cytochrome-c-t and Syndrome

The method of treatment of an acute pulmonary damage syndrome in the injured persons, suffering traumatic disease, complicated by enteral insufficiency syndrome, using preparation cytochrom C, was depicted. The results obtained witness the high clinical efficacy of the treatment scheme proposed.

Topics: Blood Gas Analysis; Carbon Dioxide; Combined Modality Therapy; Cytochromes c; Gastrointestinal Motility; Humans; Hypoxia; Oxygen; Oxygen Inhalation Therapy; Respiratory Distress Syndrome; Syndrome; Treatment Outcome; Wounds and Injuries

P450 oxidoreductase (POR) deficiency causes disordered steroidogenesis; severe mutations cause genital ambiguity in both sexes plus the Antley-Bixler skeletal malformation syndrome, whereas mild mutations can cause adult infertility.. We describe four patients with POR deficiency and identify and characterize the activities of their mutations. A 46,XY male with micropenis and two 46,XX female infants with genital ambiguity presented with skeletal malformations, and a 46,XX adolescent presented with primary amenorrhea, elevated 17alpha-hydroxyprogesterone, and low sex steroids.. The coding regions of the POR gene were sequenced, and the identified mutations were recreated in human POR cDNA expression vectors lacking 27 N-terminal residues. POR and human P450c17 were expressed in bacteria. POR activity was measured by four assays: reduction of cytochrome c, oxidation of reduced nicotinamide adenine dinucleotide phosphate, and support of the 17alpha-hydroxylase and 17,20 lyase activities of P450c17.. All four patients were compound heterozygotes for POR mutations, including five novel mutations: L577R, N185K, delE217, and frameshift mutations 1363delC and 697-698insGAAC. N185K and delE217 lacked measurable activity in the assays based on P450c17 but retained partial activity in the assays based on cytochrome c. As assessed by V(max)/Km, L577R supported 46% of 17alpha-hydroxylase activity but only 27% of 17,20 lyase activity. Computational modeling of these novel mutants revealed the structural basis for their reduced or absent activities.. These patients illustrate the broad clinical spectrum of POR deficiency, including amenorrhea and infertility as the sole manifestation. POR assays based on P450c17 correlate well with hormonal and clinical phenotypes.

Topics: Adolescent; Adult; Bone and Bones; Catalysis; Cytochromes c; Disorders of Sex Development; DNA; Escherichia coli; Female; Genetic Vectors; Genitalia; Hormones; Humans; Infant, Newborn; Infertility; Male; Mutation; NADP; NADPH-Ferrihemoprotein Reductase; Pregnancy; Steroid 17-alpha-Hydroxylase; Syndrome

Autoantibodies against recoverin, a Ca2+-binding protein found in patients with cancer-associated retinopathy (CAR syndrome), penetrate retinal cells and induce their apoptosis via a mitochondrial pathway. The goal of this study was to investigate whether the entry of anti-recoverin antibody into E1A.NR3 retinal cells causes a change in intracellular Ca2+. Intracellular Ca2+ was measured using the Ca2+-sensitive fluorescent dye Fura-2 AM in living E1A.NR3 retinal cells treated with anti-recoverin antibody Rec-1, patients' autoantibodies, and control rat and human IgG. The exposure of retinal cells to Rec-1 antibody and to the CAR patients' autoantibodies in vitro caused a significant increase in intracellular Ca2+, while non-specific antibodies did not induce such an effect. Co-treatment of the E1A.NR3 cells with Rec-1 in the presence of nifedipine, a L-type Ca2+ channel blocker, significantly suppressed the increase of Ca2+. Treatment with nifedipine also blocked changes in the anti-apoptotic protein bcl-xL and in expressions of the pro-apoptotic protein bax. Nifedipine-treated cells also showed a decrease in cytosolic cytochrome c release and a decrease in caspase 3 activation, compared to cells treated only with Rec-1 antibody. The increase in the antibody-induced Ca2+ is at least in part dependent on extracellular Ca2+. Nifedipine was found to inhibit the entry of Ca2+ into the cells and to protect them from Rec-1-induced apoptosis. Increased levels of intracellular Ca2+ may lead to retinal dysfunction and degeneration in the CAR syndrome. Our results provide a molecular basis for the use of Ca2+ blockers in the treatment of the CAR syndrome.

