• Title/Summary/Keyword: tissue samples

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Transcriptomic profiles and their correlations in saliva and gingival tissue biopsy samples from periodontitis and healthy patients

  • Jeon, Yoon-Sun;Cha, Jae-Kook;Choi, Seong-Ho;Lee, Ji-Hyun;Lee, Jung-Seok
    • Journal of Periodontal and Implant Science
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    • v.50 no.5
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    • pp.313-326
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    • 2020
  • Purpose: This study was conducted to analyze specific RNA expression profiles in gingival tissue and saliva samples in periodontitis patients and healthy individuals, and to determine their correlations in light of the potential use of microarray-based analyses of saliva samples as a periodontal monitoring tool. Methods: Gingival tissue biopsies and saliva samples from 22 patients (12 with severe periodontitis and 10 with a healthy periodontium) were analyzed using transcriptomic microarray analysis. Differential gene expression was assessed, and pathway and clustering analyses were conducted for the samples. The correlations between the results for the gingival tissue and saliva samples were analyzed at both the gene and pathway levels. Results: There were 621 differentially expressed genes (DEGs; 320 upregulated and 301 downregulated) in the gingival tissue samples of the periodontitis group, and 154 DEGs (44 upregulated and 110 downregulated) in the saliva samples. Nine of these genes overlapped between the sample types. The periodontitis patients formed a distinct cluster group based on gene expression profiles for both the tissue and saliva samples. Database for Annotation, Visualization and Integrated Discovery analysis revealed 159 enriched pathways from the tissue samples of the periodontitis patients, as well as 110 enriched pathways In the saliva samples. Thirty-four pathways overlapped between the sample types. Conclusions: The present results indicate the possibility of using the salivary transcriptome to distinguish periodontitis patients from healthy individuals. Further work is required to enhance the extraction of available RNA from saliva samples.

Detection of KRAS mutations in plasma cell-free DNA of colorectal cancer patients and comparison with cancer panel data for tissue samples of the same cancers

  • Min, Suji;Shin, Sun;Chung, Yeun-Jun
    • Genomics & Informatics
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    • v.17 no.4
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    • pp.42.1-42.6
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    • 2019
  • Robust identification of genetic alterations is important for the diagnosis and subsequent treatment of tumors. Screening for genetic alterations using tumor tissue samples may lead to biased interpretations because of the heterogeneous nature of the tumor mass. Liquid biopsy has been suggested as an attractive tool for the non-invasive follow-up of cancer treatment outcomes. In this study, we aimed to verify whether the mutations identified in primary tumor tissue samples could be consistently detected in plasma cell-free DNA (cfDNA) by digital polymerase chain reaction (dPCR). We first examined the genetic alteration profiles of three colorectal cancer (CRC) tissue samples by targeted next-generation sequencing (NGS) and identified 11 non-silent amino acid changes across six cancer-related genes (APC, KRAS, TP53, TERT, ARIDIA, and BRCA1). All three samples had KRAS mutations (G12V, G12C, and G13D), which were well-known driver events. Therefore, we examined the KRAS mutations by dPCR. When we examined the three KRAS mutations by dPCR using tumor tissue samples, all of them were consistently detected and the variant allele frequencies (VAFs) of the mutations were almost identical between targeted NGS and dPCR. When we examined the KRAS mutations using the plasma cfDNA of the three CRC patients by dPCR, all three mutations were consistently identified. However, the VAFs were lower (range, 0.166% to 2.638%) than those obtained using the CRC tissue samples. In conclusion, we confirmed that the KRAS mutations identified from CRC tumor tissue samples were consistently detected in the plasma cfDNA of the three CRC patients by dPCR.

