Uncoupling proteins 2 and 3 are highly active H+ transporters and highly nucleotide sensitive when activated by coenzyme Q (ubiquinone)

Based on the discovery of coenzyme Q (CoQ) as an obligatory cofactor for H+ transport by uncoupling protein 1 (UCP1) [Echtay, K. S., Winkler, E. & Klingenberg, M. (2000) Nature (London) 408, 609–613] we show here that UCP2 and UCP3 are also highly active H+ transporters and require CoQ and fatty acid for H+ transport, which is inhibited by low concentrations of nucleotides. CoQ is proposed to facilitate injection of H+ from fatty acid into UCP. Human UCP2 and 3 expressed in Escherichia coli inclusion bodies are solubilized, and by exchange of sarcosyl against digitonin, nucleotide binding as measured with 2′-O-[5-(dimethylamino)naphthalene-1-sulfonyl]-GTP can be restored. After reconstitution into vesicles, Cl− but no H+ are transported. The addition of CoQ initiates H+ transport in conjunction with fatty acids. This increase is fully sensitive to nucleotides. The rates are as high as with reconstituted UCP1 from mitochondria. Maximum activity is at a molar ratio of 1:300 of CoQ:phospholipid. In UCP2 as in UCP1, ATP is a stronger inhibitor than ADP, but in UCP3 ADP inhibits more strongly than ATP. Thus UCP2 and UCP3 are regulated differently by nucleotides, in line with their different physiological contexts. These results confirm the regulation of UCP2 and UCP3 by the same factors CoQ, fatty acids, and nucleotides as UCP1. They supersede reports that UCP2 and UCP3 may not be H+ transporters.

RNA replication by Q beta replicase: a working model.

Two classes of RNA ligands that bound to separate, high affinity nucleic acid binding sites on Q beta replicase were previously identified. RNA ligands to the two sites, referred to as site I and site II, were used to investigate the molecular mechanism of RNA replication employed by the four-subunit replicase. Replication inhibition by site I- and site II-specific ligands defined two subsets of replicatable RNAs. When provided with appropriate 3' ends, ligands to either site served as replication templates. UV crosslinking experiments revealed that site I is associated with the S1 subunit, site II with elongation factor Tu, and polymerization with the viral subunit of the holoenzyme. These results provide the framework for a three site model describing template recognition and product strand initiation by Q beta replicase.

The role of DT-diaphorase in the maintenance of the reduced antioxidant form of coenzyme Q in membrane systems.

The experiments reported here were designed to test the hypothesis that the two-electron quinone reductase DT-diaphorase [NAD(P)H:(quinone-acceptor) oxidoreductase, EC 1.6.99.2] functions to maintain membrane-bound coenzyme Q (CoQ) in its reduced antioxidant state, thereby providing protection from free radical damage. DT-diaphorase was isolated and purified from rat liver cytosol, and its ability to reduce several CoQ homologs incorporated into large unilamellar vesicles was demonstrated. Addition of NADH and DT-diaphorase to either large unilamellar or multilamellar vesicles containing homologs of CoQ, including CoQ9 and CoQ10, resulted in the essentially complete reduction of the CoQ. The ability of DT-diaphorase to maintain the reduced state of CoQ and protect membrane components from free radical damage as lipid peroxidation was tested by incorporating either reduced CoQ9 or CoQ10 and the lipophylic azoinitiator 2,2'-azobis(2,4-dimethylvaleronitrile) into multilamellar vesicles in the presence of NADH and DT-diaphorase. The presence of DT-diaphorase prevented the oxidation of reduced CoQ and inhibited lipid peroxidation. The interaction between DT-diaphorase and CoQ was also demonstrated in an isolated rat liver hepatocyte system. Incubation with adriamycin resulted in mitochondrial membrane damage as measured by membrane potential and the release of hydrogen peroxide. Incorporation of CoQ10 provided protection from adriamycin-induced mitochondrial membrane damage. The incorporation of dicoumarol, a potent inhibitor of DT-diaphorase, interfered with the protection provided by CoQ. The results of these experiments provide support for the hypothesis that DT-diaphorase functions as an antioxidant in both artificial membrane and natural membrane systems by acting as a two-electron CoQ reductase that forms and maintains the antioxidant form of CoQ. The suggestion is offered that DT-diaphorase was selected during evolution to perform this role and that its conversion of xenobiotics and other synthetic molecules is secondary and coincidental.

