• Biomolecules & Therapeutics
• /
• 제11권4호
• /
• pp.224-231
• /
• 2003

The aim of this study was to investigate the effect of resveratrol on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes. Anti-oxidative activity of resveratrol was measured by $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl assay and dichlorodihydrofluorescein diacetate oxidation assay. Gap junctional intercellular communication in HaCaT keratinocytes was assessed using the scrape loading/dye transfer technique. Western blots and reverse transcription-polymerase chain reaction were also analyzed for connexin 43 protein and mRNA expression, respectively. Resveratrol scavenged directly the stable $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl radical over a concentration range of 4 mg/ml ($78.2{\pm}2.7$% of control) to 500 mg/ml ($29.9{\pm}4.2$% of control) and decreased the intracellular reactive oxygen species induced by ultraviolet A (UVA) irradiation ($89.3{\pm}1.1$% of UVA group), ultraviolet B (UVB) irradiation ($70.9{\pm}1.7$% of UVB group) and 12-0-tet-radecanoylphorbol-13-acetate (TPA, $48.3{\pm}1.1$% of TPA group), respectively. UVA irradiation and TPA markedly reduced gap junctional intercellular communication, which was restored by resveratrol. There were no significant differences in the level of connexin 43 protein and mRNA expression among any of the experimental groups. Our data suggests that resveratrol has the protective effect on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes, and this protection is likely due to the scavenging of reactive oxygen species.

• Journal of Microbiology and Biotechnology
• /
• 제28권7호
• /
• pp.1112-1121
• /
• 2018

Jeotgal is a Korean traditional fermented seafood with a high concentration of salt. In this study, we isolated lactic acid bacteria (LAB) from galchi (Trichiurus lepturus, hairtail) and myeolchi (Engraulis japonicas, anchovy) jeotgal on MRS agar and MRS agar containing 5% NaCl (MRS agar+5% NaCl), and identified them by using 16S rRNA gene sequencing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as culture-dependent methods. We also performed polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as a culture-independent method to identify bacterial communities. Five samples of galchi-jeotgal and seven samples of myeolchi-jeotgal were collected from different regions in Korea. A total of 327 and 395 colonies were isolated from the galchi- and myeolchi-jeotgal samples, respectively. 16S rRNA gene sequencing and MALDI-TOF MS revealed that the genus Pediococcus was predominant on MRS agar, and Tetragenococcus halophilus on MRS agar+5% NaCl. PCR-DGGE revealed that T. halophilus, Tetragenococcus muriaticus, and Lactobacillus sakei were predominant in both types of jeotgal. T. halophilus was detected in all samples. Even though the same species were identified by both culture-dependent and -independent methods, many species identified by the culture-dependent methods were not in the bacterial list identified by the culture-independent methods. The distribution of bacteria in galchi-jeotgal was more diverse than in myeolchi-jeotgal. The diverse LAB in galchi- and myeolchi-jeotgals can be further studied as candidates for starter cultures to produce fermented foods.

• Journal of Nutrition and Health
• /
• 제54권5호
• /
• pp.448-458
• /
• 2021

본 연구는 뽕나무 열매 착즙 분말 (MJ)이 지방세포 분화과정에서 염증에 관여하는 유전자 발현 및 miR-132/143 조절에 미치는 효과에 중점을 두고 조사하였다. MJ는 3T3-L1 지방세포 분화 동안 지질축적이 억제되었고 PPAR-γ, CEBP-α 및 aP2와 같은 지방형성 유전자 발현을 억제하였다. 또한 염증 조절 매개체인 TNF-α, IL-6, MCP-1 및 iNOS 유전자 발현이 MJ 처리에 의해 감소되었다. MJ의 지질 축적 억제 효과는 염증과 지방분화에 관여하는 miR-132/143의 발현 감소와 관련이 있음을 확인하였다. 따라서 MJ는 염증을 동반하는 비만 예방 및 치료에 도움이 될 수 있는 식품 소재로서 가능성이 있음을 시사하고 있다. 그러나 MJ의 주요 성분인 안토시아닌의 생체이용율은 1-2% 미만으로 추정되며 예상되는 표적 조직이나 혈류에서 미량 검출되는 것으로 알려져 있다 [33]. 이를 해결하기 위해서는 생체 내 동물실험을 통한 다각적인 분석이 향후 진행되어야 할 것으로 판단된다.

