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Статті в журналах з теми "CTC Isolation"

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Cha, Jiwon, Hyungseok Cho, Jae-Seung Chung, Joon Seong Park, and Ki-Ho Han. "Effective Circulating Tumor Cell Isolation Using Epithelial and Mesenchymal Markers in Prostate and Pancreatic Cancer Patients." Cancers 15, no. 10 (May 18, 2023): 2825. http://dx.doi.org/10.3390/cancers15102825.

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Circulating tumor cells (CTCs) display antigenic heterogeneity between epithelial and mesenchymal phenotypes. However, most current CTC isolation methods rely on EpCAM (epithelial cell adhesion molecule) antibodies. This study introduces a more efficient CTC isolation technique utilizing both EpCAM and vimentin (mesenchymal cell marker) antibodies, alongside a lateral magnetophoretic microseparator. The effectiveness of this approach was assessed by isolating CTCs from prostate (n = 17) and pancreatic (n = 5) cancer patients using EpCAM alone, vimentin alone, and both antibodies together. Prostate cancer patients showed an average of 13.29, 11.13, and 27.95 CTCs/mL isolated using EpCAM alone, vimentin alone, and both antibodies, respectively. For pancreatic cancer patients, the averages were 1.50, 3.44, and 10.82 CTCs/mL with EpCAM alone, vimentin alone, and both antibodies, respectively. Combining antibodies more than doubled CTC isolation compared to single antibodies. Interestingly, EpCAM antibodies were more effective for localized prostate cancer, while vimentin antibodies excelled in metastatic prostate cancer isolation. Moreover, vimentin antibodies outperformed EpCAM antibodies for all pancreatic cancer patients. These results highlight that using both epithelial and mesenchymal antibodies with the lateral magnetophoretic microseparator significantly enhances CTC isolation efficiency, and that antibody choice may vary depending on cancer type and stage.
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Lee, Jae Hyuk, Sung Ho Park, Jihoon Kang, Jihyun Lee, Soee Kim, Jungwon Kim, Young Woong Sohn, and Jong Kil Lee. "Abstract 6692: Identification of circulating tumor cells based on machine learning." Cancer Research 83, no. 7_Supplement (April 4, 2023): 6692. http://dx.doi.org/10.1158/1538-7445.am2023-6692.

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Abstract Circulating tumor cells (CTC) are tumor cells that have shed into the bloodstream from primary tumor and circulate in the blood. CTC could provide better understanding of tumor metastasis and show the possibility as promising biomarker for early detection of cancer, cancer prognosis, noninvasive monitoring of treatment response, and personalized medicine. However, the isolation and characterization has been a major technological challenge due to their rareness among the massive blood cells. CytoGen’s Smart BiopsyTM CTC platform comprised of CTC isolator, IF (Immunofluorescence) Stainer, and Cell Image Analyzer is a unique and competitive technology platform for the isolation and analysis of CTC from blood. Intact live CTC could be isolated gravity-based filtration via high-density micro-porous (HDM) chip with minimized cellular damage and high retrieval rate. For the more accurate and quick analysis of CTC after isolation with Smart BiopsyTM CTC platform, we are developing the analysis method of CTC identification using machine learning technologies. Preliminary results of machine learning showed the accurate separation of CTC and PBMC (peripheral blood mononuclear cells) based on the specific markers of them. CTC identification based on machine learning technologies might be reliable analysis method and give secure information of CTC. Citation Format: Jae Hyuk Lee, Sung Ho Park, Jihoon Kang, Jihyun Lee, Soee Kim, Jungwon Kim, Young Woong Sohn, Jong Kil Lee. Identification of circulating tumor cells based on machine learning. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6692.
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Macaraniag, Celine, Jian Zhou, Ian Papautsky, Jing Li, William Putzbach, and Nissim Hay. "Abstract 5598: Microfluidic isolation and capture of circulating tumor cells and clusters from mouse blood." Cancer Research 83, no. 7_Supplement (April 4, 2023): 5598. http://dx.doi.org/10.1158/1538-7445.am2023-5598.

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Abstract Background. Circulating tumor cells (CTCs) and their clusters have been regarded as potential targets for therapies and subjects for identifying certain mechanisms of cancer metastasis. Mouse models have been especially useful tools to investigate the process of metastasis, since they are easier to control and are less variable compared to clinical patient samples. Therefore, it is of interest to extract and study CTCs and CTC clusters from such mouse models. However, most isolation systems are designed and tested for human blood and cell lines. Sized-based methods of CTC isolation have been optimized for human cell lines, but mouse CTCs are generally smaller in size (i.e., ~14 µm EO771 breast cancer cells). Therefore, mouse cell separation necessitates a varied protocol for CTC isolation. We used mouse blood and EO771, a mouse breast cancer cell line, to demonstrate the isolation of mouse CTCs from mouse blood using a sized-based microfluidic device. We also validated the presence of endogenous CTCs and CTC-neutrophil clusters in a tumor bearing mouse model. Methods. Our microfluidic device employs inertial migration in a straight channel segment (150 µm × 50 µm × 24 mm) to isolate CTCs from blood samples. We spiked 100 and 1000 ZsGreen-expressing EO771 cells in 5 × diluted blood containing tdTomato-expressing neutrophils, easily distinguishing CTCs from the red-colored neutrophils. To enumerate the recovered cells, we used Cytospin to concentrate the cells into a glass slide. Lastly, we used hydrodynamic traps to capture CTC clusters from tumor-bearing mouse blood. The blood was extracted from the posterior vena cava of the mice at the endpoint experiment. Results. We achieved an overall recovery rate of 22-29%, which combines the recovery of the isolation device and the Cytospin. Here, we enumerate cells only after their extraction from the collection tube outside of the isolation device. Other methods with higher capture efficiency enumerate the recovered CTCs before they are harvested from the isolation device which is not always representative of the number of cells that are analyzed. We were also able to discover endogenous CTC-neutrophil clusters from tumor-bearing mice which we distinguished apart with anti-EpCAM staining of CTCs (green) and anti-Ly6G (red) staining of neutrophils. Conclusion. We demonstrated and optimized a procedure for isolating mouse-derived CTCs from naïve mice blood and CTC-neutrophil clusters from tumor-bearing mice with our microfluidic isolation devices, as opposed to the isolation of human CTCs as in other sized-based isolation systems. In future studies, we aim to continue to use our microfluidic devices to capture and study CTC and CTC clusters. Citation Format: Celine Macaraniag, Jian Zhou, Ian Papautsky, Jing Li, William Putzbach, Nissim Hay. Microfluidic isolation and capture of circulating tumor cells and clusters from mouse blood. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5598.
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Cheng, Jie, Yang Liu, Yang Zhao, Lina Zhang, Lingqian Zhang, Haiyang Mao, and Chengjun Huang. "Nanotechnology-Assisted Isolation and Analysis of Circulating Tumor Cells on Microfluidic Devices." Micromachines 11, no. 8 (August 14, 2020): 774. http://dx.doi.org/10.3390/mi11080774.

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Circulating tumor cells (CTCs), a type of cancer cell that spreads from primary tumors into human peripheral blood and are considered as a new biomarker of cancer liquid biopsy. It provides the direction for understanding the biology of cancer metastasis and progression. Isolation and analysis of CTCs offer the possibility for early cancer detection and dynamic prognosis monitoring. The extremely low quantity and high heterogeneity of CTCs are the major challenges for the application of CTCs in liquid biopsy. There have been significant research endeavors to develop efficient and reliable approaches to CTC isolation and analysis in the past few decades. With the advancement of microfabrication and nanomaterials, a variety of approaches have now emerged for CTC isolation and analysis on microfluidic platforms combined with nanotechnology. These new approaches show advantages in terms of cell capture efficiency, purity, detection sensitivity and specificity. This review focuses on recent progress in the field of nanotechnology-assisted microfluidics for CTC isolation and detection. Firstly, CTC isolation approaches using nanomaterial-based microfluidic devices are summarized and discussed. The different strategies for CTC release from the devices are specifically outlined. In addition, existing nanotechnology-assisted methods for CTC downstream analysis are summarized. Some perspectives are discussed on the challenges of current methods for CTC studies and promising research directions.
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Theil, Gerit, Joanna Bialek, Christine Weiß, Felix Lindner, and Paolo Fornara. "Strategies for Isolating and Propagating Circulating Tumor Cells in Men with Metastatic Prostate Cancer." Diagnostics 12, no. 2 (February 15, 2022): 497. http://dx.doi.org/10.3390/diagnostics12020497.

