• 제목/요약/키워드: in vitro techniques

검색결과 355건 처리시간 0.027초

Structural Bioinformatics Analysis of Disease-related Mutations

  • Park, Seong-Jin;Oh, Sang-Ho;Park, Dae-Ui;Bhak, Jong
    • Genomics & Informatics
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    • 제6권3호
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    • pp.142-146
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    • 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.

Permanent Mycoplasma Removal Removel from Tissue Culture Cells: A Genetic Approach

  • Motr, Gabriele;Preininger, Alexandra;Himmelspach, Michele;Plaimauer, Barbara;Arbesser, Christine;York, Heinz;Dorner, Friedrich;Schlokat, Use
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제5권2호
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    • pp.84-91
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    • 2000
  • Mycopasma contamination of tissue culture cells easily evades detection and, thus, represents a continous therat to cell biologists. In case where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplacma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains ; however, cell associated mycoplasma are often protected from antibiotics at concentrations shown to be effective in vitro. Antibiotic concentrations high enough to be lethal to cell as sociated mycoplasmas frequently are also detrimentrations to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistanct even to high concentrations of the antibiotis applied. Hare, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes thens limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent, Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycopalsma while leaving the host cells unharmed. Upon successful mycoplasma eradicated, cultvation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibillity to the toxic agent. Cressation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chrosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.

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Candida non albicans with a High Amphotericin B Resistance Pattern Causing Candidemia among Cancer Patients

  • Kalantar, Enayatollah;Assadi, Mojan;Pormazaheri, Helen;Hatami, Shiva;Barari, Maryam Agha;Asgari, Esfandiar;Mahmoudi, Elaheh;Kabir, Kourosh;Marashi, Seyed Mahmoud Amin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권24호
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    • pp.10933-10935
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    • 2015
  • Background: Many scientists have reported Candida species to be of great concern because of the high frequency that they colonize and infect human hosts, particularly cancer patients. Moreover, in the last decades Candida species have developed resistance to many antifungal agents. Based on this, we aimed to identify and determine the prevalence of Candida spp from blood culture bottles among cancer patients and their antifungal resistance pattern. Materials and Methods: From the blood culture bottles isolation and identification of the Candida spp were performed by conventional microbiological techniques. The in vitro antibiotic resistance pattern of the isolates was determined by CLSI guidelines. Genomic DNA was isolated and amplified. Each gene was separated by agar gel electrophoresis. Results: Identification of Candida spp was based on the presence of yeast cells in direct examination, culture and DNA extraction. Of the 68 blood samples collected during the study period (April 2013 to October 2013), five (7.35%) were positive for the presence of Candida spp, 2 (40%) of which were identified as Candida albicans and 3 (60%) were Candida non-albicans. Conclusions: High resistance to amphotricin B was observed among all the Candida non-albicans isolates. Regular investigations into antifungal resistance will help us to get an updated knowledge about their antibiotic resistance pattern which may help the physician in selecting the antibiotics for empirical therapy.

Ceromer (Targis)에 의한 법랑질 마모에 관한 연구 (AN IN-VITRO WEAR STUDY OF CEROMER AND HUMAN ENAMEL)

  • 김정민;이선형;양재호
    • 대한치과보철학회지
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    • 제38권4호
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    • pp.438-445
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    • 2000
  • The ideal restorative material should mimic the properties of the tissues it replaces. Dental composite resins have been used widely as restorative materials due to its advantages such as excellent esthetics and ease of manipulation. But inadequate wear resistance has been a major factor limiting the use of composite restorative materials. Improved manufacturing techniques have allowed the development of hybrid composites, with a greater percentage volume filler loading, which have improved physical and mechanical properties. However they are lacking in the study of wear resistance. The purpose of this study was to evaluate the wear of human enamel against ceromer by the use of a pin-on-disk type wear testers. Discs of ceromer(Targis ; lvoclar Vivadent, Amherst. NY) and discs of type III gold alloy as a control were used f9r test specimens. Intact cusp of premolar and molar were used for enamel specimens. The wear of enamel was determined by weigh-ing the cusp before and after each test, and the weight converted to volumes by average densi-ty of enamel. Surface profilometer was used to quantify wear of the ceromer and gold specimens. Vicker's hardness tester was used to evaluate the surface hardness of test specimens. The SEM was used to evaluate the wear surfaces The results were as follows; 1. Ceromer produced less enamel wear than gold(p<0.05) 2. The wear volume of ceromer was greater than that of gold(p<0.01) 3. The hardess of ceromer was lower than that of gold, but there was no correlation between the hardness and wear of the ceromer and gold. 4. SEM analysis revealed that there were many voids and microcracks in the wear tract of ceromer In gold group, many minute V-shaped grooves were examined.

