• 대한의생명과학회지
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• 제10권3호
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• pp.195-202
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• 2004

Lung cancer is a leading cause of cancer death worldwide; however, despite major advances in cancer treatment during the past two decades, the prognostic outcome of lung cancer patients has improved only minimally. This is largely due to the inadequacy of the traditional screening approach of diagnosis in lung cancer, which detects only well­established overt cancers and fails to identify precursor lesions in premalignant conditions of the bronchial tree. In recent years this situation has fundamentally changed with the identification of molecular abnormalities characteristic of premalignant changes; these concern tumour suppressor genes, loss of heterozygosity at crucial sites and activation of oncogenes. Basic knowledge at the molecular level has extremely important clinical implications with regard to early diagnosis, risk assessment and prevention, and therapeutic targets. In this study we used a 'cap-finder' subtractive hybridization method, 'long distance' polymerase chain reaction (PCR), streptavidin magnetic beads mediated subtraction, and spin column chromatography to detect differential expression genes of human small cell lung carcinoma. We have now isolated ninety two genes that expressed differentially in the human small cell lung carcinoma cells and analyzed of 12 clones with sequencing, nine cDNAs include tapasin (NGS-17) mRNA, BC200 alpha scRNA, chromosome 12q24 PAC RPCI3-462E2, protein phosphatase 1 (PPPICA), translocation protein 1 (TLOC1), ribosomal protein S24 (RPS24) mRNA, protein phosphatase (PPEF2), cathepsin Z, MDM2 gene and three novel genes. They may be oncogenesis­related proteins.

• 생명과학회지
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• 제33권11호
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• pp.944-949
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• 2023

이 연구의 목적은 122종의 고세균 종에 보존된 대사 경로와 보존된 유전자를 확인하는 것이었다. 각각의 122개 고세균이 63개의 COG 대사 경로, 이를 구성하는 822개의 COG, 총 4,877개의 COG를 보유하고 있는지 분석했다. 대사경로에서는 archaeal ribosomal proteins만이 가장 보존적이었다. 122종의 고세균 모두에 공통적인 COG는 7개의 COG pathways에서 46개, 그리고 그 외가 20개였다. COG pathways에서는 ribosome을 구성하는 29개, tRNA synthetase와 전사인자가 각각 5개, RNA polymerase를 구성하는 3개, 그리고 tRNA modification에 관련된 2개의 COG가 공통적이었다. COG pathways에 속하지 않고 122종의 고세균에 공통적인 보존적 유전자까지 고려하면 외부와 세포질을 구분 짓는 세포벽과 세포외기질의 합성, 복제, 전사, 번역, 단백질 대사에 관련된 유전자들 중에서 일부가 공통적이었다. 계통수에서 구한 각 고세균의 distance value를 분류단위로 보면 Euryarchaeota 문의 Halobacteria강의 평균이 가장 낮았고 표준편차는 Thaumarchaeota 문의 Nitosospharia강, 강을 알 수 없는 Thaumarchaeota문의 고세균, Euryarchaeota 문의 Halobacteria 강, Crenarchaeota 문의 Thermoprotei 강, 기타 고세균(OA)이 높았다. 계통수 분석으로 6가지의 공통점을 찾았다. 본 연구결과는 보존된 유전자에 관한 자료 외에도 의약품 개발, 균주 개선을 위한 유전자의 선택 등에 활용될 수 있을 것이다.

• Tuberculosis and Respiratory Diseases
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• 제69권5호
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• pp.331-336
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• 2010

Background: Recently, the rate of infections with non-tuberculous mycobacteria (NTM) has been increasing in Korea. Precise identification of NTM is critical to determination of the pathogen and to target treatment of NTM patients. Methods: Sixty-eight unclassified mycobacteria isolates by rpoB PCR-RFLP assay (PRA) collected in 2008 were analyzed by National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) search after sequencing of 16S rRNA, hsp65, rpoB genes. Results: Nineteen strains of 68 isolates were specified as species after sequencing analysis of 3 gene types. We found 3 M. lentifulavum, 5 M. arupense, 4 M. triviale, 4 M. parascrofulaceum, and one M. obuense. One M. tuberculosis and another M. peregrinum were mutated at the Msp I recognition site needed for rpoB PRA. The remaining 49 isolates did not coincide with identical species at the 3 kinds genes. Conclusion: Sequencing analysis of 16S rRNA, hsp65, rpoB was useful for identification of NTM unclassified by rpoB PRA.

• Toxicological Research
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• 제22권3호
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• pp.283-291
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• 2006

The survival and growth of epithelial cells depends on adhesion to the extracellular matrix. An adhesion signal may regulate the initiation of differentiation, since epidermal keratinocytes differentiate as they leave the basement membrane. A metabolically dead cornified cell envelope is the end point of epidermal differentiation so that this process may be viewed as a specialized form of programmed cell death. In order to investigate the precise cellular signaling events loading to terminal differentiation of keratinocytes, we have utilized HaCaT cells to monitor the biological consequences of $Ca^{2+}$ stimulation and numerous downstream signaling pathways, including activation of the extracellular signal-regulated protein kinase(ERK) pathway and activation of phosphatidylinositol 3-kinase(PI3K). The results presented in this study show that $Ca^{2+}$ function as potent agents for the differentiation of HaCaT keratinocytes, and this differentiation depends or the activation of ERK, Protein kinase B(PKB) and p70 ribosomal protein S6 kinase(p70S6K). Finally, the results show that the expression of Activator protein 1(AP-1; c-Jun and c-Fos) increased following $Ca^{2+}$-mediated differentiation of HaCaT cells, suggesting that ERK-mediated AP-1 expression is critical for initiating the terminal differentiation of keratinocytes.