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Selecting a well-suited method for isolating/characterizing circulating tumor cells (CTCs) is challenging. Evaluating sensitive and specific markers for prostate cancer (PCa)-specific CTC identification and analysis is crucial. We used the CellCollector EpCAM-functionalized system (CC-EpCAM) and evaluated and developed a PCa-functionalized version (CC-PCa); we then compared CTC isolation techniques that exploit the physical and biological properties of CTCs. We established two cohorts of metastatic PCa patients (mPCa; 15 in cohort 1 and 10 in cohort 2). CTC cultivation experiments were conducted with two capturing methods (Ficoll and ScreenCell). The most sensitive detection rates and highest CTC counts were reached with the CC-PCa and ScreenCell system. Patients with ≥5 CTCs isolated with CC-EpCAM had an overall survival (OS) of 0.93 years, and patients with ≥5 CTCs isolated with CC-PCa had an OS of 1.5 years in cohort 1. Nevertheless, we observed the highest sensitivity and specificity for 24-month survival by the Ficoll with CD45 depletion and ScreenCell system with May-Grunwald Giemsa (MGG) staining. The EpCAM molecule is an essential factor related to OS for CTC isolation based on biological properties in mPCa patients. The best-suited CTC capture system is not limited to one characteristic of cells but adapted to downstream analysis.
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Francescangeli, Federica, Valentina Magri, Maria Laura De Angelis, Gianluigi De Renzi, Orietta Gandini, Ann Zeuner, Paola Gazzaniga, and Chiara Nicolazzo. "Sequential Isolation and Characterization of Single CTCs and Large CTC Clusters in Metastatic Colorectal Cancer Patients." Cancers 13, no. 24 (December 18, 2021): 6362. http://dx.doi.org/10.3390/cancers13246362.

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Circulating tumor cells (CTCs) detach from a primary tumor or its metastases and circulate in the bloodstream. The vast majority of CTCs are deemed to die into the bloodstream, with only few cells representing viable metastatic precursors. Particularly, single epithelial CTCs do not survive long in the circulation due to the loss of adhesion-dependent survival signals. In metastatic colorectal cancer, the generation of large CTC clusters is a very frequent occurrence, able to increase the aptitude of CTCs to survive in the bloodstream. Although a deepened analysis of large-sized CTC clusters might certainly offer new insights into the complexity of the metastatic cascade, most CTC isolation techniques are unfortunately not compatible with large-sized CTC clusters isolation. The inappropriateness of standard CTC isolation devices for large clusters isolation and the scarce availability of detection methods able to specifically isolate and characterize both single CTCs and CTC clusters finally prevented in-depth studies on the prognostic and predictive value of clusters in clinical practice, unlike that which has been described for single CTCs. In the present study, we validated a new sequential filtration method for the simultaneous isolation of large CTC clusters and single CTCs in patients with metastatic colorectal cancer at failure of first-line treatments. The new method might allow differential downstream analyses for single and clustered CTCs starting from a single blood draw, opening new scenarios for an ever more precise characterization of colorectal cancer metastatic cascade.
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Jiang, Xiaocheng, Keith H. K. Wong, Aimal H. Khankhel, Mahnaz Zeinali, Eduardo Reategui, Matthew J. Phillips, Xi Luo, et al. "Microfluidic isolation of platelet-covered circulating tumor cells." Lab on a Chip 17, no. 20 (2017): 3498–503. http://dx.doi.org/10.1039/c7lc00654c.

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Ahmed, Shakera, Atul Bharde, Muhammad Mosaraf Hossain, Ramendu Parial, Nusrat Jahan Nayeema, Manisha Das, Mizanur Rahman, et al. "Circulating tumor cells (CTCs) detection and isolation in different subtypes of early-stage breast cancer patients from Bangladesh." Journal of Clinical Oncology 41, no. 16_suppl (June 1, 2023): e12529-e12529. http://dx.doi.org/10.1200/jco.2023.41.16_suppl.e12529.

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e12529 Background: Breast cancer is a highly heterogeneous pathophysiology characterized by poor outcomes. Due to the increasing incidence and disease progression rates and undefined relapse periods, reliable disease monitoring is a challenge and has remained an unmet need. Advancements in liquid biopsy have significantly enhanced our understanding of clinical oncology. CTC-based liquid biopsy is emerging as a reliable prognostic tool to predict various clinical indicators. Although extensively investigated in metastatic breast cancers, little is known about CTCs in early-stage breast cancers. CTCs with respect to different molecular subtypes of breast cancer in early-stage breast cancer patients is evaluated. Methods: In this prospective clinical trial (CMC 59.27.0000.013.19 PG.009.2022/262) 40 early-stage patients with luminal (A +B, 33.33%), HER 2 positive (12.8%), triple-negative (12.8%) and undetermined (41.07%) subtype were recruited. CTCs were isolated in 1.5 ml blood using the Drug Controller General of India approved OncoDiscover CTC test. This platform contains affinity-based magnetic nanoparticles to mediate EpCAM-based CTC isolation. CTCs were detected as CK18+, DAPI+, and CD45- cells using a fluorescence detection-based automated digital imaging platform. Results: CTCs were detected in 60 % of patients with a mean CTC count of 1 cell / 1.5ml blood. Among total positive patients, the luminal subtype was the least positive (46 %), followed by TNBC (60 %) and undetermined (62.5 %) subgroup, while all HER2-positive patients showed the presence of CTCs. Besides individual cells, CTC clusters were detected in 12.5 % of patients and they were equally distributed in luminal and HER2-positive subpopulations. When analyzed on the scale of tumor grade, grade I patients did not show the presence of CTC, while 58.33 % of grade II patients had ≥ 1 CTC. All grade III patients showed the presence of ≥ 1 CTC. CTC count was high among CTC-positive grade II patients (average 2 CTCs) and correlated well with the presence of CTC clusters in these patients. Patients who had surgical intervention had a low CTC burden compared to patients who did not have a surgical resection. 75 % of treatment naïve patients showed the presence of CTC while 58 % of patients receiving chemotherapy alone showed the presence of 1 CTC. 50 % of patients who had surgery followed by CT+RT showed the presence of 1 CTC. Conclusions: The presence of CTCs may suggest for biological progression of disease in early-stage BC patients. CTC detected in all HER2-positive patients suggested the high shedding nature of these tumors, which correlates well with their reported migratory tendency. The presence of CTCs did not show a clear correlation with the treatment regimen. However, this data is based on a single time point and needs longitudinal correlation with CTC on a larger sample size. Clinical trial information: 59.27.0000.013.19 PG.009.2022/262 .
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Liao, Chia-Jung, Chia-Hsun Hsieh, Feng-Chun Hung, Hung-Ming Wang, Wen-Pin Chou, and Min-Hsien Wu. "The Integration of a Three-Dimensional Spheroid Cell Culture Operation in a Circulating Tumor Cell (CTC) Isolation and Purification Process: A Preliminary Study of the Clinical Significance and Prognostic Role of the CTCs Isolated from the Blood Samples of Head and Neck Cancer Patients." Cancers 11, no. 6 (June 6, 2019): 783. http://dx.doi.org/10.3390/cancers11060783.

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Conventional positive and negative selection-based circulating tumor cell (CTC) isolation methods might generally ignore metastasis-relevant CTCs that underwent epithelial-to- mesenchymal transition and suffer from a low CTC purity problem, respectively. To address these issues, we previously proposed a 2-step CTC isolation method integrating a negative selection CTC isolation and subsequent spheroid cell culture. In addition to its ability to isolate CTCs, more importantly, the spheroid cell culture used could serve as a cell culture model mimicking the process of new tumor tissue formation during cancer metastasis. Therefore, it is promising not only to selectively isolate metastasis-relevant CTCs but also to test the potential of cancer metastasis and thus the prognosis of disease. To explore these issues, experiments were performed. The key findings of this study demonstrated that the method was able to harvest both epithelial (E)- and mesenchymal (M)-type CTCs without selection bias. Moreover, both the M-type CTC count and the information obtained from the multidrug resistance-associated protein 2 (MRP2) and MRP5 gene expression analysis of the CTCs isolated via the 2-step CTC isolation method might be able to serve as prognostic factors for progression-free survival in head and neck squamous cell carcinoma.
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Tretyakova, Maria S., Maxim E. Menyailo, Anastasia A. Schegoleva, Ustinia A. Bokova, Irina V. Larionova, and Evgeny V. Denisov. "Technologies for Viable Circulating Tumor Cell Isolation." International Journal of Molecular Sciences 23, no. 24 (December 15, 2022): 15979. http://dx.doi.org/10.3390/ijms232415979.

