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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
11

Geometry and nonlinear dynamics underlying excitability phenotypes in biophysical models of membrane potential

Herrera-Valdez, Marco Arieli January 2014 (has links)
The main goal of this dissertation was to study the bifurcation structure underlying families of low dimensional dynamical systems that model cellular excitability. One of the main contributions of this work is a mathematical characterization of profiles of electrophysiological activity in excitable cells of the same identified type, and across cell types, as a function of the relative levels of expression of ion channels coded by specific genes. In doing so, a generic formulation for transmembrane transport was derived from first principles in two different ways, expanding previous work by other researchers. The relationship between the expression of specific membrane proteins mediating transmembrane transport and the electrophysiological profile of excitable cells is well reproduced by electrodiffusion models of membrane potential involving as few as 2 state variables and as little as 2 transmembrane currents. Different forms of the generic electrodiffusion model presented here can be used to study the geometry underlying different forms of excitability in cardiocytes, neurons, and other excitable cells, and to simulate different patterns of response to constant, time-dependent, and (stochastic) time- and voltage-dependent stimuli. In all cases, an initial analysis performed on a deterministic, autonoumous version of the system of interest is presented to develop basic intuition that can be used to guide analyses of non-autonomous or stochastic versions of the model. Modifications of the biophysical models presented here can be used to study complex physiological systems involving single cells with specific membrane proteins, possibly linking different levels of biological organization and spatio-temporal scales.
12

Ion selectivity and membrane potential effects of two scorpion pore-forming peptides / D. Elgar

Elgar, Dale January 2005 (has links)
Parabutoporin (PP) and opistoporin 1 (OP1) are cation, a-helical antimicrobial peptides isolated from the southern African scorpion species, Parabuthus schlechteri and Opistophthalmus carinatus, respectively. Along with their antimicrobial action against bacteria and fungi, these peptides show pore-forming properties in the membranes of mammalian cells. Pore-formation and ion selectivity in cardiac myocytes were investigated by measuring the whole cell leak current by means of the patch clamp technique. Pore-formation was observed as the induction of leak currents. Ion selectivity of the pores was indicated by the shift of the reversal potential (E,,,) upon substitution of intra (K' with CS' and CI- with aspartate) and extracellular (Na' with NMDG') ions. Results were compared with the effect of gramicidin A used as a positive control for monovalent cation selective pores. PP and OP I induced a fluctuating leak current and indicate non-selectivity of PP and OP1-induced pores. An osmotic protection assay to determine estimated pore size was performed on the cardiac myocytes. PP and OP1-induced pores had an estimate pore size of 1.38-1.78 nm in diameter. The effect of PP and OP1 on the membrane potential (MP) of a neuroblastoma cell line and cardiac myocytes was investigated. TMRM was used to mark the MP fluorescently and a confocal microscope used to record the data digitally. The resting membrane potential (RMP) of the neuroblastoma cells was calculated at -38.3 f 1.9 mV. PP (0.5 uM) and OP1 (0.5-1 uM) depolarized the entire cell uniformly to a MP of -1 1.9 k 3.9 mV and -9.4 k 1.9 mV, respectively. This occurred after 20-30 min of peptide exposure. In the case of the cardiac myocytes depolarization was induced to -39.7 f 8.4 mV and -32.6 f 5.2 mV by 0.5-1 uM PP and 1.5-2.5 uM OPl, respectively. / Thesis (M.Sc. (Physiology))--North-West University, Potchefstroom Campus, 2006.
13

C type natriuretic peptide facilitates autonomous Ca²⁺ entry in growth plate chondrocytes for stimulating bone growth / C型ナトリウム利尿ペプチドは自発的なCa²⁺流入を介して骨伸長を促進する

Miyazaki, Yuu 23 March 2022 (has links)
京都大学 / 新制・課程博士 / 博士(薬科学) / 甲第23834号 / 薬科博第149号 / 新制||薬科||16(附属図書館) / 京都大学大学院薬学研究科薬科学専攻 / (主査)教授 竹島 浩, 教授 金子 周司, 教授 土居 雅夫 / 学位規則第4条第1項該当 / Doctor of Pharmaceutical Sciences / Kyoto University / DFAM
14