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레이저를 이용한 임프란트 이차수술시 온도변화에 관한 실험적 연구 (TEMPERATURE CHANGES OF IMPLANT SURFACE IN SECOND STAGE SURGERY WITH DETAL LASER : IN VITRO STUDY)

  • 안현정;김현철;최병갑;송언희;김래경
    • 대한치과보철학회지
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    • 제37권2호
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    • pp.256-268
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    • 1999
  • Submerged implants require secondary surgical uncovering of implants after healing period of 3-6 months. In surgical methods, there are surgical scalpel, tissue punch, electro-surgical, and laser-used uncovering, and so forth The objectives of this study are investigation and assessment of 1) thermal change in clinical application for uncovering of HA-coated implant and pure titanium implant irradiated by pulsed Nd-YAG, $CO_2$, and Er-YAG laser. 2) surface change of cover screws aaer irradiation using laser energy. The temperature of apex & side wall of implants were recorded at 10sec, 20sec, 30sec after 30sec irradiation to implant healing screw; 1) pulsed Nd-YAG laser; 2W, 20pps, contact mode 2) $CO_2$ laser; water-infused & non-water infused state, 2.5-3.5W, contibuous mode, noncontact mode 3) $CO_2$ laser ; non-water infused state, 3W, superpulse, noncontact. mode 4) Er-YAG laser; (1) non-water infused state, 10pps, 60mj, contact mode (2) water-infused state, 10pps, 60mj, 80mj, 101mj, contact mode. According to the results of this study, pulsed Nd-YAG laser is not indicated because of increased thermal change and pitting of metal surface of implant cover screw. By contrast, $CO_2$ laser & Er-YAG laser are presumed to indicate because of narrow range of thermal change & near abscence of thermal damage of metal surface. Dental laser is thought to be much helpful to surgical procedure when it is used as optimal power and time condition considering characteristics and indications of each laser. Further research is needed to verify that these techniques are safe and beneficial to implant success.

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Efficacy of various cleaning solutions on saliva-contaminated zirconia for improved resin bonding

  • Kim, Da-Hye;Son, Jun-Sik;Jeong, Seong-Hwa;Kim, Young-Kyung;Kim, Kyo-Han;Kwon, Tae-Yub
    • The Journal of Advanced Prosthodontics
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    • 제7권2호
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    • pp.85-92
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    • 2015
  • PURPOSE. This study aimed to investigate the efficacy of cleaning solutions on saliva-contaminated zirconia in comparison to air-abrasion in terms of resin bonding. MATERIALS AND METHODS. For saliva-contaminated air-abraded zirconia, seven cleaning methods)-no contamination (NC), water-spray rinsing (WS), additional air-abrasion (AA), and cleaning with four solutions (Ivoclean [IC]; 1.0 wt% sodium dodecyl sulfate [SDS], 1.0 wt% hydrogen peroxide [HP], and 1.0 wt% sodium hypochlorite [SHC])-were tested. The zirconia surfaces for each group were characterized using various analytical techniques. Three bonded resin (Panavia F 2.0) cylinders (bonding area: $4.5mm^2$) were made on one zirconia disk specimen using the Ultradent jig method [four disks (12 cylinders)/group; a total of 28 disks]. After 5,000 thermocycling, all specimens were subjected to a shear bond strength test with a crosshead speed of 1.0 mm/minute. The fractured surfaces were observed using an optical and scanning electron microscope (SEM). RESULTS. Contact angle measurements showed that groups NC, AA, IC, and SHC had hydrophilic surfaces. The X-ray photoelectron spectroscopy (XPS) analysis showed similar elemental distributions between group AA and groups IC and SHC. Groups IC and SHC showed statistically similar bond strengths to groups NC and AA (P>.05), but not groups SDS and HP (P<.05). For groups WS, SDS, and HP, blister-like bubble formations were observed on the surfaces under SEM. CONCLUSION. Within the limitations of this in vitro study, some of the cleaning solutions (IC or SHC) were effective in removing saliva contamination and enhancing the resin bond strength.

