• Title/Summary/Keyword: in vitro detection system

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Comparison of high-resolution and standard zoom imaging modes in cone beam computed tomography for detection of longitudinal root fracture: An in vitro study

  • Taramsari, Mehran;Kajan, Zahra Dalili;Bashirzadeh, Parinaz;Salamat, Fatemeh
    • Imaging Science in Dentistry
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    • v.43 no.3
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    • pp.171-177
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    • 2013
  • Purpose: The purpose of this study was to compare the efficacy of two imaging modes in a cone beam computed tomography (CBCT) system in detecting root fracture in endodontically-treated teeth with fiber posts or screw posts by selecting two fields of view. Materials and Methods: In this study, 78 endodontically-treated single canal premolars were included. A post space was created in all of them. Then the teeth were randomly set in one of 6 artificial dental arches. In 39 of the 78 teeth set in the 6 dental arches, a root fracture was intentionally created. Next, a fiber post and a screw post were cemented into 26 teeth having equal the root fractures. High resolution (HiRes) and standard zoom images were provided by a CBCT device. Upon considering the reconstructed images, two observers in agreement with each other confirmed the presence or absence of root fracture. A McNemar test was used for comparing the results of the two modes. Results: The frequency of making a correct diagnosis using the HiRes zoom imaging mode was 71.8% and in standard zoom was 59%. The overall sensitivity and specificity in diagnosing root fracture in the HiRes mode were 71.79% and 46.15% and in the standard zoom modes were 58.97% and 33.33%, respectively. Conclusion: There were no significant differences between the diagnostic values of the two imaging modes used in the diagnosis of root fracture or in the presence of root canal restorations. In both modes, the most true-positive results were reported in the post space group.

In-Vitro Thrombosis Detection of Mechanical Valve using Artificial Neural Network (인공신경망을 이용한 기계식 판막의 생체외 모의 혈전현상 검출)

  • 이혁수;이상훈
    • Journal of Biomedical Engineering Research
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    • v.18 no.4
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    • pp.429-438
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    • 1997
  • Mechanical valve is one of the most widely used implantable artificial organs of which the reliability is so important that its failure means the death of patient. Therefore early noninvasive detection is essentially required, though mechanical valve failure with thrombosis is the most common. The objective of this paper is to detect the thrombosis formation by spectral analysis and neural network. Using microphone and amplifier, we measured the sound from the mechanical valve which is attached to the pneumatic ventricular assist device. The sound was sampled by A/D converter(DaqBook 100) and the periodogram is the main algorithm for obtaining spectrum. We made the thrombosis models using pellethane and silicon and they are thrombosis model on the valvular disk, around the sewing ring and fibrous tissue growth across the orifice of valve. The performance of the measurment system was tested firstly using 1 KHz sinusoidal wave. The measurement system detected well 1KHz spectrum as expected. The spectrum of normal and 5 kinds of thrombotic valve were obtained and primary and secondary peak appeared in each spectrum waveform. We find that the secondary peak changes according to the thrombosis model. So to distinguish the secondary peak of normal and thrombotic valve quantatively, 3 layer back propagation neural network, which contains 7, 000 input node, 20 hidden layer and 1 output was employed The trained neural network can distinguish normal and valve with more than 90% probability. As a conclusion, the noninvasive monitoring of implanted mechanical valve is possible by analysing the acoustical spectrum using neural network algorithm and this method will be applied to the performance evaluation of other implantable artificial organs.

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Validation of Methods for Isolation and Culture of Alpaca Melanocytes: A Novel Tool for In vitro Studies of Mechanisms Controlling Coat Color

