• Title/Summary/Keyword: Single-Cell Analysis

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Single Cell Dissociation Methods for Flow Cytometric Cell Death Analysis of Hypoxia-Ischemia Injured Newborn Rat Pup Brain (저산소성 허혈성 뇌손상이 유발된 신생백서에서 단일세포의 분리)

  • Hwang, Jong Hee;Sung, Dong Kyung;Choi, Chang Won;Kang, Saem;Chang, Yun Sil;Park, Won Soon;Lee, Munhyang
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.545-550
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    • 2005
  • Purpose : Newborn brain tissue has to be dissociated into a single cell suspension for flow cytometric analysis of cell death during hypoxia-ischemia. Thus the development of a method to dissociate cells from the brain tissue with least damage and maintenance of membrane and antigen integrity remains the challenge for the in vivo application of this technique. We evaluated the efficacy of mechanical or enzymatic (collagenase or tryspin) methods of brain tissue disaggregation. Methods : The extent of the damage to the plasma membrane and loss of the characteristics of the membrane induced with each dissociation method was determined by comparing the flow cytometric results labeled with both fluorescent annexin V and propidium iodide of the newborn rat pup brain tissue in the control group (n=10) and in the 48-hour after hypoxia-ischemia group (n=10). Results : In the control group, the cell percentage of damaged, apoptotic and necrotic cells of both hemispheres with the mechanical dissociation method was significantly increased compared to the trypsin or collagenase method. In the 48-hour after hypoxia-ischemia group, the cell percentage of apoptotic and necrotic cells of the right hemisphere with the collagenase method significantly increased, and live cells significantly decreased compared to the left hemisphere, control group. Although the same trend was observed, the extent of alterations made with the trypsin method was significantly less compared to the collagenase method. Conclusion : The dissociation of neonatal brain tissue for flow cytometric analysis with collagenase was most efficacious with the least cell damage and preservation of the plasma membrane characteristics.

Effect of Ionomer Content on the Anode Catalyst Layers of PEM Fuel Cells (고분자 전해질 연료전지용 수소극 촉매층의 이오노머 함량 영향)

  • PAK, BEOMJUN;LEE, SEONHO;WOO, SEUNGHEE;PARK, SEOK-HEE;JUNG, NAMGEE;YIM, SUNG-DAE
    • Journal of Hydrogen and New Energy
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    • v.30 no.6
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    • pp.523-530
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    • 2019
  • For the low-Pt electrodes for polymer electrolyte fuel cells (PEMFCs), the optimization of ionomer content for anode catalyst layers was carried out. A commercial catalyst of 20 wt.% Pt/C was used instead of 50 wt.% Pt/C which is commonly used for PEMFCs. The ionomer content varies from 0.6 to 1.2 based on ionomer to carbon ratio (I/C) and the catalyst layer is formed over the electrolyte by the ultrasonic spray process. Evaluation of the prepared MEA in the unit cell showed that the optimal ionomer content of the air electrode was 0.8 on the I/C basis, while the hydrogen electrode was optimal at the relatively high ionomer content of 1.0. In addition, a large difference in cell performance was observed when the ionomer content of the hydrogen electrode was changed. Increasing the ionomer content from 0.6 to 1.0 by I/C in a hydrogen electrode with 0.05 mg/㎠ platinum loading resulted in more than double cell performance improvements on a 0.6 V. Through the analysis of various electrochemical properties in the single cell, it was assumed that the change in ionomer content of the hydrogen electrode affects the water flow between the hydrogen and air electrodes bounded by the membrane in the cell, which affects the overall performance of the cell. A more specific study will be carried out to understand the water flow mechanism in the future, and this study will show that the optimization process of hydrogen electrode can also be a very important cell design variable for the low-Pt and high-performance MEA.

Assessment of dental age estimation using dentinal translucency in ground sections of single rooted teeth: a digital image analysis

  • Abelene Maria Durand;Madhu Narayan;Raghavendhar Karthik;Rajkumar Krishnan;Narasimhan Srinivasan;Dinesh Kumar
    • Anatomy and Cell Biology
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    • v.57 no.2
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    • pp.271-277
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    • 2024
  • Human dentition is unique to individuals and helps in identification of individuals in forensic odontology. This study proposes to study the manually ground sections of single rooted teeth using digital methods for dental age estimation. To assess the dentinal translucency from the scanned digital images of manually ground section of teeth using commercially available image edition software. Corroborating the root dentinal translucency length and region of interest (ROI) of translucency zone in pixels (as a marker of dental age) with the chronological age of the subject, as stratified by different age groups. Twenty single-rooted extracted teeth from 20 patients each from 6 groups divided as per age. Manual sectioning of the teeth followed by scanning the sections was done. Root area in pixels and ROI of translucency zone were measured. From the observed values, translucency length percentage (TLP) and percentage of ROI in pixels (TPP) was calculated and tabulated. Pearson's correlation coefficients were obtained for age with TLP and TPP. Positive correlation existed between age and TLP and also between age and TPP. With the obtained data, multilinear regression equations for specific age groups based on 10-year intervals were derived. By a step-down analysis method, age was estimated with an average error of around ±7.9 years. This study gives a novel method for age-estimation that can be applied in real-time forensic sciences.