Overexpression of EGFR Protein in Bruneian Lung Cancer Patients

  • Han, Yu Hao;Hamid, Mas R.W. Abdul;Telisinghe, Pemasiri Upali;Hussin, Juniadah Binti Haji;Mabruk, Mohamed
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.1
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    • pp.233-237
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    • 2015
  • Background: Lung cancer is the leading cause of cancer death in Brunei Darussalam, accounting for almost 20% of the total. The epidermal growth factor receptor (EGFR) is a member of the erbB family of tyrosine kinase receptor proteins, which includes c-erbb2(HER2/neu), erb-B3, and erb-B4. EGFR overexpression is found in a third of all epithelial cancers, often associated with a poor prognosis. Materials and Methods: Protein expression of EGFR in 27 cases of lung cancer tissue samples and 9 cases of normal lung tissue samples was evaluated using an immunohistochemical approach. Results: The results demonstrated significant increase and overexpression of EGFR in Bruneian lung cancer tissue samples in comparison to normal lung tissue. However, there was no significant relationship between clinicopathologic variables (age and sex) of patients and EGFR protein expression. Conclusions: EGFR is overexpressed in Bruneian lung cancer patient tissue samples in comparison to normal lung tissue samples. This may indicate that EGFR protein over expression plays an important role in the genesis of this type of cancer in Brunei Darussalam.

Application of X-ray Computer Tomography (CT) in Cattle Production

  • Hollo, G.;Szucs, E.;Tozser, J.;Hollo, I.;Repa, I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.12
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    • pp.1901-1908
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    • 2007
  • The aim of this series of experiments was to examine the opportunity for application of X-ray computer tomography (CT) in cattle production. Firstly, tissue composition of M. longissimus dorsi (LD) cuts between the $11-13^{th}$ ribs (in Exp 1. between the $9-11^{th}$ ribs), was determined by CT and correlated with tissue composition of intact half carcasses prior to dissection and tissue separation. Altogether, 207 animals of different breeds and genders were used in the study. In Exp. 2 and 3, samples were taken from LD cuts, dissected and chemical composition of muscle homogenates was analysed by conventional procedures. Correlation coefficients were calculated among slaughter records, tissues in whole carcasses and tissue composition of rib samples. Results indicated that tissue composition of rib samples determined by CT closely correlated with tissue composition results by dissection of whole carcasses. The findings revealed that figures obtained by CT correlate well with the dissection results of entire carcasses (meat, bone, fat). Close three-way coefficients of correlation (r = 0.80-0.97) were calculated among rib eye area, volume of cut, pixel-sum of adipose tissue determined by CT and intramuscular fat or adipose tissue in entire carcasses. Estimation of tissue composition of carcasses using equations including only CT-data as independent variables proved to be less reliable in prediction of lean meat and bone in carcass ($R^2 = 0.51-0.86$) than for fat (($R^2 = 0.83-0.89$). However, when cold half carcass weight was also included in the equation, the coefficient of determination exceeded $R^2 = 0.90$. In Exp. 3 tissue composition of rib samples by CT were compared to the results of EUROP carcass classification. Findings revealed that CT analysis has higher predictive value in estimation of actual tissue composition of cattle carcasses than EUROP carcass classification.

A Basic Study on the Variation of Temperature Characteristics for Attenuation Coefficient and Sound Velocity in Biological Tissues

  • Park, Heung-Ho
    • Journal of Biomedical Engineering Research
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    • v.14 no.3
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    • pp.273-282
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    • 1993
  • This study is concerned with the temperature dependence characteristics of ultrasound parameters in biological tissues, which are basic on the noninvasive deep body temperature estimation. Used parameters are ultrasonic attenuation coefficient and sound velocity In order to accomplishment our purpose, several signal processing methods were used. Attenua4iorl coefficient was estimated by spectral difference method and sound velocity was estimated by P-P method. And we also examined these methods through a series of IN VITRO experi mentis that used tissue-mimicking phantom samples and biological tissue samples. In order to imitate the biological soft tissue two kinds of phantom samples are used, one is agar phantom sample which is composed of agar, graphite, N-propyl alcohol and distilled water, and the other is fat phantom sample which is composed of pure animal fat. And the ultrasound transmission mode and reflection mode experiments are performed on the pig's spleen, kidney and fat. As a result, it is found that the temperature characteristics are uniform in case of phan- tom samples but not in biological tissues because of complicate wave propagation within them. Consequently, the possibility of temperature measurement using ultrasound on biological tissue is confirmed and its results may contribute to the establishment of reference values of internal temperature measurement of biological tissues.

