Mitochondria are double membraned organelles responsible for the majority of ATP production in eukaryotic cells. The mitochondrial inner membrane is folded into cristae structures and is the site of the electron transport chain which terminates in ATP generation. ATP is produced by ATP synthase, a protein complex that has also been shown to have a role in the maintenance of cristae folding. This activity is dependent on Tim11p, a subunit required for the dimerization of ATP synthase super-complexes. Additional proteins located within the inner membrane that are important to mitochondrial morphology include Pcp1p, a serine protease, and its substrate, Mgm1p. Mgm1p is required for mitochondrial fusion and cells deleted for MGM1 do not contain detectable Tim11p. Using biochemical assays and transmission electron microscopy, this study characterized pcp1 mutants in order to analyze the link between Pcp1p functionality, ATP synthase super-complex assembly, and mitochondrial morphology.

No description available.

Assignment (MMed)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: Lipoprotein X (LpX) is an abnormal cholesterol-containing particle that may be present in the serum of subjects with cholestasis, lecithin:cholesterol acyltransferase (LCAT) deficiency and parenteral nutrition. The biochemistry, metabolism, clinical significance and laboratory analysis of LpX is discussed in this study. This laboratory-based project investigated icteric samples received at the Chemical Pathology laboratory, Tygerberg Hospital, for serum predictors of LpX and the use of a modified non-denaturing polyacrylamide gradient gel electrophoresis system in the detection of LpX. The study showed that the non-denaturing polyacrylamide gradient gel electrophoresis system (2-8%) is a useful test in demonstrating LpX in icteric plasma and has potential for a screening test in LCAT deficiency. Serum concentration of conjugated bilirubin, alkaline phosphatase, gamma glutamyltransferase, free cholesterol, phospholipid, free cholesterol: total cholesterol ratio and conjugated bilirubin: total bilirubin ratio are all good predictors of LpX. The ratio of free cholesterol to total cholesterol (FC/TC > 0.6) was the best predictor of LpX. In the setting of obstructive liver disease LpX is seen in 66% of patients if total cholesterol is > 7.5 mmol/L. / AFRIKAANSE OPSOMMING: Lipoproteien X (LpX) is ‘n abnormale cholesterol-bevattende partikel wat teenwoordig mag wees in die serum van persone met cholestase, lesitien:cholesterol asieltransferase (LCAT) gebrek en parenterale voeding. Die biochemie, metabolisme, kliniese belang en laboratorium analise van LpX word bespreek in hierdie werkstuk. Hierdie laboratorium-gebaseerde projek het geelsugtige monsters ondersoek wat ontvang is by die Chemiese Patologie laboratorium, Tygerberg Hospitaal, vir serum voorspellers van LpX en die gebruik van ‘n gemodifiseerde nie-denaturerende polie-akrielamied gradiënt gel elektroforese sisteem in die demonstrasie van LpX. Die bevindinge was dat die nie-denaturerende polie-akrielamied gradient gel elektroforese sisteem (2-8%) is ‘n nuttige toets om LpX te demonstreer in geelsugtige plasma en het potensiaal as ‘n siftingstoets in LCAT gebrek. Serum konsentrasie van gekonjugeerde bilirubien, alkaliese fosfatase, gamma glutamieltransferase, vry cholesterol, fosfolipied, vry cholesterol:totale cholesterol verhouding en gekonjugeerde bilirubien:totale bilirubien verhouding is alles goeie voorspellers van LpX. Die verhouding van vry cholesterol tot totale cholesterol (VC/TC > 0.6) was die beste voorspeller van LpX. In gevalle van obstruktiewe lewersiekte word LpX gesien in 66% van pasiente as die totale cholesterol meer as 7.5 mmol/l is.

No description available.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 106-113).

Thesis (Ph. D.)--University of Washington, 1999. / Vita. Includes bibliographical references (leaves 82-88).

Made available in DSpace on 2015-04-09T12:28:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-08Bitstream added on 2015-04-09T12:48:01Z : No. of bitstreams: 1 000814587.pdf: 233099 bytes, checksum: 86c73283cfc6d5c2954be91ce17d3f29 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Endotoxins are often found in high concentration in the peritoneal fluid from horses with acute gastrointestinal disease and blood of septicemic foals. The conditions mentioned earlier can lead to inflammatory processes and cause changes in the concentrations of acute phase proteins. The objectives of this study were to determine by means of polyacrylamide gel electrophoresis (SDS-PAGE), the concentrations of acute phase proteins (transferrin, ceruloplasmin, albumin, haptoglobin and acid glycoprotein) in serum and peritoneal fluid of horses submitted to experimental endotoxemia and evaluate the possible effects of the administration of lidocaine on the production of these proteins after intraperitoneal injection of endotoxin in these animals. To this end, 12 animals were distributed in two groups, control which was treated with 0.9% NaCl solution and the experimental which was treated with continuous infusion for 6 hours of 2% lidocaine solution. It is concluded that in this experimental model the intraperitoneal administration of endotoxin caused an increase in the concentrations of peritoneal transferrin, ceruloplasmin and haptoglobin in the control group, of albumin in both groups, and the acid glycoprotein in the group treated with infusion of lidocaine. However, the absence of changes in the concentrations of serum ceruloplasmin, serum albumin, haptoglobin and acidic glycoprotein indicate that the amount of the circulatory endotoxin was not enough to alter serum concentrations of these proteins. The continuous infusion of lidocaine interfered in peritoneal acute phase proteins values and had an anti-endotoxemic effect / FAPESP: 2009/15683-0

