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Dissertations / Theses on the topic 'M-cels'

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Published: 4 June 2021

Last updated: 1 February 2022

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1

Tyrer, Peter Charles, and n/a. "Targeting M-cells for oral vaccine delivery." University of Canberra. Health Sciences, 2004. http://erl.canberra.edu.au./public/adt-AUC20060427.122012.

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An in vitro model of the follicle-associated epithelia that overlie the Peyer's patches of the small intestine was developed and validated to examine the mechanisms of mucosal antigen sampling. This model displays many phenotypic and physiological characteristics of M cells including apical expression of [alpha]5[beta]l integrin and enhanced energy dependent participate transport. CD4+ T-cells were shown to be an important influence on the development of Mlike cells. The model was used to examine the M cell mediated uptake of several putative whole-cell killed bacterial vaccines. Greater numbers of non-typeable Haemophilus influenzae NTHi 289, NTHi 2019, Escherichia coli 075 HMN and Streptococcus pneumoniae were transported by model M cells compared to control Caco-2 enterocyte-like cells. Studies in isolated murine intestine segments confirmed the selective uptake of NTHi 289 and Escherichia coli demonstrating that intestinal mucosal sampling of these antigens is performed by M cells. Pseudomonas aeruginosa was not absorbed as whole cell bacteria but as soluble antigen, as indicated by the presence of bacterial DNA in the cytoplasm of epithelial cells. These results suggest that bacteria such as NTHi and E. coli are sampled by the mucosal immune system in a different manner to that of bacteria such as Pseudomonas. A number of potential cell surface receptors were investigated to identify which molecules are responsible for intestinal uptake whole-cell killed bacteria. Immunofluorescence studies detected the presence of toll-like receptor-2, toll-like receptor-4, PAF-R and [alpha]5[beta]l integrin on in vitro M-like cell cultures. Examinations of murine intestine confirmed the presence of TLR-4 and PAF-R. TLR-4 was found in small quantities and on M cells. In contrast to the M cell model, TLR-2 expression in the murine intestine was sparse. Receptor inhibition experiments provided evidence for the involvement of TLR-4, PAF-R and [alpha]5[beta]l integrin in M cell uptake of killed bacteria both in vitro and in vivo. This thesis has contributed valuable information regarding the mechanisms of uptake of whole-cell killed bacteria by the intestinal mucosal immune system. For the first time, M cell sampling of whole-cell killed bacteria has been demonstrated. Furthermore, the receptors involved in these processes have been identified. This information will be of great use in the development and optimisation of new oral vaccines.

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2

Blakemore, Louise Margaret. "Curcumin-induced G2/M cell cycle arrest in colorectal cancer cells." Thesis, University of Leicester, 2011. http://hdl.handle.net/2381/9809.

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Curcumin, a diet-derived chemopreventive and chemotherapeutic agent has been shown to induce G2/M cell cycle arrest, and previous studies suggested that microtubule depolymerisation may be linked to M-phase arrest. However, mechanisms involved have not been elucidated and often non-physiological concentrations of curcumin were used. The goal of this study was to characterise in more detail curcumin-induced cell cycle arrest using a panel of human colorectal cancer cell (CRC) lines, HT-29, SW480, HCT116 p53+/+, HCT116 p53-/- and HCT116 p21-/-. Using physiologically relevant concentrations of curcumin (5-10μM), achievable in the gut tissue following oral ingestion, cell cycle analysis showed that treatment for 12 hours results in significant G2/M arrest in all five cell lines. Curcumin treatment significantly increased the number of cells in M phase in 4 out of the 5 lines tested for this duration, and those with microsatellite instability (HCT116) were found to have a higher mitotic index than those with chromosomal instability. Pre-treatment with caffeine abrogated mitotic arrest in these cell lines, indicating the involvement of the ATM/ATR kinases. Activating phosphorylation of the Chk1 kinase was increased and total protein levels of CDC25C reduced, further implicating the DNA damage pathway in the induction of arrest. Higher levels of HSP70 were also found, indicating proteotoxic stress such as proteasomal inhibition. Image analysis revealed impaired mitotic progression, and significantly higher levels of mitotic spindle abnormalities following curcumin treatment. Aurora B mislocalisation and significantly lower levels of centrosomal separation were found in the HCT116 p53+/+ line. Furthermore, the high levels of pH2A.X staining seen in curcumin-treated mitotic but not interphase cells suggest that aberrant mitosis may result in DNA damage. This proteotoxic and genotoxic stress incurred following curcumin treatment may contribute to the upregulation of NKG2D ligands on the cell surface, leading to CRC lysis and enhancement of the anti-cancer immune response.

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3

Mairopoulos, Dimitrios. "M-Cell assembly." Thesis, Massachusetts Institute of Technology, 2015. http://hdl.handle.net/1721.1/99294.

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Thesis: S.M., Massachusetts Institute of Technology, Department of Architecture, June 2015.
Cataloged from PDF version of thesis. "June 2015."
Includes bibliographical references (page 47).
In this thesis I propose a self- assembly procedure called the Morphocell(M-Cell) assembly. This procedure is based on an assembly unit called the M-Cell. The M-Cell is comprised out of two components, the M-Block and the M-Clay (in which the M-Block is embedded). During the assembly procedure the M-Clay acts as the environment of the assembly for the M-Blocks. This allows a global, parallel assembly that is highly autonomous and has large error correcting capacities. When the assembly procedure is complete the M-Blocks have assembled into a spatial lattice. Then the M-Clay surrounds this lattice thus creating a solid object, the M-Object. The M-Object, which is the goal of this procedure, is a dynamic object that can be easily modified, expanded or dismantled. Furthermore, it can respond in various ways to its environment. This system was optimized though a feedback loop that was informed by constant digital and physical simulations. The findings of this thesis can have important applications in construction of structures in extreme-remote environments and in the fabrication-rapid prototyping field.
by Dimitrios Mairopoulos.
S.M.

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4

Mason, Caroline Margaret. "Characterisation of intestinal M cells using monoclonal antibodies." Thesis, University of Newcastle Upon Tyne, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.260952.

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5

Sehgal, Anuj. "Investigating the development and function of M cells." Thesis, University of Edinburgh, 2017. http://hdl.handle.net/1842/28738.