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Expression of Extracellular Superoxide Dismutase Protein in Diabetes

  • Kim, Chul Han
    • Archives of Plastic Surgery
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    • v.40 no.5
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    • pp.517-521
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    • 2013
  • Background Diabetes is characterized by chronic hyperglycemia, which can increase reactive oxygen species (ROS) production by the mitochondrial electron transport chain. The formation of ROS induces oxidative stress and activates oxidative damage-inducing genes in cells. No research has been published on oxidative damage-related extracellular superoxide dismutase (EC-SOD) protein levels in human diabetic skin. We investigated the expression of EC-SOD in diabetic skin compared with normal skin tissue in vivo. Methods The expression of EC-SOD protein was evaluated by western blotting in 6 diabetic skin tissue samples and 6 normal skin samples. Immunohistochemical staining was also carried out to confirm the EC-SOD expression level in the 6 diabetic skin tissue samples. Results The western blotting showed significantly lower EC-SOD protein expression in the diabetic skin tissue than in the normal tissue. Immunohistochemical examination of EC-SOD protein expression supported the western blotting analysis. Conclusions Diabetic skin tissues express a relatively small amount of EC-SOD protein and may not be protected against oxidative stress. We believe that EC-SOD is related to the altered metabolic state in diabetic skin, which elevates ROS production.

Expression of Survivin and Caspase 3 in Oral Squamous Cell Carcinoma and Peritumoral Tissue

  • Li, S.X.;Chai, L.;Cai, Z.G.;Jin, L.J.;Chen, Y.;Wu, H.R.;Sun, Z.
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5027-5031
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    • 2012
  • This study was conducted to investigate the expression of survivin and caspase 3 in oral squamous cell carcinoma and peritumoral tissue, and possible pathogenesis mechanisms. We used ELISA and western blotting to detect the protein expression levels of survivin and caspase 3 in tissue. In situ hybridization and real-time PCR were applied to assess mRNA expression levels. In this study, 13 tumor samples and 13 peritumoral tissue samples were collected from oral squamous cell carcinoma patients and 10 normal tissue samples obtained from patients without tumor. The result showed that the protein and mRNA expression of survivin in carcinoma was the highest among three types of tissue; following was that in peritumoral tissue. No difference in caspase 3 zymogen between peritumoral tissue and normal tissue could be found, while it was evidently decreased in carcinoma tissue. Activated caspase 3 was detected in normal tissue but could not be identified in peritumoral or carcinoma tissue. Our results indicate that the expression of survivin is apparently elevated in tumoral and peritumoral tissue. Expression of activated caspase 3 was not detected in tumoral tissue and the expression of caspase 3 zymogen was decreased in tumoral tissue. Our findings suggest that survivin may inhibit both synthesis and activation of caspase 3, hence inhibiting cell apoptosis and facililitating eventual development of oral squamous cell carcinoma.

The expressions of C-reactive protein and macrophage colony-stimulating factor in gingival tissue of human chronic periodontitis with hypertension (고혈압을 동반한 만성 치주염 환자의 치은 조직에서 C-reactive protein과 macrophage colony-stimulating factor의발현)