In Vitro Intrinsic Radiosensitivity Of Human Squamous Cell Carcinoma in Primary Culture (인체 상피 세포암의 일차 배양을 이용한 방사선 민감도 측정)

  • Choi Eun Kyung;Yang Kwang Mo;Yi Byong Yong;Chang Hyesook;Kim Sang-Yoon;Nam Joo-Hyun;Yu Eunsil;Lee Inchul
    • Radiation Oncology Journal
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    • v.12 no.1
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    • pp.27-31
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    • 1994
  • There are a number of reports suggesting that there may be a correlation between the clinical response to radiotherapy in various tumors and the clonogenic survival of cell lines derived from these tumors following exposure to 2 Gy(SF2). Authors conducted this study to determine SF2 for cells in primary culture from surgical specimens. The tumor tissues with squamous cell carcinoma of uterine cervix and head and neck were obtained. The tumor tissues were disaggregated to single cells by incubating with collagenase type w for 2 hours with constant stirring. Single cell suspensions were inoculated in four 24-well plates precoated with cell adhesive matrix. After 24 hours of incubation at 37$ ^{\circ}C $, rows of four wells were then irradiated, consisting of control set and five other sets each receiving doses of 1,2,3,4, and 6 Gy. After incubation for a total of 13 days, the cultures were stained with crystal violet and survival at each dose was determined by quantitative image analysis system, To determine whether cell growth was of epithelial origin, immunocytochemical staining with a mixture of cytokeratin and epithelial monoclonal antibodies were performed on cell cultures. During the period of this study, we received 5 squamous cell carcinoma specimens of head and neck and 20 of uterine cervical carcinoma. Of these, 15 yielded enough cells for radiosensitivity testing. This resulted an overall success rate of 60$ \% $. The mean SF2 value for 15 tumours was 0.55$\pm$0.17 ranging from 0.20 to 0.79. These results indicate that there is a broad range of sensitivities to radiation in same histologic type. So with a large patient population, we plan to determine whether a different SF2 value is associated with tumours that are controlled with radiotherapy than those that are not.

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Proteomic Analysis of Circadian Clock Mutant Mice

  • Lee Joon-Woo;Kim Han-Gyu;Bae Kiho
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.493-501
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    • 2005
  • Circadian rhythms, time on a scale of about 24 hours, are present in a number of organisms including animals, plants, and bacteria. The control of the biochemical, physiological and behavioral processes is regulated by endogenous clocks in the suprachiasmatic nucleus (SCN). At the core of this timing mechanism is molecular machinery that are present both in the brain and in the peripheral tissues throughout the body, and even in a single cultured cell. In this study, we performed two-dimensional gel electrophoresis to figure out any correlation between protein expression patterns and the requirement of two canonical clock proteins, either mPER1 or mPER2, by comparing global protein expression profiles in livers from wildtype or mPer1/mPer2 double mutant mice. We could identify several differentially expressed protein candidates with respect to time and genotypes. Further analysis of these candidate proteins in detail in vivo will lead us to the better understanding of how circadian clock functions in mammals.

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A Study on the Effect of Energy Dissipation in Extruding Clad Rod (복합봉재 압출에 의한 에너지 소산의 영향에 관한 연구)

  • Kim, Chang-Hoon
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.5 no.2
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    • pp.56-64
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    • 2006
  • Rapid progress in many branches of technology has led to a demand on new materials such as high strength light weight alloys, powdered alloys and composite materials. The hydrostatic extrusion is essentially a method of extruding a clad rod through a die. In order to investigate the effect of the process conditions such as friction heat, deformation and clad thickness on the clad extrusion process, viscoplastic finite element simulations were conducted. A specific model for theoretical analysis used in this study is The single scalar variable version of Hart's model. An experiment also has been carried out using 1.5MN hydrostatic extruder with variable speed ram, LVDT and load cell for comparison. It is found that the hydrostatic extrusion pressure considering the effect of heat dissipation in this theoretical work was closer to the experimental pressure than the isothermal hydrostatic extrusion pressure.