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Gut-associated lymphoid tissues such as Peyer’s patches (PP) are inductive sites for immune response in the intestine. Unlike other peripheral lymphoid tissues, gut-associated lymphoid tissues lack afferent lymphatics and can directly sample mucosal antigens by specialized epithelial cells in the follicular associated epithelia (FAE), known as M cells. M cells derive from Lgr5+ intestinal stem cells in intestinal crypts, where the daughter cells of Lgr5+ cells differentiate into M cells after stimulation from the cytokine receptor activator of nuclear factor-κB ligand (RANKL). RANKL is produced by stromal cells within the sub-epithelial dome (SED) residing below the FAE. The transcytosis of antigens across the FAE by M cells is an important initial step in the induction of efficient mucosal immune responses against certain pathogenic bacteria as well as the commensal bacterial flora. However some pathogens, for example orally-acquired prions, may also exploit M cells to infect the host. M cells have been implicated in the uptake of orally acquired prions from the gut lumen. After oral exposure, the accumulation of prions in PP is important for their efficient spread to the nervous system. Previous studies have also shown that pathogen-induced inflammation increases M cell density and this effect can be mimicked by exogenous administration of RANKL. This has led to the hypothesis tested in this thesis that inflammation-related enhancement of M cell differentiation aids the delivery of prions into the lamina propria of villi. The administration of RANKL resulted in increased M cell density in the gut epithelium of mice. Consequently, RANKL treatment enhanced the accumulation of orally-administered prions in PP, decreased disease incubation time and increased prion disease susceptibility. These data indicate the importance of M cells in prion disease pathogenesis and highlight the potential of M cells as vaccine targets against prion disease. The fate and terminal differentiation of distinct intestinal epithelial cell lineages from their uncommitted precursors is dependent on their intrinsic expression of one or more specific transcription factors during their development. Alongside inducing M cell differentiation, RANKL stimulation can also induce the nuclear translocation of the NF-κB transcription factor subunit c-Rel. A comparison of the genes encoding the individual NF-κB subunits c-Rel, Rel-A and Rel-B revealed that they were expressed at the mRNA level in the FAE and by M cells. A c-Rel-deficiency in mice did not influence the expression of RANKL or RANK in PP. The subsequent induction of M cell maturation in the FAE was also unaffected in, indicating that c-Rel is dispensable for the RANKL-mediated differentiation and functional maturation of M cells. The factors implicated in Lgr5+ intestinal stem cell proliferation and their differentiation into M cells are poorly understood. Some reports have indicated that crypt-associated macrophages may provide extrinsic factors that assist Lgr5+ intestinal stem cell proliferation. In this thesis, the ablation of macrophages in the gut resulted in dysregulation of crypt microarchitecture, depleting Paneth cells and the Lgr5+ stem cells. This adversely affected the subsequent differentiation of intestinal epithelial cell lineages and impeded the functional development of M cells. These data reveal a previously unknown role for macrophages in the maintenance of intestinal crypts and intestinal stem cell proliferation and differentiation.

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6

Schäkel, Knut, Claudia Poppe, Elfriede Mayer, Christine Federle, Gert Riethmüller, and Ernst Peter Rieber. "M-DC8+ Leukocytes – A Novel Human Dendritic Cell Population." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-135252.

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Dendritic cells (DC) constitute a heterogeneous leukocyte population having in common a unique capacity to induce primary T cell responses and are therefore most attractive candidates for immunomodulatory strategies. Two populations of blood DC (CD11c+ CD123dim and CD11c– CD123high) have been defined so far. However, their direct isolation for experimental purposes is hampered by their low frequency and by the lack of selective markers allowing large scale purification from blood. Here we describe the monoclonal antibody (mAb) M-DC8, which was generated by immunizing mice with highly enriched blood DC. This mAb specifically reacts with 0.2–1% of blood leukocytes and enables their direct isolation by a one-step immunomagnetic procedure from fresh mononuclear cells. These cells can be differentiated from T cells, B cells, NK cells and monocytes using lineage-specific antibodies. M-DC8+ cells express HLA class II molecules, CD33 and low levels of the costimulatory molecules CD86 and CD40. Upon in vitro culture M-DC8+ cells spontaneously mature into cells with the phenotype of highly stimulatory cells as documented by the upregulation of HLA-DR, CD86 and CD40; in parallel CD80 expression is induced. M-DC8+ cells display an outstanding capacity to present antigen. In particular, they proved to be excellent stimulators of autologous mixed leukocyte reaction and to activate T cells against primary antigens such as keyhole limpet hemocyanin. Furthermore, they induce differentiation of purified allogeneic cytotoxic T cells into alloantigen-specific cytotoxic effector cells. While the phenotypical analysis reveals similarities with the two known blood DC populations, the characteristic expression of Fc=γRIII (CD16) and the M-DC8 antigen clearly defines them as a novel population of blood DC. The mAb M-DC8 might thus be a valuable tool to determine circulating DC for diagnostic purposes and to isolate these cells for studies of antigen-specific T cell priming
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich

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7

Baker, Peter Kenneth. "The role of M-CSF in hairy-cell leukaemia." Thesis, University of Liverpool, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.367653.

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8

Sansom, Nigel P. "Antigen sampling by porcine intestinal Peyer's patch M-cells." Thesis, University of Aberdeen, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.322637.

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9

Schäkel, Knut, Claudia Poppe, Elfriede Mayer, Christine Federle, Gert Riethmüller, and Ernst Peter Rieber. "M-DC8+ Leukocytes – A Novel Human Dendritic Cell Population." Karger, 1999. https://tud.qucosa.de/id/qucosa%3A27632.

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Dendritic cells (DC) constitute a heterogeneous leukocyte population having in common a unique capacity to induce primary T cell responses and are therefore most attractive candidates for immunomodulatory strategies. Two populations of blood DC (CD11c+ CD123dim and CD11c– CD123high) have been defined so far. However, their direct isolation for experimental purposes is hampered by their low frequency and by the lack of selective markers allowing large scale purification from blood. Here we describe the monoclonal antibody (mAb) M-DC8, which was generated by immunizing mice with highly enriched blood DC. This mAb specifically reacts with 0.2–1% of blood leukocytes and enables their direct isolation by a one-step immunomagnetic procedure from fresh mononuclear cells. These cells can be differentiated from T cells, B cells, NK cells and monocytes using lineage-specific antibodies. M-DC8+ cells express HLA class II molecules, CD33 and low levels of the costimulatory molecules CD86 and CD40. Upon in vitro culture M-DC8+ cells spontaneously mature into cells with the phenotype of highly stimulatory cells as documented by the upregulation of HLA-DR, CD86 and CD40; in parallel CD80 expression is induced. M-DC8+ cells display an outstanding capacity to present antigen. In particular, they proved to be excellent stimulators of autologous mixed leukocyte reaction and to activate T cells against primary antigens such as keyhole limpet hemocyanin. Furthermore, they induce differentiation of purified allogeneic cytotoxic T cells into alloantigen-specific cytotoxic effector cells. While the phenotypical analysis reveals similarities with the two known blood DC populations, the characteristic expression of Fc=γRIII (CD16) and the M-DC8 antigen clearly defines them as a novel population of blood DC. The mAb M-DC8 might thus be a valuable tool to determine circulating DC for diagnostic purposes and to isolate these cells for studies of antigen-specific T cell priming.
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

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10

Messerly, Erin. "M-CSF Stress-Induced Priming of the Macrophage Cell." Ohio Dominican University Honors Theses / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=oduhonors1449484244.