Influence of heat treatment on the microstructure and the physical and mechanical properties of dental highly translucent zirconia

  • Dimitriadis, Konstantinos;Sfikas, Athanasios Konstantinou;Kamnis, Spyros;Tsolka, Pepie;Agathopoulos, Simeon
    • The Journal of Advanced Prosthodontics
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    • 제14권2호
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    • pp.96-107
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    • 2022
  • PURPOSE. Microstructural and physico-mechanical characterization of highly translucent zirconia, prepared by milling technology (CAD-CAM) and repeated firing cycles, was the main aim of this in vitro study. MATERIALS AND METHODS. Two groups of samples of two commercial highly-translucent yttria-stabilized dental zirconia, VITA YZ-HTWhite (Group A) and Zolid HT + White (Group B), with dimensions according to the ISO 6872 "Dentistry - Ceramic materials", were prepared. The specimens of each group were divided into two subgroups. The specimens of the first subgroups (Group A1 and Group B1) were merely the sintered specimens. The specimens of the second subgroups (Group A2 and Group B2) were subjected to 4 heat treatment cycles. The microstructural features (microstructure, density, grain size, crystalline phases, and crystallite size) and four mechanical properties (flexural strength, modulus of elasticity, Vickers hardness, and fracture toughness) of the subgroups (i.e. before and after heat treatment) were compared. The statistical significance between the subgroups (A1/A2, and B1/B2) was evaluated by the t-test. In all tests, P values smaller than 5% were considered statistically significant. RESULTS. A homogenous microstructure, with no residual porosity and grains sized between 500 and 450 nm for group A and B, respectively, was observed. Crystalline yttria-stabilized tetragonal zirconia was exclusively registered in the X-ray diffractograms. The mechanical properties decreased after the heat treatment procedure, but the differences were not statistically significant. CONCLUSION. The produced zirconia ceramic materials can be safely (i.e., according to the ISO 6872) used in extensive fixed prosthetic restorations, such as substructure ceramics for three-unit prostheses involving the molar restoration and substructure ceramics for prostheses involving four or more units. Consequently, milling technology is an effective manufacturing technology for producing zirconia substructures for dental fixed all-ceramic prosthetic restorations.

The effect of individualization of fiberglass posts using bulk-fill resin-based composites on cementation: an in vitro study

  • Lins, Rodrigo Barros Esteves;Cordeiro, Jairo Matozinho;Rangel, Carolina Perez;Antunes, Thiago Bessa Marconato;Martins, Luis Roberto Marcondes
    • Restorative Dentistry and Endodontics
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    • 제44권4호
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    • pp.37.1-37.10
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    • 2019
  • Objectives: This study evaluated the bond strength of various fiberglass post cementation techniques using different resin-based composites. Materials and Methods: The roots from a total of 100 bovine incisors were randomly assigned to 5 treatment groups: G1, post + Scotchbond Multi-Purpose (SBMP) + RelyX ARC luting agent; G2, relined post (Filtek Z250) + SBMP + RelyX ARC; G3, individualized post (Filtek Z250) + SBMP; G4, individualized post (Filtek Bulk-Fill) + SBMP; G5, individualized post (Filtek Bulk-Fill Flow) + SBMP. The samples were subjected to the push-out (n = 10) and pull-out (n = 10) bond strength tests. Data from the push-out bond strength test were analyzed using 2-way analysis of variance (ANOVA) with the Bonferroni post hoc test, and data from the pull-out bond strength test were analyzed using 1-way ANOVA. Results: The data for push-out bond strength presented higher values for G2 and G5, mainly in the cervical and middle thirds, and the data from the apical third showed a lower mean push-out bond strength in all groups. No significant difference was noted for pull-out bond strength among all groups. The most frequent failure modes observed were adhesive failure between dentine and resin and mixed failure. Conclusions: Fiberglass post cementation using restorative and flowable bulk-fill composites with the individualization technique may be a promising alternative to existing methods of post cementation.