  • Bai, Rui;Sen, Aritro;Yu, Zhihui;Yang, Gang;Wang, Haidong;Fan, Ruiwen;Lv, Lihua;Lee, Kyung-Bon;Smith, George W;Dong, Changsheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.430-436
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    • 2010
  • The objective of the present studies was to develop and validate a system for isolation, purification and extended culture of pigment-producing cells in alpaca skin (melanocytes) responsible for coat color and to determine the effect of alpha melanocyte stimulating hormone treatment on mRNA expression for the melanocortin 1 receptor, a key gene involved in coat color regulation in other species. Skin punch biopsies were harvested from the dorsal region of 1-3 yr old alpacas and three different enzyme digestion methods were evaluated for effects on yield of viable cells and attachment in vitro. Greatest cell yields and attachment were obtained following dispersion with dispase II relative to trypsin and trypsin-EDTA treatment. Culture of cells in medium supplemented with basic fibroblast growth factor, bovine pituitary extract, hydrocortisone, insulin, 12-O-tetradecanolphorbol-13-acetate and cholera toxin yielded highly pure populations of melanocytes by passage 3 as confirmed by detection of tyrosinase activity and immunocytochemical localization of melanocyte markers including tyrosinase, S-100 and micropthalmia-associated transcription factor. Abundance of mRNA for tyrosinase, a key enzyme in melanocyte pigment production, was maintained through 10 passages showing preservation of melanocyte phenotypic characteristics with extended culture. To determine hormonal responsiveness of cultured melanocytes and investigate regulation of melanocortin 1 receptor expression, cultured melanocytes were treated with increasing concentrations of ${\alpha}$-melanocyte stimulating hormone. Treatment with ${\alpha}$-melanocyte stimulating hormone increased melanocortin receptor 1 mRNA in a dose dependent fashion. The results demonstrated culture of pure populations of alpaca melanocytes to 10 passages and illustrate the potential utility of such cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in fiber-producing species.

Molecular Imaging Using Sodium Iodide Symporter (NIS) (Sodium Iodide Symporter (NIS)를 이용한 분자영상)

  • Cho, Je-Yoel
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.2
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    • pp.152-160
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    • 2004
  • Radioiodide uptake in thyroid follicular epithelial cells, mediated by a plasma membrane transporter, sodium iodide symporter (NIS), provides a first step mechanism for thyroid cancer detection by radioiodide injection and effective radioiodide treatment for patients with invasive, recurrent, and/or metastatic thyroid cancers after total thyroidectomy. NIS gene transfer to tumor cells may significantly and specifically enhance internal radioactive accumulation of tumors following radioiodide administration, and result in better tumor control. NIS gene transfers have been successfully performed in a variety of tumor animal models by either plasmid-mediated transfection or virus (adenovirus or retrovirus)-mediated gene delivery. These animal models include nude mice xenografted with human melanoma, glioma, breast cancer or prostate cancer, rats with subcutaneous thyroid tumor implantation, as well as the rat intracranial glioma model. In these animal models, non-invasive imaging of in vivo tumors by gamma camera scintigraphy after radioiodide or technetium injection has been performed successfully, suggesting that the NIS can serve as an imaging reporter gene for gene therapy trials. In addition, the tumor killing effects of I-131, ReO4-188 and At-211 after NIS gene transfer have been demonstrated in in vitro clonogenic assays and in vivo radioiodide therapy studies, suggesting that NIS gene can also serve as a therapeutic agent when combined with radioiodide injection. Better NIS-mediated imaging and tumor treatment by radioiodide requires a more efficient and specific system of gene delivery with better retention of radioiodide in tumor. Results thus far are, however, promising, and suggest that NIS gene transfer followed by radioiodide treatment will allow non-invasive in vivo imaging to assess the outcome of gene therapy and provide a therapeutic strategy for a variety of human diseases.

Expression Profiling of Lipopolysaccharide Target Genes in RAW264.7 Cells by Oligonucleotide Microarray Analyses

  • Huang, Hao;Park, Cheol-Kyu;Ryu, Ji-Yoon;Chang, Eun-Ju;Lee, Young-Kyun;Kang, Sam-Sik;Kim, Hong-Hee
    • Archives of Pharmacal Research
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    • v.29 no.10
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    • pp.890-897
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    • 2006
  • In inflammatory responses, induction of cytokines and other immune regulator genes in macrophages by pathogen-associated signal such as lipopolysaccharide (LPS) plays a crucial role. In this study, the gene expression profile changes by LPS treatment in the macrophage/monocyte lineage cell line RAW264.7 was investigated. A 60-mer oligonucleotide microarray of which probes target 32381 mouse genes was used. A reverse transcription-in vitro translation labeling protocol and a chemileuminescence detection system were employed. The mRNA expression levels in RAW264.7 cells treated for 6 h with LPS and the control vehicle were compared. 747 genes were up-regulated and 523 genes were down-regulated by more than 2 folds. 320 genes showing more than 4-fold change by LPS treatment were further classified for the biological process, molecular function, and signaling pathway. The biological process categories that showed high number of increased genes include the immunity and defense, the nucleic acid metabolism, the protein metabolism and modification, and the signal transduction process. The chemokine-cytokine signaling, interleukin signaling, Toll receptor signaling, and apoptosis signaling pathways involved high number of genes differentially expressed in response to LPS. These expression profile data provide more comprehensive information on LPS-target genes in RAW264.7 cells, which will be useful in comparing gene expression changes induced by extracts and compounds from anti-inflammatory medicinal herbs.