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11

Frick, Florian Verfasser], John M. [Akademischer Betreuer] Sullivan, and Imre [Akademischer Betreuer] [Bárány. "Combinatorial restrictions on cell complexes / Florian Frick. Gutachter: Imre Bárány ; John M. Sullivan. Betreuer: John M. Sullivan." Berlin : Technische Universität Berlin, 2015. http://d-nb.info/1076082246/34.

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12

Fowler, Daniel William. "The anti-tumour immune responses of γδ T cells activated with BCG, M. vaccae and M. obuense." Thesis, St George's, University of London, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.559387.

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Introduction: BCG and heat-killed M. vaccae and M. obuense are currently being used as cancer immunotherapeutic agents; however, the induced immune response is poorly characterised and potential biomarkers of response ill-defined. γδ T cells have recently emerged as key players in immunity against both bacterial infections and malignant transformations, yet their role in cancer immunotherapies such as BCG, M. vaccae and M. obuense has been largely over-looked. This project investigates whether these three bacterial preparations can elicit anti-tumour responses in human peripheral blood γδ T cells. Methods: In this thesis, in vitro proof-of-concept experiments were conducted on human peripheral blood γδ T cells in order to test the activatory capacity of BCG, M. vaccae and M. obuense. To determine whether γδ T cells are activated by these bacterial preparations, activation marker expression and proliferation responses were assessed. Furthermore, the anti-tumour responses were also investigated by measuring cytokine production, expression of granzyme B and cytotoxicity against tumour target cells. In addition, the mechanism by which BCG, M. vaccae and M. obuense can activate these γδ T cell responses was assessed. Results: Results show that γδ T cells are activated by these three bacterial preparations, as suggested by upregulation of activation marker expression and proliferation responses. It was found that γδ T cells produce the T H1 cytokines IFN-y and TNF-a, and upregulate granzyme B expression, which correlates with an enhanced capacity to kill Daudi target cells and zoledronate-treated A549 cells. Results also suggest that γδ T cell responses are induced by IL-12, IL-1~ and TNF-a from a subpopulation of peripheral blood myeloid dendritic cells, and that M. obuense- but not BCG- or M. vaccae-induced cytokine production by these cells is, at least in part, dependent on TLR2 signalling. Conclusion: Taken together, data suggest a potential mechanism for the anti-cancer effects of BCG, M. vaccae and M. obuense, which could ultimately guide clinicians in harnessing the full potential of these bacterial preparations in cancer immunotherapy.

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13

Frick, Florian [Verfasser], John M. Akademischer Betreuer] Sullivan, and Imre [Akademischer Betreuer] [Bárány. "Combinatorial restrictions on cell complexes / Florian Frick. Gutachter: Imre Bárány ; John M. Sullivan. Betreuer: John M. Sullivan." Berlin : Technische Universität Berlin, 2015. http://d-nb.info/1076082246/34.

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14

Petris, Carisa Kay. "Identification and characterization of M cells in the mammalian conjunctiva." Diss., Columbia, Mo. : University of Missouri-Columbia, 2007. http://hdl.handle.net/10355/4888.

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Thesis (Ph. D.)--University of Missouri-Columbia, 2007.
The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on December 12, 2007) Vita. Includes bibliographical references.

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15

Mcshane, Helen Irene. "Immunisation strategies for enhancing T cell responses against M. tuberculosis." Thesis, University of London, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.271722.

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16

Montanez-Wiscovich, Marjorie E. "Discerning the Role of LMO4 as a Global Modulator of G2/M Cell Cycle Progression and Centrosome Cycle in Breast Cancer Cells." Case Western Reserve University School of Graduate Studies / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=case1259899890.

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17

LeaÌƒo, Maria JoaÌƒo de Lemos Pinto Estrela. "Modulation of G2/M cell cycle checkpoints by Epstein-Barr virus." Thesis, Imperial College London, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.414473.

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18

CHEN, WENDY YUNTIEN. "SYNTHESIS AND OLIGOSACCHARIDE PROCESSING OF IMMUNOGLOBULIN-M DURING B-CELL DIFFERENTIATION." Diss., The University of Arizona, 1987. http://hdl.handle.net/10150/184207.

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In order to understand glycoprotein biosynthesis and processing, we have studied the glycosylation and intracellular assembly kinetics of murine IgM which are expressed functionally only at specific stages of B-cell differentiation by the corresponding tumor cell lines. We have shown that the majority of carbohydrate chains on intracellular IgM contain predominantly Man₈GlcNac₂ rate limiting step in the carbohydrate processing is the transport from the RER to the Golgi apparatus. We made comparisons of carbohydrate structures on secretory and membrane-bound u chains produced by different cell lines. Our results show that the carbohydrates on WEHI231 membrane-bound IgM are less processed, and the processing at individual glycosylation sites is different for IgMs produced by plasmacytoma (MOPC104E) and hybridoma (MPC11xW279.2) cell lines. In addition, we also show that the glycosylation and processing are dramatically altered by lowering the glucose concentration in the cell culture medium. These results are a beginning for our understanding of the influence of the polypeptide on the final glycosylation patterns of a glycoprotein, and the genetic and environmental control over the carbohydrate processing during intracellular transport. The kinetic studies on IgM synthesis and maturation in WEHI231 as well as WEHI279.1/12 cells have led to the conclusion that membrane bound IgM and soluble IgM are segregated and processed individually even in the same cell. These differences appear to lead to the changes in carbohydrate/processing for membrane-bound and soluble IgM.

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19

Quinn, Jennifer E. "BRCA1 mediated G2/M cell cycle arrest in response to taxol." Thesis, Queen's University Belfast, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.326034.

