Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

An analysis of safety data from five phase III clinical trials on the use of botulinum neurotoxin type A-ABO for the treatment of glabellar lines

A new formulation of a botulinum neurotoxin type A (BoNTA-ABO; Dysport [abobotulinumtoxinA]; Medicis Aesthetics, Scottsdale, AZ) was recently approved by the US Food and Drug Administration for the treatment of moderate to severe glabellar lines. This article summarizes the safety data from five phase III clinical trials investigating the use of BoNTA-ABO in the treatment of glabellar lines. Of the five phase III studies conducted, three were multicenter, randomized, placebo-controlled, double-blind studies and two were multicenter, open-label, repeat-dose studies (one of which was an extension trial). Two fixed-dose, placebo-controlled studies randomized a total of 416 patients to receive one 50-unit dose of BoNTA-ABO. The single variable-dose study randomized 544 patients to receive 50 to 80 units of BoNTA-ABO, as determined by gender and patient muscle mass. A substudy of this variable-dose study on QT/QTc prolongation included 50 patients randomized to BoNTA-ABO. One open-label repeat-dose study administered 50 units of BoNTA-ABO to 1200 patients. The single extension study (N = 1415) included both fixed (5423 treatments) and variable dosing (1337 treatments) following a protocol amendment. The extension study included patients from the four previously mentioned studies. Safety endpoints were adverse events (AE), laboratory data, and changes in vital signs. Of 2485 healthy adult patients with moderate to severe glabellar lines enrolled in the trials, 2160 received at least one cycle of BoNTA-ABO. Treatment of glabellar lines with 50 units of BoNTA-ABO was well tolerated, with similar rates of treatment-emergent adverse events (TEAE) observed in the active treatment and placebo groups in terms of type, frequency, severity, and relatedness-with the exception of injection site reactions and ptosis. In the variable-dose, single-treatment study, BoNTA-ABO was well tolerated, with an incidence of active TEAE (31%) only slightly greater than that observed for placebo (28%). In the repeat-dose studies, there was no evidence of cumulative safety issues, the incidence of TEAE decreased over time, and patients did not drop out because of TEAE. The most frequently reported AE were nasopharyngitis, sinusitis, upper respiratory tract infection, headache, and injection site reactions. The majority of TEAE were considered unlikely to be related or were not related to BoNTA-ABO treatment. In all studies, the TEAE that were considered possibly related to treatment were primarily headaches (with rates comparable to those observed for placebo), injection site reactions, and eye disorders (such as blepharospasm and eyelid ptosis). There were no clinically significant changes in hematologic or biochemical parameters or in vital signs. The cardiovascular substudy revealed that BoNTA-ABO had no effect on QT/QTc prolongation. Treatment of glabellar lines with BoNTA-ABO is well tolerated. Overall, the safety profile of BoNTA-ABO is comparable to that of placebo in terms of type, frequency, severity, and relatedness of AE

Formation of a Cyclopropyl Epoxide via a Leukotriene A Synthase-related Pathway in an Anaerobic Reaction of Soybean Lipoxygenase-1 with 15S-Hydroperoxyeicosatetraenoic Acid: EVIDENCE THAT OXYGEN ACCESS IS A DETERMINANT OF SECONDARY REACTIONS WITH FATTY ACID HYDROPEROXIDES*

The further conversion of an arachidonic acid hydroperoxide to a leukotriene A (LTA) type epoxide by specific lipoxygenase (LOX) enzymes constitutes a key step in inflammatory mediator biosynthesis. Whereas mammalian 5-LOX transforms its primary product (5S-hydroperoxyeicosatetraenoic acid; 5S-HPETE) almost exclusively to LTA4, the model enzyme, soybean LOX-1, normally produces no detectable leukotrienes and instead further oxygenates its primary product 15S-HPETE to 5,15- and 8,15-dihydroperoxides. Mammalian 15-LOX-1 displays both types of activity. We reasoned that availability of molecular oxygen within the LOX active site favors oxygenation, whereas lack of O2 promotes LTA epoxide synthesis. To test this, we reacted 15S-HPETE with soybean LOX-1 under anaerobic conditions and identified the products by high pressure liquid chromatography, UV, mass spectrometry, and NMR. Among the products, we identified a pair of 8,15-dihydroxy diastereomers with all-trans-conjugated trienes that incorporated 18O from H218O at C-8, indicative of the formation of 14,15-LTA4. A pair of 5,15-dihydroxy diastereomers containing two trans,trans-conjugated dienes (6E,8E,11E,13E) and that incorporated 18O from H218O at C-5 was deduced to arise from hydrolysis of a novel epoxide containing a cyclopropyl ring, 14,15-epoxy-[9,10,11-cyclopropyl]-eicosa-5Z,7E,13E-trienoic acid. Also identified was the δ-lactone of the 5,15-diol, a derivative that exhibited no 18O incorporation due to its formation by intramolecular reaction of the carboxyl anion with the proposed epoxide intermediate. Our results support a model in which access to molecular oxygen within the active site directs the outcome from competing pathways in the secondary reactions of lipoxygenases