• Title/Summary/Keyword: In vivo detection

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Cellular Mechanism of Newly Synthesized Indoledione Derivative-induced Immunological Death of Tumor Cell

  • Oh, Su-Jin;Ryu, Chung-Kyu;Baek, So-Young;Lee, Hyun-Ah
    • IMMUNE NETWORK
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    • v.11 no.6
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    • pp.383-389
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    • 2011
  • Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell. Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC. Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and $100{\mu}M$) with carleticulin induction. And EY-6 induced the secretion of IFN-${\gamma}$ but not of TNF-${\alpha}$ from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation). Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

Numerical Design of Double Quantum Coherence Filter for the Detection of Myo-Inositol In vivo (인체 내 myo-Inositol 검출을 위한 수치해석적 이중양자 필터 디자인)

  • Lee, Yun-Jung;Jung, Jin-Young;Noh, Hyung-Joon;Yu, Ung-Sik;Kim, Hyeon-Jin
    • Investigative Magnetic Resonance Imaging
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    • v.13 no.2
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    • pp.117-126
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    • 2009
  • Purpose : A numerical method of designing a multiple quantum filter (MQF) is presented for the optimum detection of myo-inositol (mI), an important brain metabolite, by using in vivo proton nuclear magnetic resonance spectroscopy ($^1$-HMRS). Materials and Methods : Starting from the characterization of the metabolite, the filter design includes the optimization of the sequence parameters such as the two echo times (TEs), the mixing time (TM), and the flip angle and offset frequency of the 3rd $90^{\circ}$ pulse which converts multiple quantum coherences (MQCs) back into single quantum coherences (SQCs). The optimized filter was then tested both in phantom and in human brains. Results : The results demonstrate that the proposed MQF can improve the signal-to-background ratio of the target metabolite by a factor of more than three by effectively suppressing the signal from the background metabolites. Conclusion : By incorporating a numerical method into the design of MQFs in $^1$-HMRS the spectral integrity of a target metabolite, in particular, with a complicated spin system can be substantially enhanced.

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Direct detection of cariogenic streptococci in metal brackets in vivo using polymerase chain reaction (교정용 메탈 브라켓에서 자가중합효소연쇄반응을 통한 치아우식증 원인균의 탈출)

  • Ahn, Sug-Joon;Lee, Shin-Jae;Baek, Seung-Hak;Kim, Tae-Woo;Chang, Young-Il;Nam, Dong-Seok;Lim, Bum-Soon
    • The korean journal of orthodontics
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    • v.35 no.4 s.111
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    • pp.312-319
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    • 2005
  • Streptococcus mutans and Streptococcus sobrinus are major etiological agents in enamel demineralization around orthodontic appliances. This study was designed to examine the prevalence of these streptococci on orthodontic brackets in vivo using polymerase chain reaction. Four incisor brackets in the upper and lower arches were removed and collected from 80 patients at the time of debonding. The genomic DMA of adhered bacteria was extracted and each dextranase gene of S. mutans and S. sobrinus was amplified using the specific oligonucleotide primers. The results showed that the maxillary incisor brackets were colonized by both cariogenic streptococci to a somewhat higher degree than that taken from the mandible. The prevalence of S. mutans was $50.0\%$ on the maxillary incisor brackets and $33.8\%$ on the mandibular incisor brackets, and that of S. sobrinus was $17.5\%$ and $15.0\%$, respectively. Both species were detected on the maxillary incisor brackets of 7 patients $(8.8\%)$ and the mandibular incisor brackets of 5 patients $(6.3\%)$. These results suggest that cariogenic streptococci can adhere to the incisor brackets and may be resident species on the incisor brackets.

Notoginseng leaf triterpenes ameliorates mitochondrial oxidative injury via the NAMPT-SIRT1/2/3 signaling pathways in cerebral ischemic model rats

  • Weijie, Xie;Ting, Zhu;Ping, Zhou;Huibo, Xu;Xiangbao, Meng;Tao, Ding;Fengwei, Nan;Guibo, Sun;Xiaobo, Sun
    • Journal of Ginseng Research
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    • v.47 no.2
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    • pp.199-209
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    • 2023
  • Background: Due to the interrupted blood supply in cerebral ischemic stroke (CIS), ischemic and hypoxia results in neuronal depolarization, insufficient NAD+, excessive levels of ROS, mitochondrial damages, and energy metabolism disorders, which triggers the ischemic cascades. Currently, improvement of mitochondrial functions and energy metabolism is as a vital therapeutic target and clinical strategy. Hence, it is greatly crucial to look for neuroprotective natural agents with mitochondria protection actions and explore the mediated targets for treating CIS. In the previous study, notoginseng leaf triterpenes (PNGL) from Panax notoginseng stems and leaves was demonstrated to have neuroprotective effects against cerebral ischemia/reperfusion injury. However, the potential mechanisms have been not completely elaborate. Methods: The model of middle cerebral artery occlusion and reperfusion (MCAO/R) was adopted to verify the neuroprotective effects and potential pharmacology mechanisms of PNGL in vivo. Antioxidant markers were evaluated by kit detection. Mitochondrial function was evaluated by ATP content measurement, ATPase, NAD and NADH kits. And the transmission electron microscopy (TEM) and pathological staining (H&E and Nissl) were used to detect cerebral morphological changes and mitochondrial structural damages. Western blotting, ELISA and immunofluorescence assay were utilized to explore the mitochondrial protection effects and its related mechanisms in vivo. Results: In vivo, treatment with PNGL markedly reduced excessive oxidative stress, inhibited mitochondrial injury, alleviated energy metabolism dysfunction, decreased neuronal loss and apoptosis, and thus notedly raised neuronal survival under ischemia and hypoxia. Meanwhile, PNGL significantly increased the expression of nicotinamide phosphoribosyltransferase (NAMPT) in the ischemic regions, and regulated its related downstream SIRT1/2/3-MnSOD/PGC-1α pathways. Conclusion: The study finds that the mitochondrial protective effects of PNGL are associated with the NAMPT-SIRT1/2/3-MnSOD/PGC-1α signal pathways. PNGL, as a novel candidate drug, has great application prospects for preventing and treating ischemic stroke.