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The spread of tumor cells throughout the body by traveling through the bloodstream is a critical step in metastasis, which continues to be the main cause of cancer-related death. The detection and analysis of circulating tumor cells (CTCs) is important for understanding the biology of metastasis and the development of antimetastatic therapy. However, the isolation of CTCs is challenging due to their high heterogeneity and low representation in the bloodstream. Different isolation methods have been suggested, but most of them lead to CTC damage. However, viable CTCs are an effective source for developing preclinical models to perform drug screening and model the metastatic cascade. In this review, we summarize the available literature on methods for isolating viable CTCs based on different properties of cells. Particular attention is paid to the importance of in vitro and in vivo models obtained from CTCs. Finally, we emphasize the current limitations in CTC isolation and suggest potential solutions to overcome them.
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Дисертації з теми "CTC Isolation"

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Zeinali, Mina [Verfasser], and Mathias [Akademischer Betreuer] Hafner. "Isolation, characterization, and expansion of heterogeneous circulating tumor cell (CTC) populations from cancer patients using microfluidic technologies / Mina Zeinali ; Betreuer: Mathias Hafner." Heidelberg : Universitätsbibliothek Heidelberg, 2019. http://d-nb.info/1201088259/34.

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Broncy, Lucile. "Isolement et caractérisation moléculaire de cellules rares circulantes individuelles : développement de nouvelles approches méthodologiques en oncologie prédictive et diagnostic prénatal." Thesis, Université Paris-Saclay (ComUE), 2017. http://www.theses.fr/2017SACLS391/document.

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L’objectif principal de ce projet de recherche doctorale est la mise au point d’approches méthodologiques fiables et reproductibles permettant la caractérisation génétique de cellules rares circulantes isolées par la méthode de filtration ISET® (Rarecells®, France). La première application développée consiste en la détection des mutations du gène suppresseur de tumeur VHL (Von Hippel Lindau) dans les cellules rares circulantes (CRC) uniques isolées du sang de 30 patients atteints de carcinome rénal à cellules claires (CRCC), et réalisée comparativement à l’analyse cytopathologique. L’étude génétique a également été conduite en parallèle dans les 30 tissus tumoraux correspondants. Les résultats ont mis en lumière une potentielle complémentarité de l’approche de génétique moléculaire sur cellules uniques avec l’analyse cytomorphologique de référence et suggèrent que combiner ces approches permettrait d’obtenir une plus grande sensibilité de détection des cellules cancéreuses circulantes chez les patients atteints de CRCC. Une deuxième application a consisté en le développement d’une approche innovante pour le diagnostic prénatal non-invasif des maladies génétiques récessives par analyse de cellules trophoblastiques rares collectées au niveau du col de l’utérus. Enfin, des développements supplémentaires ont permis d’optimiser les analyses de séquençage à haut débit et de les appliquer à des CRC individuelles isolées par ISET®. Cette nouvelle approche, associée à l’isolement de CRC non fixées, est en mesure de fournir des données génétiques élargies à l’exome entier pour des applications à la fois en oncologie prédictive et en diagnostic prénatal non invasif
The aim of this doctoral research project is the development of reliable and reproducible methodological approaches enabling the genetic characterization of circulating rare cells (CRC) isolated by ISET® filtration (Rarecells®, France). The first application developed consists in detecting mutations of the VHL (Von Hippel Lindau) tumor suppressor gene in single CRC isolated from the blood of 30 patients patients with clear cell renal cell carcinoma (ccRCC), assessed according to the results obtained by cytopathological analysis. In parallel, genetic analysis of VHL mutations was conducted in the corresponding tumor tissues. Results revealed a potential complementarity of the molecular genetic approach targeted to single cells with the reference method of cytopathological analysis and suggested that combining both strategies could improve the sensitivity of circulating cancer cells’ detection in patients with ccRCC. A second application consisted in the development of an innovative approach for non-invasive prenatal diagnosis of recessive genetic diseases by analysis of rare trophoblastic cells collected from the cervix. Finally, further developments allowed to optimize high-throughput sequencing analyses and to apply them to single CRC isolated by ISET®. This approach, combined with the isolation of living CRC, enabled us to obtain broader genetic data from the whole exome and should foster innovative applications to both predictive oncology and non-invasive prenatal diagnosis
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Zheng, Xiangjun. "Selective Isolation of Circulating Tumor Cells in Antibody-Functionalized Microsystems." Diss., The University of Arizona, 2011. http://hdl.handle.net/10150/203438.

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Attachment of circulating tumor cells in microfluidic devices functionalized with proper antibodies was studied. Under static experimental conditions, microchambers were utilized to study the parameters such as cell suspension concentration, incubation time or ambient temperature that may affect the binding of cell to the functionalized surfaces. Specific capture of cells from suspensions increases exponentially with incubation time and linearly with concentration within the tested range. Functionalizing a surface with counter-receptors enables capture of almost 100% of cells within 15 minutes incubation time at ambient temperature higher than 25°C. Suspending cells with different receptors, changing the counter receptors immobilized on the surface, or incubation the cell suspension at low ambient temperature result in a poor capture ratio. To illustrate the specific binding of target cells, various binary mixtures of target cancer and blood cells were incubated in the microchambers. The microsystem sensitivity, specificity and accuracy were determined as a function of the incubated cell concentrations. In general, the system specificity increases while the sensitivity decreases with increasing cell concentration; the accuracy of the system depends weakly on cell concentration within the tested range. The cell attachment dynamics in shear flow was studied by driving the MDA-MB- 231 or BT-20 cells through microchannels functionalized with EpCAM antibodies. The cell attachment ratio was experimentally determined at different flow rates. A modeling system based on Stokesian as well as cell-adhesive dynamics is adopted to analyze the cell motion. The cell motion is modeled as a rigid sphere, with receptors on its surface, moving under shear flow above a surface immobilized with ligands. The system is described mathematically by the Langevin equation, in which the receptor-ligand bonds are modeled as linear springs. Primarily depending on the applied flow rate, three distinct dynamic states of cell motion have been observed: free motion, rolling adhesion, and firm adhesion. The fraction of cells captured due to firm adhesion, defined as attachment ratio, depends on the applied flow rate with a characteristic value that increases with either cellreceptor or surface-ligand density. Utilizing this characteristic flow rate as a scaling parameter, all measured and calculated attachment ratios for different receptor and ligand densities collapse onto a single exponential curve. Binary mixtures of target MDA-MB-231 cells and non-target BT-20 cells were driven through anti-cadherin-11 functionalized microchannels to study the selective isoaltion of target cells from binary mixtures. The system sensitivity is very high, above 0.95, while the specificity is only moderately high, about 0.85, essentially independent of the relative concentration of the target and non-target cells in the binary mixture. An attachment/detachment flow field pattern is proposed to enhance the system specificity. Utilizing this flow pattern with a 1:1,000 MDA-MB-231:BT-20 binary cell mixture, the microfluidic system specificity increased to about 0.95 while the sensitivity remained above 0.95. In order to obtain high experimental throughput allowing lower relative concentration of target cells, a microchannel array which enables processing samples containing about 510⁵ cells with a minimum target cell concentration ratio of 1/100,000 was designed and fabricated. To demonstrate selective isolation of target cells, binary mixtures of BT-20 cells and MIA PaCa-2 cells were driven through microchannel arrays functionalized with EpCAM antibodies; the EpCAM positive BT-20 cells served as target cells and the EpCAM negative MIA PaCa-2 cells as non-target cells. The relative concentration ratio of target/non-target cells varied from 1:1 to 1:100,000. The sensitivity was close to 1.0 while the specificity was also high, about 0.95. The additional detachment step, with a faster flow rate, enhanced the specificity to about 0.985. Initial results of two sets of experiments are reported as preliminary studies for future work. In the first set of experiments, whole blood samples from healthy donors were spiked with a known number of BT-20 cells at a concentration of 500 CTCs per milliliter blood or 50 CTCs per milliliter blood. After a pretreatment to enrich the CTCs, the samples were driven through microchannel arrays functionalized with anti-EpCAM. For both samples, around 55% of the target CTCs were captured in the microchannel arrays. The second set of experiments was dedicated to characterization of target cells exposed to applied shear stress. BT-20 or MDA-MB-231 cells were driven through microchannels functionalized with EpCAM antibodies to allow target cell attachment; then, a high flow rate was applied to detach the captured cells. The detached cells were collected and cultured in an incubator to test their viability. For both cell lines, the majority of the captured CTCs collected from the microchannels were viable. The images taken after three and seven days of culture demonstrate continuous cell growth and division.
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Marsavela, Anda-Gabriela. "Optimisation of the isolation and identification of circulating melanoma cells." Thesis, Edith Cowan University, Research Online, Perth, Western Australia, 2017. https://ro.ecu.edu.au/theses/1993.

