• Title/Summary/Keyword: single cell imaging

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Advances in higher-order chromatin architecture: the move towards 4D genome

  • Jung, Namyoung;Kim, Tae-Kyung
    • BMB Reports
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    • v.54 no.5
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    • pp.233-245
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    • 2021
  • In eukaryotes, the genome is hierarchically packed inside the nucleus, which facilitates physical contact between cis-regulatory elements (CREs), such as enhancers and promoters. Accumulating evidence highlights the critical role of higher-order chromatin structure in precise regulation of spatiotemporal gene expression under diverse biological contexts including lineage commitment and cell activation by external stimulus. Genomics and imaging-based technologies, such as Hi-C and DNA fluorescence in situ hybridization (FISH), have revealed the key principles of genome folding, while newly developed tools focus on improvement in resolution, throughput and modality at single-cell and population levels, and challenge the knowledge obtained through conventional approaches. In this review, we discuss recent advances in our understanding of principles of higher-order chromosome conformation and technologies to investigate 4D chromatin interactions.

Radio-Sensitization by Piper longumine of Human Breast Adenoma MDA-MB-231 Cells in Vitro

  • Yao, Jian-Xin;Yao, Zhi-Feng;Li, Zhan-Feng;Liu, Yong-Biao
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3211-3217
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    • 2014
  • Background: The current study investigated the effects of Piper longumine on radio-sensitization of human breast cancer MDA-MB-231 cells and underlying mechanisms. Materials and Methods: Human breast cancer MDA-MB-231 cells were cultured in vitro and those in logarithmic growth phase were selected for experiments divided into four groups: control, X-ray exposed, Piper longumine, and Piper longumine combined with X-rays. Conogenic assays were performed to determine the radio-sensitizing effects. Cell survival curves were fitted by single-hit multi-target model and then the survival fraction (SF), average lethal dose ($D_0$), quasi-threshold dose ($D_q$) and sensitive enhancement ratio (SER) were calculated. Cell apoptosis was analyzed by flow cytometry (FCM). Western blot assays were employed for expression of apoptosis-related proteins (Bc1-2 and Bax) after treatment with Piper longumine and/or X-ray radiation. The intracellular reactive oxygen species (ROS) level was detected by FCM with a DCFH-DA probe. Results: The cloning formation capacity was decreased in the group of piperlongumine plus radiation, which displayed the values of SF2, D0, Dq significantly lower than those of radiation alone group and the sensitive enhancement ratio (SER) of D0 was1.22 and 1.29, respectively. The cell apoptosis rate was increased by the combination treatment of Piper longumine and radiation. Piper longumine increased the radiation-induced intracellular levels of ROS. Compared with the control group and individual group, the combination group demonstrated significantly decreased expression of Bcl-2 with increased Bax. Conclusions: Piper longumine at a non-cytotoxic concentration can enhance the radio-sensitivity of MDA-MB-231cells, which may be related to its regulation of apoptosis-related protein expression and the increase of intracellular ROS level, thus increasing radiation-induced apoptosis.

Effects of low dose irradiation on the calcific nodule formation in MC3T3-E1 osteoblastic cell line (저선량 방사선이 MC3T3-E1 골모세포주의 석회화결절 형성에 미치는 영향)

  • Kim Kyung-A;Koh Kwang-Joon
    • Imaging Science in Dentistry
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    • v.34 no.3
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    • pp.137-144
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    • 2004
  • Purpose: To investigate the effects of low dose irradiation on the calcium content and calcific nodule formation of the MC3T3-El osteoblastic cell line. Materials and Methods: Cells were irradiated with a single dose of 0.2, 0.4 and 0.6 Gy at a dose rate of 5.38 Gy/min using Cs-137 irradiator. After irradiation, the calcium content and calcific nodule formation were examined on the 1st, 2nd, 3rd and 4th week. Results: We did not find any significant difference of total calcium content after irradiation of 0.2, 0.4 and 0.6 Gy when compared with the unirradiated control group. There was no significant difference of total calcium content between 0.2, 0.4 and 0.6 Gy irradiated groups. We found an increased tendency of the calcific nodule formation after irradiation of 0.2, 0.4 and 0.6 Gy when compared with the unirradiated control group without significant difference of calcific nodule formation between 0.2, 0.4 and 0.6 Gy irradiated groups. Conclusion : The results showed an increased tendency of the calcific nodule formation after low dose irradiation. However, this tendency did not increase with the increase of irradiation dose.

