• Title/Summary/Keyword: Cell labeling

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Rapid and exact molecular identification of the PSP (paralytic shellfish poisoning) producing dinoflagellate genus Alexandrium

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.132-133
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    • 2003
  • The marine dinoflagellate genus Alexandrium comprise PSP producing A. acatenella, A. angustitabuzatum, A. catenella, A. fundyense, A. minutum, A. ostenfezdii, A. tamiyavanichii and A. tamarense. In monitoring toxic Alexandrium, rapid and exact species identification is one of the significant prerequisite work, however we have suffered confusion of species definition in Alexandrium. To surmount this problem, we chose DNA probing, which has long been used as an alternative for conventional identification methods, primarily relying on morphological approaches using microscope in microbial field. Oligonucleotide DNA probes targeting rRNA or rDNA have been commonly used in diverse studies to detect and enumerate cells concerned as a culture-indetendent powerful tool. Despite of the massive literature on the HAB species containing Alexandrium, application of DNA probing for species identification and detection has been limited to a few documents. DNA probes of toxic A. tamarense, A. catenella and A. tamiyavanichii, and non-toxic A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax were designed from LSU rDNA D1-D2, and applied to whole cell-FISH. Each DNA probes reacted only the targeted Alexandrium cells with very high species-specificity within Alexandrium. The probes could detect each targeted cells obtained from the natural sea water samples without cross-reactivity. Labeling intensity varied in the growth stage, this showed that the contents of probe-targeted cellular rRNA decreased with reduced growth rate. Double probe TAMID2S1 achieved approximately two times higher fluorescent intensity than that with single probe TAMID2. This double probe did not cross-react with any kinds of microorganisms in the natural sea waters. Therefore we can say that in whole-cell FISH procedure this double DNA probe successfully labeled targeted A. tamiyavanichii without cross-reaction with congeners and diverse natural bio-communities.

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Protective Effects of Rehmannia Glutinosa Extract and Rehmannia Glutinosa Vinegar against b-amyloid-induced Neuronal Cell Death (베타아밀로이드로 유도된 신경세포사멸에 대한 지황(地黃) 및 지황식초(地黃食醋)의 보호효과)

  • Song, Hyo-In;Kim, Kwang-Joong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.190-198
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    • 2007
  • Alzheimer's disease, a representative neurodegenerative disorder, is characterized by the presence of senile plaques and neurofibrillary tangles accompanied by neuronal damages. b-Amyloid peptide is considered to be responsible for the formation of senile plagues that accumulate in the brains of patients with Alzheimer's disease. There has been compelling evidence supporting that b-amyloid-induced cytotoxicity is mediated through generation of reactive oxygen species. In this study, we have investigated the possible protective effect of Rehmannia glutihosaagainst b-amyloid-induced oxidative ceil death in cultured human neuroblastoma SH-SY5Y cells. SH-SY5Y cells treated with b-amyloid underwent apoptotic death as determined by morphological features and positive in situterminal end-labeling (TUNEL staining). Rehmannia glutinosawater extract, wine, and vinegar pretreatments attenuated b-amyloid-induced cytotoxicity and apoptosis. Rehmannia glutinosa vinegar exhibited maximum protective effect by increasing the expression of anti-apoptotic protein, Bcl-2. in addition to oxidative stress, b-amyloid-treatment caused nitrosative stress via marked increase in the levels of nitric oxide, which was effectively blocked by Rehmannia glutinosa. To further explore the possible molecular mechanisms underlying the protective effect of Rehmannia glutinosa, we assessed the mRNA expression of cellular antioxidant enzymes. Treatment of Rehmannia glutinosa vinegar led to up-regulation of heme oxygemase-1 and catalase. These results suggest that Rehmannia glutinosa could modulate oxidative neuronal cell death caused by b-amyloid and may have preventive or therapeutic potential in the management of Alzheimer's disease. Particularly, Rehmannia glutinosa vinegar can augment cellular antioxidant capacity, there by exhibiting higher neuroprotective potential.

