Mitochondrial ROS Produced via Reverse Electron Transport Extend Animal Lifespan.

Increased production of reactive oxygen species (ROS) has long been considered a cause of aging. However, recent studies have implicated ROS as essential secondary messengers. Here we show that the site of ROS production significantly contributes to their apparent dual nature. We report that ROS increase with age as mitochondrial function deteriorates. However, we also demonstrate that increasing ROS production specifically through respiratory complex I reverse electron transport extends Drosophila lifespan. Reverse electron transport rescued pathogenesis induced by severe oxidative stress, highlighting the importance of the site of ROS production in signaling. Furthermore, preventing ubiquinone reduction, through knockdown of PINK1, shortens lifespan and accelerates aging; phenotypes that are rescued by increasing reverse electron transport. These results illustrate that the source of a ROS signal is vital in determining its effects on cellular physiology and establish that manipulation of ubiquinone redox state is a valid strategy to delay aging.

Human Mitochondrial DNA-Protein Complexes Attach to a Cholesterol-Rich Membrane Structure.

The helicase Twinkle is indispensable for mtDNA replication in nucleoids. Previously, we showed that Twinkle is tightly membrane-associated even in the absence of mtDNA, which suggests that Twinkle is part of a membrane-attached replication platform. Here we show that this platform is a cholesterol-rich membrane structure. We fractionated mitochondrial membrane preparations on flotation gradients and show that membrane-associated nucleoids accumulate at the top of the gradient. This fraction was shown to be highly enriched in cholesterol, a lipid that is otherwise low abundant in mitochondria. In contrast, more common mitochondrial lipids, and abundant inner-membrane associated proteins concentrated in the bottom-half of these gradients. Gene silencing of ATAD3, a protein with proposed functions related to nucleoid and mitochondrial cholesterol homeostasis, modified the distribution of cholesterol and nucleoids in the gradient in an identical fashion. Both cholesterol and ATAD3 were previously shown to be enriched in ER-mitochondrial junctions, and we detect nucleoid components in biochemical isolates of these structures. Our data suggest an uncommon membrane composition that accommodates platforms for replicating mtDNA, and reconcile apparently disparate functions of ATAD3. We suggest that mtDNA replication platforms are organized in connection with ER-mitochondrial junctions, facilitated by a specialized membrane architecture involving mitochondrial cholesterol.

Basic diffraction phenomena in time domain.

Using a recently developed technique (SEA TADPOLE) for easily measuring the complete spatiotemporal electric field of light pulses with micrometer spatial and femtosecond temporal resolution, we directly demonstrate the formation of theo-called boundary diffraction wave and Arago's spot after an aperture, as well as the superluminal propagation of the spot. Our spatiotemporally resolved measurements beautifully confirm the time-domain treatment of diffraction. Also they prove very useful for modern physical optics, especially in micro- and meso-optics, and also significantly aid in the understanding of diffraction phenomena in general.

Direct spatiotemporal measurements of accelerating ultrashort Bessel-type light bullets.

We measure the spatiotemporal field of ultrashort pulses with complex spatiotemporal profiles using the linear-optical, interferometric pulse-measurement technique SEA TADPOLE. Accelerating and decelerating ultrashort, localized, nonspreading Bessel-X wavepackets were generated from a approximately 27 fs duration Ti:Sapphire oscillator pulse using a combination of an axicon and a convex or concave lens. The wavefields are measured with approximately 5 microm spatial and approximately 15 fs temporal resolutions. Our experimental results are in good agreement with theoretical calculations and numerical simulations.

Measuring the spatiotemporal field of ultrashort Bessel-X pulses.

We present direct measurements of the spatiotemporal electric field of an ultrashort Bessel-X pulse generated using a conical lens (axicon). These measurements were made using the linear-optical interferometric technique SEA TADPOLE, which has micrometer spatial resolution and femtosecond temporal resolution. From our measurements, both the superluminal velocity of the Bessel pulse and the propagation invariance of the central spot are apparent. We verified our measurements with simulations.

Determination of thyreostats in urine and thyroid gland by ultra high performance liquid chromatography tandem mass spectrometry.

Thyreostatic compounds could be illegally administered to animals in order to obtain a weight gain due to a higher retention of water in the edible tissue and the gastro-intestinal tract. In the European Union their use for animal production is banned since 1981. Recently a highly sensitive method exploiting the determination of thyreostats with 3-iodobenzylbromide prior to purification to determine thyreostats in urine and other matrices was reported. For the first time, the UPLC instrumentation was used to separate the 3-iodobenzyl derivatives of various thyreostats. The deuterated internal standards tapazole-d(3) and propylthiouracil-d(5) were for the first time used for the quantification of tapazole, thiouracil, methylthiouracil, propylthiouracil, phenylthiouracil and mercaptobenzimidazole. The confirmative quantitative liquid chromatographic tandem mass spectrometric (LC-MS/MS) method was validated according to Commission Decision 2002/657/EC. The decision limit (CCalpha) and the detection capability (CCbeta) were found to be for all compounds below the recommended value of 10 microg kg(-1).

Determination of gestagens in kidney fat by liquid chromatography tandem mass spectrometry.

The use of gestagens in animal fattening is prohibited within the European Union. Recently, the use of spectrometric methods for the detection and confirmation of banned substances was made obligatory. Therefore, conventional high-performance liquid chromatographic (HPLC) methods have been superseded. It has been possible to couple a previously described HPLC method for the determination of acetyl-gestagens in kidney fat to tandem mass spectrometry (LC-MS/MS). The decision limits CCalpha and the detection capability CCbeta are found to be below the minimum required performance limit (MRPL) established for medroxyprogesterone acetate (MPA) at 1 microg kg(-1). The calculated values for CCalpha are as follows: megestrol acetate (MGA)--0.15 microg kg(-1), melengesterol acetate (MLA)--0.15 microg kg(-1), chlormadinone acetate (CMA)--0.37 microg kg(-1) and for medroxyprogesterone acetate (MPA)--0.24 microg kg(-1). The CCbeta values for these compounds have been determined as 0.19, 0.19, 0.47 and 0.32 microg kg(-1), respectively.