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20

Gunn, Lena Elizabeth. "Validation of the M-Vac cell collection system for forensic purposes." Thesis, Boston University, 2013. https://hdl.handle.net/2144/21161.

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Thesis (M.S.F.S.)
There is need for further development of cellular collection techniques in the field of forensic science. Currently, forensic analysts are limited to the use of swabs, taping, cutting, and scraping methods to collect cellular material. Each of these methods has its own benefits and drawbacks, however, none of them result in 100% recovery of the cells. The Microbial Vacuum system (M-Vac®), developed by MSI, is a liquid based cellular collection system that was originally developed to collect microbes in the food-processing industry from various surfaces. This research represents a detailed study into the feasibility of utilizing the M-Vac® system for forensic purposes. Specifically, the phosphate buffer used with the M-Vac® was tested to confirm that it does not have a detrimental effect on cellular retrieval. Further, the ability of the M-Vac® to collect cellular material from a variety of substrates was tested. It was determined the M-Vac® can successfully collect both blood and semen from tile, denim, carpet, and brick materials in sufficient quantity for downstream PCR analysis. Additionally, examination into whether DNA was dispersed during collection due to the significant force of impact of the liquid striking the surface was conducted. Specifically, areas surrounding the sample collection region were swabbed after collection with the M-Vac® and tested. Quantitative PCR analysis showed that DNA was retrieved up to 4 inches away from the collection area. This indicates that the M-Vac® system is a viable cell collection technique for forensic purposes, but only for samples which are isolated (i.e. where there is not another probative sample adjacent to it). If there are two probative samples within the same vicinity, then swabbing or taping is the recommended method of collection.

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21

Samaraweera, Preminda. "Oligosaccharides of mouse immunoglobulin-M: Structural variations in hybridoma and myeloma cells." Diss., The University of Arizona, 1988. http://hdl.handle.net/10150/184496.

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Many protein-linked oligosaccharides are believed to impart biological specificities to the molecules. The knowledge of detailed structural characteristics of oligosaccharides is essential for understanding their functions. In order to develop methodology for characterization of oligosaccharides of glycoproteins, and to compare glycosylation patterns of different immunoglobulins, oligosaccharides of IgM from two cell lines, MOPC 104E and PC 700, were analyzed. Homogeneous preparations of glycopeptides carrying individual glycosylation sites of the heavy chain were obtained from the two IgM's. The oligosaccharides of these glycopeptides were prepared by hydrazinolysis, and fractionated by HPLC under conditions that resolve oligosaccharides by charge and size, and by affinity chromatography on Concavalin A-Sepharose. Structures of some of these oligosaccharides were determined by 400 MHz NMR spectroscopy. HPLC fractionation by charge resolved oligosaccharides with zero, one, two, and three sialic acids. As indicated by HPLC analyses, oligosaccharides at all the glycosylation sites of both the IgM's were highly heterogeneous. A comparative study on oligosaccharides prepared by peptide-N-glycosidase F digestion of glycopeptides showed a similar degree of heterogeneity. Therefore, it was concluded that the observed heterogeneity of oligosaccharides was not an artefact caused by hydrazinolysis. Major differences between the glycosylation patterns of the two IgM's were evident from analyses of the oligosaccharides by both chromatographic techniques and NMR spectroscopy. MOPC IgM contained a high proportion of sialylated oligosaccharides when compared to PC IgM. Furthermore, the major oligosaccharide structures of MOPC IgM were of triantennary type whereas PC IgM contained biantennary oligosaccharides as its major species. In both the IgM's, a decreased trend of oligosaccharide processing was observed from the N-terminus to the C-terminus.

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22

Hirabayashi, Shigeki. "APOBEC3B is preferentially expressed at the G2/M phase of cell cycle." Doctoral thesis, Kyoto University, 2021. http://hdl.handle.net/2433/264663.

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京都大学
新制・課程博士
博士(医学)
甲第23382号
医博第4751号
新制||医||1052(附属図書館)
京都大学大学院医学研究科医学専攻
(主査)教授伊藤貴浩, 教授滝田順子, 教授江藤浩之
学位規則第4条第1項該当
Doctor of Medical Science
Kyoto University
DFAM

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23

Alsadoon, Hamadah S. "Use of Cell Phones in Education at King Saud University in the Kingdom of Saudi Arabia." Ohio University / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1353509728.

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24

Roberts, Carol Louise. "The interaction of adherent invasive escherichia coli with intestinal epithelial cells, in vitro derived M-cells and macrophages." Thesis, University of Liverpool, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.501692.

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The initial lesions observed in Crohn's disease typically occur over lymphoid aggregates in tne large bowel and Heyers patches in the small bowel where M-cells are present. An increased presence of Adherent and Invasive Escherichia coli (AlEC) has been reported in Crohn's disease mucosa. Crohn's disease is common in westernised countries where low levels of soluble fibre are eaten and where the use of emulsifiers in processed food is common. It is possible that dietary factors may be implicated in the pathogenesis of the disease. The aims of this work were to establish if AIEC interact with intestinal M-cells, and to establish the effect that dietary factors have on these interactions.

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25

Pröller, Stephan [Verfasser], Eva M. [Akademischer Betreuer] Herzig, Peter [Gutachter] Müller-Buschbaum, and Eva M. [Gutachter] Herzig. "Morphology Formation and Manipulation in Printed Organic Solar Cells / Stephan Pröller ; Gutachter: Peter Müller-Buschbaum, Eva M. Herzig ; Betreuer: Eva M. Herzig." München : Universitätsbibliothek der TU München, 2018. http://d-nb.info/1155303512/34.

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Pröller, Stephan Verfasser], Eva M. [Akademischer Betreuer] [Herzig, Peter [Gutachter] Müller-Buschbaum, and Eva M. [Gutachter] Herzig. "Morphology Formation and Manipulation in Printed Organic Solar Cells / Stephan Pröller ; Gutachter: Peter Müller-Buschbaum, Eva M. Herzig ; Betreuer: Eva M. Herzig." München : Universitätsbibliothek der TU München, 2018. http://d-nb.info/1155303512/34.

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27

Schwingel, Melanie [Verfasser], and M. [Akademischer Betreuer] Bastmeyer. "Multiple trap optical tweezers for cell force measurements / Melanie Schwingel. Betreuer: M. Bastmeyer." Karlsruhe : KIT-Bibliothek, 2012. http://d-nb.info/1023081792/34.