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Although melanoma is largely curable when detected in its earliest stages, it can metastasise to other tissues, drastically reducing survival rates. The most recent therapies used to treat metastatic melanoma are effective long-term in only 11 to 33% of patients. Our ability to monitor treatment failure is limited. New prognostic markers are urgently required to allow monitoring of treatment response and disease progression. Circulating tumour cells (CTCs) are released into the bloodstream by the tumours within a patient, this being a key step in melanoma spread. Since CTCs can be detected in the blood of metastatic melanoma patients, these cells can be used as a “liquid biopsy”, providing critical insight into each person’s melanoma biology. Melanoma CTCs have been described as very heterogeneous, hindering their isolation via commonly used CTC capturing methods. To address this, microfluidic devices have been developed to isolate viable CTCs from blood, independently of their marker expression. This study aimed to determine the effectiveness of two different microfluidic devices (Slanted and A5) in recovering melanoma cell lines, and their potential use in the isolation of CTCs from the blood of metastatic melanoma patients. It also aimed to study additional cancer or melanoma specific markers to be used in immunostaining protocols for detection of CTCs after microfluidic enrichment. The optimal isolation procedure was identified as two rounds of enrichment with the Slanted spiral device, after which we obtained a 3-log depletion of white blood cells and a recovery of over 60% when cells from two melanoma cell lines were spiked into blood samples from healthy volunteers. In addition, we optimised the detection of CTCs using four melanoma markers (gp100, Melan-A, s100 and MCSP) combined in a multimarker immunocytochemistry staining protocol. The optimised enrichment and detection procedures were validated in a cohort of ten metastatic melanoma patients. Results showed that 40% of the patients had one or more CTCs in their blood (1-4 CTCs/8 mL of blood). Furthermore, three additional markers (Vimentin, RANK, and ABCB5) were trialled so as to increase detection of highly heterogeneous melanoma CTCs in samples that have been processed through the Slanted microfluidic device. The improved enrichment and detection of CTCs in the blood of melanoma patients using the methods developed as part of this study will facilitate the molecular, genomic and functional characterisation of melanoma CTCs. This will ultimately improve our understanding of the biology of melanoma CTCs and their role in metastatic spread and treatment response.
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5

Arraki, Kamel. "Les stilbénoïdes chez les Cypéracées : isolation, identification et étude de leurs activités biologiques : identification et dosage des stilbènes dans des vins Tunisiens." Thesis, Bordeaux, 2014. http://www.theses.fr/2014BORD0481/document.

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Les stilbènes sont des composés phénoliques issus du métabolisme secondairevégétal, dont la distribution au sein du règne végétal est limitée aux espèces qui ont acquis aucours de l’évolution la capacité à synthétiser ces molécules. Leurs impacts et leurs activitésbiologiques tels que les effets neuroprotecteurs, anticancérigènes, antioxydants ont déjàconcerné plusieurs sujets d’étude. C’est dans ce contexte que l’objectif de notre travail a prisnaissance. Dans un premier temps, nous avons isolé et identifié ces molécules chez quelquesespèces de la famille des Cypéracées. Cette étude phytochimique a été réalisée en utilisant unensemble de stratégies analytiques et préparatives faisant appel à la CLHP analytique etpréparative et la CPC (Chromatographie de Partage Centrifuge) pour l’obtention desmolécules pures et à la LC-Masse et la RMN pour l’identification des composées isolés. Dansun second temps, nous avons étudié les activités biologiques in vitro de ces produits telles queles activités antioxydantes par trois méthodes (ORAC, DPPH et MCA) et l’effet de cesstilbènes sur la cytotoxicité neuronale induite par le peptide β-amyloïde avec des cellulesPC12. Nous avons isolé une nouvelle molécule, le carexinol A, pour la première fois, qui amontré une forte activité anti-amyloïde. La dernière partie de cette thèse fait état de l’analyseet du dosage des stilbènes dans des vins tunisiens dont le vin Sidi Zahia qui a donné lesmeilleurs résultats
Stilbenes are phenolic compounds of plant secondary metabolism, whosedistribution within the plant kingdom is limited to species that have acquired during theevolution the ability to synthesize these molecules. Their impacts and their biologicalactivities such as neuroprotective, anticarcinogenic, antioxidant effects have already touchedseveral topics. It is in this context that the purpose of our work arose. First, we have isolatedand identified these molecules in some species of the sedge family. Phytochemical studieswere performed using a set of analytical and preparative strategies by means of analytical andpreparative HPLC and CPC (Centrifugal Partition Chromatography) for obtaining puremolecules and LC-Mass and NMR for identification of compound isolated. In a second step,we investigated the in vitro biological properties of these products such as antioxidantactivities by three methods (ORAC, DPPH and MCA) and their effect on neuronalcytotoxicity induced by β-amyloid peptide with PC12 cells. We isolated a new molecule, forthe first time, carexinol A, that showed strong anti-amyloid activity. The last part of this thesisrefers to the analysis and determination of stilbenes in Tunisian wines including Sidi Zahiawine that gave the best results
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Almutawa, Qamar E. B. A. "Impact of Chromosomal Translocations (CTs) on reproductive isolation and fitness in natural yeast isolates." Thesis, University of Manchester, 2017. https://www.research.manchester.ac.uk/portal/en/theses/impact-of-chromosomal-translocations-cts-on-reproductive-isolation-and-fitness-in-natural-yeast-isolates(2a16e0ae-0e02-4e67-9075-bac626e81a7e).html.

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Identifying the molecular mechanisms behind reproductive isolation between closely related yeast species provides avaluable understanding of their evolution. Sequence divergence and chromosomal rearrangements are the main post-zygotic barriers behind reproductive isolation within Saccharomyces „sensu strico’ species, where hybrids are readily formed but sterile upon meiosis. Saccharomyces paradoxus and Saccharomyces cariocanus have an almost identical genome in terms of sequence, and therefore provide good model systems to explore the impact of karyotypic rearrangements on reproductive isolation. According to the biological species concept they are considered two different species despite having low sequence divergence. Since the karyotypic analysis revealed that the genomic differences are restricted to four chromosomal translocations, we hypothesized that such rearrangements may be the cause of low spore viability between them. To test this expectation, we engineered two chromosomal translocations in S. paradoxus YPS138, via Cre-loxP mediated recombination event, to render those parts of genome collinear to S.cariocanus UFRJ50816. Our analysis revealed that hybrids between S. cariocanus and engineered S. paradoxus harbouring two translocations showed a significant increase in spore viability (12.7%) compared to control hybrids harbouring five translocations (3.4%) (P=0.0031and P=0.0125, respectively, Two-sample t-test). Consequently, fitness in meiosis was improved four fold by undoing two translocations. Given this result, the prediction for spore viability in complete collinear crossing would be around 50.8 %, which is still far from the value of ca. 100%, which would be expected for strains with very low sequence divergence and belonging to the same species. This indicates that other factors may contribute to meiotic fitness in these hybrids. Further investigation was carried to determine the genome structures by using the PacBio sequencing approach. Our DNA sequencing data revealed other, previously undetected, rearrangements in S. cariocanus strain: one new reciprocal translocation between chromosomes XIII and XIV and 11 inversions distributed in 6 chromosomes. The variations in meiotic viability observed in the engineered hybrids could be because of these 5 chromosomal translocations. Further experiments were also carried out to evaluate the impact of translocations on mitotic fitness and gene expression; we observed a significant drop in the mitotic fitness of engineered translocant strains under different nutritional and temperature stresses. These changes were also accompanied with alteration in genes expression throughout the genome. Our RNA- seq data revealed that many genes were up- or down- regulated because of the translocation. Several genes with altered expression in translocant strains are correlated with morphology changes when they are up- or down- regulated. Therefore, the cell morphology was evaluated under light microscopy and different abnormal cells were detected compared to the wild type. Irregular cell morphology included elongated and clumped cells. Overall, these data confirmed that chromosomal translocations were the cause of reproductive isolation between S. paradoxus and S. cariocanus and play an important role in altering the phenotype and gene expression.
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Clément, Chami Mélissa. "Développement de méthodologies analytiques innovantes dans le domaine des compléments alimentaires à base de plantes : séparation, purification et caractérisation de marqueurs spécifiques." Thesis, Université Côte d'Azur (ComUE), 2019. http://www.theses.fr/2019AZUR4113.