X-정자와 Y-정자의 분류에 관한 연구 IV. Sephadex Gel여과법과 Percoll중층원심분류법의 병용에 의한 우 정자의 분류 (Separation of X-and Y-Bearing Spermatozoa IV. Separtion of bull spermatozoa by the combination of density Gradient Centrifugation and Sephadex Gel filtration)

  • 이주영;정길생;김종배
    • 한국가축번식학회지
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    • 제12권3호
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    • pp.141-147
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    • 1988
  • These experiments were carried out to develop new techniques for in vitro separation of x-and Y-bearing spermatozoa. The results obtained in these experiments were summarized as follows: 1. Following centrifugation of discontinuous percoll density gradient, populatin of spermatozoa increased progressively from low to high density. The highest concentration of spermatozoa was observed at the 4th fraction which included 36.6% of spermatozoa. 2. As increasing percoll concentration, the higher motility index was obtained and the highest motility index(74.2) was obtained at the 5th fraction. 3. The percentage of B-body bearing spermatozoa following percoll density gradient centrifugation was decreased from 39.7% to 25.6%. 4. The sperm population following chromatography by sephadex gel and percoll density gradient centrifugation was decreased in 1st, 5th and 6th fractions but the reverse was turn for 2nd, 3rd and 7th fractions, and the highest sperm concentration was observed at the 7th fraction which included 37.4% of spermatozoa. 5. Motility index of spermatozoa was increased from 77.6 to 79.4 after the sephadex gel filtration, however it was decreased at all fractions after percoll density gradient centrifugation. The lowest motility index(33.2) was obtained from the 7th fraction. 6. The rate of B-body bearing spermatozoa was shown the trend to decrease by the sephadex gel filtration and the trend was accelerated by the percoll density gradient centrifugation. The lowest percentage of B-body bearing spermatozoa, 12.0% was obtained from the 5th fraction.

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Y 염색체 특이성 DNA분리와 단일 H-Y 항체 개발에 의한 토끼의 수정란 성 감별에 관한 연구 II. PCR을 이용한 Y 염색체 특이성 DNA의 증폭에 의한 토끼 수정란의 성 감별 (Studies on Isolaton of Y-specific DNA Marker and Development of Monoclonal H-Y Antibody for Embryo Sexing in Rabbit II. Sex Determination of Rabbit Embryo by PCR Amplified Y-specific DNA)

  • 박영일;임경순;한재용;남경우;황규춘;박화춘
    • 한국가축번식학회지
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    • 제20권2호
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    • pp.89-99
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    • 1996
  • The purpose of this study was to develop the diagnosis techniques for sex determination of rabbit embryos at preimplantation stage. To detect male specific sequences using polymerase chain reaction, two genes functional on sex determination including SRY and ZFX/Y genes were targeted using multiple oligonucleotide primer sets. Three of them for conserved SRY gene were used for appropriate amplification pattern, and then only one primer set #3 proved to be most efficient, showing male-specific strong signal ofamplified sequences. Using this male specific bandsfrom human, cattle, pig and mouse, the gender of rabbit was determined. As an another system for sex determination system, amplified 910bp fragment from ZFX/Y was digested with several restriction endonuclease and showed gender specific restriction fragments only by Hinf I. Using two different system for sex identification of rabbit in this study, blind tests for 17 samples was conducted and showed identical results from two different methods. And then, amplification limit of PCR reaction for template DNA was estimated using various amounts of DNA for both SRY and ZFX/Y systems, resulted as 20pg and 800pg, respectively. With this results, test for gender identification of rabbit embryos were performed using SRY derived amplification system. From total 22 embryos selected for its developmental state 18 were identified as male embryos, showing significant difference from expected sex ratio 1:1. This biased sex ratio was interpreted as to have been caused by the fact, reported by the fact, reported by several researchers, that male embryos develop more rapidly and are more resistant against the in vitro manipulation than female embryos.

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