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28

Brearley, Madelaine C. "Mitochondrial phosphoenolpyruvate carboxykinase (PEPCK-M/PCK2), phosphoglycerate dehydrogenase (PHGDH) and muscle cell growth." Thesis, University of Nottingham, 2018. http://eprints.nottingham.ac.uk/52138/.

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Our group reported upregulation of a novel group of genes was associated with beta-adrenergic agonist (BA)-induced muscle hypertrophy in pigs. The aim of this PhD was to investigate the expression of these genes, and particularly the role of mitochondrial phosphoenolpyruvate carboxykinase (PEPCK-M/PCK2) and phosphoglycerate dehydrogenase (PHGDH), in muscle cell growth. A significant (p < 0.01) increase in mRNA transcript abundance was detected at day 2 of differentiation in C2C12 cells for PEPCK-M, PHGDH, phosphoserine aminotransferase-1, phosphoserine phosphatase, asparagine synthetase, sestrin-2 and activating transcription factor-5. This novel peak coincided with the peak in myogenin mRNA, connecting these genes with a crucial point of myogenic differentiation. Hypertrophy was induced in C2C12 myotubes treated with dibutyryl-cAMP (dbcAMP), mimicking the BA response in vivo, however mRNA expression of these genes were unaffected. The porcine myosin heavy chain (MyHC)-IIB promoter-reporter C2C12 cell assay demonstrated similar in vivo responses to known anabolic and catabolic agents. Thus, C2C12 cells were utilised to determine the role of PEPCK-M and PHGDH in myogenic differentiation. Firstly, C2C12 cells were treated with a PEPCK inhibitor, 3-Mercaptopicolinic acid (3-MPA). 3-MPA induced differentiation, resulting in a hypertrophic response comparable to dbcAMP treatment. However, it was unclear whether 3-MPA inhibited PEPCK-M enzyme activity as 3-MPA interfered with the in vitro assay. Next, C2C12 cells were transfected with either PCK2 or PHGDH overexpression construct. No obvious phenotype was observed, but PHGDH and PEPCK-M overexpression both increased MyHC-IIB mRNA. The reoccurring induction of the same group of genes along with MyHC-IIB supports the hypothesis that co-ordinated upregulation of these genes may drive hypertrophic growth. To conclude, PEPCK-M, along with other genes upregulated with BA-induced hypertrophy and C2C12 differentiation, show co-ordinate regulation in times of high biosynthetic demand. PEPCK-M appears to sit at an intersection that allows metabolic flux to be largely altered by diverting intermediates during energy metabolism.

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Bryson, Benjamin Levi. "The Paradoxical Roles of Oncostatin M in Mammary Epithelial Cell Senescence and Transformation." Case Western Reserve University School of Graduate Studies / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=case1510584483133814.

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30

Smigiel, Jacob. "ONCOSTATIN M & TRANSFORMING GROWTH FACTOR SIGNALING CONVERGE TO REGULATE CANCER CELL PLASTICITY." Case Western Reserve University School of Graduate Studies / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=case152891618991579.

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31

Managh, Amy J. "Single-cell tracking of therapeutic cells using Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry." Thesis, Loughborough University, 2014. https://dspace.lboro.ac.uk/2134/16723.

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Cellular therapy is emerging as a clinically viable strategy in the field of solid organ transplantation, where it is expected to reduce the dependency on conventional immunosuppression. This has produced a demand for highly sensitive methods to monitor the persistence and tissue distribution of administered cells in vivo. However, tracking cells presents significant challenges. In many cases transplanted cells are autologous with the immune system of the transplant recipient, and hence are invisible to typical methods of detection. To enable their differentiation, the cells must be labelled with a suitable, non-toxic and long lifetime label, prior to their administration to patients. In addition, administered cells represent only a small fraction of the recipient's endogenous cells, which necessitates the use of an extremely sensitive detection method. Laser ablation – inductively coupled plasma – mass spectrometry (LA-ICP-MS) is an exquisitely sensitive analytical technique, capable of imaging trace elements in complex samples, at high spatial resolution.

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32

Mei, Lin. "The M(1) muscarinic receptor and its second messenger coupling in human neuroblastoma cells and transfected murine fibroblast cells." Diss., The University of Arizona, 1989. http://hdl.handle.net/10150/184759.

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The data of this study indicate that pirenzepine(PZ)-high affinity muscarinic receptors (mAChRs) are coupled to the hydrolysis of inositol lipids and not to the adenylate cyclase system in human neuroblastoma SH-SY5Y cells. The maximal carbachol(CCh)-stimulated [³H]IP₁ accumulation in the SH-SY5Y cells was decreased in the presence of 1 μg/ml pertussis toxin, suggesting that a pertussis toxin sensitive G-protein may be involved in the coupling. Several cell clones which express only M₁ mAChR were generated by transfecting the murine fibroblast B82 cells with the cloned rat genomic m₁ gene. The transfected B82 cells (cTB10) showed specific [³H](-)QNB binding activity. The mAChRs in these cells are of the M₁ type defined by their high affinity for PZ and low affinity for AF-DX 116 and coupled to hydrolysis of inositol lipids, possibly via a pertussis toxin sensitive G protein. The relationship between the M₁ mAChR density and the receptor-mediated hydrolysis of inositol lipids was studied in 7 clones. The M₁ mAChR densities in these cells characterized by [³H](-)MQNB binding ranged from 12 fmol/10⁶ cells in LK3-1 cells to 260 fmol/10⁶ cells in the LK3-8 cells. The Hill coefficients of the CCh/[³H](-)MQNB competition curves were close to unity for the LK3-1 cells and were less than one in the higher receptor clones. The percentage of the M₁ mAChRs which had high affinity for CCh decreased as the receptor density increased, suggesting the presence of endogenous factors in these cells which may be important for the agonist affinity state of the receptor. A significant correlation was observed between the density of the M₁ mAChR with high affinity for CCh and the maximum [³H]IP₁ accumulation in these cells. There is no significant difference among the CCh EC₅₀ values, its K(A) values and the K(H) values of the CCh/[³H](-)MQNB competition curves. These results suggest that the high affinity state for CCh may be the functional state of the M₁ mAChRs in these cells. Although a linear correlation between the total M₁ mAChR density and the maximum [³H]IP₁ accumulation was observed, there was evidence for the existence of spare M₁ receptors in the clones with high receptor densities but not in those with low receptor densities.