• 동의생리병리학회지
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• 제17권1호
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• pp.241-246
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• 2003

Bambusae Caulis in Liquamen(BCL) has been used to relieve the cough and asthma, and remove the phlegm in traditional Oriental medicine. In recent years, it was studied for its antiinflammatory, antiallergenic, immune-modulating, and anticarcinogenic capabilities. This experiment was performed to evaluate the microarray profiles of CD genes in human mast cells before and after BCL treatment. The results are as follows: The expression of 51 of the genes studied was up-regulated in the Bel-treated group; they include the genes coding L apoferritin, beta-2-microglobulin, ferritin light polypeptide, CD63, monocyte chemotactic and activating fact, heme oxygenase 1, CD140a, integrin alpha M, colony stimulating factor 2 receptor, eukaryotic translation elongation factor, CD37, interleukin 18, NADH dehydrogenase 1 beta, CD48, 5-lipoxygenase activating protein, interleukin 4, ribosomal protein L5, GABA(A) receptor-associated protein, beta-tubulin, integrin beta 1, CD162, CD32, lymphotoxin beta, alpha-tublin, integrin alpha L, CD2, CD151, CD331, 90 kDa heat shock protein, CD59, CD3Z, microsomal glutathione S-transferase 2, CD33, CD162R, cyclophilinA, CD84, interleukin 9 receptor, interleukin 11, CD117, CD39-Like 2, and so forth. The expression of 7 of the genes studied was down-regulated in the BCL-treated group; they include the genes coding con, CD238, SCF, CD160, CD231, CD24, and CD130. Consequently, the treatment of BCL on the human mast cells increased the expression of 51 genes and decreased the expression of 7 genes. These data would provide a fundamental basis to the traditional applications of Bambusae Caulis in Liquamen.

• 대한의생명과학회지
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• 제6권1호
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• pp.11-17
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• 2000

SNAP-25는 presynaptic plasma membrane에 위치하는 단백질로서 synaptic vesicle의 docking과 fusion에 있어서 매우 중요한 역할을 한다. 생쥐 SNAP-25$^{2)}$ 유전자와 99%의 높은 homology를 갖고 있는 Z2 cDNA를 probe로 사용하여 쥐의 뇌 cDNA library에서 SNAP-25유전자를 screening하였다. 그 결과 6개 의 양성 클론을 분리 해 냈으며, 이들 각각을 S1, S2, S3, S4, S5, S6으로 명명하였다. 이 중에서 생쥐 SNAP-25와 가장 높은 homology를 보여 주고 있는 S5 클론을 선택하여 염기서열을 분석하였다. 2,100 bp의 염기서열로 구성된 쥐 SNAP-25 cDNA는 206개의 아미노산을 coding하는 618 bp의 open reading frame을 가지고 있으며, ORF는 209~211 bp에 위치하는 AUG codon에서 시작하여 827~829 bp에 위치하는 stop codon TAA에서 끝난다. 3' untranslated region에서 는 28과 19개 의 CA 반복 염기서열을 보여주고 있었으며, SNAP-25 peptide sequence에서 4개의 cystein residues는 84~91에 위치하고 있었으며, amino terminus 부분에서 amphipathic $\alpha$-helix를 형성하고 있는 것을 볼 수 있었다. 사람과 쥐의 SNAP-25 유전자는 88%, 생쥐와 쥐의 경우는 97%의 homology를 보여 주고 있었다. 그리고 사람과 쥐의 ORF에서 염기서열은 94%,생쥐와 쥐의 ORF에서 염기서열은 100%의 homology를 보여주고 있었으며 사람, 생쥐, 그리고 쥐의 ORF에서 아미노산 서열은 100%의 homology를 보여주고 있었다.

• 생명과학회지
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• 제15권2호
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• pp.261-266
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• 2005

생명체의 본질적 기능에 중요한 역할을 담당하는 유전자들을 밝히기 위해 미생물 유전체들 사이의 공통적 유전자를 밝히는 COG알고리듬을 이용하였다. 진핵생물 3종을 포함한 66종의 미생물에서 63개의 유전자가 보존적이었으며, 단백질 합성에 관여하는 유전자들이 총 52개로 생명현상에서의 단백질의 중요성을 알 수 있었다. 각 보존적 유전자들의 distance value를 이용하여 종간의 유전자 변이의 정도를 보면, ribosomal protein S12 (COG0048)와 ribosomal protein L14 (COG0093)의 보존성이 가장 높았다. 보존적 유전자들의 평균과 분산으로 유전체 분석을 수행한 결과, 고세균과 진정세균의 각 그룹 등 근연종들이 독자적 그룹을 형성하였다. 하지만 각 그룹내의 속 및 유전체의 수와 유전체 변이의 정도는 비례하지 않는 것을 알 수 있었다.