Comparison of the Sensitivity of Two Micronucleus Assays for Detection of Micronucleus Induction by Cigarette-Smoke Condensate (담배연기응축물의 소핵생성 측정시 두가지 방법간의 민감성 비교)

  • Sohn Hyung-Ok;Lee Young-Gu;Han Jung-Ho;Hur Jae-Yeon;Lee Dong-Wook;Hyun Hak-Chul;Shin Han-Jae
    • Journal of the Korean Society of Tobacco Science
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    • v.26 no.2 s.52
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    • pp.152-158
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    • 2004
  • Among short-term in vitro genotoxicity assays, micronucleus assays are rapid, inexpensive, and less labor-intensive system. We have undertaken a comparative study of sensitivity of cigarette smoke condensate(CSC) by general micronucleus(MN) assay and cytokinesis-block micronucleus(CBMN) assay. In this study, V79 Chinese hamster cells were employed to evaluate and compare the genotoxicity of CSC of Kentucky Reference Cigarette 2R4F by 2 kinds of in vitro MN assay methods. To determine the optimum concentration of cytochalasin B(CYB) to obtain the maximal number of binucleated cells for CBMN assay, triplicate cultures of growing cells were treated with CYB for 15 h. CYB treatments caused a concentration-dependent increase in cytotoxicity($1\~4{\mu}g/mL$) and proportion($0.25\~1\;{\mu}g/mL$) of binucleated cells. These data suggested that 1 ug/mL of CYB is as an optimum dose for CBMN assay in binucleated V79 cells. Short treatment(4 h) of CSC induced a micronucleated cells with a concentration-dependent response in the presence or absence of CYB, but CSC-induced MNs were weakened when S9 was present. Long treatments(19 h) of CSC also induced a significant increase MN formation with a concentration-dependent response. At a concentration of 75 ${mu}g/mL$, the MN cell frequencies of general MN assay and CBMN assay were $6.5\%\;and\;11.7\%$, respectively. Linear regression analysis revealed a good correlation in CBMN assay between a concentration of CSC and MN cell frequency. All these data indicated that CBMN assay is more sensitive to the induction CSC-induced MN than general MN assay.

QUANTITATIVE ANALYSIS OF THE ALVEOLAR BONE CHANGE BY THE DIGITAL SUBTRACTION RADIOGRAPHY (Digital subtraction radiography를 이용한 치조골 변화의 정략적 분석)

  • Ryue, Myung-Girl;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.25 no.1
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    • pp.67-75
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    • 1995
  • The progress of periodontal disease and the wound healing process after treatment result in alveolar bone bone change. So, detection of it is very important in the diagnosis and the radiograph of periodontal disease. Various effects have been made to assess the subtle alveolar bone change and digital subtraction radiography (DSR) has been reported to be the best method in evaluating it qualitatively and quantitatively. The present study was performed to estimate the detectable alveolar bone change qualitatively with digital subtraction radiography. For the in vitro study, 10 intraoral standard radiographs were taken from porcine dry mandible which a rectangular cortical bone chip of 0.1mm to 1.0mm thickness with 0.1mm increment was attached on the buccal surface. The radiographs without and with bone plates were reviewed at the same time by 10 observers and requested to detect the presence of cortical bone plates. Digital Subtraction radiograph was reviewed subsequently by using the DSR system(digital converter-256 grey-levels,DT 2851,Data Translation Co., U.S.A;IBM 386 ; CCD camera, FOTOVIX, Tamrom Co., Japan). The detectable thickness of cortical bone plate was O.4mm on the intraoral radiograph and 0.2mm on the subtaction images. For the human study, radiographs were taken from patients by using intraoral film holding device and aluminum reference wedge before and 3 month after bone graft and 1 week after osteoplasty. The grey level change was estimated in the subtraction images and calculated to aluminum equivalent thickness. The grey level of the grafted site was higher that that of healthy controls. Average grey levels of change on healthy controls were O.48mm aluminum equivalent. However, the amount of changes in grafted sites were 1.87mm aluminum thickness equivalent and in the site of osteoplasty were -1.49mm aluminum thickness equivalent. In conclusion, digital subtraction radiography was more effective in detecting as subtle change of alveolar bone than intraoral standard radiography. With the aid of quantitative analysis of digital subtraction radiography, alveolar bone resorption of apposition can be estimated during diagnosis and treatment of periodontally diseased patients.

