• Title/Summary/Keyword: Molecular techniques

Search Result 882, Processing Time 0.026 seconds

Cytotoxic effects of different self-adhesive resin cements: Cell viability and induction of apoptosis

  • Sismanoglu, Soner;Demirci, Mustafa;Schweikl, Helmut;Ozen-Eroglu, Gunes;Cetin-Aktas, Esin;Kuruca, Serap;Tuncer, Safa;Tekce, Neslihan
    • The Journal of Advanced Prosthodontics
    • /
    • v.12 no.2
    • /
    • pp.89-99
    • /
    • 2020
  • PURPOSE. The effects of four different self-adhesive resin cement materials on cell viability and apoptosis after direct and indirect exposure were evaluated using different cell culture techniques. MATERIALS AND METHODS. Self-adhesive cements were applied to NIH/3T3 mouse fibroblasts by the extract test method, cell culture inserts, and dentin barrier test method. After exposure periods of 24 h and 72 h, the cytotoxicity of these self-adhesive materials was evaluated using the MTT assay (viability) and the Annexin-V-FITC/PI staining (apoptosis). RESULTS. The lowest cell viability was found in cells exposed to BeautiCem SA for 24 h in the extract test method. Cell viability was reduced to 70.6% compared to negative controls. After the 72 h exposure period, viability rate of cell cultures exposed to BeautiCem SA decreased more than 2- fold (29.5%) while cells exposed to RelyX U200 showed the highest viability rate of 71.4%. In the dentin barrier test method, BeautiCem SA induced the highest number of cells in apoptosis after a 24 h exposure (4.1%). Panavia SA Cement Plus was the material that caused the lowest number of cells in apoptosis (1.5%). CONCLUSION. The used self-adhesive cements have showed different cytotoxic effects based on the evaluation method. As exposure time increased, the materials showed more cytotoxic and apoptotic effects. BeautiCem SA caused significantly more severe cytotoxic and apoptotic effects than other cements tested. Moreover, cements other than BeautiCem SA have caused necrotic cell death rather than apoptotic cell death.

Structural Bioinformatics Analysis of Disease-related Mutations

  • Park, Seong-Jin;Oh, Sang-Ho;Park, Dae-Ui;Bhak, Jong
    • Genomics & Informatics
    • /
    • v.6 no.3
    • /
    • pp.142-146
    • /
    • 2008
  • In order to understand the protein functions that are related to disease, it is important to detect the correlation between amino acid mutations and disease. Many mutation studies about disease-related proteins have been carried out through molecular biology techniques, such as vector design, protein engineering, and protein crystallization. However, experimental protein mutation studies are time-consuming, be it in vivo or in vitro. We therefore performed a bioinformatic analysis of known disease-related mutations and their protein structure changes in order to analyze the correlation between mutation and disease. For this study, we selected 111 diseases that were related to 175 proteins from the PDB database and 710 mutations that were found in the protein structures. The mutations were acquired from the Human Gene Mutation Database (HGMD). We selected point mutations, excluding only insertions or deletions, for detecting structural changes. To detect a structural change by mutation, we analyzed not only the structural properties (distance of pocket and mutation, pocket size, surface size, and stability), but also the physico-chemical properties (weight, instability, isoelectric point (IEP), and GRAVY score) for the 710 mutations. We detected that the distance between the pocket and disease-related mutation lay within $20\;{\AA}$ (98.5%, 700 proteins). We found that there was no significant correlation between structural stability and disease-causing mutations or between hydrophobicity changes and critical mutations. For large-scale mutational analysis of disease-causing mutations, our bioinformatics approach, using 710 structural mutations, called "Structural Mutatomics," can help researchers to detect disease-specific mutations and to understand the biological functions of disease-related proteins.

A novel pattern recognition protein of the Chinese oak silkmoth, Antheraea pernyi, is involved in the pro-PO activating system

  • Wang, Xialu;Zhang, Jinghai;Chen, Ying;Ma, Youlei;Zou, Wenjun;Ding, Guoyuan;Li, Wei;Zhao, Mingyi;Wu, Chunfu;Zhang, Rong
    • BMB Reports
    • /
    • v.46 no.7
    • /
    • pp.358-363
    • /
    • 2013
  • In this paper, we firstly reported a C-type lectin cDNA clone of 1029 bps from the larvae of A. Pernyi (Ap-CTL) using PCR and RACE techniques. The full-length cDNA contains an open reading frame encoding 308 amino acid residues which has two different carbohydrate-recognition domains (CRDs) arranged in tandem. To investigate the biological activities in the innate immunity, recombinant Ap-CTL was expressed in E. coli with a 6-histidine at the amino-terminus (Ap-rCTL). Besides acted as a broad-spectrum recognition protein binding to a wide range of PAMPs and microorganisms, Ap-rCTL also had the ability to recognize and trigger the agglutination of bacteria and fungi. In the proPO activation assay, Ap-rCTL specifically restored the PO activity of hemolymph blocked by anti-Ap-rCTL antibody in the presence of different PAMPs or microorganisms. In summary, Ap-rCTL plays an important role in insect innate immunity as an pattern recognition protein.