• International Journal of Oral Biology
• /
• 제41권2호
• /
• pp.97-103
• /
• 2016

Mammals have 3 pairs of major salivary glands i.e., the parotid, submandibular, and sublingual glands. Saliva secretion of these glands is modulated by taste perception. Salivary glands are composed mainly of acinar and ductal cells. Primary saliva is secreted by acinar cells and modified during ductal flow. Recently, of the murine 35 bitter taste receptors, Tas2r108 was expressed at highest levels in the submandibular gland by qPCR. Further, Tas2r108-transfected cells respond to a range of bitter compounds, such as denatonium, quinine, colchicine, diphenidol, caffeine and dapson. The objective of the present study was to characterize the expression of Tas2r108 mRNA in acinar and/or ductal cells of the submandibular gland using in situ hybridization (ISH). Male 42-60 days old DBA2 mice were used in the study. Messenger RNAs were extracted from the submandibular gland for generating digoxigenin (DIG) labeled-cRNA probes. These probes were transcribed in anti-sense and sense orientation using T7 RNA polymerase. Dot blot hybridization was performed using DIG labeled-cRNA probes, in order to estimate integrity and optimal diluting concentration of these probes. Subsequently, ISH was performed on murine submandibular gland to detect Tas2r108 mRNA. Dot blot hybridization data demonstrated that Tas2r108 DIG labeled-cRNA anti-sense probes specifically detected Tas2r108 cDNA. ISH results showed that the anti-sense probes labeled acinar and ductal cells in the submandibular gland, whereas no staining was visible in sense controls. Interestingly, the Tas2r108 expression levels were higher in acinar than ductal cells. These results suggested that Tas2r108 might be more associated with primary saliva secretion than with ductal modification of saliva composition.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권7호
• /
• pp.3369-3375
• /
• 2016

MicroRNAs, a novel class of small non-coding RNAs, are key players in many cellular processes, including cell proliferation, differentiation, invasion and regeneration. Tissue and circulatory microRNAs could serve as useful clinical biomarkers and deregulated expression levels have been observed in various cancers. Gene variants may alter microRNA processing and maturation. Thus, we aimed to investigate the association of MIR-196a2 rs11614913 (C/T), MIR-499a rs3746444 (A/G) polymorphisms and their combination with cancer susceptibility in an Egyptian population. Sixty five renal cell carcinoma (RCC) and 60 hepatocellular carcinoma (HCC) patients and 150 controls were enrolled in the study. They were genotyped using real-time polymerase chain reaction technology. Both $miR-196a2^*T$ and $miR-499a*G$ were associated with RCC risk, but only $miR-196a^*T$ was associated with HCC development. Carriage of the homozygote combinations ($MIR196a2^*TT+MIR499a^*AA$) and ($MIR196a2^*CC+MIR499a^*GG$) was associated with 25 and 48 fold elevation of likelhood to develop RCC, respectively. The miR-196a2 SNP was also linked with larger tumor size in RCC and advanced tumor stage in HCC. miR-196a2 and miR-499a combined genotypes were associated with RCC and HCC. Further functional analysis of SNPs is required to confirm relationships between genotypes and phenotypes.

• Food Science and Biotechnology
• /
• 제17권6호
• /
• pp.1171-1177
• /
• 2008

This study was undertaken to classify Enterobacter sakazakii isolates from 13 powdered infant formula products, 25 powdered weaning diet products, and 33 weaning diet ingredients on polymerse chain reaction (PCR) methods. The numbers of the isolates from 1 powdered infant formula product, 7 powdered weaning diet products, and 6 weaning diet ingredients were 1, 14, and 8, respectively. The contaminated ingredients were 1 rice powder, 2 millet powders, 2 vegetable powders, and 1 fruit and vegetable premix. PCR with the primer of repetitive extragenic palindromic element (REP-PCR) and random amplification of polymorphic DNA(RAPD) were effective in discriminating among the isolates, but tRNA-PCR and PCR with the primer of l6S-23S internal transcribed spacer (ITS-PCR) were not. Some of E. sakazakii isolates from vegetable powders, fruit and vegetable premix, and millets powders were classified into the clonal groups based on the DNA patterns in the REP-PCR and RAPD analysis. A close genetic relationship among the isolates from some of the powdered weaning diet products and the rice powder was also detected in the cluster analysis based on the DNA patterns in RAPD.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권5호
• /
• pp.3011-3015
• /
• 2013

Objective: This study aimed to investigate the expression of B7-H4 in human thyroid cancer and determine any association with patient clinicopathological parameters and survival. Methods: B7-H4 expression in 64 clinical thyroid cancer specimens was assessed with immunohistochemistry. Moreover, B7-H4 mRNA expression in 10 fresh resected specimens were evaluated by the reverse transcription-polymerase chain reaction (RT-PCR). Immunohistochemical staining of CD3 was performed to assess the number of tumor infiltrating T lymphocytes (TILs) in thyroid cancers. Results: Positive B7-H4 immunohistochemical staining was observed in 61 out of 64 (95.3%) specimens of thyroid cancer tissues. Significantly more B7-H4 mRNA copies were found in thyroid cancer tissue than that adjacent normal tissue. Moreover, B7-H4 expression in human thyroid cancer tissues was significantly correlated with patient TNM stages and extrathyroidal extension (P<0.05), being inversely correlated with the number of TILs (P<0.05). The overall survival rate of the patients with higher B7-H4 expression was significantly worse than that of the patients with lower B7-H4 expression. Conclusions: This present study suggests that high B7-H4 expression is associated with cancer progression, reduced tumor immunosurveillance and worse patient outcomes in human thyroid cancer.