Metody měření elektrického napětí na buněčné membráně / Methods of measurement of electrical voltage on the cell membrane

DIVOKÝ, Karel January 2013 (has links)
The aim of this work was to compile a comprehensive list of methods that deal with the research of voltage on the cell membrane. The work should help in orientation in these methods, as well as understanding of their physical and biological principle. In the first part is analysis of physical properties of membrane potential; in the second part are the most important methods for measuring membrane potential. Very important is the comparison of these methods in terms of their focus, localization, physical demands or in terms degree of damage of object under examination.
15

Efeito da deformação mecânica no transporte iônico em filmes poliméricos. / Effect of mechanical deformation on ionic transport in polimeric films.

Walter Katsumi Sakamoto 10 December 1990 (has links)
Recentemente um efeito piezoelétrico foi encontrado quando um filme polimérico, separando duas soluções de eletrólitos em diferentes concentrações, foi deformado. Este efeito foi atribuído à modulação da mobilidade dos íons no polímero. Foi observado que o efeito diminui com o aumento da concentração de eletrólitos. Para entender mais profundamente este último efeito, foram realizados estudos sobre o transporte de água e sal através do polímero, com e sem deformação, e os resultados podem ser resumidos como segue: a) O aumento da hidratação acarreta um aumento na porosidade e um decréscimo na cristalinidade do polímero. b) O aumento da concentração de sal, no interior do polímero acarreta um aumento na permeabilidade, sendo mais pronunciado na presença de deformação. Esses efeitos tornam mais difícil a modulação dos íons, diminuindo o sinal piezoelétrico observado. / Recently a piezoelectric effect was found when a polymeric film, separating two electrolyte solutions at different concentrations, was deformed. This effect was attributed to the modulation of the mobility of the íons in the polymer. It was observed that the effect decreases as the electrolyte concentrations increases. In order to understand more deeply this last effect a study on water and salt transport through the polymer with and without deformation was carried out, and the results may be summarized as follows: a) An increase in hydration of the polymer leads to an increase in porosity and a decrease in cristallinity. b) An increase in the concentration of the salt leads to an increase in permeability, which is more pronounced in the presence of deformation. The effects make more difficult the modulation of the íons, so lower piezoelectric signals are observed.
16

Metabolismo energético mitocondrial na proliferação de células de glioblastoma U-87MG e T98G em cultura / Mitochondrial energy metabolism in proliferation of cultured U-87MG and T98G glioblastoma cells

