Transcription profiling suggests that mitochondrial topoisomerase IB acts as a topological barrier and regulator of mitochondrial DNA transcription

Mitochondrial DNA (mtDNA) is essential for cell viability because it encodes subunits of the respiratory chain complexes. Mitochondrial topoisomerase IB (TOP1MT) facilitates mtDNA replication by removing DNA topological tensions produced during mtDNA transcription, but it appears to be dispensable. To test whether cells lacking TOP1MT have aberrant mtDNA transcription, we performed mitochondrial transcriptome profiling. To that end, we designed and implemented a customized tiling array, which enabled genome-wide, strand-specific, and simultaneous detection of all mitochondrial transcripts. Our technique revealed that KO mouse cells process the mitochondrial transcripts normally but that protein-coding mitochondrial transcripts are elevated. Moreover, we found discrete long noncoding RNAs produced by H-strand transcription and encompassing the noncoding regulatory region of mtDNA in human and murine cells and tissues. Of note, these noncoding RNAs were strongly up-regulated in the absence of TOP1MT. In contrast, 7S DNA, produced by mtDNA replication, was reduced in the KO cells. We propose that the long noncoding RNA species in the D-loop region are generated by the extension of H-strand transcripts beyond their canonical stop site and that TOP1MT acts as a topological barrier and regulator for mtDNA transcription and D-loop formation

Alteration of Extracellular Nucleotide Metabolism in Pseudoxanthoma Elasticum

Pseudoxanthoma elasticum (PXE) is a rare genetic condition primarily caused by hepatic ABCC6 transporter dysfunction. Most clinical manifestations of PXE are due to premature calcification of elastic fibers. However, the vascular impact of PXE is pleiotropic and remains ill defined. ABCC6 expression has recently been associated with cellular nucleotide export. We studied the impact of ABCC6 deficiency on blood levels of adenosine triphosphate and related metabolites and on soluble nucleotidase activities in PXE patients and Abcc6 mice. In addition, we investigated the expression of genes encoding ectocellular purinergic signaling proteins in mouse liver and aorta. Plasma adenosine triphosphate and pyrophosphate levels were significantly reduced in PXE patients and in Abcc6 mice, whereas adenosine concentration was not modified. Moreover, 5\u27-nucleotidase/CD73 activity was increased in the serum of PXE patients and Abcc6 mice. Consistent with alterations of purinergic signaling, the expression of genes involved in purine and phosphate transport/metabolism was dramatically modified in Abcc6 mouse aorta, with much less impact on the liver. ABCC6 deficiency causes impaired vascular homeostasis and tissue perfusion. Our findings suggest that these alterations are linked to changes in extracellular nucleotide metabolism that are remote from the liver. This opens new perspectives for the understanding of PXE pathophysiology

Metastase axillaire d'un carcinome papillaire de la thyroïde: À propos d'un cas

Les métastases axillaires dans le cadre d'un cancer de la thyroïde sont extrêmement rares. Plusieurs hypothèses expliquent ce drainage. Nousrapportons le cas d'un patient de 78 ans ayant pour antécédents  chirurgicaux une lobo-isthmectomie en 1987, puis une totalisation chirurgicale en 1997 non documentés, admis en septembre 2008 pour exploration d'une masse latéro-cervicale gauche. Le reste de l'examen clinique révèle la présence de multiples adénopathies axillaires bilatérales. La biopsie exérèse d'une adénopathie axillaire droite (côté controlatéral) retrouve une métastase ganglionnaire d'un carcinome papillaire de la thyroïde avec effraction capsulaire. La tomodensitométrie cervico-thoracique note la présence d'un processus tumoral latéro-cervical gauche, un lobe thyroïdien droit siège de multiples nodules hypodenses, des adénopathies cervicales et axillaires et des lésions suspectes au niveau du parenchyme pulmonaire. Une thyroïdectomie totale avec curage  ganglionnaire cervical est décidée, complétée par une ablation des ganglions axillaires macroscopiquement atteints. Des cures d'iode  radioactif (IRA-thérapie) sont indiquées. Bien qu'exceptionnelle, la  présence de métastases axillaires d'un carcinome thyroïdien est de  pronostic péjoratif. On se demande alors si ces patients ne nécessitent pas une prise en charge particulière. Une réflexion à une stratégie  thérapeutique est donc nécessaire