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Evaluation of Hepatic Hemangioma by Tc-99m Red Blood Cell Hepatic Blood Pool Scan (간 혈관종의 Tc-99m 표지 적혈구 혈액풀 스캔)

  • Sohn, Myung-Hee
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.3
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    • pp.151-162
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    • 2005
  • Hemangioma is the most common benign tumor of the liver, with a prevalence estimated as high as 7%. Tc-99m red blood cell (RBC) hepatic blood pool scan with single photon omission computed tomography (SPECT) imaging is extremely useful for the confirmation or exclusion of hepatic hemangiomas. The classic finding of absent or decreased perfusion and increased blood pooling ("perfusion/blood pool mismatch") is the key diagnostic element in the diagnosis of hemangiomas. The combination of early arterial flow and delayed blood pooling ("perfusion/blood pool match") is shown uncommonly. In giant hemangioma, filling with radioactivity appears first in the periphery, with progressive central fill-in on sequential RBC blood pool scan. However, the reverse filling pattern, which begins first in the center with progressive peripheral filling, is also rarely seen. Studies with false-positive blood pooling have been reported infrequently in nonhemangiomas, including hemangiosarcoma, hepatocellular carcinoma, hepatic adenoma, and metastatic carcinomas (adenocarcinoma of the colon, small cell carcinoma of the lung, neruroendocrine carcinoma). False-negative results have been also reported rarely except for small hemagniomas that are below the limits of spatial resolution of gamma camera.

A Study on the PET/CT Fusion Imaging (PET/CT 결합영상진단 검사에 관한 연구)

  • Kim, Jong Gyu
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.2
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    • pp.193-198
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    • 2004
  • PET/CT combines the functional information from a positron emission tomography (PET) exam with the anatomical information from a computed tomography (CT) exam into one single exam. A CT scan uses a combination of x-rays and computers to give the radiologist a non-invasive way to see inside your body. One advantage of CT is its ability to rapidly acquire two-dimensional pictures of your anatomy. Using a computer these 2-D images can be presented in 3-D for in-depth clinical evaluation. A PET scan detects changes in the cellular function - how your cells are utilizing nutrients like sugar and oxygen. Since these functional changes take place before physical changes occur, PET can provide information that enables your physician to make an early diagnosis. The PET exam pinpoints metabolic activity in cells and the CT exam provides an anatomical reference. When these two scans are fused together, your physician can view metabolic changes in the proper anatomical context of your body. PET/CT offers significant advantages including more accurate localization of functional abnormalities, and the distinction of pathological from normal physiological uptake, and improvements in monitoring treatment. A PET/CT scan allows physicians to measure the body's abnormal molecular cell activity to detect cancer (such as breast cancer, lung cancer, colorectal cancer, lymphoma, melanoma and other skin cancers), brain disorders (such as Alzheimer's disease, Parkinson's disease, and epilepsy), and heart disease (such as coronary artery disease).

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Management and rehabilitation of moderate-to-severe diabetic foot infection: a narrative review

  • Chi Young An;Seung Lim Baek;Dong-Il Chun
    • Journal of Yeungnam Medical Science
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    • v.40 no.4
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    • pp.343-351
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    • 2023
  • Diabetic foot is one of the most devastating consequences of diabetes, resulting in amputation and possibly death. Therefore, early detection and vigorous treatment of infections in patients with diabetic foot are critical. This review seeks to provide guidelines for the therapy and rehabilitation of patients with moderate-to-severe diabetic foot. If a diabetic foot infection is suspected, bacterial cultures should be initially obtained. Numerous imaging studies can be used to identify diabetic foot, and recent research has shown that white blood cell single-photon emission computed tomography/computed tomography has comparable diagnostic specificity and sensitivity to magnetic resonance imaging. Surgery is performed when a diabetic foot ulcer is deep and is accompanied by bone and soft tissue infections. Patients should be taught preoperative rehabilitation before undergoing stressful surgery. During surgical procedures, it is critical to remove all necrotic tissue and drain the inflammatory area. It is critical to treat wounds with suitable dressings after surgery. Wet dressings promote the formation of granulation tissues and new blood vessels. Walking should begin as soon as the patient's general condition allows it, regardless of the wound status or prior walking capacity. Adequate treatment of comorbidities, including hypertension and dyslipidemia, and smoking cessation are necessary. Additionally, broad-spectrum antibiotics are required to treat diabetic foot infections.