Ruas, Juliana Silveira, 1989- 26 August 2018 (has links)
Orientador: Roger Frigério Castilho / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-26T14:26:13Z (GMT). No. of bitstreams: 1 Ruas_JulianaSilveira_M.pdf: 1871713 bytes, checksum: a20e3cd08d0b770aed3b059541e382e3 (MD5) Previous issue date: 2015 / Resumo: A maioria das células tumorais depende da glicólise para a ressíntese de ATP durante um processo de rápida proliferação, mesmo que haja disponibilidade de oxigênio para a transdução de energia mitocondrial (Efeito Warburg). O objetivo do presente estudo foi avaliar o papel do metabolismo oxidativo mitocondrial na proliferação de células de glioblastoma humano U-87MG e T98G. Quando as células foram cultivadas na presença de oligomicina (um inibidor da ATP sintase) ou antimicina A (um inibidor do complexo III da cadeia transportadora de elétrons), observou-se apenas uma inibição parcial da proliferação das células. Notadamente, a incubação dessas células com ambos os inibidores causou uma inibição quase completa na proliferação celular. Resultados semelhantes foram observados em cultura primária de astrócitos, havendo uma queda na proliferação celular somente quando ambos os inibidores mitocondriais estavam presentes. Medidas de consumo de oxigênio indicaram que células de glioblastoma utilizam parcialmente a fosforilação oxidativa para a ressíntese de ATP e apresentam uma respiração bem acoplada. Quando se inibiu, nestas células, a fosforilação oxidativa do ADP com oligomicina ou antimicina A, houve um pequeno aumento no consumo de glicose e na produção de lactato. No entanto, o tratamento com ambos os inibidores mitocondriais promoveu um menor consumo de glicose e produção de lactato, em comparação com os efeitos que a antimicina A promoveu. Isso indica que a cadeia transportadora de elétrons quando inibida pela presença de antimicina A, promove um funcionamento inverso da ATP sintase, promovendo a hidrólise de ATP para que haja um bombeamento de prótons para o espaço intermembranar mitocodrial. De acordo com os resultados acima descritos, uma queda quase completa do potencial de membrana mitocondrial foi observada apenas quando as células de glioblastoma foram incubadas na presença de ambos os inibidores mitocondriais, oligomicina e antimicina A. Quando a análise do ciclo celular foi realizada, observou-se uma diminuição da percentagem das células em G0-G1 e um aumento nas fases S e G2-M quando tratadas com oligomicina. Quando as células foram tratadas com antimicina A e oligomicina mais antimicina A foi constatado uma diminuição significativa nas fases G0-G1 e G2-M, e um aumento na fase S. Em conclusão, estes resultados indicam que a rápida proliferação de células de glioblastoma depende da existência do potencial de membrana mitocondrial, mas não da fosforilação oxidativa ou do transporte de elétrons na cadeia respiratória / Abstract: Most tumor cells rely on glycolysis for ATP resynthesis during rapid proliferation, despite the availability saturating levels of oxygen for mitochondrial energy transduction (Warburg effect). The aim of the present study was to evaluate the role of mitochondrial oxidative metabolism on proliferation of human glioblastoma cells U-87MG and T98G. When cells were cultured in the presence of oligomycin (ATP synthase inhibitor) or antimycin A (inhibitor of complex III of the electron transport chain), we observed only a partial inhibition of cell proliferation. Remarkably, incubation of cells with both inhibitors caused an almost complete inhibition of cell proliferation. Similar results were observed in primary culture of astrocytes, with a decrease in cell proliferation only when both mitochondrial inhibitors were present. Oxygen consumption measurements indicated that glioma cells partially rely on oxidative phosphorylation for ATP turnover and exhibit a well-coupled respiration. In fact, shutting down mitochondrial ADP phosphorylation in these glioma cells with either oligomycin or antimycin inhibitors slightly increased glucose consumption and lactate release. However, the treatment with both mitochondrial inhibitors promoted lower glucose consumption and lactate release as compared with the effects of antimycin alone, which indicates that ATP synthase is operating reversely and thus hydrolyzing ATP and pumping H+ out when the respiratory chain is inhibited by antimycin. In agreement, an almost complete collapse of mitochondrial membrane potential was only observed when the glioma cells were incubated in the presence of both antimycin and oligomycin, but not of only antimycin. When cell cycle analyses were performed in oligomycin-treated cells, a decrease in the percentage of cells in G0-G1 phase and an increase in S and G2-M phases were observed. When cells were treated with antimycin A or oligomycin plus antimycin A, it was observed a significant decrease in G0-G1 and G2-M cell phases and an increase in S phase. Overall, our results suggest that the rapid proliferation of glioblastoma cells is dependent on the mitochondrial membrane potential, but not on oxidative phosphorylation or electron transport in the respiratory chain / Mestrado / Biologia Estrutural, Celular, Molecular e do Desenvolvimento / Mestra em Ciências
17

Effects of glutamine deprivation on oxidative stress and cell survival in breast cell lines

