• Title/Summary/Keyword: Differential screening

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Differential expression of microRNAs in the saliva of patients with aggressive periodontitis: a pilot study of potential biomarkers for aggressive periodontitis

  • Lee, Nam-Hun;Lee, Eunhye;Kim, Young-Sung;Kim, Won-Kyung;Lee, Young-Kyoo;Kim, Su-Hwan
    • Journal of Periodontal and Implant Science
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    • v.50 no.5
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    • pp.281-290
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    • 2020
  • Purpose: The aim of this study was to compare microRNA (miRNA) gene expression in saliva using miRNA polymerase chain reaction (PCR) arrays in healthy and aggressive periodontitis (AP) patients. Methods: PCR arrays of 84 miRNAs related to the human inflammatory response and autoimmunity from the saliva samples of 4 patients with AP and 4 healthy controls were performed. The functions and diseases related to the miRNAs were obtained using TAM 2.0. Experimentally validated targets of differentially expressed miRNAs were obtained from mirTarBase. Gene ontology terms and pathways were analyzed using ConsensusPathDB. Results: Four downregulated miRNAs (hsa-let-7a-5p, hsa-let-7f-5p, hsa-miR-181b-5p, and hsa-miR-23b-3p) were identified in patients with AP. These miRNAs are associated with cell death and innate immunity, and they target genes associated with osteoclast development and function. Conclusions: This study is the first analysis of miRNAs in the saliva of patients with AP. Identifying discriminatory human salivary miRNA biomarkers reflective of periodontal disease in a non-invasive screening assay is crucial for the development of salivary diagnostics. These data provide a first step towards the discovery of key salivary miRNA biomarkers for AP.

Factors associated with depressive mood among problem drinkers by gender (성인 문제음주자의 성별에 따른 우울경험과 관련 요인)

  • Lee, Eun-Ju;Lee, Eun-Sook
    • Journal of the Korea Convergence Society
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    • v.11 no.2
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    • pp.375-384
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    • 2020
  • The purpose of this study was to assess the relationship between drinking patterns and depressive mood, and to understand the factors associated with depressive mood among problem drinkers. Data of 19,878 adults over 19 years of age and older from the third Korea National Health and Nutrition Examination Survey were evaluated by multivariate logistic regression analysis. The risk of depressive mood was greater with higher level of problem drinking in both genders. For male problem drinkers, the factors associated with depressive mood include older age, lower education level, unemployment, high perceived stress, bad subjective health status, and non-married status. For female, higher risk was found in groups with the age of 40-49, lower education level, unemployment, high perceived stress, and bad subjective health status. By early screening of the identified risk factors, progression to mental health problem can be avoided. For preventative measures, differential approach depending on gender is suggested.

Screening of Antimicrobial Activity from Differential Extracts of Allii sativi Bulbus (대산의 분획별 추출물에서 항균활성 검색)

  • Kim Hee Seok;Bae Heung Mo;Kim Shin Moo;Lee Hyun Ok;Kim Ki young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.6
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    • pp.1184-1189
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    • 2002
  • Allii sativi Bulbus(garlic) have been shown to possess medicinal value, in particular, antimicrobial activity. In this study, we compared the efficacy on some pathogenic bacteria and fungus among several different extracts(water, hexane, ethyl acetate, methanol, chloroform) of Allii sativi Bulbus. Animal pathogenic bacteria and fungus(S. gallinarium: KCTC 2441, S. flexneri: KCTC 2361, E. cloacae: KCTC 2006, K. pneumonia: KCTC 2208, C. albicans: KCTC 1940) were used to test by measurement of minimum inhibitory concentrations(MIC) and disc diffusion. Allii sativi Bulbus were cut and mixed with water at 37℃ about 24 h and filtered, and several different solvents(hexane, chloroform, ethyl acetate, methanol) were respectively added to separate the fraction of each solvent. The antimicrobial(bacteriocidal) and antifungal effect were apparently shown from water extract, hexane and ethyl acetate extract against using strains(Staphylococcus gallinarium, Shigella flexneri, Enterobacter doacae, Klebsiella pneumonia, Candida albicans). Especially, the water extract showed the superior efficacy. And the clear zone size of water extract (11~27 mm) was greater than that of gentamycin, hexane extract and ethyl acetate extract against S. gallinarium. S. flexneri, K. pneumonia and C. albicans. Minimum inhibitory concentrations(MIC) of water extract appeared to around 2.0~7.5 ㎎/㎖ against S. gallinarium, S. flexneri, E. cloacae and K. pneumonia. The greater activity was shown by water extract because the MIC of water extract for C. albicans observed in very low concentration(<1.0 ㎎/㎖) compared to hexane(5.0 ㎎/㎖) and ethyl acetate(10.0 ㎎/㎖). Therefore, these results exhibited that water extract of Allii sativi Bulbus have stronger antimicrobial activity than hexane and ethyl acetate extract, and may be useful as topical medicine of superficial infections causing C. albicans as well as antifungal agents.