Epidemiological Investigation and Antibiotic Sensitivity of Salmonellosis in Goats at the Selected Areas of Bangladesh

  • Saha, Gobindha Kumar;Paul, Ashit Kumar;Abdussamad, Abdussamad;Islam, M. Ariful;Khan, M. Shahidur Rahman
    • Journal of Embryo Transfer
    • /
    • v.28 no.4
    • /
    • pp.337-342
    • /
    • 2013
  • Salmonellosis is one of the life-threating diseases of goat in Bangladesh. Therefore, the present study was designed to study the prevalence of Salmonellosis, and isolation and characterizations of the Salmonella spp. from apparently healthy and diarrheic goat. A total of 47 faces samples were collected from selected place and cultured onto different prescribed medium to isolate it. In this study, 12.76% (6/47) samples were found to be positive for Salmonella spp. During culture on SS agar medium, all of the Salmonella isolates produced round, smooth, opaque, translucent and black color colonies on SS agar media. All of the isolated Salmonella spp. fermented dextrose, maltose and mannitol with production of acid and gas but did not ferment sucrose and lactose. However, these isolates had showed Indole and Voges-Proskauer test negative, Methyl-Red test positive. All of these isolates were subjected to rapid plate agglutination test with polyvalent "O" (Poly 'O') and polyvalent "H" (poly 'H') antisera where positive agglutination were observed. They were highly sensitive to ciprofloxacin, spiramycin and gentamycin; moderately sensitive to oxytetracyline, streptomycin and amoxicillin; less sensitive to sulphamethoxazole and resistant to penicillin-G. Based on the present findings, it may be concluded that the investigated Salmonella spp. from goats might be S. typhimurium, S. enteritidis, S. brandenburg, S. salford, S. newbrunswick, S. newport or S. dublin. Further study will be needed, therefore it requires further characterization using other serological and molecular techniques.

Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel;Davies, Cristina Veronica;Solda, Carina Alejandra;Corrado, Maria Belen;Fernandez, Maria Veronica
    • Microbiology and Biotechnology Letters
    • /
    • v.48 no.2
    • /
    • pp.193-204
    • /
    • 2020
  • Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

Molecular Cloning and Alternative Splicing of Growth Hormone Transcripts in Greenling, Hexagrammos otakii (쥐노래미 (Hexagrammos otakii) 성장호르몬 cDNA유전자의 염기서열 변이 및 발현 특성)

  • Nam Yoon Kwon;Kim Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.35 no.6
    • /
    • pp.676-681
    • /
    • 2002
  • Different types of transcripts encoding growth hormone (GH) were identified from cDNA libraries constructed with pituitaries of a marine fish species, greenling (Hexagrammos otakii). GH-homologous cDNA clones were isolated using the high-density filter hybridization and the expressed sequence tag techniques. Of 39 full-length positive cDNA clones, 31 clones ($79\%$) displayed an identical sequence, however, remaining 8 clones exhibited several polymorphisms in their sequences including (1) the length and sequence variability in the 5' upstream region, (2) insertional sequences in open reading frame, and (3) deletion and/or single nucleotide polymorphism in the untranslated 3' region. Based on RT-PCT and RNA dot blot analyses, these transcripts were proven to be expressed in a pituitary-specific manner.

Culture-Independent Methods of Microbial Community Structure Analysis and Microbial Diversity in Contaminated Groundwater with Major Pollutants (주요 오염물질로 오염된 지하수에서 미생물의 무배양식 군집분석방법과 미생물상에 대한 조사방법 연구)

  • Kim Jai-Soo
    • Journal of Soil and Groundwater Environment
    • /
    • v.11 no.3
    • /
    • pp.66-77
    • /
    • 2006
  • This review inquired the recently applied molecular biological and biochemical methods analyzing the microbial community structure of groundwater and, as a result, summarized the functional or taxonomic groups of active microorganisms with major contaminants in groundwater. The development of gene amplification through PCR has been possible to figure out microbial population and identification. Active microbial community structures have been analyzed using a variety of fingerprinting techniques such as DGGE, SSCP, RISA, and microarray and fatty acid analyses such as PLFA and FAME, and the activity of a specific strain has been examined using FISH. Also, this review included the dominant microflora in groundwater contaminated with fuel components such as n-alkanes, BTEX, MTBE, and ethanol and chlorinated compounds such as TCE, PCE, PCB, CE, carbon tetrachloride, and chlorobenzene.