  • Kim, Chul-Woo;Park, Jin-Woo;Suh, Jo-Young;Cho, Je-Yoel;Lee, Jae-Mok
    • Journal of Periodontal and Implant Science
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    • v.39 no.4
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    • pp.391-398
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    • 2009
  • Purpose: The purpose of this study was to quantify and compare the expressions of CRP and M-CSF in the gingival tissues of the patients with chronic periodontitis associated to hypertension. Methods: Gingival tissue samples were obtained during periodontal surgery or tooth extraction. Clinically healthy gingival tissue samples from systemically healthy 12 patients were categorized as group 1 (n=12). Inflammatory gingival tissue samples from patients with chronic periodontitis were categorized as group 2 (n=12). Inflammatory gingival tissue samples from patients with chronic periodontitis associated with hypertension were categorized as group 3 (n=12). Tissue samples were prepared and analyzed by Western blotting. The quantification of CRP and M-CSF were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. Results: There were significant differences between group 1 and group 2 and between group 1 and group 3 in both CRP and M-CSF. The differences between group 2 and group 3 were not statistically significant in both proteins. However, the expression levels of CRP and M-CSF in hypertensive inflammatory gingiva showed increased tendency compared to non-hypertensive inflammatory gingiva. Conclusions: It is suggested that CRP and M-CSF might be used as inflammatory and bone resorption markers in periodontal diseased tissue. It is assumed that hypertension may be associated with the progression of periodontal inflammation and alveolar bone resorption.

Isolation and characterization of Brucella abortus isolates from wildlife species in South Korea

  • Truong, Quang Lam;Kim, Kiju;Kim, Jong-Taek;Her, Moon;Jung, Suk-Chan;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.56 no.3
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    • pp.147-153
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    • 2016
  • A total of 782 blood and 465 tissue samples from 1,039 wild animals and 127 dairy goats were collected from January 2011 to December 2013 in 10 provinces of South Korea and tested for the presence of brucellosis. The Rose Bengal test revealed that 8.0% (52/650) of the serum samples were seropositive, while 4.2% (33/782) of the serum samples were positive for Brucella antibodies by competitive enzyme-linked immunosorbent assay. Of the 650 sera examined, only 16 (2.5%) were positive by both serological tests. Direct polymerase chain reaction (PCR) assay using B4/B5 primers for Brucella abortus (BCSP31) revealed the prevalence of Brucella to be 26.5% (129/487) in blood samples and 21% (98/465) in tissue samples while, 16S rRNA PCR detected Brucella DNA in 6.8% (33/487) and 2.6% (12/465) in blood and tissue samples, respectively. Of PCR-positive samples, only 6.2% (30/487) of blood samples and 2.4% (11/465) of tissue samples were found to be positive by both BCSP31 and 16S rRNA PCRs. However, Brucella strains were isolated by blood culture from only two out of 487 blood samples (0.4%). This characterization and identification of pathogenic Brucella isolates is the first to clearly indicate that the organisms were Brucella abortus biovar 1.

Enzyme Immunoassay for the Sulfamethazine Residues in Pork Tissue

  • Park, Jun-Hong;Lim, Yoon-Kyu
    • Journal of Food Hygiene and Safety
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    • v.11 no.4
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    • pp.287-290
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    • 1996
  • To control the maximum residue level (MRL) for sulfamethazine (SMZ) residues in pork tissue, a microbial inhibition method is a regulatory screening assay method in Korea. Microwell plate-based competitive enzyme immunoassay (ELISA) kit is avalable for routine screening of SMZ residues in pork tissue. One ELISA kit is evaluated. Phosphate buffer extracts of samples fortified with SMZ at 0, 1, 5, and 10 ng/g were used in a recovery test of the kit. Market pork samples were assayed by the kit. Recovery of sulfamethazine was 104% at 10 ng/g. Intraassay variations and interassay variations for the kit were 7.70% and 5.76%, respectively. Concentration causing 50% inhibition of color development compared with blanks was 16.4ng. The violative pork samples with over MRL (0.1 $\mu\textrm{g}$/g) was 4 of 32 cases (12.5%) by used ELISA kit. This result indicates a possibility of the ELISA kit for screening test of SMZ residues in pork tissue, and still needs a comfirmatory assay for mandatory purposes.

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