Resumo:As endotoxinas são frequentemente detectadas em alta concentração no líquido peritoneal de equinos com doença gastrintestinal aguda e no sangue de potros septicêmicos. As condições citadas anteriormente podem levar a processos inflamatórios e provocar alterações nas concentrações de proteínas de fase aguda. Os objetivos deste estudo foram determinar, por meio de eletroforese em gel de poliacrilamida (SDS-PAGE), as concentrações de proteínas de fase aguda (transferrina, ceruloplasmina, albumina, haptoglobina e glicoproteína ácida) no soro e no líquido peritoneal de equinos submetidos à endotoxemia experimetal, e avaliar os possíveis efeitos da administração da lidocaína sobre a produção destas proteínas depois da injeção intraperitoneal de endotoxina nestes animais. Para tanto, foram utilizados 12 animais distribuídos em dois grupos, o controle, tratado com solução de NaCl a 0,9%, e o experimental, tratado com infusão contínua de solução de lidocaína a 2% por 6 horas. Conclui-se que neste modelo experimental a administração intraperitoneal de endotoxina causou o aumento nas concentrações peritoneais de transferrina, ceruloplasmina e haptoglobina no grupo controle, da albumina em ambos os grupos, e da glicoproteína ácida no grupo tratado com infusão de lidocaína. Porém a ausência de alterações nas concentrações séricas de ceruloplasmina, albumina, haptoglobina e glicoproteína ácida indicam que a quantidade de endotoxina que chegou à corrente circulatória não foi suficiente para alterar as concentrações séricas destas proteínas. A infusão contínua de lidocaína interferiu nos valores peritoneais das proteínas mensuradas, tendo efeito anti-endotoxêmico / Abstract:Endotoxins are often found in high concentration in the peritoneal fluid from horses with acute gastrointestinal disease and blood of septicemic foals. The conditions mentioned earlier can lead to inflammatory processes and cause changes in the concentrations of acute phase proteins. The objectives of this study were to determine by means of polyacrylamide gel electrophoresis (SDS-PAGE), the concentrations of acute phase proteins (transferrin, ceruloplasmin, albumin, haptoglobin and acid glycoprotein) in serum and peritoneal fluid of horses submitted to experimental endotoxemia and evaluate the possible effects of the administration of lidocaine on the production of these proteins after intraperitoneal injection of endotoxin in these animals. To this end, 12 animals were distributed in two groups, control which was treated with 0.9% NaCl solution and the experimental which was treated with continuous infusion for 6 hours of 2% lidocaine solution. It is concluded that in this experimental model the intraperitoneal administration of endotoxin caused an increase in the concentrations of peritoneal transferrin, ceruloplasmin and haptoglobin in the control group, of albumin in both groups, and the acid glycoprotein in the group treated with infusion of lidocaine. However, the absence of changes in the concentrations of serum ceruloplasmin, serum albumin, haptoglobin and acidic glycoprotein indicate that the amount of the circulatory endotoxin was not enough to alter serum concentrations of these proteins. The continuous infusion of lidocaine interfered in peritoneal acute phase proteins values and had an anti-endotoxemic effect / Orientador: Francisco Leydson Formiga Feitosa / Banca: Marcos Pinheiro Franque / Banca: Taciana Rabelo Ramalho Ramos / Banca: Paulo Sérgio Patto dos Santos / Banca: Juliana Regina Peiró / Doutor

京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第18824号 / 理博第4082号 / 新制||理||1587(附属図書館) / 31775 / 京都大学大学院理学研究科化学専攻 / (主査)教授 杉山 弘, 教授 三木 邦夫, 教授 秋山 芳展 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM

Mass spectrometry coupled with liquid chromatography and gel electrophoresis enables separation and detection of components in a complex mixture. During the last two decades, its applications were dramatically extended and remarkable progress has been made in many fields, in particular, environmental and biological analyses. This dissertation focuses on identification and characterization of biologically active compounds and comparative analysis of protein expression changes. The first two projects (Chapters 2 and 3) focus on the application of LC/MS approach to profile the bioactivated intermediates of 4, 4'-methylenedianiline (DAPM) from rat vascular smooth muscle cells (VSMCs) and bile. In our study, several DAPM metabolites were detected and characterized in detail by liquid chromatography-electrospray tandem mass spectrometry. The structural assignments of these metabolites from VSMCs and rat bile significantly improve our understanding of DAPM biotransformations and toxicity. The third project described in Chapter 4 focuses on using electrospray tandem mass spectrometry (ES-MS/MS) and theoretical calculation (GAUSSIAN 03 program) to investigate the unusual methyl radical loss and consecutive fragment ions that dominate the low-energy collision induced dissociation (CID) mass spectra of prodiginine compounds. Structures of the fragment ions are proposed and explanations are given to rationalize the observed competition between the formation of even-electron ions and radical ions. Our study shows that the lower apparent threshold associated with methyl radical loss points to a lower kinetic barrier. In Chapter 5, hypoxia-induced changes of zebrafish skeletal muscle were studied using two-dimensional difference in-gel electrophoresis (2D-DIGE) in vivo after 48 h in hypoxia vs. normoxia. The results showed that proteins involved in mitochondrial oxidative metabolism are down-regulated, whereas glycolytic enzymes are up-regulated to compensate for the loss of ATP synthesis in aerobic metabolism. The up-regulation of two spots identified as hemoglobin variants was also observed. These protein expression changes are consistent with a hypoxic response that enhances anaerobic metabolism or O2 transport to tissues.