• Journal of Veterinary Science
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• 제24권3호
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• pp.23.1-23.16
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• 2023

Background: Irritable bowel syndrome (IBS) is a functional bowel disorder (FBD). Objectives: To assess the therapeutic effects of paeoniflorin (PF) on IBS in rats. Method: Sixty male Sprague-Dawley rats were randomly divided into normal, model, positive drug, low-dose PF, medium-dose PF and high-dose PF groups (n = 10). After gavage for 2 consecutive weeks, the effect of PF on abdominal pain symptoms was assessed based on the abdominal withdrawal reflex (AWR) score, fecal water content and pathological changes in colon tissues. D-lactate, interleukin-1β (IL-1β), transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay, and phosphorylated nuclear factor kappa B (p-NF-κB) p65 was detected by Western blotting. The abundance and diversity changes of intestinal flora were explored using 16S ribosomal RNA sequencing. Result: In PF groups, the mucosal morphology of colon tissues was intact, and the glands were arranged neatly and structured clearly, without obvious inflammatory cell infiltration. Compared with the model group, PF groups had significantly elevated pain threshold, and mRNA and protein levels of zonula occludens-1 (ZO-1) and occludin, decreased AWR score at 20 mmHg pressure, fecal water content, mRNA levels of IL-1β, TGF-β, and TNF-α, protein level of p-NF-κB p65 and level of serum D-lactate, and reduced levels of serum IL-1β, TGF-β, and TNF-α (p < 0.05, p < 0.01). PF groups had higher abundance of Lactobacillus, Akkermansia, Alistipes, and Bacteroides, but lower abundance of Desulfovibrio, Parasutterella, and Enterococcus than those of the model group. Conclusions: PF exerts therapeutic effects on IBS in rats probably by regulating the intestinal flora, and then up-regulating the expressions of ZO-1 and occludin in colon tissue while down-regulating the levels of IL-1β, TGF-β, TNF-α, D-lactate and p-NF-κB p65.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1230-1237
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• 2009

The selective plugging strategy of Microbial Enhanced Oil Recovery (MEOR) involves the use of microbes that grow and produce exopolymeric substances, which block the high permeability zones of an oil reservoir, thus allowing the water to flow through the low permeability zones leading to increase in oil recovery. Bacillus licheniformis TT33, a hot water spring isolate, is facultatively anaerobic, halotolerant, and thermotolerant. It produces EPS as well as biosurfactant and has a biofilm-forming ability. The viscosity of its cell-free supernatant is $120\;mPa{\cdot}s$ at $28^{\circ}C$. Its purified EPS contained 26% carbohydrate and 3% protein. Its biosurfactant reduced the surface tension of water from 72 to 34 mN/m. This strain gave $27.7{\pm}3.5%$ oil recovery in a sand pack column. Environmental scanning electron microscopy analysis showed bacterial growth and biofilm formation in the sand pack. Biochemical tests and Amplified Ribosomal DNA Restriction Analysis confirmed that the oil recovery obtained in the sand pack column was due to Bacillus licheniformis TT33.

• Biomolecules & Therapeutics
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• 제25권6호
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• pp.593-598
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• 2017

The $Na^+/H^+$ exchanger-1 (NHE-1) is a ubiquitously expressed pH-regulatory membrane protein that functions in the brain, heart, and other organs. It is increased by intracellular acidosis through the interaction of intracellular $H^+$ with an allosteric modifier site in the transport domain. In the previous study, we reported that glutamate-induced NHE-1 phosphorylation mediated by activation of protein kinase C-${\beta}$ (PKC-${\beta}$) in cultured neuron cells via extracellular signal-regulated kinases (ERK)/p90 ribosomal s6 kinases (p90RSK) pathway results in NHE-1 activation. However, whether glutamate stimulates NHE-1 activity solely by the allosteric mechanism remains elusive. Cultured primary cortical neuronal cells were subjected to intracellular acidosis by exposure to $100{\mu}M$ glutamate or 20 mM $NH_4Cl$. After the desired duration of intracellular acidosis, the phosphorylation and activation of PKC-${\beta}$, ERK1/2 and p90RSK were determined by Western blotting. We investigated whether the duration of intracellular acidosis is controlled by glutamate exposure time. The NHE-1 activation increased while intracellular acidosis sustained for >3 min. To determine if sustained intracellular acidosis induced NHE-1 phosphorylation, we examined phosphorylation of NHE-1 induced by intracellular acidosis by transient exposure to $NH_4Cl$. Sustained intracellular acidosis led to activation and phosphorylation of NHE-1. In addition, sustained intracellular acidosis also activated the PKC-${\beta}$, ERK1/2, and p90RSK in neuronal cells. We conclude that glutamate stimulates NHE-1 activity through sustained intracellular acidosis, which mediates NHE-1 phosphorylation regulated by PKC-${\beta}$/ERK1/2/p90RSK pathway in neuronal cells.