{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/72407"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/72407","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Development of EPR Methods for Simultaneous Measurements of Intra- and Extra-Cellular Oxygen Concentration in Vitro","abstract":"In this thesis, the electron paramagnetic resonance (EPR) methods for simultaneous measurement of intra- and extracellular concentration of oxygen ( (O$\\sb2$)) in viable cells have been developed.","abstract_html":"In this thesis, the electron paramagnetic resonance (EPR) methods for simultaneous measurement of intra- and extracellular concentration of oxygen ( (O$\\sb2$)) in viable cells have been developed.","abstract_has_math":true,"creators":["Hu, Huping"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Biophysics","degree_department":null,"school":null,"contributors":["Swartz, Harold M."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":1993,"date_issued":"1993","date_published":"1993","updated_at":"2026-07-22T22:26:06Z","subjects":["Biology, Animal Physiology","Biophysics, Medical"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(UMI)AAI9329063"],"render_values":[{"text":"(UMI)AAI9329063","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/72407","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Swartz, Harold M."]},{"key":"dc:creator","label":"Author","values":["Hu, Huping"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["1993","2014-12-17T22:22:51Z","10000-01-01"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biophysics"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biology, Animal Physiology","Biophysics, Medical"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/72407","(UMI)AAI9329063"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["In this thesis, the electron paramagnetic resonance (EPR) methods for simultaneous measurement of intra- and extracellular concentration of oxygen ( (O$\\sb2$)) in viable cells have been developed.","In the presence of Fe(CN)$\\sb6\\sp{-3}$ in the medium, intracellular (O$\\sb2$) is measured by a neutral $\\sp $N-nitroxide and extracellular (O$\\sb2$) is measured by a negatively charged $\\sp $N, since charged species do not enter cells and the EPR spectrum of a $\\sp $N-nitroxide does not overlap with that of a $\\sp $N-nitroxide. The method is based in part on the minimal broadening of negatively charged nitroxides by Fe(CN)$\\sb6\\sp{-3}$ and the very effective broadening of neutral nitroxides by the same paramagnetic ions. Results with this method confirm the existence of gradients in (O$\\sb2$) between the extracellular and intracellular compartment in CHO cells and M5076 tumor cells, even without stimulation of cellular resiration by carbonyl cyanide m-chlorophenylhydrazone (CCCP). The nature of the barrier that needs to be involved to account for the experimental results raises some significant questions.","Due to the potential toxic effect of Fe(CN)$\\sb6\\sp{-3}$ to cells after long time exposure and the difficulties to extend the measurement of intracellular (O$\\sb2$) to in vivo in the presence of Fe(CN)$\\sb6\\sp{-3},$ a new method without the use of Fe(CN)$\\sb6\\sp{-3}$ was also studied. The study has been focused on the feasibility of developing nitroxides that localize selectively in the intracellular compartment so that the intracellular (O$\\sb2$) could be measured without the use of Fe(CN)$\\sb6\\sp{-3}$ and extracellular (O$\\sb2$) could be measured simultaneously by a charged $\\sp $N-nitroxide. It is based on the use of readily hydrolysed ester linkages, such that the nitroxides become converted intracellularly to ionic derivatives that do not cross cell membranes readily. Up to 120-fold increased concentrations of intracellular nitroxides and their one electron reduction product, the hydroxylamines, were obtained. The EPR spectra of the intracellular nitroxides and their partitions into water and CHCl$\\sb3$ were consistent with their conversion to the ionic species. Further studies indicate that these nitroxides have the properties needed for their use as probes of intracellular (O$\\sb2$) and that it should be feasible to synthesize nitroxides that will be even more effective for this purpose.","Made available in DSpace on 2014-12-17T22:22:51Z (GMT). No. of bitstreams: 1 9329063.pdf: 4353930 bytes, checksum: 5ee3388e66ca3396285e7589572b8a3c (MD5) Previous issue date: 1993","Embargo set by: Seth Robbins for item 72575 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","129 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1993."]},{"key":"dc:title","label":"Title","values":["Development of EPR Methods for Simultaneous Measurements of Intra- and Extra-Cellular Oxygen Concentration in Vitro"]}]}],"canonical_facts":{"dc:contributor":["Swartz, Harold M."],"dc:creator":["Hu, Huping"],"dc:date":["1993","2014-12-17T22:22:51Z","10000-01-01"],"dc:description":["In this thesis, the electron paramagnetic resonance (EPR) methods for simultaneous measurement of intra- and extracellular concentration of oxygen ( (O$\\sb2$)) in viable cells have been developed.","In the presence of Fe(CN)$\\sb6\\sp{-3}$ in the medium, intracellular (O$\\sb2$) is measured by a neutral $\\sp $N-nitroxide and extracellular (O$\\sb2$) is measured by a negatively charged $\\sp $N, since charged species do not enter cells and the EPR spectrum of a $\\sp $N-nitroxide does not overlap with that of a $\\sp $N-nitroxide. The method is based in part on the minimal broadening of negatively charged nitroxides by Fe(CN)$\\sb6\\sp{-3}$ and the very effective broadening of neutral nitroxides by the same paramagnetic ions. Results with this method confirm the existence of gradients in (O$\\sb2$) between the extracellular and intracellular compartment in CHO cells and M5076 tumor cells, even without stimulation of cellular resiration by carbonyl cyanide m-chlorophenylhydrazone (CCCP). The nature of the barrier that needs to be involved to account for the experimental results raises some significant questions.","Due to the potential toxic effect of Fe(CN)$\\sb6\\sp{-3}$ to cells after long time exposure and the difficulties to extend the measurement of intracellular (O$\\sb2$) to in vivo in the presence of Fe(CN)$\\sb6\\sp{-3},$ a new method without the use of Fe(CN)$\\sb6\\sp{-3}$ was also studied. The study has been focused on the feasibility of developing nitroxides that localize selectively in the intracellular compartment so that the intracellular (O$\\sb2$) could be measured without the use of Fe(CN)$\\sb6\\sp{-3}$ and extracellular (O$\\sb2$) could be measured simultaneously by a charged $\\sp $N-nitroxide. It is based on the use of readily hydrolysed ester linkages, such that the nitroxides become converted intracellularly to ionic derivatives that do not cross cell membranes readily. Up to 120-fold increased concentrations of intracellular nitroxides and their one electron reduction product, the hydroxylamines, were obtained. The EPR spectra of the intracellular nitroxides and their partitions into water and CHCl$\\sb3$ were consistent with their conversion to the ionic species. Further studies indicate that these nitroxides have the properties needed for their use as probes of intracellular (O$\\sb2$) and that it should be feasible to synthesize nitroxides that will be even more effective for this purpose.","Made available in DSpace on 2014-12-17T22:22:51Z (GMT). No. of bitstreams: 1 9329063.pdf: 4353930 bytes, checksum: 5ee3388e66ca3396285e7589572b8a3c (MD5) Previous issue date: 1993","Embargo set by: Seth Robbins for item 72575 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","129 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1993."],"dc:identifier":["http://hdl.handle.net/2142/72407","(UMI)AAI9329063"],"dc:subject":["Biology, Animal Physiology","Biophysics, Medical"],"dc:title":["Development of EPR Methods for Simultaneous Measurements of Intra- and Extra-Cellular Oxygen Concentration in Vitro"],"dc:type":["text"],"thesis:degree_discipline":["Biophysics"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:26:06Z"}