Lack of mitochondrial topoisomerase I (TOP1mt) impairs liver regeneration

The liver has an exceptional replicative capacity following partial hepatectomy or chemical injuries. Cellular proliferation requires increased production of energy and essential metabolites, which critically depend on the mitochondria. To determine whether Top1mt, the vertebrate mitochondrial topoisomerase, is involved in this process, we studied liver regeneration after carbon tetrachloride (CCl4) administration. TOP1mt knockout (KO) mice showed a marked reduction in regeneration and hepatocyte proliferation. The hepatic mitochondrial DNA (mtDNA) failed to increase during recovery from CCl4 exposure. Reduced glutathione was also depleted, indicating increased reactive oxygen species (ROS). Steady-state levels of ATP, O2 consumption, mtDNA, and mitochondrial mass were also reduced in primary hepatocytes from CCl4-treated KO mice. To further test whether Top1mt acted by enabling mtDNA regeneration, we tested TOP1mt KO fibroblasts and human colon carcinoma HCT116 cells and measured mtDNA after 3-d treatment with ethidium bromide. Both types of TOP1mt knockout cells showed defective mtDNA regeneration following mtDNA depletion. Our study demonstrates that Top1mt is required for normal mtDNA homeostasis and for linking mtDNA expansion with hepatocyte proliferation

Warburg-like effect is a hallmark of complex I assembly defects

Due to its pivotal role in NADH oxidation and ATP synthesis, mitochondrial complex I (CI) emerged as a crucial regulator of cellular metabolism. A functional CI relies on the sequential assembly of nuclear- and mtDNA-encoded subunits; however, whether CI assembly status is involved in the metabolic adaptations in CI deficiency still remains largely unknown. Here, we investigated the relationship between CI functions, its structure and the cellular metabolism in 29 patient fibroblasts representative of most CI mitochondrial diseases. Our results show that, contrary to the generally accepted view, a complex I deficiency does not necessarily lead to a glycolytic switch, i.e. the so-called Warburg effect, but that this particular metabolic adaptation is a feature of CI assembly defect. By contrast, a CI functional defect without disassembly induces a higher catabolism to sustain the oxidative metabolism. Mechanistically, we demonstrate that reactive oxygen species overproduction by CI assembly intermediates and subsequent AMPK-dependent Pyruvate Dehydrogenase inactivation are key players of this metabolic reprogramming. Thus, this study provides a two-way-model of metabolic responses to CI deficiencies that are central not only in defining therapeutic strategies for mitochondrial diseases, but also in all pathophysiological conditions involving a CI deficiency

CLUH couples mitochondrial distribution to the energetic and metabolic status

Mitochondrial dynamics and distribution are critical for supplying ATP in response to energy demand. CLUH is a protein involved in mitochondrial distribution whose dysfunction leads to mitochondrial clustering, the metabolic consequences of which remain unknown. To gain insight into the role of CLUH on mitochondrial energy production and cellular metabolism, we have generated CLUH-knockout cells using CRISPR/Cas9. Mitochondrial clustering was associated with a smaller cell size and with decreased abundance of respiratory complexes, resulting in oxidative phosphorylation (OXPHOS) defects. This energetic impairment was found to be due to the alteration of mitochondrial translation and to a metabolic shift towards glucose dependency. Metabolomic profiling by mass spectroscopy revealed an increase in the concentration of some amino acids, indicating a dysfunctional Krebs cycle, and increased palmitoylcarnitine concentration, indicating an alteration of fatty acid oxidation, and a dramatic decrease in the concentrations of phosphatidylcholine and sphingomyeline, consistent with the decreased cell size. Taken together, our study establishes a clear function for CLUH in coupling mitochondrial distribution to the control of cell energetic and metabolic status

Melissopalynological, physicochemical and antioxidant properties of honey from West Coast of Malaysia

Stingless bees are native to tropical region and produce honey which are high in moisture content. Compared to honey from honeybees, there are limited studies on honey derived from stingless bees. Hence, the aim of this study was to evaluate the chemical composition and antioxidant activities of stingless bee honey. Fifteen types of honey were collected from six states in West Coast of Malaysia and pollen analyses were carried out. Four types of unifloral honey samples produced by stingless bees were selected to determine their physicochemical and antioxidant activities including total phenolic, total flavonoid and ascorbic acid contents. Melissopalynological study of 15 honey samples collected from different states showed presence of both unifloral and multifloral origins. Honey samples collected from Apis mellifera (honeybee) combs had lower number of total pollen compared to samples collected from Heterotrigona itama and Geniotrigona thoracica (stingless bees). Jambul Merak honey contains the highest phenolic and flavonoid contents with greatest color intensity and has the highest antioxidant potential. This study highlights the chemical composition and biological activity of honey from stingless bees which may increase its commercial value or to be utilised as potential functional food ingredient