A Segmentation Method for Counting Microbial Cells in Microscopic Image

  • Kim, Hak-Kyeong;Lee, Sun-Hee;Lee, Myung-Suk;Kim, Sang-Bong
    • Transactions on Control, Automation and Systems Engineering
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    • v.4 no.3
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    • pp.224-230
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    • 2002
  • In this paper, a counting algorithm hybridized with an adaptive automatic thresholding method based on Otsu's method and the algorithm that elongates markers obtained by the well-known watershed algorithm is proposed to enhance the exactness of the microcell counting in microscopic images. The proposed counting algorithm can be stated as follows. The transformed full image captured by CCD camera set up at microscope is divided into cropped images of m$\times$n blocks with an appropriate size. The thresholding value of the cropped image is obtained by Otsu's method and the image is transformed into binary image. The microbial cell images below prespecified pixels are regarded as noise and are removed in tile binary image. The smoothing procedure is done by the area opening and the morphological filter. Watershed algorithm and the elongating marker algorithm are applied. By repeating the above stated procedure for m$\times$n blocks, the m$\times$n segmented images are obtained. A superposed image with the size of 640$\times$480 pixels as same as original image is obtained from the m$\times$n segmented block images. By labeling the superposed image, the counting result on the image of microbial cells is achieved. To prove the effectiveness of the proposed mettled in counting the microbial cell on the image, we used Acinetobacter sp., a kind of ammonia-oxidizing bacteria, and compared the proposed method with the global Otsu's method the traditional watershed algorithm based on global thresholding value and human visual method. The result counted by the proposed method shows more approximated result to the human visual counting method than the result counted by any other method.

Effect of Chungpaesagan-tang on Ischemic Damage in Organotypic Hippocampal Slice Culture (청폐사간탕(淸肺瀉肝湯)이 뇌해마 조직배양의 신경세포 자연사에 미치는 영향)

  • Lee, Min-Young;Ku, Ja-Seung;Kim, Sung-Hoon;Kim, Yoon-Bum;Kim, Sun-Yeou;Choi, Hyeon;Sohn, Young-Joo;Jung, Hyuk-Sang;Sohn, Nak-Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.771-777
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    • 2008
  • Chungpaesagan-tang which is used for treating patients of brain in cerebrovascular disease frequently from clinical doctor has not reported about the effect of neuronal aptosis caused of brain ischemia. The aim of this study is to investigate effect of Chungpaesagan-tang protecting neuronal cells from being damaged by brain ischemia through using organotypic hippocampal slice cultures. We caused ischemic damage to organotypic hippocampal slice cultures by oxygen and glucose deprivation. And added Chungpaesagan-tang extract to cultures. thereafter we measured area percentage of propidium iodide (PI)-stained neuronal cell, lactate dehydrogenase (LDH) levels in culture media and Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells. Area percentage of PI-stained neuronal cells and count of TUNEL-positive cells in CA1 and DG area of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Chungpaesagan-tang extract. LDH levels in culture media of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Chungpaesagan-tang extract. Within pertinent density level, Chungpaesagan-tang has cell protection effect that prevents brain ischemia damaging neuronal cells and apoptosis increasing.

Chemical Imaging Analysis of the Micropatterns of Proteins and Cells Using Cluster Ion Beam-based Time-of-Flight Secondary Ion Mass Spectrometry and Principal Component Analysis

  • Shon, Hyun Kyong;Son, Jin Gyeong;Lee, Kyung-Bok;Kim, Jinmo;Kim, Myung Soo;Choi, Insung S.;Lee, Tae Geol
    • Bulletin of the Korean Chemical Society
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    • v.34 no.3
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    • pp.815-819
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    • 2013
  • Micropatterns of streptavidin and human epidermal carcinoma A431 cells were successfully imaged, as received and without any labeling, using cluster $Au_3{^+}$ ion beam-based time-of-flight secondary ion mass spectrometry (TOF-SIMS) together with a principal component analysis (PCA). Three different analysis ion beams ($Ga^+$, $Au^+$ and $Au_3{^+}$) were compared to obtain label-free TOF-SIMS chemical images of micropatterns of streptavidin, which were subsequently used for generating cell patterns. The image of the total positive ions obtained by the $Au_3{^+}$ primary ion beam corresponded to the actual image of micropatterns of streptavidin, whereas the total positive-ion images by $Ga^+$ or $Au^+$ primary ion beams did not. A PCA of the TOF-SIMS spectra was initially performed to identify characteristic secondary ions of streptavidin. Chemical images of each characteristic ion were reconstructed from the raw data and used in the second PCA run, which resulted in a contrasted - and corrected - image of the micropatterns of streptavidin by the $Ga^+$ and $Au^+$ ion beams. The findings herein suggest that using cluster-ion analysis beams and multivariate data analysis for TOF-SIMS chemical imaging would be an effectual method for producing label-free chemical images of micropatterns of biomolecules, including proteins and cells.