Radiation-induced Apoptosis, Necrosis and G2 Arrest in Fadu and Hep2 Cells

  • Lee Sam-Sun;Kang Beom-Hyun;Choi Hang-Moon;Jeon In-Seong;Heo Min-Suk;Choi Soon-Chul
    • Imaging Science in Dentistry
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    • v.30 no.4
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    • pp.275-279
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    • 2000
  • Purpose: Radiation damage is produced and viable cell number is reduced. We need to know the type of cell death by the ionizing radiation and the amount and duration of cell cycle arrest. In this study, we want to identified the main cause of the cellular damage in the oral cancer cells and normal keratinocytes with clinically useful radiation dosage. Materials and Methods: Human gingival tissue specimens obtained from healthy volunteers were used for primary culture of the normal human oral keratinocytes (NHOK). Primary NHOK were prepared from separated epithelial tissue and maintained in keratinocyte growth medium containing 0.15 mM calcium and a supplementary growth factor bullet kit. Fadu and Hep-2 cell lines were obtained from KCLB. Cells were irradiated in a /sup 137/Cs γ-irradiator at the dose of 10 Gy. The dose rate was 5.38 Gy/min. The necrotic cell death was examined with Lactate Dehydrogenase (LDH) activity in the culture medium. Every 4 day after irradiation, LDH activities were read and compared control group. Cell cycle phase distribution and preG1-incidence after radiation were analyzed by flow cytometry using Propidium Iodine staining. Cell cycle analysis were carried out with a FAC Star plus flowcytometry (FACS, Becton Dickinson, USA) and DNA histograms were processed with CELLFIT software (Becton Dickinson, USA). Results: LDH activity increased in all of the experimental cells by the times. This pattern could be seen in the non-irradiated cells, and there was no difference between the non-irradiated cells and irradiated cells. We detected an induction of apoptosis after irradiation with a single dose of 10 Gy. The maximal rate of apoptosis ranged from 4.0% to 8.0% 4 days after irradiation. In all experimental cells, we detected G2/M arrest after irradiation with a single dose of 10 Gy. Yet there were differences in the number of G2/M arrested cells. The maximal rate of the G2/M ranges from 60.0% to 80.0% 24h after irradiation. There is no significant changes on the rate of the G0/G1 phase. Conclusion: Radiation sensitivity was not related with necrosis but cell cycle arrest and apoptosis. These data suggested that more arrested cell is correlated with more apoptosis.

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Single-Cell-Imaging-Based Analysis of Focal Adhesion Kinase Activity in Plasma Membrane Microdomains Under a Diverse Composition of Extracellular Matrix Proteins (다양한 ECM 조건하에서의 세포막 미세영역 부위 국소접착인산화효소 활성의 단일세포 이미징 기반 분석)

  • Choi, Gyu-Ho;Jang, Yoon-Kwan;Suh, Jung-Soo;Kim, Heon-Su;Ahn, Sang-Hyun;Han, Ki-Seok;Kim, Eunhye;Kim, Tae-Jin
    • Journal of Life Science
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    • v.32 no.2
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    • pp.148-154
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    • 2022
  • Focal adhesion kinase (FAK) is known to regulate cell adhesion, migration, and mechanotransduction in focal adhesions (FAs). However, studies on how FAK activity is regulated in the plasma membrane microdomains according to the composition of extracellular matrix (ECM) proteins are still lacking. A genetically encoded fluorescence resonance energy transfer (FRET)-based biosensor can provide useful information on the activity of intracellular signals with high spatiotemporal resolution. In this study, we analyzed the FAK activities in lipid raft (detergent-resistant membrane) and non-lipid raft (non-detergent-resistant membrane) microdomains using FRET-based membrane targeting FAK biosensors (FAK-Lyn and FAK-KRas biosensors) under four different ECM protein compositions: glass, type 1 collagen, fibronectin, and laminin. Interestingly, FAK activity in response to laminin in a lipid raft microdomain was lower than that in other ECM conditions. Cells subjected to fibronectin showed higher FAK activity in a lipid raft microdomain than that in a non-lipid raft microdomain. Therefore, this study demonstrates that the FAK activity can be distinctively regulated according to the ECM type and the environment of the plasma membrane microdomains.