• Journal of Periodontal and Implant Science
• /
• 제28권4호
• /
• pp.659-678
• /
• 1998

본 연구에서는 한국인 성인성 치주염의 관련세균 중에서 구강 스피로헤타의 분포를 조사하기 위하여 배양하지 않고도 구강 스피로헤타를 분리할 수 있는 16S rRNA에 의거한 올리고뉴클레오타이드 소식자를 사용하였다. 성인성 치주염 환자 29명을 대상으로 한 사람당 6mm 이상의 탐침 깊이를 보이는 부위 4곳(실험군)과 3mm 이하의 탐침 깊이를 보이는 건강한 부위 1곳(대조 1군), 건강한 치주조직을 가진 학생 20명을 대상으로 한 사람당 5부위로부터 치은연하 치태를 채취한 뒤(대조 2군) 중합효소연쇄반응과 점상블롯보합결합법을 시행하였다. 소식자로는 구강 스피로헤타의보편 소식자 및 현재 배양이 되는 구강 스피로헤타중에서 T. denticola, T. pectinovorum, T. socranskii, T. vincentii, T. maltophilum에 대한 종 특이 소식자 TDEN, TPEC, TSOC, TVIN, TMAL과 현재 배양이 되지않은 구강스피로헤타중에서 I-VII군에 대한 군 특이 소식자 TRE I-TRE VII을 이용하여 다음과 같은 결론을 얻었다. 1. 위상차 현미경으로 본 결과는 실험군, 대조 1군에서 각기 91.37%, 14.28%의 구강스피로헤타가 관찰되었으며 대조 2군에서는 관찰되지 않았다. 2. 보편 소식자를 사용한 경우는 실험군, 대조 1군, 대조 2군에서 각기 98.27%, 46.42%, 22.0%의 구강 스피로헤타가 관찰되었다. 3. 특이 소식자를 사용한 경우는 실험군, 대조 1군, 대조 2군에서 각기 95.68%, 35.71%, 19.0%의 구강 스피로헤타가 관찰되었다. 4. 종 특이 소식자를 사용한 경우는 T. socranskii가 가장 많이 관찰되었으며 (81.89%), 그다음이 T. maltophilum(50.0%), T. vincentii(36.20%), T. denticola(13.79%)순이었고, 군 특이 소식자를 사용한 경우는 TREIV(85.34%), TRE II(77.58%), TREI(56.89%), TRE III(25.86%), TREVI(5.17%), TRE V(2.58%) 순이었다. 5. T. vincentii는 치주염에 이환된 부위에서만 관찰되었고, 건강한 부위에서는 관찰되지 않았다. 6. T. pectinovorum과 VII군에 속하는 구강스피로헤타는 어느 표본에서도 관찰되지 않았다. 이상의 결과에서 16S rRNA에 의거한 올리고뉴클레오타이드 소식자로 구강 스피로헤타의 성인성 치주염과의 연관성과 분리되지 않은 구강 스피로헤타를 확인하였으며, 인종 및 치주염의 형태에 따른 구강 스피로헤타의 분포 차이, T. vincentii의 병원성, 치료 전후의 구강 스피로헤타의 분포 변화등의 보다 세분화된 연구가 필요하다고 생각된다.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
• /
• pp.65-65
• /
• 2003

The aim of this study was to investigate the effect of resveratrol on the oxidative stress-induced inhibition of gap junctional intercellular communication (GJIC) in HaCaT keratinocyte. Anti-oxidative activity of resveratrol was measured by a,a-diphenyl-b-picrylhydrazyl (DPPH) assay and dichlorodihydrofluorescein diacetate oxidation assay. GJIC of HaCaT keratinocyte was assessed using the scrape loading/dye transfer technique. Western blots and reverse transcription-polymerase chain reaction were also analyzed for Connexin 43 protein and mRNA expression, respectively. Resveratrol scavenged directly the stable DPPH radical over a concentration range of 4 mg/$\mell$ (78.2 $\pm$ 2.7% of control) to 500 mg/$\mell$ (29.9 $\pm$ 4.2% of control) and prevent to increase the intracellular fluorescence induced by oxidative stress significantly. Ultraviolet A irradiation (UVA) and 12-O-tetradecanoylphorbol-13-acetate markedly reduced GJIC, which was restored by resveratrol. There were no significant differences in the level of Connexin 43 protein and mRNA expression among any of the experimental groups. Our data suggests that resveratrol has the protective effect on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocyte and this protection is likely due to the scavenging of reactive oxygen species.

• Animal cells and systems
• /
• 제3권2호
• /
• pp.181-185
• /
• 1999

Xenopus oocyte is one of the widely used heterologous expression systems of ion channels for electrophysiological studies. Here we describe a new method in which cRNA produced by polymerase chain reaction (PCR) and in vitro transcription is injected to express ion channels in oocytes. This method enables us (1) to eliminate all or a part of the untranslated region of the cDNA and to replace it with a known sequence which helps increase the expression level in oocytes, and (2) to use the PCR product for in vitro transcription without subcloning. Using this method, the expression level of one of the neuronal nicotinic acetylcholine receptors (nAChRs) $\alpha$$_{6}$ subtype in oocytes was systematically increased by more than 100-fold, which was confirmed both by the $\alpha$-Bungarotoxin ($\alpha$,/TEX>Bgt) binding assay and the current measurement.t.