Improved Lung and Pulmonary Vessels Segmentation and Numerical Algorithms of Necrosis Cell Ratio in Lung CT Image (흉부 CT 영상에서 개선된 폐 및 폐혈관 분할과 괴사 세포 비율의 수치적 알고리즘)

  • Cho, Joon-Ho;Moon, Sung-Ryong
    • Journal of Digital Convergence
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    • v.16 no.2
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    • pp.19-26
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    • 2018
  • We proposed a numerical calculation of the proportion of necrotic cells in pulmonary segmentation, pulmonary vessel segmentation lung disease site for diagnosis of lung disease from chest CT images. The first step is to separate the lungs and bronchi by applying a three-dimensional labeling technique from a chest CT image and a three-dimensional region growing method. The second step is to divide the pulmonary vessels by applying the rate of change using the first order polynomial regression, perform noise reduction, and divide the final pulmonary vessels. The third step is to find a disease prediction factor in a two-step image and calculate the proportion of necrotic cells.

Gold nanoparticles enhance anti-tumor effect of radiotherapy to hypoxic tumor

  • Kim, Mi Sun;Lee, Eun-Jung;Kim, Jae-Won;Chung, Ui Seok;Koh, Won-Gun;Keum, Ki Chang;Koom, Woong Sub
    • Radiation Oncology Journal
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    • v.34 no.3
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    • pp.230-238
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    • 2016
  • Purpose: Hypoxia can impair the therapeutic efficacy of radiotherapy (RT). Therefore, a new strategy is necessary for enhancing the response to RT. In this study, we investigated whether the combination of nanoparticles and RT is effective in eliminating the radioresistance of hypoxic tumors. Materials and Methods: Gold nanoparticles (GNPs) consisting of a silica core with a gold shell were used. CT26 colon cancer mouse model was developed to study whether the combination of RT and GNPs reduced hypoxia-induced radioresistance. Hypoxia inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) was used as a hypoxia marker. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were conducted to evaluate cell death. Results: Hypoxic tumor cells had an impaired response to RT. GNPs combined with RT enhanced anti-tumor effect in hypoxic tumor compared with RT alone. The combination of GNPs and RT decreased tumor cell viability compare to RT alone in vitro. Under hypoxia, tumors treated with GNPs + RT showed a higher response than that shown by tumors treated with RT alone. When a reactive oxygen species (ROS) scavenger was added, the enhanced antitumor effect of GNPs + RT was diminished. Conclusion: In the present study, hypoxic tumors treated with GNPs + RT showed favorable responses, which might be attributable to the ROS production induced by GNPs + RT. Taken together, GNPs combined with RT seems to be potential modality for enhancing the response to RT in hypoxic tumors.

Cyclin D1, Retinoblastoma and p16 Protein Expression in Carcinoma of the Gallbladder

  • Srivastava, Vineeta;Patel, Brijesh;Kumar, Mohan;Shukla, Mridula;Pandey, Manoj
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.2711-2715
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    • 2013
  • Background: Cancer of the gallbladder is a relatively rare neoplasm with a poor prognosis. The exact mechanisms of its genesis are not known and very little information is available on molecular events leading to labeling this as an orphan cancer. Materials and Methods: In this prospective case control study we evaluated the expression of p16, pRb and cyclin D1 by immunohistochemistry to study the G1-S cell-cycle check point and its possible role in gallbladder carcinogenesis. A total of 25 patients with gallbladder carcinoma (group I), 25 with cholelithiasis (group II) and 10 normal controls. were enrolled Results: Cyclin D1 expression was seen in 10 (40%) patients each with carcinoma and cholelithiasis while only in 2 (20%) of the normal gallbladders but differences were not statistically significant (p value=0.488). p16 was expressed in 12% patients of carcinoma of the gallbladder and 28% of cholelithiasis, however this difference was not statistically significant (p value=0.095). Retinoblastoma protein was found to be expressed in 50% of normal gallbladders and 6 (24%) of carcinoma and 8 (32%) of gallstones. The present study failed to demonstrate any conclusive role of cyclin D1/RB/ p16 pathway in carcinoma of the gallbladder. Conclusions: The positive relation observed between tumor metastasis and cyclinD1 expression and p16 with nodal metastasis suggested that higher cyclin D1/p16 expression may act as a predictive biomarker for aggressive behavior of gallbladder malignancies.