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VAZQUEZ, MORENO LUZ. "BIOCHEMICAL AND GENETIC STUDIES OF ANTIBODY (IMMUNOGLOBULIN-M) PRODUCING CELLS (GLYCOPROTEINS, U-CHAIN, HYBRIDOMAS)." Diss., The University of Arizona, 1985. http://hdl.handle.net/10150/187920.

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We have chosen the murine immunoglobulin M (IgM) as system to study glycoprotein biosynthesis and carbohydrate processing. Secreted IgM heavy chain (m) has five glycosylation sites which location and structures have been determined. m chain variable region (VH) is involved in antigen binding, while the constant region (CH) is responsible for the effector functions in which the carbohydrate plays an important role. We have determined the carbohydrate structures present at each glycosylation site of IgM produced by a hybridoma cell line (PC 700) and its derived mutants and compared them to IgM from myeloma cell MOPC 104E. PC 700 mutants secrete altered IgM. The alterations include: deletion of one or more constant domains (mutants: 128, 313, and 562) and m chain hyperglycosylation (mutants 21 and 38). Gene analysis indicated that deletions can arise from two different mechanisms. One of these involve a major gene change (mutant 128), while others come from base point mutations (mutants 313 and 562). Cells 21 and 38 did not appear to have m gene insertions. Determination of purified single glycosylation site structures show that PC 700 m chain is processed only to biantennary. Heavy chain protein fragmentation and carbohydrate studies indicate that mutants 21 and 38 alterations are due to an increase in oligosaccharide processing and reduction of unprocessed structures. There is a trend of processing going from PC 700 < 21 < 38. In addition, our results show how growth cell conditions can affect the carbohydrate processing without altering the determinants of m chain oligosaccharide structures. Studies on the IgM molecule illustrate the need for precisely define structure-function relationships. This would allow the selection of the best antibodies for studies such as those involved in immunotherapy.

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Brennecke, Anne-M. [Verfasser], and Gerhard [Akademischer Betreuer] Gross. "Kinetics of B cell development in adult mice / Anne-M. Brennecke ; Betreuer: Gerhard Gross." Braunschweig : Technische Universität Braunschweig, 2014. http://d-nb.info/1175821411/34.

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35

Cervigni, Romina Ines. "Analysis of the molecular mechanisms of the Golgi-based G2/M cell cycle checkpoint." Thesis, Open University, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.580685.

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This thesis is focused on the role of Golgi fragmentation in the regulation of the G2/M transition of the cell cycle, and it is based on previous findings that Golgi fragmentation is required to enter into mitosis. The Golgi complex is composed of many cisternal stacks that are interconnected by tubules, to form a continuous 'ribbon-like' structure. During mitosis, the Golgi ribbon undergoes extensive fragmentation through a multi stage process that promotes its correct partitioning and inheritance by the daughter cells. The first part of my work is focused on the understanding of the mechanisms which block cells in G2 when Golgi fragmentation is inhibited. I show that the Golgi-dependent G2 arrest is mediated by a failure of centrosome maturation, an event that is essential to achieve activation of the CdkllCyclinB (Cdkl/CycB) complex, the master regulator of mitosis. Indeed, the failure of Golgi fragmentation inhibits the recruitment to and activation at the centrosome of the kinase Aurora-A. This kinase is essential for the activation of Cdkl/CycB at the centrosome. This part of the thesis contributes to the definition of a previously unidentified point of dialogue between the Golgi apparatus and the centrosome in the regulation of G2/M transition. The second part of the thesis describes the development of three novel experimental approaches to induce the block of Golgi fragmentation. They integrate a previously developed assay that is based on the microinjection of blockers of Golgi fragmentation, a reliable but demanding approach. The assays that I have developed are based on the ability of the GRASP65 protein to regulate Golgi fragmentation. As well as being essential for inducing the Golgi checkpoint in a wide cell population, they are also useful for the unravelling of the mechanism through which GRASP65 acts in the Golgi checkpoint.

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Weber, Patrick [Verfasser], M. Cristina [Akademischer Betreuer] Cardoso, and Bodo [Akademischer Betreuer] Laube. "DNA replication dynamics in embryonic stem cells / Patrick Weber ; M. Cristina Cardoso, Bodo Laube." Darmstadt : Universitäts- und Landesbibliothek Darmstadt, 2018. http://d-nb.info/1171988176/34.

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37

Nassra, Merian. "Effets anti-inflammatoires des stilbènes sur des cultures cellulaires de microglies et mécanismes d'action." Thesis, Bordeaux 2, 2012. http://www.theses.fr/2012BOR22018/document.

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Les processus neuro-inflammatoires sont observés dans de nombreux troubles neurodégénératifs. Les cellules microgliales étant les principales cellules immunitaires du système nerveux central, plusieurs recherches ont été menées afin de déterminer des molécules possédant des propriétés anti-inflammatoires au niveau de ces cellules. Une famille de polyphénols, les stilbènes (des dérivés du resvératrol), présentent des activités anti-inflammatoires au niveau périphérique et central. Dans notre étude, nous avons premièrement évalué les propriétés anti-inflammatoires de 25 stilbènes en déterminant leur capacité à inhiber la libération de NO dans un modèle de cellules microgliales BV-2 activées par le LPS. Dix stilbènes inhibent significativement cette production avec des IC50 comprises entre 3,9 ± 0,7 et 23,4 ±1,0 µM. Parmi ces composés, 1 monomère (moracine M) et 2 tétramères du resvératrol (vitisines A et B) diminuent la synthèse d’iNOS, enzyme responsable à la libération de NO, au niveau transcriptionnel et traductionnel. Nous avons ensuite mis en évidence que la moracine M inhibe la phosphorylation d’ERK1/2 et JNK (deux MAPK) et d’Akt (la voie PI3K/Akt) dans des cellules BV-2 activées. Ces enzymes étant impliquées dans la signalisation de la réponse inflammatoire, elles induisent la production de plusieurs médiateurs inflammatoires. La moracine M inhibe significativement la production de certains d'entre eux (NO, TNF-α, IL-1β et PGE2). Ce stilbène attenue également la synthèse de la protéine de mPGEs-1 (enzyme impliquée à la production de PGE2). Ainsi, la moracine M pourrait être un candidat potentiel pour prévenir l’inflammation impliquée dans les maladies neurodégénératives
Chronic neuro-inflammatory processes observed in many neurodegenerative disorders. Microglia are the main immune cells of the central nervous system. Many studies have been conducted to find molecules with anti-inflammatory properties in the central nervous system. A family of polyphénols, Stilbenoids (resveratrol derivatives), showed anti-inflammatory effects in peripheral and central levels.In this study, we have first evaluated anti-inflammatory effects of 25 stilbenes for their potential to inhibit NO release by LPS-activated BV-2 microglial cells. Ten stilbenes significantly reduced LPS-induced NO production with IC50 ranging from 3.9 ± 0.7 to 23.4 ±1.0 µM. Among these molecules 1 monomer (moracin M) and two tetramers (vitisins A and B) attenuated the expression of iNOS, a responsible enzyme for NO release on transcriptional and translational levels. Then, we have demonstrated that moracin M inhibits ERK1/2 and JNK phosphorylation of MAPK pathway and Akt phosphorylation of PI3K/Akt pathway, two signaling pathways involved in the inflammatory response in activated BV-2 cells. Indeed, the activation of these pathways leads to the production of several inflammatory mediators. We have shown that moracin M significantly inhibits the production of certain mediators such as NO, TNF-α, IL-1β and PGE2. This stilbene reduces the synthesis of mPGES-1 protein (an enzyme involved in the production of PGE2). In conclusion, we suggest that moracine M could be a potential candidate prevents the inflammation which involved in the progress of neurodegenerative diseases