Q/R site editing in kainate receptor GluR5 and GluR6 pre-mRNAs requires distant intronic sequences.

RNA editing by adenosine deamination in brain-expressed pre-mRNAs for glutamate receptor (GluR) subunits alters gene-specified codons for functionally critical positions, such as the channel's Q/R site. We show by transcript analysis of minigenes transiently expressed in PC-12 cells that, in contrast to GluR-B pre-mRNA, where the two editing sites (Q/R and R/G) require base pairing with nearby intronic editing site complementary sequences (ECSs), editing in GluR5 and GluR6 pre-mRNAs recruits an ECS located as far as 1900 nucleotides distal to the Q/R site. The exon-intron duplex structure of the GluR5 and GluR6 pre-mRNAs appears to be a substrate of double-stranded RNA-specific adenosine deaminase. This enzyme when coexpressed in HEK 293 cells preferentially targets the adenosine of the Q/R site and of an unpaired position in the ECS which is highly edited in brain.

Operational RNA code for amino acids: species-specific aminoacylation of minihelices switched by a single nucleotide.

The genetic code is based on aminoacylation reactions where specific amino acids are attached to tRNAs bearing anticodon trinucleotides. However, the anticodon-independent specific aminoacylation of RNA minihelix substrates by bacterial and yeast tRNA synthetases suggested an operational RNA code for amino acids whereby specific RNA sequences/structures in tRNA acceptor stems correspond to specific amino acids. Because of the possible significance of the operational RNA code for the development of the genetic code, we investigated aminoacylation of synthetic RNA minihelices with a human enzyme to understand the sequences needed for that aminoacylation compared with those needed for a microbial system. We show here that the species-specific aminoacylation of glycine tRNAs is recapitulated by a species-specific aminoacylation of minihelices. Although the mammalian and Escherichia coli minihelices differ at 6 of 12 base pairs, two of the three nucleotides essential for aminoacylation by the E. coli enzyme are conserved in the mammalian minihelix. The two conserved nucleotides were shown to be also important for aminoacylation of the mammalian minihelix by the human enzyme. A simple interchange of the differing nucleotide enabled the human enzyme to now charge the bacterial substrate and not the mammalian minihelix. Conversely, this interchange made the bacterial enzyme specific for the mammalian substrate. Thus, the positional locations (if not the actual nucleotides) for the operational RNA code for glycine appear conserved from bacteria to mammals.

Coenzyme Q reductase from liver plasma membrane: purification and role in trans-plasma-membrane electron transport.

A specific requirement for coenzyme Q in the maintenance of trans-plasma-membrane redox activity is demonstrated. Extraction of coenzyme Q from membranes resulted in inhibition of NADH-ascorbate free radical reductase (trans electron transport), and addition of coenzyme Q10 restored the activity. NADH-cytochrome c oxidoreductase (cis electron transport) did not respond to the coenzyme Q status. Quinone analogs inhibited trans-plasma-membrane redox activity, and the inhibition was reversed by coenzyme Q. A 34-kDa coenzyme Q reductase (p34) has been purified from pig-liver plasma membranes. The isolated enzyme was sensitive to quinone-site inhibitors. p34 catalyzed the NADH-dependent reduction of coenzyme Q10 after reconstitution in phospholipid liposomes. When plasma membranes were supplemented with extra p34, NADH-ascorbate free radical reductase was activated but NADH-cytochrome c oxidoreductase was not. These results support the involvement of p34 as a source of electrons for the trans-plasma-membrane redox system oxidizing NADH and support coenzyme Q as an intermediate electron carrier between NADH and the external acceptor ascorbate free radical.