Gwangwa, Mokgadi Violet January 2019 (has links)
Tumourigenic cells utilize aberrant metabolic process that supports the biosynthetic requirements for hyperproliferation, survival and prolonged maintenance characterised by glucose metabolism to lactate dehydrogenase independent of oxygen availability (Warburg effect). In addition, tumourigenic cells exert increased glycolytic- and glutaminolytic activity in order to provide increased quantities of adenosine triphosphate. The aim of this research project was to investigate the influence of glutamine deprivation on proliferation, morphology, oxidative stress, mitochondrial membrane potential, cell cycle progression, antioxidant defences, deoxyribonucleic acid (DNA) damage, energy status, cell survival signaling and cell death induction in tumourigenic- and non-tumourigenic breast cell lines. In this study it was found that glutamine deprivation results in differential antiproliferative activity where the MCF-7 cell line was the most affected with decreased cell growth to 61% after 96 h of glutamine deprivation. Aberrant redox activity was most prominently observed in the MCF-7 cell line accompanied with biphasic mitochondrial membrane potential- and reactive oxygen species production. The MCF-7 cell line showed significant mitochondrial membrane depolarisation after 24 h and 96 h deprivation from glutamine (1.5- and 1.37 fold). Cell cycle progression analysis illustrated an increase in the amount of cells present in the S-phase in the MCF-7 cell line after 72 h of glutamine deprivation. The MDA-MB-231 cell line resulted in a significant increase in cells occupying the G2/M phase after 24 h of glutamine deprivation. Glutamine deprivation in the BT-20 cell line resulted in a significant increase in cells occupying G1 phase after 72 h of glutamine deprivation. The MCF-7 cell line demonstrated the least amount of viable cells when analysing apoptosis induction, when compared to the MDA-MB-231-, MCF-10A- and BT-20 cell lines after glutamine deprivation suggesting that the MCF-7 cell line is the most affected cell line. Analysis of antioxidant mechanism via superoxide dismutase (SOD) inhibition illustrated increased SOD activity in the MCF-7 cell line (9.1%) after 72 h of glutamine deprivation. Evaluation of catalase protein concentration indicated that the MCF-7 catalase expression increased to 1.28 fold after 24 h of glutamine deprivation when compared to cell propagated in complete growth medium. DNA damage was demonstrated by visualising the presence of fluorescent 8-hydroxydeoxyguanosine and showed that the MCF-7 cell line presented with significant 8-hydroxydeoxyguanosine staining. Survival signaling was also evaluated through visualising extracellular signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3K) signaling which demonstrated increased ERK activation in the non-tumourigenic MCF-10A cell line and decreased PI3K activation. This study provides evidence that there are differential- and time-dependent responses in breast tumourigenic cells versus non-tumourigenic cells, to glutamine deprivation thus unraveling the crosstalk between glutamine deprivation, oxidative stress and cell death and different cell types will enable us to better understand the basics of tumour cell metabolism and thus develop therapeutics that provide promising pre-sensitization potential for chemotherapeutic agents. / Dissertation (MSc)--University of Pretoria, 2020. / Physiology / MSc / Unrestricted
18

Investigating alternative sperm preservation methods for assisted reproductive technologies

Slabbert, Marisa January 2013 (has links)
Introduction: Cryopreservation of human sperm is considered a routine practice in assisted reproduction laboratories. Semen samples are mainly cryopreserved as a back-up for procedures, donor sperm, and validation of samples from human immunodeficiency virus-positive patients. Human immunodeficiency virus semen samples generally result in a low yield of purified spermatozoa after decontamination. These samples need to be cryopreserved for later use. Unlike conventional cryopreservation, vitrification does not use harmful cryoprotectants, thereby potentially reducing sperm damage. Vitrification is not yet common practice for sperm cryopreservation in assisted reproduction. The aim of this study was to establish the feasibility of utilising vitrification as an alternative to current conventional cryopreservation of spermatozoa. Methods: Semen samples were collected from human immunodeficiency virus-negative patients seeking diagnostic assistance from the unit. All samples were processed according to the unit’s standard protocol. For Study 1A (n=10) washed samples were divided and cryopreserved using three different cryopreservation media, and two different freezing protocols. In Study 1B (n=15), washed samples were divided and preserved using cryoprotectant-free vitrification in 100 μl, 300 μl and 500 μl volumes. For Study 2 (n=35) washed samples were split and cryopreserved using cryoprotectant-free vitrification (utilizing the volume that resulted in the highest quality spermatozoa in Study 1B) and conventional slow freezing (using the medium and protocol that resulted in superior quality spermatozoa in Study 1A). Post thawing, motility and kinetic parameters (Studies 1 and 2), viability (Study 1), mitochondrial membrane potential (Study 2), and DNA fragmentation (Study 2) of the two groups were compared. vi Results: Study 1A indicated that cryopreserving spermatozoa using Freezing Medium resulted in the highest quality spermatozoa with regards to motility and viability (p<0.05). Comparing the two preservation protocols, no conclusion could be reached on which protocol yielded superior results (p>0.05). The RBL freezing method is shorter, simpler and requires less equipment, and was therefore deemed the preferred method. Study 1B showed that the larger vitrification volumes (300 μl and 500 μl) yielded better spermatozoa in terms of motility and viability (p<0.05). No significant difference was observed with respect to the 300 μl and 500 μl vitrification volume groups. For practical reasons, 300 μl volumes will provide sufficient sperm for any procedure and, the intermediate volume ensures that more than one straw can be preserved. Study 2 found that cryoprotectant-free vitrification resulted in spermatozoa with significantly higher mitochondrial membrane potential and significantly lower apoptosis post thawing (p<0.05). Discussion: Conventional cryopreservation methods may compromise various sperm parameters and final yield. In this study, cryopreservation and cryoprotectant-free vitrification had equivalent outcomes with respect to sperm motility. However, the latter method yielded superior results in terms of ΔΨ and DNA sperm fragmentation. In conclusion, vitrification is an easy, rapid and more affordable technique that requires no special equipment. Using vitrification for purified sperm samples of patients could potentially result in a better post thaw quality for ART procedures. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Obstetrics and Gynaecology / unrestricted
19