Inhibitory Effects of Low-Dose Aloe-Emodin on the Development of Colorectal Tumors in Min Mice

  • Shimpo, Kan;Chihara, Takeshi;Kaneko, Takaaki;Beppu, Hidehiko;Wakamatsu, Kazumasa;Shinzato, Masanori;Yukitake, Jun;Sonoda, Shigeru
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5587-5592
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    • 2014
  • Aloe-emodin (AE), a natural anthraquinone compound, has been reported to exhibit anticancer activity in various cancer cell lines and anti-inflammatory effects in murine macrophages. In the present study, we investigated the cancer chemopreventive effects of AE in an Apc-deficient Min mouse model. In the first experiment, male Min mice were fed a basal diet or diets containing 5 ppm AE and 10 ppm AE for 12 weeks. The dietary administration of 5 ppm AE significantly reduced the number of colorectal tumors. In a second experiment, we investigated the effects of AE on colitis-related colon carcinogenesis in Min mouse treated with dextran sodium sulfate (DSS). Female Min mice were administered 1% DSS in their drinking water for 7 days. AE was given to mice in their diet at a dose of 5 or 50 ppm for 5 weeks. Feeding with AE significantly reduced the number of colorectal tumors. When proliferation of cells in normal-appearing colonic mucosa was assessed by monoclonal anti-rat Ki-67 antibody (MIB-5) immunohistochemistry in experiments 1 and 2, the AE treatment significantly decreased the mean MIB-5-labeling index. These results suggest that the dietary administration of low-dose AE may have chemopreventive effects against development of colorectal tumors in Min mice, possibly in part by reducing cell proliferation in colorectal mucosa.

Effect of Yanggyuksanhwa-tang on Ischemic Damage in Organotypic Hippocampal Slice Culture (양격산화탕(凉膈散火湯)이 뇌해마 조직배양의 허혈손상에 따른 신경세포손상에 미치는 영향)

  • Lee, Hwan-Sung;Park, Sung-Joon;Jung, Kwang-Sik;Sohn, Young-Joo;Jung, Hyuk-Sang;Park, Dong-Il;Sohn, Nak-Won
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.231-242
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    • 2008
  • Objectives : We can find out the experimental reports of Yanggyuksanhwa-tang, which has the function of regulating blood pressure related with cerebral disease, and increasing local cerebral blood stream volume, also has the recoveries for the damage of vessel endothelium, and endothelium hypertrophy caused by angiospasm after subarachnoid hemorrhage, and reduces the contraction of smooth muscle, so simultaneously improves necrosis. The aim of this study is to investigate effect of Yanggyuksanhwa-tang protecting neuronal cells from being damaged by brain ischemia through using organotypic hippocampal slice cultures. Methods : We caused ischemic damage to organotypic hippocampal slice cultures by oxygen and glucose deprivation, and Yanggyuksanhwa-tang extract was added to cultures. Thereafter we measured area percentage of propidium iodide (PI)-stained neuronal cell, lactate dehydrogenase (LDH) levels in culture media and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells. Results : Area percentage of PI-stained neuronal cells and count of TUNEL-positive cells in CA1 and DG area of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Yanggyuksanhwa-tang extract. LDH levels in culture media of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Yanggyuksanhwa-tang extract. Conclusions : Within pertinent density level, Yanggyuksanhwa-tang has cell protection effect that prevents brain ischemia damaging neuronal cells and apoptosis increasing.

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