Screening of Ecotoxicant Responsive Genes and Expression Analysis of Benzo[a]pyrene-exposed Rockfish (Sebastes schlgeli)

  • Yum, Seung-Shic;Woo, Seon-Ock;Lee, Taek-Kyun
    • Molecular & Cellular Toxicology
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    • v.2 no.2
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    • pp.114-119
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    • 2006
  • Benzo[a]pyrene is a representative ecotoxicant in marine environment and a model compound of polycyclic aromatic hydrocarbons, which has an ability to bioaccumulate in aquatic organisms. This study aimed to identify molecular biomarkers suitable for assessing environmental pollution using a microarray technique. We examined the effects of benzo[a]pyrene on gene expressions in the rockfish, Sebastes schlegeli. We constructed the subtractive cDNA library with hepatic RNA from benzo[a]pyrene-exposed and non-exposed control fish. From the library 10,000 candidate clones were selected randomly and cDNA microarray was constructed. We determined benzo[a]pyrene-responsive genes using a high-density microarray. Statistical analysis showed that approximately 400 genes are significantly induced or reduced by benzo[a]pyrene treatment ($2\;{\mu}m$). Especially gene expression changes of 4 candidate clones among the up- or down-regulated genes were investigated in 6, 12 and 24 hr BaP-exposed fish groups. Many methods have been developed to monitor marine environmental status, which depend on quantifying the levels of the toxic components in polluted seawater or on ecological accessing, such as species diversity or richness. However, those methods could not provide information on physiological or genetic changes induced by such environmental stresses. Comparing with the conventional methods, these data will propose that benzo[a]pyrene-responsive genes can be useful for biological risk assessment of polycyclic aromatic hydrocarbons on marine organism at molecular level.

Suppression of ADAM 10-induced Delta-1 Shedding Inhibits Cell Proliferation During the Chondro-Inhibitory Action of TGF-β3

  • Jin, Eun-Jung;Choi, Young-Ae;Sonn, Jong-Kyung;Kang, Shin-Sung
    • Molecules and Cells
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    • v.24 no.1
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    • pp.139-147
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    • 2007
  • Although transforming growth factors (TGFs) are implicated in the process of endochondral ossification, which is initiated by the differentiation of mesenchymal cells into chondrocytes, it is not clear how $TGF-{\beta}3$ regulates the chondrogenic differentiation of limb bud mesenchymal cells. Here, differential display polymerase chain reaction (DD-PCR) screening and RT-PCR analysis revealed that transcripts of A Disintegrin And Metalloprotease 10 (ADAM 10) decreased during the chondro-inhibitory action of $TGF-{\beta}3$ on cultured chick leg bud mesenchymal cells. Electroporation of ADAM 10 morpholino antisense oligonucleotides inhibited the ectodomain shedding of delta-1, and cell proliferation and subsequent precartilage condensation, in a manner similar to that caused by $TGF-{\beta}3$. The suppression of mesenchymal cell proliferation induced by $TGF-{\beta}3$ and ADAM 10 morpholino antisense oligonucleotides was reversed by activation of ADAM 10 with phorbol 12-myristate 13-acetate (PMA) or knockdown of Notch-1 with siRNA. Collectively, these data indicate that, in cultured chick leg bud mesenchyme cells, $TGF-{\beta}3$ downregulates ADAM 10 and inhibits cell proliferation and subsequent precartilage condensation by inhibiting the ectodomain shedding of delta-1, and that this results in the activation of Notch signaling.