Calidad y rentabilidad: análisis del certificado Q en las cadenas hoteleras

La implantación de sistemas de calidad aporta a las empresas beneficios internos y externos. Este artículo analiza la relación entre la certificación de calidad del Instituto para la Calidad Turística Española y los resultados y el tamaño de las cadenas hoteleras con presencia en España. Los análisis muestran que la certificación tiene efectos positivos en los resultados y que el tamaño de la cadena no es un factor importante para certificarse.

Nile River and Birkat Qārūn Regions, Egypt, 1854 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Carte hydrographique de la Moyenne Égypte, où sont indiqués les travaux exécutés ou à exécuter, d'après les ordres de Son Altesse Mehémet-Ali, Vice-Roi d'Egypte. Avec le projet de communication directe des deux mers au travers de l'Isthme, par M. Linant de Bellefonds ; gravée par Schwaerzlé et Erhard. It was published by Dépôt de la Guerre in 1854. Scale 1:250,000. Covers a portion of the Nile River region near Birkat Qārūn, Banī Suwayf, Al Fayyūm, and Al Minyā. Map in French.The image inside the map neatline is georeferenced to the surface of the earth and fit to the Egypt Red Belt projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as drainage, canals, roads, cities and other human settlements, administrative boundaries, ruins and historical sites, and more. Relief shown by hachures. Includes also text and notes.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Hādhihi Taqrīrāt rāʼiqah wa-taḥqīqāt fāʼiqah

nammaqahā Muḥammad al-Azharī najl al-Ḥasan al-ʻAdawī al-Ḥamzāwī. Washshaḥa bihā ḥawāshī Muḥammad al-Ḥifnī ʻalá Sharḥ al-Risālah al-waḍʻīyah al-ʻAḍudīyah / li-Abī al-Qāsim al-Samarqandī wa-bi-hāmishihā al-ḥawāshī al-madhkūrah maʻa al-sharḥ al-madhkūr.

Talfīq al-akhbār wa-talqīḥ al-āthār fī waqāʻiʻ Qāzān wa-Bulghār wa-mulūk al-Tatār

athar M. M. al-Ramzī.

Kitāb Sullam al-ʻulūm wa-ḥāshiyatihi al-mashhūrah bi-al-Qāḍī maʻa minhīyātih

[Muḥibb Allāh ibn ʻAbd al-Shakūr al-Bihārī].

Qānūn dīwān al-rasāʼil: manqūl ʻan nuskhah khaṭṭīyah bi-Maktabat Kambritsh

li-Tāj al-Riyāsah Abī al-Qāsim ʻAlī ibn Munjib ibn Sulaymān al-shahīr bi-Ibn al-Ṣayrafī ; ʻuniya bi-nashrihi wa-al-taʻlīq ʻalayhi ʻAlī Bahjat.

al- Khulāṣah al-naqīyah fī umarāʼ Ifrīqīyah: wa-maʻahu ʻUqūd al-farāʼid fī tadhyīl al-Khulāṣah wa-fawāʼid al-rāʼid lil-muʼallif ayḍan

taʼlīf Abī ʻAbd Allāh Muḥammad al-Bājī al-Masʻūdī.

Nāmūs al-īqān

Kharbūtī Yūsuf Afandī.

Kitāb Qāmūs al-ʻāshiqīn fī akhbār al-Sayyid Ḥusayn Burhān al-Dīn

taʼlīf ʻAbd al-Munʻim al-ʻĀnī thumma al-Rāwī ; [muṣaḥḥiḥuhu Aḥmad ibn Saʻd al-Dīn ibn Muṣṭafá al-Qabbānī].