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38

Ghai, Kanika. "Notch-Signaling in Retinal Regeneration and Müller glial Plasticity." The Ohio State University, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=osu1256322124.

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39

Thanasoula, Maria. "ATM/ATR-dependent responses to dysfunctional telomeres at the G2/M transition." Thesis, University of Oxford, 2012. http://ora.ox.ac.uk/objects/uuid:b9f806e3-88e5-4dc4-b2e9-8ecf854249d1.

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Mammalian telomeres are nucleoprotein complexes at the end of chromosomes containing a specific protein complex, called shelterin. Shelterin protects chromosome ends from the DNA damage response (DDR), by facilitating the formation of a telomeric capping structure, called the T-loop. During their elongation in S phase, telomeres become transiently uncapped and can be sensed as DNA damage in G2 phase. This leads to the recruitment of DDR factors, such as phosphorylated histone H2AX (γH2AX), to the telomeres forming the so-called, telomere dysfunction-induced foci (TIFs). My PhD work described here, indicates that DNA damage occurring during interphase can persist after entry into mitosis, indicated by the detection of γH2AX at a subset of mitotic telomeres in human and mouse cells. This accumulation of γH2AX to mitotic telomeres is ATM-dependent and the γH2AX-labelled uncapped telomeres that persist, are shorter than the average telomere length for the entire cell population. Most importantly, my work suggests that telomere uncapping, naturally occurring or artificially induced, is detected by two parallel ATM/ATR-dependent pathways at the G2/M transition: a p53/p21-dependent pathway through the ATM/ATR-mediated phosphorylation of p53 at Ser15 and a CHK1/CHK2-dependent pathway that acts through negative regulation of CDC25 phosphatases. In particular, telomere uncapping triggered by TRF2 depletion leads to CHK2-dependent CDC25A degradation, while POT1 depletion results in CHK1-mediated CDC25A and CDC25C degradation. Both pathways act as sensors of unprotected telomeres at the G2/M transition and block cell cycle progression through inhibition of CDK1/Cyclin B complex, allowing telomere re-capping before entry into mitosis. This mechanism protects telomere integrity by the maintenance of a cell cycle stage conducive for capping reactions and thereby prevents genomic instability induced by telomere dysfunction. Finally, I studied the cellular functions of 3 poorly characterised shelterin components, TRF1, RAP1 and TPP1, in telomere protection. TRF1 and to a lesser extent RAP1 were shown to be important for telomere protection by suppressing DDR at the telomeres, while TPP1 was shown to be mainly responsible for the recruitment of the catalytic subunit of telomerase, TERT , to the chromatin, contributing to telomere maintenance. In conclusion, my work on both human and mouse models, reveals an important part of the DDR pathways activated by dysfunctional telomeres, as well as the molecular mechanisms underlying the cell cycle specific regulation of telomere capping, which ensures that only cells with intact telomeres enter mitosis.

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40

Rytkönen, J. (Jani). "Effect of heat denaturation of bovine milk beta-lactoglobulin on its epithelial transport and allergenicity." Doctoral thesis, University of Oulu, 2006. http://urn.fi/urn:isbn:9514281209.

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Abstract Beta-lactoglobulin (β-lg) is the main whey protein in bovine milk. It belongs to the lipocalin protein family, and it is one of the main milk allergens. Resistance to hydrolysis is a particular feature of β-lg making it possible that β-lg reaches the small intestine in its native form. Heat treatments during milk processing may change the native structure of bovine β-lg and change its intestinal transport properties. Heat induced conformational alterations may also expose new antigenic sites. However, there have been no previous studies on the effects of heat treatment on the transport of β-lg or on its sensitizing properties. Cow's milk allergy is one of the most important food allergies affecting about 2.4% of infants. Milk proteins, including β-lg, in breast milk substitute formulas are often the earliest foreign antigens in the diet of newborns. According to the hygiene hypothesis, natural infections and vaccinations may modify the immunological balance and decrease the risk of allergy. Isoelectric precipitations followed by anion exchange and gel filtration were used to purify bovine milk β-lg in its native form. Transport of native and heat-denatured β-lg was compared in two in vitro cell models, Caco-2 and M-cells. Sensitization properties of native and heat-denatured β-lg were studied with an animal model using Hooded-Lister rats. Effects of BCG vaccination in combination with the native β-lg were also studied. Effects of different sensitizations were assessed by antibody levels in serum and inflammation locally in the gastrointestinal tract. Heat denaturation of β-lg made its transport slower in both enterocytes and M-cells. M-cells were more effective transporters of both native and heat-denatured β-lg than caco-2 cells. Animals generated higher levels of IgE when sensitized with native β-lg, but heat-denatured β-lg induced a more intense inflammatory cell reaction in the gastrointestinal tract. Vaccination with BCG decreased serum IgE concentration and modified the predominant site of the inflammatory cell response in intestine. The results indicate that, heat denaturation of β-lg and BCG vaccination, change both the systemic and the mucosal response to bovine milk β-lg. The reasons for this remain speculative. The effect of BCG vaccination is consistent with the hygiene hypothesis. The observed alteration of transport properties could be one mechanism by which heat denaturation modifies the allergenic properties of this protein, but additional studies are necessary to assess whether other mechanisms, such as exposure of new antigenic determinants are also relevant.

