.9$ and $66.9{\pm}5.0%$ in homologous PPD antigens, while $27.4{\pm}7.4$ and $24.4{\pm}7.1%$ in heterologous PPD antigens, showing the prominently high value of LAI in the homologous system (p<0.01). 4. Comparing with the specificity of LAI and skin tests on the basis of the value obtained from the homologous system, deviation of reaction was revealed to be 49.5 to 100.2 in LAI test, and -15.9 to 52.0 in skin test.

• 한국동물학회지
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• 제39권3호
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• pp.257-265
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• 1996

참개구리를 사용하여 난소내 여포들이 성장시기에 획즉하게 되는 스테로이드들의 생성능력을 조사하였다. 개구리로부터 중간(0.7-1.11mm) 크기 이하의 여포들을 분리한 다음 전구스테로이들이나 뇌하수체추출물(FPH)을 포함한 배양액에서 6시간 배양 후 이들에 의해 산물스테로이드로 전환된 양을 방사면역 측정법으로 조사하였다. FPH의 처리는 중간 크기의 여포만 스테로이드들의 생성을 현저히 촉진하고 작은 여포들(0.35-0.7mm)에서는 촉진효과가 매우 미약하였다. Progenenolone의 첨가는 가장 작은 여포 (0.22-0.4mm)들에 의해서도 progesterone(P$_4$)으로 전환되었다. 그러나 가장 작은 여포에서는 P$_4$를 첨가했을 때에도 역시 같은 현상이 나타났다. 작은 여포와 중간 여포들에 의한 전환된 스테로이드들의 양을 단위표면적의 전환량으로 계산 비교해 본 결과 전환된 양은 거의 같았다. 이는 작은 여포의 여포세포들 (granulosa cells)이 이미 E$_2$에 이르는 모든 중간 산물들을 생성할 능력을 가졌다는 것을 의미한다. 그러나 이들이 FPH에 의해서는 스테로이드를 조금밖에 생성하지 못하는 것으로 보아 아직 부족한 또다른 요인들이 있는것으로 추정된다.

• 동의생리병리학회지
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• 제23권3호
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• pp.685-694
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• 2009

This experimental study was designed to investigate the inhibitory effects of ethanol extract of modified Yukgunja-tang(mYGJT) on high-fat diet-induced obesity and hyperlipidemia in Sprague-Dawley rats, Animals were divided into normal, control, mYGJT(100 mg/kg and 200 mg/kg) treated groups. Obesity with hyperlipidemia was induced by high fat diet treatment for 6 weeks. mYGJT was given to the amimals by oral gavage for 4 weeks, starting at the high-fat diet regimen, The effect of mYGJT on the differentiation of 3T3 L1 adipocytes in vitro and serological paramamters for obesity and hyperlipidemia in vivo were evaluated, mYGJT significnatly inhibited the differentiation of 3T3 L1 adipocytes in a concentration dependent manner. mYGJT treatment siginficantly reduced body weight, abdominal and epididymal fat weight, and FER(Food Efficiency Ratio) compared with control group in a dose dependent manner. It also signficantly inhibited the levels of serum total lipid, triglyceride, phospholipid, total cholesterol, LDL, AI(Atherosclerosis Index) and returned the serum HDL to normal. Total lipids, triglycerides and cholesterols in the liver, as well as malondialdehyde(MDA) and hydroxy radical in the serum were significantly reduced. However, superoxide dismutase(SOD) activity was significantly increased in mYGJT treated group compared with control group. Finally, mYGJT treatment signficantly decreased the MDA and protein carbonyl concentrations of the hepatic homogenate but signficantly increased the activities of SOD, GSH-Px and Catalase. Taken together, these results suggest that mYGJT can be clinically useful in inhibiting high-fat diet-induced obesity and hyperlipidemia.