Screening for Metastatic Osteosarcoma Biomarkers with a DNA Microarray

  • Diao, Chun-Yu;Guo, Hong-Bing;Ouyang, Yu-Rong;Zhang, Han-Cong;Liu, Li-Hong;Bu, Jie;Wang, Zhi-Hua;Xiao, Tao
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1817-1822
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    • 2014
  • Objective: The aim of this study was to screen for possible biomarkers of metastatic osteosarcoma (OS) using a DNA microarray. Methods: We downloaded the gene expression profile GSE49003 from Gene Expression Omnibus database, which included 6 gene chips from metastatic and 6 from non-metastatic OS patients. The R package was used to screen and identify differentially expressed genes (DEGs) between metastatic and non-metastatic OS patients. Then we compared the expression of DEGs in the two groups and sub-grouped into up-regulated and down-regulated, followed by functional enrichment analysis using the DAVID system. Subsequently, we constructed an miRNA-DEG regulatory network with the help of WebGestalt software. Results: A total of 323 DEGs, including 134 up-regulated and 189 down-regulated, were screened out. The up-regulated DEGs were enriched in 14 subcategories and most significantly in cytoskeleton organization, while the down-regulated DEGs were prevalent in 13 subcategories, especially wound healing. In addition, we identified two important miRNAs (miR-202 and miR-9) pivotal for OS metastasis, and their relevant genes, CALD1 and STX1A. Conclusions: MiR-202 and miR-9 are potential key factors affecting the metastasis of OS and CALD1 and STX1A may be possible targets beneficial for the treatment of metastatic OS. However, further experimental studies are needed to confirm our results.

Screening of Differentially Expressed Genes among Various TNM Stages of Lung Adenocarcinoma by Genomewide Gene Expression Profile Analysis

  • Liu, Ming;Pan, Hong;Zhang, Feng;Zhang, Yong-Biao;Zhang, Yang;Xia, Han;Zhu, Jing;Fu, Wei-Ling;Zhang, Xiao-Li
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6281-6286
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    • 2013
  • Background: To further investigate the molecular basis of lung cancer development, we utilize a microarray to identify differentially expressed genes associated with various TNM stages of adenocarcinoma, a subtype with increasing incidence in recent years in China. Methods: A 35K oligo gene array, covering about 25,100 genes, was used to screen differentially expressed genes among 90 tumor samples of lung adenocarcinoma in various TNM stages. To verify the gene array data, three genes (Zimp7, GINS2 and NAG-1) were confirmed by real-time RT-PCR in a different set of samples from the gene array. Results: First, we obtained 640 differentially expressed genes in lung adenocarcinomas compared to the surrounding normal lung tissues. Then, from the 640 candidates we identified 10 differentially expressed genes among different TNM stages (Stage I, II and IIIA), of which Zimp7, GINS2 and NAG-1 genes were first reported to be present at a high level in lung adenocarcinoma. The results of qRT-PCR for the three genes were consistent with those from the gene array. Conclusions: We identified 10 candidate genes associated with different TNM stages in lung adenocarcinoma in the Chinese population, which should provide new insights into the molecular basis underlying the development of lung adenocarcinoma and may offer new targets for the diagnosis, therapy and prognosis prediction.