Topics: Animals; Antibodies, Monoclonal; Apoptosis; Autoantibodies; bcl-2-Associated X Protein; bcl-X Protein; Calcium; Calcium Channel Blockers; Calcium-Binding Proteins; Cell Line; Cytochromes c; Humans; Neoplasms; Nifedipine; Rats; Recoverin; Retina; Retinal Diseases; Syndrome

Barth syndrome (BTHS) is an X-linked recessive disorder that is biochemically characterized by low cellular levels of the mitochondrial phospholipid cardiolipin (CL). Previously, we discovered that the yeast disruptant of the TAZ ortholog in Saccharomyces cerevisiae not only displays CL deficiency but also accumulates monolysocardiolipins (MLCLs), which are intermediates in CL remodeling. Therefore, we set out to investigate whether MLCL accumulation also occurs in BTHS. Indeed, we observed MLCL accumulation in heart, muscle, lymphocytes, and cultured lymphoblasts of BTHS patients; however, only very low levels of these lysophospholipids were found in platelets and fibroblasts of these patients. Although the fatty acid composition of the MLCLs was different depending on the tissue source, it did parallel the fatty acid composition of the (remaining) CLs. The possible implications of these findings for the two reported CL remodeling mechanisms, transacylation and deacylation/reacylation, are discussed. Because MLCLs have been proposed to be involved in the initiation of apoptosome-mediated cell death by the sequestration of the proapoptotic protein (t)BH3-interacting domain death agonist (Bid) to the mitochondrial membrane, we used control and BTHS lymphoblasts to investigate whether the accumulation of MLCLs results in higher levels of apoptosis. We found no differences in susceptibility to death receptor-mediated apoptosis or in cellular distribution of Bid, cytochrome c, and other parameters, implying that MLCL accumulation does not lead to enhanced apoptosis in cultured BTHS lymphoblasts.

Topics: Acylation; Apoptosis; BH3 Interacting Domain Death Agonist Protein; Cardiolipins; Cardiomyopathies; Carrier Proteins; Cell Death; Chromatography, High Pressure Liquid; Chromosomes, Human, X; Cytochromes c; fas Receptor; Genes, Recessive; Genetic Linkage; Growth Disorders; Humans; Immunoblotting; Jurkat Cells; Lipid Metabolism; Lymphocytes; Lysophospholipids; Mass Spectrometry; Mitochondria; Muscles; Muscular Diseases; Myocardium; Phospholipids; Poly(ADP-ribose) Polymerases; Protein Structure, Tertiary; Subcellular Fractions; Syndrome

Kostmann syndrome, or severe congenital neutropenia (SCN), is an autosomal recessive disorder of neutrophil production. To investigate the potential role of apoptosis in SCN, bone marrow aspirates and biopsies were obtained from 4 patients belonging to the kindred originally described by Kostmann and 1 patient with SCN of unknown inheritance. An elevated degree of apoptosis was observed in the bone marrow of these patients, and a selective decrease in B-cell lymphoma-2 (Bcl-2) expression was seen in myeloid progenitor cells. Furthermore, in vitro apoptosis of bone marrow-derived Kostmann progenitor cells was increased, and mitochondrial release of cytochrome c was detected in CD34(+) and CD33(+) progenitors from patients, but not in controls. Administration of granulocyte colony-stimulating factor (G-CSF) restored Bcl-2 expression and improved survival of myeloid progenitor cells. In addition, cytochrome c release was partially reversed upon incubation of progenitor cells with G-CSF. In sum, these studies establish a role for mitochondria-dependent apoptosis in the pathogenesis of Kostmann syndrome and yield a tentative explanation for the beneficial effect of growth factor administration in these patients.

Topics: Apoptosis; Bone Marrow; Case-Control Studies; Child, Preschool; Cytochromes c; Family Health; Female; Granulocyte Colony-Stimulating Factor; Humans; Infant; Infant, Newborn; Male; Mitochondria; Myeloid Progenitor Cells; Neutropenia; Proto-Oncogene Proteins c-bcl-2; Syndrome

Topics: Animals; Cytochromes c; Disease; Ectromelia; Extremities; Leukemia; Mice; Rabbits; Syndrome; Vaccination; Virus Diseases