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L’une des principales contraintes dans l’analyse de produits naturels est l’accès au standard analytique de molécules considérées comme marqueurs d’une plante ou faisant l’objet de restrictions car potentiellement dangereuses. Les problématiques qui en découlent ont tissé le fil conducteur de l’ensemble des travaux de cette thèse. Le premier objectif a été d’étudier les performances qu’offre le couplage entre l’HPTLC et la spectrométrie de masse via l’Interface MS 2 dans le domaine de l’identification de marqueurs de plantes employées dans les compléments alimentaires. Il apparaît que dans le cadre de contrôle de routine, ce couplage a toute sa place car les informations obtenues pourraient permettre l’identification de plantes selon des monographies issues de la pharmacopée européenne. Cependant, dans le cadre de la recherche de nouveaux composés, les résultats obtenus restent malgré tout aléatoires. Le deuxième objectif a été d’étudier la CPC dans le cadre de l’isolement de marqueurs à l’échelle préparative. Pour cela, les valépotriates, molécules soumises à diverses réglementations à travers le monde et dont l’accès au standard analytique est difficile ont été choisie comme analytes modèles. Après un développement de méthode à l’échelle analytique, un transfert d’échelle non linéaire se basant sur le concept de l’« espace libre entre les pics » a été évalué. Pour la première fois ce modèle a été utilisé pour isoler non pas une, mais deux molécules qui en plus, coéluent. La CPC a permis d’isoler en une étape, deux molécules structurellement proches à un niveau de pureté supérieur à 95% et avec un taux de recouvrement de 90%. De plus, cette étude a permis de confirmer la structure du 7-homovaltrate qui faisait, jusqu’alors, l’objet de discussion. Le troisième et dernier objectif de cette thèse a été d’évaluer l’apport de l’HRMS pour la caractérisation et le dosage de familles de composés. Pour illustrer cette étude, les alcaloïdes pyrrolizidiniques (AP), qui sont des molécules responsables de dizaines de milliers d’intoxication à travers le monde, ont été sélectionnés. L’emploi d’une HRMS a permis de mettre en perspective la pertinence des résultats issus d’un dosage des seuls AP dont le standard analytique est disponible car elle a permis d’identifier d’autres AP dans une mauvaise herbe fréquemment incriminée dans la contamination de denrées alimentaires
Principal limitation to natural product analysis is the access to the analytical standards of molecules considered as markers or potentially dangerous. Their emerging issues are the common thread of this doctoral work. The first objective was to study the performances of HPTLC-MS coupling via Interface MS 2 in the field of plants markers identifications that are commonly used in dietary supplements elaboration. It seems that this coupling is really usefull for routine control, because resulting mass spectrometric informations may permit plants identifications in accordance with pharmacopoeias monographs. But, in case of structural elucidation of new active compounds, the obtained results are extremely uncertains. The second objective was to evaluate CPC performances in the field of markers isolation at an industrial scale. To illustrate this study, valepotriates have been chosen. These secondary metabolites are regulated all over the world but the access to their analytical standards remains problematic. After a chromatographic method development at an analytical scale, a non lineary scale up based on the « free space between peak » concept has been performed. For the first time, this concept has been applied to two molecules which, in addition, are co-eluting. CPC allowed a one step isolation of two structurally close molecules at over 95% purity and with 90% recovery. More over, this study permitted to confirm the structure of 7-homovaltrate which was ambiguous. The third and last objectif was to evaluate HRMS for the characterization and the quantification of family of compounds. To illustrate this study, pyrrolizidine alkaloids family (PA) has been selected. These molecules are responsible of tens of thousands deaths around the world. HRMS afforded identification of AP which analytical standard is not available in weed commonly incriminated in food contamination. This put into perspective the relevance of the results obtained through a method that only quantifies available analytical standards of AP. Furthermore, HRMS informations allowed to discriminate a molecule wich had the same fragment ions as the AP but which wasn’t an AP
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Abdellatif, Meriem. "Continuité de service des entraînements électriques pour une machine à induction alimentée par le stator et le rotor en présence de défauts capteurs." Thesis, Toulouse, INPT, 2010. http://www.theses.fr/2010INPT0107/document.

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Le développement de commandes en boucle fermée pour des entraînements électriques nécessite l'installation de capteurs pour avoir l'information de la rétroaction. Cependant, un éventuel défaut survenant sur l'un des capteurs installés (de courant, de vitesse/position,…) implique un disfonctionnement de la commande conduisant dans la plupart du temps à la mise hors service du système. Ces conséquences sont contraires aux exigences des industriels qui demandent des degrés de fiabilité du système de plus en plus élevés. Des statistiques montrent que le défaut capteur est fréquent. Il est donc impératif de trouver des solutions pour assurer la continuité de service des systèmes électriques dans le cas de présence de ce type de défaut. Tout d'abord, l'étude présentée dans ce manuscrit présente les technologies des différents capteurs installés et ce pour comprendre les raisons et le type de pannes qui pourraient survenir. Ensuite, le système sur lequel la validation des algorithmes développés est décrit. Il s'agit d'un entraînement électrique basé sur une machine à Double Alimentation (MADA) fonctionnant en mode moteur et connectée au réseau via deux convertisseurs. La commande associée est une Commande Directe de Couple (CDC). Elle est validée en mode sain aussi bien par simulation qu'expérimentalement. Après, les études réalisées prennent en considération les défauts capteurs de courants alternatifs et de vitesse/position. Les algorithmes développés, permettant une continuité de service, utilisent une redondance analytique et sont basés sur l'estimation et aussi sur la Détection et l'Isolation d'un éventuel Défaut (DID). Ils sont caractérisés par leur simplicité. Aussi, ils ne sont pas gourmands en termes de consommation en ressources matérielles et leur temps d'exécution est très court. Enfin, la validation expérimentale de ces algorithmes montre bien leur efficacité en cas de défaut, vu que le système s'avère insensible au défaut et continue à fonctionner sans interruption. La commande obtenue est alors tolérante aux défauts capteurs
The development of closed loop controls for electrical drives requires the sensor installations in order to get feed back information. Nevertheless, any occurred sensor fault (current sensor,speed/position sensor,…) shows an operation system deterioration which leads in most cases to its shut down. This consequence is in contrast to industrial expectations especially concerning the system high accuracy that they are asking for. Statistic studies point out the sensor faults as frequent. So, it is necessary to find out solutions ensuring the system service continuity in case of any sensor fault. Firstly, the study presented in this work shows the used sensor technologies in order to understand both of the reason and the kind of occurred faults. Secondly, the studied system is presented which is an electrical drive based on a Doubly Fed Induction Machine (DFIM) operating in motor mode and connected to the grid by two inverters. The control developed is a Direct Torque Control (DTC). The control validation, in healthy operating mode, is realised throw simulation and experimentally. After, a study considering alternative current sensor and speed/position sensor faults are achieved. The developed algorithms are based on signal estimation, on a Fault Detection Isolation (FDI) and reconfiguration algorithms. In fact, they are simple to carry out, they don't need much hardware resources for implementation and their execution time is short. Finally, the experimental validation of the developed algorithms shows their efficiency. The system continues working even in presence of a sensor fault. Thus, the obtained control becomes a fault tolerant control thanks to these algorithms
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Po, Joseph W. "Beyond epithelial circulating tumour cells (CTCs) : establishing important methods for CTC isolation and analysis." Thesis, 2019. http://hdl.handle.net/1959.7/uws:56104.