Characterization of the Rosellinia necatrix Transcriptome and Genes Related to Pathogenesis by Single-Molecule mRNA Sequencing

  • Kim, Hyeongmin;Lee, Seung Jae;Jo, Ick-Hyun;Lee, Jinsu;Bae, Wonsil;Kim, Hyemin;Won, Kyungho;Hyun, Tae Kyung;Ryu, Hojin
    • The Plant Pathology Journal
    • /
    • v.33 no.4
    • /
    • pp.362-369
    • /
    • 2017
  • White root rot disease, caused by the pathogen Rosellinia necatrix, is one of the world's most devastating plant fungal diseases and affects several commercially important species of fruit trees and crops. Recent global outbreaks of R. necatrix and advances in molecular techniques have both increased interest in this pathogen. However, the lack of information regarding the genomic structure and transcriptome of R. necatrix has been a barrier to the progress of functional genomic research and the control of this harmful pathogen. Here, we identified 10,616 novel full-length transcripts from the filamentous hyphal tissue of R. necatrix (KACC 40445 strain) using PacBio single-molecule sequencing technology. After annotation of the unigene sets, we selected 14 cell cycle-related genes, which are likely either positively or negatively involved in hyphal growth by cell cycle control. The expression of the selected genes was further compared between two strains that displayed different growth rates on nutritional media. Furthermore, we predicted pathogen-related effector genes and cell wall-degrading enzymes from the annotated gene sets. These results provide the most comprehensive transcriptomal resources for R. necatrix, and could facilitate functional genomics and further analyses of this important phytopathogen.

keV and MeV Ion Beam Modification of Polyimide Films

  • Lee, Yeonhee;Seunghee Han;Song, Jong-Han;Hyuneui Lim;Moojin Suh
    • Proceedings of the Korean Vacuum Society Conference
    • /
    • 2000.02a
    • /
    • pp.170-170
    • /
    • 2000
  • Synthetic polymers such as polyimide, polycarbonate, and poly(methyl methacrylate) are long chain molecules which consist of carbon, hydrogen, and heteroatom linked together chemically. Recently, polymer surface can be modified by using a high energy ion beam process. High energy ions are introduced into polymer structure with high velocity and provide a high degree of chemical bonding between molecular chains. In high energy beam process the modified polymers have the highly crosslinked three-dimensionally connected rigid network structure and they showed significant improvements in electrical conductivity, in hardness and in resistance to wear and chemicals. Polyimide films (Kapton, types HN) with thickness of 50~100${\mu}{\textrm}{m}$ were used for investigations. They were treated with two different surface modification techniques: Plasma Source Ion Implantation (PSII) and conventional Ion Implantation. Polyimide films were implanted with different ion species such as Ar+, N+, C+, He+, and O+ with dose from 1 x 1015 to 1 x 1017 ions/cm2. Ion energy was varied from 10keV to 60keV for PSII experiment. Polyimide samples were also implanted with 1 MeV hydrogen, oxygen, nitrogen ions with a dose of 1x1015ions/cm2. This work provides the possibility for inducing conductivity in polyimide films by ion beam bombardment in the keloelectronvolt to megaelectronvolt energy range. The electrical properties of implanted polyimide were determined by four-point probe measurement. Depending on ion energy, doses, and ion type, the surface resistivity of the film is reduced by several orders of magnitude. Ion bombarded layers were characterized by Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS), XPS, and SEM.

  • PDF

Comparison of the volatile flavor compounds in different tobacco types by different extraction methods (추출방법에 따른 잎담배 종류별 휘발성 향기성분 특성비교)

  • Lee, Jang-Mi;Lee, Jeong-Min;Lee, Chang-Gook;Bock, Jin-Young;Hwang, Keon-Joong
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.32 no.2
    • /
    • pp.77-87
    • /
    • 2010
  • Traditional simultaneous distillation extraction(SDE) and solid-phase micro extraction(SPME) methods using GC/MS were compared for their effectiveness in the extraction of volatile flavor compounds from different tobacco leaves types(flue-cured, burley, oriental). The major volatile flavor compounds of flue-cured and burley tobacco were similar such as neophytadiene, solanone, megastigmatrienone isomers, ${\beta}$-damascenone and ${\beta}$-ionone. On the other hand, volatile flavor compounds such as norambreinolide, sclareolide were specifically identified in oriental tobacco. Each method was used to evaluate the responses of some analytes from real samples and standards in order to provide sensitivity comparisons between two techniques. Among three types of SPME fibers such as PDMS(Polydimethylsiloxane), PA(Polyacrylate) and PDMS/DVB (Polydimethylsiloxane/Divinylbenzene) which were investigated to determine the selectivity and adsorption efficiency, PDMS/DVB fiber was selected for the extractions of the volatile flavor compounds due to its effectiveness. The qualitative analysis showed that the total amount of volatile flavor compounds in SDE method(130 species) was much more than that in SPME method(85 species). SPME method was more efficient for all the highly volatile compounds than SDE method, but on the other hand, low-volatile compounds such as fatty acids or high-molecular hydrocabons were detected in SDE method. SPME method based on a short-time sampling can be adjusted to favor a selected group compounds in tobacco. Furthermore this results could be used to estimate the aroma characteristics of cigarette blending by using a different type of tobacco with more effectiveness and convenience.