Optimal Conditions of Single Cell Gel Electrophoresis (Comet) Assay to detect DNA single strand breaks in Mouse Lymphoma L5178Y cells

  • Ryu, Jae-Chun;Kwon, Oh-Seung;Kim, Hyung-Tae
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.89-94
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    • 2001
  • Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

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Regulation of Phthalates and Their Alternatives in Children's Products and Their Toxicity Data (어린이제품 내 프탈레이트류 및 대체제의 규제와 독성자료에 대한 연구)

  • Lee, Inhyae;Ra, Jinsung;Ji, Kyunghee
    • Journal of Environmental Health Sciences
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    • v.47 no.1
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    • pp.1-19
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    • 2021
  • Objectives: Phthalates, which are widely used as plasticizers, have been recognized as endocrine disruptors. In the present study, we provided information on the regulation of these chemicals and summarized the information available on their detection and toxicity in children's products and those of their alternatives. Methods: The regulatory frameworks related to phthalates in children's products in Korea, the United States (US), and the European Union (EU) were compared. Data on the detection concentration of 16 phthalates and seven phthalate alternatives that could be used in polyvinyl chloride (PVC) plastic products for children as well as on their toxicity classification and endocrine disruption toxicity were collected from the literature. Results: Korea adopted US and EU chemical standards for six phthalates (DEHP, BBP, DBP, DINP, DIDP, and DNOP), but not others (e.g., DIBP, DPP, DHP, and DCHP). Among the ten phthalates and seven substitutes for which regulatory standards were not determined, DIBP, DHP, DEHA, DIBA, DINA, and DEHT were detected in children's products made from PVC plastic. DIBP and DHP, which have a reproductive toxicity classification of 1B, were frequently detected in PVC toys. The reproductive toxicity, estrogenicity, and anti-androgenic activity of the unregulated phthalates and their alternatives have been reported in diverse in vitro and in vivo assays. Conclusion: The use of unregulated phthalates and their substitutes in children's products is increasing. Further monitoring and toxicological information on phthalate alternatives is required to develop proper management plans.

Cone-beam computed tomography versus digital periapical radiography in the detection of artificially created periapical lesions: A pilot study of the diagnostic accuracy of endodontists using both techniques

  • Campello, Andrea Fagundes;Goncalves, Lucio Souza;Guedes, Fabio Ribeiro;Marques, Fabio Vidal
    • Imaging Science in Dentistry
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    • v.47 no.1
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    • pp.25-31
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    • 2017
  • Purpose: The aim of this study was to compare the diagnostic accuracy of previously trained endodontists in the detection of artificially created periapical lesions using cone-beam computed tomography (CBCT) and digital periapical radiography (DPR). Materials and Methods: An ex vivo model using dry skulls was used, in which simulated apical lesions were created and then progressively enlarged using #1/2, #2, #4, and #6 round burs. A total of 11 teeth were included in the study, and 110 images were obtained with CBCT and with an intraoral digital periapical radiographic sensor (Instrumentarium dental, Tuusula, Finland) initially and after each bur was used. Specificity and sensitivity were calculated. All images were evaluated by 10 previously trained, certified endodontists. Agreement was calculated using the kappa coefficient. The accuracy of each method in detecting apical lesions was calculated using the chisquare test. Results: The kappa coefficient between examiners showed low agreement (range, 0.17-0.64). No statistical difference was found between CBCT and DPR in teeth without apical lesions (P=.15). The accuracy for CBCT was significantly higher than for DPR in all corresponding simulated lesions(P<.001). The correct diagnostic rate for CBCT ranged between 56.9% and 73.6%. The greatest difference between CBCT and DPR was seen in the maxillary teeth (CBCT, 71.4%; DPR, 28.6%; P<.01) and multi-rooted teeth (CBCT, 83.3%; DPR, 33.3%; P<.01). Conclusion: CBCT allowed higher accuracy than DPR in detecting simulated lesions for all simulated lesions tested. Endodontists need to be properly trained in interpreting CBCT scans to achieve higher diagnostic accuracy.