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This thesis has demonstrated the various applications for antibody-based CTC capture, extending beyond conventional methods. We reported the inclusion of EMT-markers for detection and characterisation of EMT-CTCs in the ovarian cancer setting. This methodological advancement may prove a critical step in understanding the role EMT plays in CTC formation, metastasis and potentially therapeutic resistance. In addition, we explored integration of electron microscopy methods into CTC sample processing, allowing for ultrastructure analysis of CTCs and improving the tools to help understand CTC biology. Finally, we explored antibody-based CTC isolation methods in the melanoma setting with additional biomarker PD-L1 detection, enabling real-time monitoring of therapy response to PD-1 inhibitors. Overall, the knowledge gained from this thesis will aid the CTC research field from three different perspectives: (1) The clinical perspective: capitalize on CTC detection by adding important biomarker detection that may indicate response to therapy; (2) The technical perspective: demonstrating feasibility of integrating electron microscopy sample preparation into CTC analyses; (3) The biological perspective: establishing EMT detection in a range of cancers.
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(10695393), Yuan Zhong. "DETECTION AND ISOLATION OF CIRCULATING TUMOR CELLS FROM WHOLE BLOOD USING A HIGH-THROUGHPUT MICROCHIP SYSTEM." Thesis, 2021.

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Circulating tumor cells (CTCs) have been proved to possess great value and potential in detection, diagnosis, and prognosis of non-haematologic cancers. Their unique characteristics in providing both phenotypic as well as genotypic information make them highly valuable in liquid biopsy assays. At the same time, though numerous studies and research have been done, identification and enumeration of CTCs is still technically challenging due to their rarity and heterogeneity. The primary goal of the thesis is to develop a CTC detection and isolation system with ultra-high sensitivity and purity, while keeping it fast and scalable. We proposed a microfluidic system that integrates positive immunomagnetic capturing, high-throughput parallel flow and size filtration. In this thesis, two generations of the system have been developed to achieve the goal, and are approved to be able to effectively detect and isolate CTCs from hundreds of breast cancer blood samples in real clinical applications.

The first-generation system is based on a sandwich-structured microfluidic chamber, which has a micro-aperture chip as the core to detect and isolate immunomagnetically targeted CTCs. The system achieves high detection yield (>95%) and purity (>99.9998% depletion of leukocytes) by streamlining the workflow and using unprocessed whole blood (without centrifuging), as well as utilizing an advanced surface coating approach to passivate the microchip surface. We first demonstrate experiments for determining the optimal detection parameters. Then we characterize the system by isolating deterministically spiked 1, 10, and 100 single MCF-7 breast cancer cells into tubes of whole blood, and show that >95% of cells were captured. A detection yield of 100% from single cell spiking experiments (n = 6) demonstrates excellent detection capability and repeatability of the system. We finally demonstrate the use of the system for CTC detection in the context of a phase II clinical trial of early-stage triple-negative breast cancer (TNBC) patients. As a part of the trial, 182 blood samples were collected from 124 early-stage TNBC patients at high-risk of relapse. We detected CTCs in 36.3% of patients who had already completed chemotherapy and surgery for curative intent and were thus nominally expected to have very few to zero CTCs. Moreover, increasing CTC count from the same patients shows good correlation with their clinical course. The ability to detect CTCs’ presence using this first-generation system illustrates its important clinical utility.

The second-generation system applies a similar detection strategy but employs an upgraded microchip and device, as well as a further streamlined process flow to achieve an even higher detection efficiency, especially for capturing the target cells with low surface marker expression level. We first did modeling and simulation of the new system to find the optimal magnet configuration and verify the detection sensitivity improvement on the first-generation system. Then we characterized the new system by detecting spiked JEG-3 and JAR cells in both cell culture medium and human blood. The result demonstrates that the detection yield increased by ~20% using the second-generation system under the same experiment condition. Next, we applied the system to a phase I clinical trial for CTC detection from metastatic triple-negative breast cancer (mTNBC) patient blood samples. CTCs of mTNBC are known to with in the low marker expression phenotype, which requires ultra-high detection sensitivity. Our system captured CTCs from 48 out of 102 (47%) blood samples, the positivity rate agrees with the conclusions from other studies and presents the reliability to the system. Finally, we explored a novel 4-marker panel for CTC detection from mTNBC patient blood samples. We conducted paired comparisons using the 4-marker panel versus a single marker for detection. The 4-marker panel yielded more CTCs in 5/8 complete paired assessments, and less CTCs in 1/8. The association missed the significance level only slightly (p = 0.08), and the result strongly illustrates the potential for using the panel to cover the mTNBC cells’ heterogeneity for enhanced CTC detection. Furthermore, the presence of CTCs from blood samples correlates well with the patient’s disease progression.

Finally, we demonstrated downstream analysis ability of the CTCs detected by the second-generation system. Captured CTCs can be readily released from our system without any loss or damage to a secondary microchip device to be further isolated as single cells, and picked up individually for downstream analysis like DNA/RNA sequencing or single-cell cultivation. Directions for future work is also discussed. We envision this versatile and efficient system to be highly beneficial in a broad range of clinical and research applications regarding CTCs.

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Книги з теми "CTC Isolation"

1

Lambe, Kevin Gerard. Isolation of cell cycle genes using yeast CDC mutants. Manchester: University of Manchester, 1993.

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2

Mazzone, Horace M. CRC handbook of viruses: Mass-molecular weight values and related properties. Boca Raton, FL: CRC Press, 1998.

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3

Bennett, Gail. Prevent Infections With Isolation Precautions: Strategies for the Cdc Guidelines. HCPro, Inc., 2007.

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4

NCCER. CT1 5-17 Inspect and Test Electrical Isolation Trainee Guide. Pearson Education, Limited, 2018.

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5

Rothstein, Mark A. Quarantine And Isolation: Lessons Learned from Sars: A Report to the Cdc. Diane Pub Co, 2003.

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6

CRC handbook of methods for oxygen radical research. Boca Raton, Fla: CRC Press, 1985.

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7

CRC handbook of methods for oxygen radical research. CRC Press in Boca Raton, Fla, 1985.

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8

Handbook of Natural Pesticides: Methods, Isolation and Identification, Volume II (Crc Series in Naturally Occurring Pesticides). CRC, 1985.

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9

What's It Mean To Quarantine? self, 2020.

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10

What's It Mean To Quarantine? 2020.

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Частини книг з теми "CTC Isolation"

1

Zhao, Mengxia, Perry G. Schiro, and Daniel T. Chiu. "Ensemble-decision Aliquot Ranking (eDAR) for CTC Isolation and Analysis." In Circulating Tumor Cells, 51–84. Hoboken, NJ, USA: John Wiley & Sons, Inc, 2016. http://dx.doi.org/10.1002/9781119244554.ch3.

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2

Rokem, J. Stefan, Astrid Schön, and Dieter Söll. "Isolation and Sequence of a tRNAGly (CCC) from Streptomyces coelicolor A3(2)." In Genetics and Product Formation in Streptomyces, 47–52. Boston, MA: Springer US, 1991. http://dx.doi.org/10.1007/978-1-4684-5922-7_7.

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3

Santiago, Llipsy, Marta Castro, Julián Pardo, and Maykel Arias. "Mouse Model of Colitis-Associated Colorectal Cancer (CAC): Isolation and Characterization of Mucosal-Associated Lymphoid Cells." In Methods in Molecular Biology, 189–202. New York, NY: Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-8885-3_13.

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4

Aktar, Sharmin, Tracie T. Cheng, Sujani M. K. Gamage, Vinod Gopalan, and Farhadul Islam. "Circulating Tumour Cells in Solid Cancer." In Current Cancer Biomarkers, 115–47. BENTHAM SCIENCE PUBLISHERS, 2023. http://dx.doi.org/10.2174/9789815079364123010010.

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Circulating tumour cells (CTCs), as 'liquid biopsy”, has a major benefit over traditional tissue biopsy and has the potential to become a less invasive and more cost.effective cancer biomarker. The presence of CTCs in the circulation indicates the presence of a tumour and the possibility of metastatic spread. Hence, the characterisation of CTCs is expected to provide crucial insights into the mechanisms of metastasis. It can also provide useful information about the future use of CTCs as a surrogate endpoint biomarker in diagnosis, prognosis, and treatment response prediction by minimizing the limitations of tissue biopsies. Also, it provides a new horizon for the development of novel targeted therapies. However, the lack of specific and effective methods is the key limitation in CTC detection and isolation in patients with cancer. Therefore, more responsive methods and approaches may be needed to improve the accuracy of CTC measurements. Herein, this book chapter will provide a current picture of CTCs as surrogate biomarkers for disease diagnosis, prognosis and predicting therapy response, along with the risk of relapse in cancers.
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Morgan, E. David, and Ian D. Wilson. "Methods and Techniques for Isolation of Pesticides." In CRC Handbook of Natural Pesticides: Methods, 3–81. CRC Press, 2019. http://dx.doi.org/10.1201/9781351072700-1.