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Analysis of Case Report Related with Mountain Ginseng Pharmacopuncture in Korean Medical Database (한국 논문 데이터베이스의 산삼약침 암 증례 논문 분석)

  • Park, Soo-jung;Joo, Jong-cheon
    • Journal of Korean Traditional Oncology
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    • v.20 no.2
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    • pp.51-59
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    • 2015
  • Objective: This paper is to investigate the current evidence and clinical effectiveness of mountain ginseng pharmacopuncture (MGP) in cancer treatment. Methods: Along with hand searches, relevant literatures were searched from Korean medical database until November 2015 without language restriction and clinical researches were selected for this review. Results: Nine articles were searched. The MGP was generally prepared in the concentration of ten percent. Also, the MGP has been used by intravenous injection much more than acupoint injection. The clinical effects were the suppression of the cancer cell growth, the improvement the quality of life, the reduction of the side effect by chemotherapy and radiotherapy. Conclusion: This analysis suggests that the MGP can be used as a single therapy or a supportive therapy with conventional therapies, which are chemotherapy or radiotherapy.

Free Radical Involvement in the DNA Damaging Activity of Fumonisin Bl

  • Lee, Wan-Hee;Lee, Kil-Soo
    • Toxicological Research
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    • v.17 no.4
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    • pp.249-253
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    • 2001
  • Fumonisin B1, a mycotoxin, is thought to induce esophageal cancer in humans and apoptosis in animal cells by inhibiting ceramide synthase. Dumonisin Bl may also generate reactive oxygen species directly or indirectly, leading to DNA damage and lipid peroxidation. In this study, a DNA fragmentation assay, dichlorofluorescein (DCF) analysis, and single cell gel electrophoresis (SCGE) were used to investigate the involvement of cellular free radicals, specifically hydrogen peroxide, in the DNA damaging activity of fumonisin B1. From an in vitro DNA fragmentation assay, E. coli DNA, damage by fumonisin Bl was increased by the addition of superxide dismutase (SOD) and decreased by catalase. SCGE and DCF analysis in vivo showed that the nuclear DNA damage and intracellular free radicals in cultured rat hepatocytes treated with fumonisin B1 were increased with the concentration of fumonisin Bl . DNA damage and free radical generation were inhibited by the addition of catalase. Fumonisin Bl , in the presence of SOD, produces hydrogen peroxide causing oxidative DNA damage and protein malfunction, leading to genotoxicity and cytotoxicity of the toxin.

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X-ray Crystal Structure of Hetaryl Leuco-TAM Dyes, (2Z,2'E)-2,2'-(2-Phenyl Propane-1,3-diylidene) Bis(1,3,3-trimethyl indoline) Derivatives

  • Keum, Sam-Rok;Roh, Se-Jung;Kim, Young-Nan;Im, Do-Hyuck;Ma, So-Young
    • Bulletin of the Korean Chemical Society
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    • v.30 no.11
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    • pp.2608-2612
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    • 2009
  • The unequivocal solid-state structure and stereochemistry of the hetaryl leuco-TAM dye, 2,2’-(2-phenyl propane- 1,3-diylidene) bis(1,3,3- trimethylindoline) derivatives were established using X-ray single crystal analysis. The X-ray crystal analysis showed that the (Z, E)-isomers only formed stereoselectively, with a so-called “threebladed propeller” conformation, from the reaction of a Fischer base and benzaldehyde derivatives. These isomers were stacked in a juxtaposition to form a dimer or a double dimer, adopting either a triclinic, with P-1, or monoclinic crystal system with a space group P21/n in the unit cell of the crystal.

Development of a Mushroom Powder Certified Reference Material for Element Analysis

  • Betru, Tegegn Gizachew;Yim, Yong-Hyeon;Lee, Kyoung-Seok
    • Mass Spectrometry Letters
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    • v.11 no.4
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    • pp.108-112
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    • 2020
  • A certified reference material (CRM) for the analysis of nutrient elements in an edible mushroom (Ganoderma lyceum) powder has been developed (KRISS CRM 108-10-011). The mass fractions of calcium (Ca), iron (Fe), and zinc (Zn) were measured by isotope dilution inductively coupled plasma mass spectrometry (ID ICP/MS). To dissolve the fungi cell wall of mushroom consisted of chitin fibers, sample preparation method by single reaction chamber type microwave-assisted acid digestion with acid mixtures was optimized. The mean measurement results obtained from 12 sample bottles were used to assign as the certified values for the CRM and the between-bottle homogeneities were evaluated from the relative standard deviations. The certified values were metrologically traceable to the definition of the kilogram in the International System of Units (SI). This CRM is expected to be used for validation of analytical methods or quality control of measurement results in analytical laboratories when they determine the mass fractions of elements in mushroom or other similar samples.