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High Prevalence of Fluoroquinolone- and Methicillin-Resistant Staphylococcus pseudintermedius Isolates from Canine Pyoderma and Otitis Externa in Veterinary Teaching Hospital

  • Yoo, Jong-Hyun;Yoon, Jang-W.;Lee, So-Young;Park, Hee-Myung
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.798-802
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    • 2010
  • Recently, a total of 74 Staphylococcus pseudintermedius isolates were collected from clinical cases of canine pyoderma and otitis externa in Korea. In this study, we examined in vitro fluoroquinolone resistance among those isolates using a standard disc diffusion technique. The results demonstrated that, except for one isolate, approximately 18.9% to 27.0% of the isolates possessed bacterial resistance to both veterinary- and human-licensed fluoroquinolones including moxifloxacin (18.9% resistance), levofloxacin (20.3% resistance), ofloxacin (24.3% resistance), ciprofloxacin (25.7% resistance), and enrofloxacin (27.0% resistance). Most surprisingly, 14 out of 74 (18.9%) isolates were resistant to all the five fluoroquinolones evaluated. Moreover, a PCR detection of the methicillin resistance gene (mecA) among the 74 isolates revealed that 13 out of 25 (52.0%) mecApositive isolates, but only 7 out of 49 (14.3%) mecA-negative isolates, were resistant to one or more fluoroquinones. Taken together, our results imply that bacterial resistance to both veterinary- and human-use fluoroquinolones becomes prevalent among the S. pseudintermedius isolates from canine pyoderma and otitis externa in Korea, as well as that the high prevalence of the mecA-positive S. pseudintermedius isolates carrying multiple fluoroquinolones resistance could be a potential public health problem.

Monoclonal antibodies against porcine group C rotavirus VP6 (돼지 group C 로타바이러스 VP6 특이 단클론항체)

  • Yoon, Young-Sim;Lee, Seung-Chul;Woo, Sang-Kyu;Cho, Kyoung-Oh;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.175-182
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    • 2012
  • Rotaviruses have been known to be a major etiological agent of gastroenteritis in both infants and young animals. Subsequently new rotaviruses, which were morphologically indistinguishable but antigenically and electrophoretically distinct with each other, were reported from several animals throughout world including Korea. These new rotaviruses were named as non-group A or group B or group C rotaviruses and so on. It has been very difficult to isolate and grow the non-group A rotaviruses in vitro, and this has greatly limited the characterizations of non-group A rotaviruses and serological studies. In this study, monoclonal antibodies (MAbs) against porcine non-group A rotavirus were produced and characterized. The VP6 gene of porcine group C rotavirus Korean isolate(#06-52-1) was cloned and expressed. For expression of VP6 gene, baculovirus expression system was applied. The VP6 gene and expressed protein in the recombinant virus were confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test and Western blot, respectively. The expressed VP6 was used for MAbs production. The MAbs produced in this study would be promising as diagnostic reagents for detection of group C rotavirus infection.

Complexation of Adiponectin-encoding Plasmid DNA with Rosiglitazone-loaded Cationic Liposomes

  • Davaa, Enkhzaya;Jeong, Ui-Hyeon;Shin, Baek-Ki;Choi, Soon-Gil;Myung, Chang-Seon;Park, Jeong-Sook
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.357-362
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    • 2010
  • To enhance therapeutic effects of insulin-sensitizing adipokine, ADN gene and potent agonists, rosiglitazone for the $PPAR{\gamma}$, cationic liposomes as non-viral vectors were formulated. The particle size and zeta potential of drug loaded and unloaded cationic liposomes were investigated. The complex formation between cationic liposomes and negatively charged plasmid DNA was confirmed and the protection from DNase was observed. In vitro transfection was investigated in HepG2, HeLa, and HEK293 cells by mRNA expression of ADN. Encapsulation efficacy of rosiglitazone-loaded liposomes was determined by UV detection. Particle sizes of cationic liposomes were in the range of 110-170 nm and those of rosiglitazone-loaded cationic liposomes were in the range of 130-180 nm, respectively. Gel retardation of complexes indicated that the complex was formed at weight ratios of cationic lipid to plasmid DNA higher than 20:1. Both complexes protected plasmid DNA from DNase either drug free or drug loading. Encapsulation efficiency of rosiglitazone-loaded emulsion was increased by drug dose. The mRNA expression levels of ADN were dose-dependently increased in cells transfected with plasmid DNA. Therefore, cationic liposomes could be potential co-delivery system for drug and gene.