Studium exprese MDR pump u kvasinek Saccharomyces cerevisiae za různých růstových podmínek: metoda s fluorescenční sondou diS-C3(3) / Studium exprese MDR pump u kvasinek Saccharomyces cerevisiae za různých růstových podmínek: metoda s fluorescenční sondou diS-C3(3)

Zahumenský, Jakub January 2011 (has links)
In this work, we studied two yeast ABC transporters, Pdr10p and Pdr15p. At the time of assignment of this thesis, it was believed that these proteins contribute to the yeast MDR phenotype (PDR) on the grounds of their high homology to another yeast MDR protein, Pdr5p. In order to study these pumps, two sets of isogenic null-mutant strains were prepared with all possible combinations of gene deletions. We report that both of the studied proteins are very important in sus- taining the normal plasma membrane microenvironment for the most abundant, and essential, yeast plasma membrane protein, H+ -ATPase and so influence the membrane potential. Pdr10p and Pdr15p thus play an as yet unknown role in reg- ulation of the activity of this enzyme. Furthermore, we report that deletion of the genes coding for these proteins severely reduces the ability of the H+ -ATPase to be activated by the protonophore CCCP which is a weak acid. Studies performed with immunosuppressant FK506 further show that this compound reduces the viability of S. cerevisiae mutant strain PLY643 lacking genes coding for Pdr5p, Snq2p and Yor1p. Further deletion of Pdr10p and Pdr15p does not increase the lethality of this compound. Neither CCCP nor FK506 are substrates of the stud- ied pumps. 1
20

Srovnání vybraných karbocyaninových fluorescenčních sond z hlediska jejich použitelnosti při měření změn membránového potenciálu kvasinek. / The comparison of the performace of selected carbocyanine dyes in fluorescent probing of yeast cell membrane potential.

Mudroňová, Kateřina January 2013 (has links)
The membrane potential is one of the most important parameters of the living cell. It can be measured using carbocyanine fluorescent probes. In this thesis we examined parameters of several dyes of this family. For further experiments three of them were chosen - diOC3(3), diIC1(3) a diIC2(5) as a supplement to diSC3(3) and diSC3(5), which represent standard probes used at biophysical department of Institut of Physics. We compared the rates of their accumulation in S. cerevisiae cells to determine if they were MDR pumps' substrates. The other goal of this work was to decide whether the results obtained using different probes are equivalent and to determine if the presence of a probe affects the spectral characteristics of another. For this purpose we have chosen diSC3(3) and diSC3(5). With those dyes we examined the influence of the acidification on membrane potencial of the yeast S. cerevisiae. We showed that the information on depolarization obtained using both probes were matching very well.

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