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41

Hadley, Jennifer Kathleen. "Identification of channels underlying the M-like potassium current in NG108-15 neuroblastoma-glioma cells." Thesis, University College London (University of London), 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.393710.

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42

Konishi, Satoshi. "Directed induction of functional multi-ciliated cells in proximal airway epithelial spheroids from human pluripotent stem cells." Kyoto University, 2016. http://hdl.handle.net/2433/215418.

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43

Mumtaz, Imtiaz M. [Verfasser]. "Effects of immunosuppressive drugs and CD4+ T-cell depletion on plasma cell survival in lupus prone (NZB/W) mice / Imtiaz M. Mumtaz." Berlin : Medizinische Fakultät Charité - Universitätsmedizin Berlin, 2009. http://d-nb.info/1023710315/34.

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44

Singh, Vibhuti [Verfasser], and M. [Akademischer Betreuer] Zöller. "Attending persistent T cell activation in alopecia areata : A therapeutic option / Vibhuti Singh. Betreuer: M. Zöller." Karlsruhe : KIT-Bibliothek, 2011. http://d-nb.info/1014279712/34.

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45

Czepukojc, Beate M. [Verfasser], and Alexandra K. [Akademischer Betreuer] Kiemer. "Investigations on IMP2-2 and Kupffer cells in steatohepatitis / Beate M. Czepukojc ; Betreuer: Alexandra K. Kiemer." Saarbrücken : Saarländische Universitäts- und Landesbibliothek, 2019. http://d-nb.info/1199933023/34.

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46

Shokri, Beshr [Verfasser], and M. [Akademischer Betreuer] Grimm. "Expression of 1,25-Dihydroxyvitamin D3 receptor in oral squamous cell carcinoma / Beshr Shokri ; Betreuer: M. Grimm." Tübingen : Universitätsbibliothek Tübingen, 2018. http://d-nb.info/1199355887/34.

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47

Jones, Matthew Dunford. "Effects of radiation on the Gâ†2/M checkpoint in human tumour cells of differing radiosensitivities." Thesis, University of Liverpool, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.387452.

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48

Hatayama, Hiroshi. "Progesterone Enhances Macrophage Colony-Stimulating Factor(M-CSF) Production in Human Endometrial Stromal Cells in vitro." Kyoto University, 1995. http://hdl.handle.net/2433/160724.

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本文データは平成22年度国立国会図書館の学位論文(博士)のデジタル化実施により作成された画像ファイルを基にpdf変換したものである
Kyoto University (京都大学)
0048
新制・課程博士
博士(医学)
甲第5973号
医博第1639号
新制||医||603(附属図書館)
UT51-95-D292
京都大学大学院医学研究科外科系専攻
(主査)教授西川伸一, 教授塩田浩平, 教授森崇英
学位規則第4条第1項該当

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49

Kieswetter, Nathan Scott. "Remodelling of Mycobacterial Peptidoglycan During Cell Division and the Epigenetics of Macrophages during M. tuberculosis infection." Doctoral thesis, Faculty of Health Sciences, 2021. http://hdl.handle.net/11427/33815.

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There are so many people who I would like to thank. If it takes a village to raise a child, it certainly takes a city to train a scientist. Firstly, I would like to thank my supervisors, A/Prof Reto Guler, Prof Frank Brombacher and Dr Mumin Ozturk for allowing me to further my studies and allowing me to work on several interesting projects. Specifically, I would like to take this opportunity to thank A/Prof Reto Guler for his insightful patient advice, training and ever willingness to talk about my work. His brilliant example has made me a better scientist. Further, I would also like to thank Dr Mumin Ozturk for his constant, patient mentorship, help, advice and friendship. His influence, guidance and example have affected me more than he'll ever know. Lastly, but certainly not least, I would like Professor Bavesh Kana and for all his advice and support. I would also like to express my gratitude to my labmates from the Brombacher group. All the conversations, laughs, celebrations and commiserations have made this journey undeniably easier. In particular, I would like to thank Shelby-Sara Jones for her constant willingness to help with lab work whilst chatting about everything under the sun. To my friends and family, there are no words to express my unending gratitude. Without their love and support along the way, I would never have gotten to this stage in my life. To my parents and sister, I would like to say a huge thank you for their constant support and love during my academic career so far. You guys have been wonderful. A huge thank you to Daniela de Almeida and the French's for their support and love from afar– you guys have been great. I would like to say a special thank you to Dustin Fischer who has always been there for a beer and good old-fashioned rant. I can only hope that my friendship and advice have been even the smallest bit as helpful to him as he has been to me during our long trek through academia. To the Cunniffes, thank you for all your support down this long road and truly making me feel like one of the family. Last but by no means least, I would like to thank my partner, Teagan Cunniffe, whose effortless grace, wit, humour, friendship and constant love have been the single greatest gifts I have ever received. I look forward to our adventures to come. Thank you for being there every step of the way and keeping me sane - This dissertation is dedicated to you.

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50

Beckta, Jason. "ATM, BRCA1, and Aurora A: Mechanisms of G2/M Checkpoint Control in Human Embryonic Stem Cells." VCU Scholars Compass, 2014. http://scholarscompass.vcu.edu/etd/3477.

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When cultured in vitro, human embryonic stem cells (hESCs) acquire genetic abnormalities that have slowed their therapeutic use. As hESCs have a “leaky” G1/S boundary, the pressure of ensuring genetic integrity falls on the G2/M checkpoint, which can be activated by failed chromosomal decatenation (among other stimuli). It is hypothesized that hESCs have a deficient decatenation checkpoint, but little data supports this. Evidence suggests that the ataxia telangiectasia mutated (ATM) kinase controls the G2/M decatenation and DNA damage checkpoints, though previous reports are conflicting on this point. My work demonstrates that inhibition of decatenation activates ATM and arrests hESCs in G2. Pharmacologic inhibition of ATM (ATMi) abrogates this arrest, allowing hESCs to enter mitosis. Live cell imaging studies reveal that ATMi increases the time it takes to complete mitosis. Culture of cells under ATMi causes a gain of DNA content, which is reversed once ATMi is relieved. BRCA1, a known target of ATM, is also involved in the G2/M checkpoint. Experimental evidence reveals that activated ATM phosphorylates BRCA1, preventing Aurora A from interacting with and phosphorylating BRCA1 on S308, a modification necessary for mitotic entry. Together, this data illuminates a novel pathway by which ATM activation mediates G2 arrest in hESCs.

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