Comparative Serum Proteomic Analysis of Serum Diagnosis Proteins of Colorectal Cancer Based on Magnetic Bead Separation and MALDI-TOF Mass Spectrometry

  • Deng, Bao-Guo;Yao, Jin-Hua;Liu, Qing-Yin;Feng, Xian-Jun;Liu, Dong;Zhao, Li;Tu, Bin;Yang, Fan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6069-6075
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    • 2013
  • Background: At present, the diagnosis of colorectal cancer (CRC) requires a colorectal biopsy which is an invasive procedure. We undertook this pilot study to develop an alternative method and potential new biomarkers for diagnosis, and validated a set of well-integrated tools called ClinProt to investigate the serum peptidome in CRC patients. Methods: Fasting blood samples from 67 patients diagnosed with CRC by histological diagnosis, 55 patients diagnosed with colorectal adenoma by biopsy, and 65 healthy volunteers were collected. Division was into a model construction group and an external validation group randomly. The present work focused on serum proteomic analysis of model construction group by ClinProt Kit combined with mass spectrometry. This approach allowed construction of a peptide pattern able to differentiate the studied populations. An external validation group was used to verify the diagnostic capability of the peptidome pattern blindly. An immunoassay method was used to determine serum CEA of CRC and controls. Results: The results showed 59 differential peptide peaks in CRC, colorectal adenoma and health volunteers. A genetic algorithm was used to set up the classification models. Four of the identified peaks at m/z 797, 810, 4078 and 5343 were used to construct peptidome patterns, achieving an accuracy of 100% (> CEA, P<0.05). Furthermore, the peptidome patterns could differentiate the validation group with high accuracy close to 100%. Conclusions: Our results showed that proteomic analysis of serum with MALDI-TOF MS is a fast and reproducible approach, which may provide a novel approach to screening for CRC.

Cloning and Expression of Lactate Dehydrogenase H Chain Gene in Adipose Tissues of Korean Cattle

  • Kim, H.H.;Seol, M.B.;Jeon, D.H.;Sun, S.S.;Kim, K.H.;Choi, Y.J.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1670-1674
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    • 2001
  • To understand molecular mechanisms that regulate deposition and release of intramuscular fat, a fasting-induced clone was identified by differential screening from cDNA library of adipose tissues of Korean cattle. The clone had a total length of 1,319 nucleotides coding for 334 amino acids. It was identified as one encoding L-lactate dehydrogenase H chain (LDH-B). Comparison of the deduced amino acid sequences of bovine LDH-B with those of pig, human, rat, and mouse showed 98%, 98%, 97%, and 96% identity, respectively. Food deprivation for 48 h increased mRNA levels of LDH-B gene in adipose tissues of Korean cattle compared to fed- and 6 h refed- tissues. The expression of obese mRNA was examined for individual adipose tissue from several fat depots. Fasting induced expression of LDH-B gene in subcutaneous adipose tissues, but it did not affect expression levels in abdominal, perirenal and intramuscular tissues. Results demonstrate that induction of LDH-B gene during fasting may represent a metabolic shift from anaerobic state to aerobic predominance in fasted adipose tissues and that its responses to fasting are different among several adipose tissues.

Differential Diagnosis of Hyperphenylalaninemias (고페닐알라닌혈증의 진단 및 평가)

  • Lee, Jeongho
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.15 no.3
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    • pp.110-117
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    • 2015
  • All infants should be screened for phenylketonuria (PKU) within the three days of life, in order to allow timely dietary intervention to protect children with PKU from neurologic damage in Korea. A commonly used cut-off level for diagnosis of PKU is $240{\mu}mol/L$ (4 mg/dL). Up to 2% of cases of hyperphenylalaninemias (HPA) detected by the screening test will account for a disorder of $BH_4$ metabolism. Therefore, analysis of blood or urinary pterins is essential, backed up with measurement of DHPR activity, as this allows differentiation of $BH_4$ disorders. A $BH_4$ loading test and measurement of neurotransmitters in CSF provide further important information to the severity of $BH_4$ deficiency and $BH_4$ loading test can detect patients with $BH_4$ deficiency and $BH_4$ responsive PKU. Several protocols for $BH_4$ loading test have been described, involving treatment with $BH_4$ for periods ranging from 1 day to 1 month, and using doses of $BH_4$ of 10-20 mg/kg. There is general agreement that a reduction on blood phenylalanine of at least 30% in response to $BH_4$ loading indicates a clinically significant effect, although in some tests a lower cut-off value may be defined for individual patients, or no specific cut-off value is proposed. The frequency of $BH_4$ responsiveness is highest in patients with mild HPA and mild to moderate PKU resulting from PAH mutations with residual activity.