Identification of urinary metabolite(s) of CKD-712 by gas chromatography/mass spectrometry in rats

  • Jeon, Hee-Kyung;Park, Hae-Yeon;Kim, Youn-Jung;Kwon, Oh-Seung;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.188-188
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    • 2003
  • Examination was made of the urinary metabolite(s) of CKD-712, which is a chiral compound, named S-YS49 derived from higenamine (one component of Aconite spp.) derivatives. First of all, to analyze the metabolite(s) of CKD-712, a simple and sensitive detection method for CKD-712 was developed by using gas chromatography-mass spectrometry GC/MS). Urine was collected from adult male Sprague-Dawley rats 250${\pm}$10g) in metabolic cage for 24hr after oral administration of 100 mg/kg of CKD-712. The recovery of CKD-712 after extraction and concentration with AD-2 resin column was above 90 % from rat urine. The detection limits of CKD-712 in urine was approximately 0.1 ng/mL. It has well been suggested that isoquinoline possessing catechol moiety such as CKD-712 should be subjected to the catechol-O-methyl kransferase activity in vivo. We detected three major peaks of presumed CKD-712 metabolites in the total ion chromatogram obtained from the rat urine sample after oral administration of CKD-712. From these results, it is assumed that the urinary metabolites are mono-methylation in the naphthyl moiety (metabolite I ), methylation at the C-6 or 7 hydroxy group in the isoquinoline moiety and hydroxylation at in the naphthyl moiety (metaboliteII), and methylation at the C-6 or 7 hydroxy group in the isoquinoline moiety (metaboliteIII).

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Comparison of two different methods of detecting residual caries

  • Vural, Uzay Koc;Kutuk, Zeynep Bilge;Ergin, Esra;Cakir, Filiz Yalcin;Gurgan, Sevil
    • Restorative Dentistry and Endodontics
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    • v.42 no.1
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    • pp.48-53
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    • 2017
  • Objectives: The aim of this study was to investigate the ability of the fluorescence-aided caries excavation (FACE) device to detect residual caries by comparing conventional methods in vivo. Materials and Methods: A total of 301 females and 202 males with carious teeth participated in this study. The cavity preparations were done by grade 4 (Group 1, 154 teeth), grade 5 (Group 2, 176 teeth), and postgraduate (Group 3, 173 teeth) students. After caries excavation using a handpiece and hand instruments, the presence of residual caries was evaluated by 2 investigators who were previously calibrated for visual-tactile assessment with and without magnifying glasses and trained in the use of a FACE device. The tooth number, cavity type, and presence or absence of residual caries were recorded. The data were analyzed using the Chi-square test, the Fisher's Exact test, or the McNemar test as appropriate. Kappa statistics was used for calibration. In all tests, the level of significance was set at p = 0.05. Results: Almost half of the cavities prepared were Class II (Class I, 20.9%; Class II, 48.9%; Class III, 20.1%; Class IV, 3.4%; Class V, 6.8%). Higher numbers of cavities left with caries were observed in Groups 1 and 2 than in Group 3 for all examination methods. Significant differences were found between visual inspection with or without magnifying glasses and inspection with a FACE device for all groups (p < 0.001). More residual caries were detected through inspection with a FACE device (46.5%) than through either visual inspection (31.8%) or inspection with a magnifying glass (37.6%). Conclusions: Within the limitations of this study, the FACE device may be an effective method for the detection of residual caries.

The study on the Fluorescence Characteristics of Several Freshwater Bloom Forming Algal Species and Its Application (수종 담수적조 원인종들의 형광특성과 적용연구)

  • Son, Moon-Ho;Zulfugarov, Ismayil S.;Kwon, O-Seob;Moon, Byoung-Young;Chung, Ik-Kyo;Lee, Choon-Hwan;Lee, Jin-Ae
    • ALGAE
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    • v.20 no.2
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    • pp.113-120
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    • 2005
  • The freshwater blooms mainly blue-green algal blooms occur frequently in the lower Naktong River in summer, which provoke many socio-economical problems; therefore, the early detection of bloom events are demanding through the quantitative and qualitative analyses of blue green algal species. The in vivo fluorescence properties of cultured strains of Microcystis aeruginosa, M. viridis, M. wesenbergii, M. ichthyoblabe, Anabaena cylindrica, A. flos-aquae, and Synedra sp. were investigated. Wild phytoplankton communities of the lower Naktong River were also monitored at four stations in terms of their standing stocks, biomass and fluorescence properties compared with its absorption spectram. The 77K fluorescence emission spectra of each cultured strains normalized at 620 nm was very specific and enabled to detect of blue green algal biomass qualitatively and quantitatively. The relative chlorophyll a concentration determined by chlorophyll fluorescence analysis method showed significant relationship with chlorophyll a concentration determined by solvent extraction method ($R^2$ = 0.906), and the blue-green algal cell number determined by microscopic observation ($R^2$ = 0.588), which gives insight into applications to early detection of blue green algal bloom.