Effect of irradiation on the Streptococcus mutans (방사선조사가 Streptococcus mutans에 미치는 영향)

  • Ahn, Ki-Dong;Kim, Gyu-Tae;Choi, Yong-Suk;Hwang, Eui-Hwan
    • Imaging Science in Dentistry
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    • v.37 no.1
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    • pp.35-43
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    • 2007
  • Purpose : To observe direct effect of irradiation on cariogenic Streptooccus mutans. Materials and Methods : S. mutans GS5 was exposed to irradiation with a single absorbed dose of 10, 20, 30, and 40Gy. Viability and changes in antibiotic sensitivity, morphology, transcription of virulence factors, and protein profile of bacterium after irradiation were examined by pour plate, disc diffusion method, transmission electron microscopy, RT-PCR, and SDS-PAGE, respectively. Results : After irradiation with 10 and 20Gy, viability of S. mutans was reduced. Further increase in irradiation dose, however, did not affect the viability of the remaining cells of S. mutans. Irradiated 5. mutans was found to have become sensitive to antibiotics. In particular, the bacterium irradiated with 40Gy increased its susceptibility to cefotaxime, penicillin, and tetracycline. Under the transmission electron microscope, number of morphologically abnormal cells was increased as the irradiation dose was increased. S. mutans irradiated with 10 Gy revealed a change in the cell wall and cell membrane. As irradiation dose was increased, a higher number of cells showed thickened cell wall and cell membrane and Iysis, and appearance of ghost cells was noticeable. In RT-PCR, no difference was detected in expression of gtfB and spap between cells with and without irradiation of 40Gy. In SDS-PAGE, proteins with higher molecular masses were gradually diminished as irradiation dose was increased. Conclusion : These results suggest that irradiation affects the cell Integrity of S. mutans, as observed by SDS-PAGE, and as manifested by the change in cell morphology, antibiotic sensitivity, and eventually viability of the bacterium.

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Primary Hepatosplenic B-cell Lymphoma: linitial Diagnosis and Assessment of Therapeutic Response with F-18 FDG PET/CT (원발성 간비장 B세포성 림프종에서 치료전과 치료 후 F-18 FDG PET/CT 소견)

  • Kang, Sung-Min;Lee, Hong-Je;Seo, Ji-Hyoung;Lee, Sang-Woo;Ahn, Byeong-Cheol;Lee, Jae-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.4
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    • pp.333-336
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    • 2008
  • A 52-year-old woman with a history of general weakness, fatigue, weight loss, elevated serum levels of liver transaminase enzyme for three months underwent an F-18 FDG PET/CT scan to evaluate a cause of the hepatosplenomegaly found on abdominal ultrasonography. Initial PET/CT revealed markedly enlarged liver and spleen with intense FDG uptake. Otherwise, there were no areas of abnormal FDG uptake in whole body image. Histological evaluation by a hepatic needle biopsy demonstrated diffuse large B cell type lymphoma and final diagnosis for this patient was hepatosplenic B-cell lymphoma. She received five cycles of CHOP chemotherapy, and second PET/CT scan was followed after then. Follow-up PET-CT revealed normal sized liver with disappearance of abnormal FDG uptake. Hepatosplenic B-cell lymphoma is relatively rare and mostly presents as single or multiple nodules.1,2 Diffuse type hepatosplenic lymphoma is extremely rare and poorly recognized entity.3 The diagnosis is very difficult and complicated by the presence of misleading symptoms.4 In this rare hepatosplenic B-cell lymphoma case, F-18 FDG PET/CT scan provided a initial diagnostic clue of hepatosplenic lymphoma and an accurate chemotherapy response.