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Phang, C. H., and K. Jeyaseelan. "ISOLATION AND CHARACTERIZATION OF citC GENE OF BACILLUS SUBTILIS." In Genetics and Biotechnology of Bacilli, 97–100. Elsevier, 1988. http://dx.doi.org/10.1016/b978-0-12-274161-6.50021-8.

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B. Oti, Victor, Isa H. Mohammed, Fatima Y. Al-Mustapha, and Salamatu B. Buhari. "Helicobacter pylori Seromarkers in a University Students Population in Central Nigeria." In Helicobacter pylori - From First Isolation to 2021. IntechOpen, 2021. http://dx.doi.org/10.5772/intechopen.96762.

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Infection due to Helicobacter pylori is a public health challenge worldwide as over 3 billion persons are infected with the bacterium globally. There is a serious need to update the knowledge on the epidemiology of this bacterial pathogen and its probable risks factors to generate intervention programs that will reduce the morbidity and mortality of infected individuals. This chapter evaluated the seromarkers of H. pylori infection and its predisposing factors among students of Nasarawa State University, Keffi, Central Nigeria. This study was done between June through August 2019; blood and stool specimens were collected from 400 students of the institution. Before the commencement of the study, ethical clearance and informed consent were retrieved and a structured questionnaire was administered to each participant. Specimens were screened for H. pylori antigen and antibody using rapid test kits (CTK Biotech, Inc., San Diego, USA and Biotest Biotech, China). Information obtained were analyzed using SSP version 2.80. P values <0.05 were reflected statistically significant. Out of the 400 students tested, 166 (41.5%) and 128 (32.0%) showed positive for anti-H. pylori IgG and Ag markers respectively. The antibody seromarker was higher in female while the H. pylori antigen was higher in males. Those students aged 21–30 years old reported the highest prevalence of the seromarkers while those of more than 41 years old had the least prevalence. Location, type of toilet facility and place of residence were statistical associated between H. pylori antigen (P < 0.05). There was a statistically significant association between anti-H. pylori IgG and the sources of water of the students (P < 0.05). This is the first public report that has successfully reported the prevalence of these seromarkers among students of a tertiary institution in Nasarawa state. The overall outcomes of this study stressed the need for student-based intervention programs to stem the transmission of this infection in Nasarawa State, Nigeria.
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Kingston, David G. I., and M. Madhusudana Rao. "Chromatographic Methods for Isolation and Identification of Naturally Occurring Pesticides." In CRC Handbook of Natural Pesticides: Methods, 83–101. CRC Press, 2019. http://dx.doi.org/10.1201/9781351072700-2.

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"Evaluation of the left atrium and pulmonary veins." In Cardiovascular Computed Tomography, edited by James Stirrup, Russell Bull, Michelle Williams, and Ed Nicol, 285–94. Oxford University Press, 2019. http://dx.doi.org/10.1093/med/9780198809272.003.0019.

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Clinical cardiac electrophysiology (EP) presupposes that, for every dysrhythmia, there is a critical anatomical region responsible for either the generation or propagation of abnormal electrical impulses. The destruction or isolation of these regions should prevent the recurrence of that dysrhythmia. Therefore, the two broad aims of EP are the identification, characterization, and localization of dysrhythmia; and ablation of the focus of the dysrhythmia. This chapter covers cardiac CT prior to EP procedures, the left atrium and pulmonary veins, other relevant anatomy, and a template report for pre-EP CCT.
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"Preparation for Follicle Aspiration and Isolation of Cumulus-Oocyte-Complexes (COC)." In Standard Operational Procedures in Reproductive Medicine, 30–31. CRC Press, 2017. http://dx.doi.org/10.1201/9781315270975-11.

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Тези доповідей конференцій з теми "CTC Isolation"

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Chen, Kangfu, Teodor Georgiev, and Z. Hugh Fan. "Interactions Between Circulating Tumor Cells and Aptamer-Functionalized Microposts in a Flow." In ASME 2017 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2017. http://dx.doi.org/10.1115/imece2017-70342.

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Circulating Tumor Cells (CTCs) have been considered as important biomarkers for cancer prognosis and treatment. However, there are only tens of CTCs in one billion of healthy blood cells. This CTC rarity challenge has been addressed by microfluidics technology that sheds light on efficient CTC detection and isolation. Using antibodies or aptamers to capture CTCs is one of the strategies for CTC isolation. A lot of work has been carried out to improve CTC capture efficiency and purity (i.e., specificity). The main consideration to optimize microfluidic device performance includes increasing surface-area-to-volume ratio and reducing shear stress, both of which are closely related to the interaction between CTCs and the microfluidic device. Here we report a detailed study on the interactions between CTCs and aptamer-functionalized microposts in a microfluidic device. We have evaluated the distribution of captured CTCs around a micropost. In addition, simulation was conducted to model CTC capture patterns around microposts. We found the simulated CTC capture pattern largely agree with the experimental results. The simulation methodology could be applicable for other affinity-based CTC isolation devices and approaches. The goal of the study is to improve the microfluidic device performance and provide a rapid and economical way to optimize the geometry design of the microfluidic devices for CTC isolation.
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2

Celik, Emrah, Nicolas Rongione, Amelia Bahamonde, Zheng Ao, and Ram Datar. "Isolation of Circulating Tumor Cells Using Stiffness-Based Filtration Platform." In ASME 2015 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2015. http://dx.doi.org/10.1115/imece2015-53241.

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Analysis of isolated cancer cells in circulation is proven to help determine the success of the cancer treatment and understand the genetic signature of cancer disease. Scarcity of these cells in blood circulation (1–10 CTC in 1ml blood) however, makes the isolation process extremely challenging. Ever improving CTC isolation methods fall into two main categories: 1.Immunomagnetic separation based on antibody binding to tumor specific biomarkers expressed on the cell 2. Physical separation based on the size of the CTCs. Efficiency in cell isolation is still low in these techniques due to the variation in expression level of tumor specific antigens and tumor cell size. Therefore, tumor cell isolation strategies using new CTC biomarkers must be explored. In this study, we investigated the feasibility of using mechanical stiffness difference in order to detect and isolate the circulating tumor cells from the blood cells. AFM nanindentation experiments revealed that cancer cells are significantly softer than the surrounding white blood cells and therefore, stiffness can be used as a biomarker for CTC isolation. In addition, finite element analysis simulations have shown that CTC isolation can be performed at high efficiency using stiffness-based isolation. Therefore, stiffness based isolation has a potential to achieve fast, label-free isolation of CTCs at high efficiency for clinical applications.
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Western, Laura T., Kuldeepsinh Rana, and Michael R. King. "Flow-Based Isolation and Neutralization of Circulating Tumor Cells." In ASME 2009 7th International Conference on Nanochannels, Microchannels, and Minichannels. ASMEDC, 2009. http://dx.doi.org/10.1115/icnmm2009-82137.

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Circulating tumor cells (CTC) have the potential to be used clinically as a diagnostic tool and a treatment tool in the field of oncology. As a diagnostic tool, CTC may be used to indicate the presence of a tumor before the tumor is large enough to cause noticeable symptoms. As a treatment tool, CTC isolated from patients may be used to test the efficacy of chemotherapy options to personalize patient treatment. One way for tumors to spread is through metastasis via the circulatory system. CTC are able to exploit the natural leukocyte recruitment process that is initially mediated by rolling on transient selectin bonds. Our capture devices take advantage of this naturally occurring recruitment step to isolate CTC from whole blood by flowing samples through selectin and antibody-coated microtubes. Whole blood was spiked with a known concentration of labeled cancer cells and then perfused through pre-coated microtubes. Microtubes were then rinsed to remove unbound cells and the number of labeled cells captured on the lumen was assessed. CTC were successfully captured from whole blood at a clinically relevant level on the order of 10 cells per mL. Combination tubes with selectin and antibody coated surface exhibited higher capture rate than tubes coated with selectin alone or antibody alone. Additionally, CTC capture was demonstrated with the KG1a hematopoietic cell line and the Du145 epithelial cell line. Thus, the in vivo process of selectin-mediated CTC recruitment to distant vessel walls can be used in vitro to target CTC to a tube lumen. The microtube device can also be used to capture CTC of hematopoietic and epithelial tumor origin and is demonstrated sensitivity down to the order of 10 CTC per mL. In a related study aimed at reducing the blood borne metastatic cancer load, we have shown that cells captured to a surface can be neutralized by a receptor-mediated biochemical signal (Rana et al. 2008). In the proposed method we have shown that using a combined selecting and TRAIL (TNF Related Apoptosis Inducing Ligand or Apo 2L) functionalized surface we are able to kill about 30% of the captured cells in a short duration of 1 hour whereas it took about 4 hours to kill the same proportion of cells without flow on a similarly functionalized. Here we have taken the approach a step further by showing that with very small doses of chemotherapeutic agents like Bortezomib, we can increase the kill rate of CTCs., thus allowing the device to function in senarios where the patient is undergoing treatment. We show here with leukemic cells that are treated with Bortezomib that we are able kill about 41% of the captured cells.
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Liu, Yang, Jungwook Park, Tong Xu, Yucheng Xu, Jay Han-Chieh Chang, Dongyang Kang, Xiaoxiao Zhang, Amir Goldkorn, and Yu-Chong Tai. "Magnesium-embedded live cell filter for CTC isolation." In 2015 28th IEEE International Conference on Micro Electro Mechanical Systems (MEMS). IEEE, 2015. http://dx.doi.org/10.1109/memsys.2015.7050958.

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5

Andree, Kiki C., Anouk Mentink, Martin Scholz, Roland Kirchner, Rui P. Neves, Christiane Driemel, Rita Lampignano, et al. "Abstract 1532: The isolation of CTC from diagnostic leukapheresis." In Proceedings: AACR 107th Annual Meeting 2016; April 16-20, 2016; New Orleans, LA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.am2016-1532.

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6

Amontree, Jacob, Kangfu Chen, Jose Varillas, and Z. Hugh Fan. "Capillary Force Driven Single-Cell Spiking Apparatus for Studying Circulating Tumor Cells." In ASME 2018 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2018. http://dx.doi.org/10.1115/imece2018-87109.

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The characterization of single cells within heterogeneous populations has great impact on both biomedical sciences and cancer research. By investigating cellular compositions on a broad scale, pertinent outliers may be lost in the sample set. Alternatively, an investigation focused on the behavior of specific cells, such as circulating tumor cells (CTCs), will reveal genetic biomarkers or phenotypic characteristics associated with cancer and metastasis. On average, CTC concentration in peripheral blood is extremely low, as few as one to two per billion of healthy blood cells. Consequently, the critical element lacking in many methods of CTC detection is accurate cell capture efficiency at low concentrations. To simulate CTC isolation, researchers usually spike small amounts of tumor cells to healthy blood for separation. However, spiking tumor cells at extremely low concentrations is challenging in a standard laboratory setting. We report our study on an innovative apparatus and method designed for low-cost, precise, and replicable single-cell spiking (SCS). Our SCS method operates solely from capillary aspiration without the reliance on external laboratory equipment. To ensure that our method does not affect the viability of each cell, we investigated the effects of surface membrane tensions induced by aspiration. Finally, we performed affinity-based CTC isolation using human acute lymphoblastic leukemia cells (CCRF-CEM) spiked into healthy whole blood with the SCS technique. The results of the isolation experiments demonstrate the reliability of our method in generating low-concentration cell samples.
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7

Jackson, Seth, Jeff Darabi, and Joseph Schober. "Fabrication and Testing of a Magnetophoretic Bioseparation Chip for Isolation and Detection of Circulating Tumor Cells From Peripheral Blood." In ASME-JSME-KSME 2019 8th Joint Fluids Engineering Conference. American Society of Mechanical Engineers, 2019. http://dx.doi.org/10.1115/ajkfluids2019-5082.

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Abstract Significant research involving the isolation and detection of circulating tumor cells (CTCs) has become prevalent in the field of biomedicine. It plays a crucial role in the diagnosis and treatment of cancer and has made substantial strides in recent years. A major event in the timeline of cancer is metastasis, a set of occurrences where cells are shed from a cancerous site, then flow through the circulatory system and seed themselves throughout the body, forming secondary tumors. There are few observable symptoms in the early stages of metastasis and this fact severely limits clinical treatment. The fabrication and preliminary testing of a magnetophoretic bioseparation chip capable of isolating and detecting CTCs from peripheral blood, which can aid in early detection of metastases, is presented in this work. MCF7 breast cancer cells along with superparamagnetic microparticles, which are specifically coated with anti-EpCAM to bind to the cancer cells, are spiked into a blood sample. After the spiked blood sample is introduced into the biochip, a locally engineered magnetic field gradient captures the magnetically labeled cancer cells while the non-target cells are allowed to pass by. Once the target cells are isolated from the blood sample, flow cytometry is used to determine the recovery rate of the magnetophoretic device. The proposed device can operate at continuous flow rates magnitudes higher than other CTC isolation devices and is fabricated using much simpler methods which make it quite unique. These properties combined with greater than 80% recovery rates make the device quite favorable for economic point of care use in clinical applications.
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Strauss, William, Alex Parker, Frank Juhn, Maureen Cronin, Emily White, Behrad Vahidi, Cong Fang, et al. "Abstract 4554: QPCR and sequence analysis of DNA template from a microfluidic CTC isolation platform." In Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL. American Association for Cancer Research, 2012. http://dx.doi.org/10.1158/1538-7445.am2012-4554.

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Connelly, Mark, David Chianese, Carrie Morano, Thai Bui, Shemeeakah Powell, Noel Ngoubilly, Renouard Sanders, et al. "Abstract 4955: Isolation and characterization of circulating tumor cells (CTCs) in breast and prostate cancer: Comparison of Harpoon CTC assay performance with the CellSearch CTC Test." In Proceedings: AACR 107th Annual Meeting 2016; April 16-20, 2016; New Orleans, LA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.am2016-4955.

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Chianese, David, Mark Connelly, Carrie Morano, Thai Bui, Steven Gross, Renouard Sanders, Tom Barber, et al. "Abstract 2422: Isolation of canonical and non-canonical circulating tumor cells (CTCs) by negative depletion using the Harpoon CTC Isolator and Chip." In Proceedings: AACR 107th Annual Meeting 2016; April 16-20, 2016; New Orleans, LA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.am2016-2422.

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Звіти організацій з теми "CTC Isolation"

1

Gui, Feng, and Sean Brossia. PR-186-073502-R01 Large-Scale Cathodic Disbondment Testing for Coal Tar Enamel(CTE). Chantilly, Virginia: Pipeline Research Council International, Inc. (PRCI), September 2010. http://dx.doi.org/10.55274/r0010646.

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Corrosion protection of installed transmission pipelines is mainly achieved by isolating the pipeline from the environment through the use of applied coatings. In areas where coating damage (e.g., holidays) exist, cathodic protection (CP), often by impressed current, is used to protect any bare areas. The amount of current output required for CP tends to increase as the damaged area (e.g. holidays, cracks) progresses over time. Although CP is necessary to protect any bare areas, it can also enhance coating disbondment. Likewise, the extent of cathodic disbondment (CD) could increase as the current from the rectifier is augmented to compensate for the extended damage. In the United Stated approximately 50 % of the transmission pipelines are coated with coal tar enamel (CTE), many of which have been in service for more than 50 years. The extent of the coating damage (in the form of holidays, open disbondments, cracks and tears) requires incremental increases in CP levels to be applied over time. The objective of this work was to investigate the effect of CP levels on the growth of CD of CTE coated pipes. The CD extent at under protected CP levels was investigated as well. Additionally, the effect of surface contaminant (chloride) on CD of the CTE coated pipes was evaluated.
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2

Carlson, Damian, Jennifer De Lurio, Andrea Druga, Randy Hulshizer, Marcus Lynch, and Misha Mehta. PCORI COVID-19 Scan: Isolation Bags for Safe CT Imaging, Population-wide Antibody Testing (June 25-July 8, 2020). Patient-Centered Outcomes Research Institute (PCORI), July 2020. http://dx.doi.org/10.25302/bcs4.2020.7.

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