A novel approach to identify Duchenne muscular dystrophy patients for aminoglycoside antibiotics therapy.

Aminoglycoside antibiotics have been found to suppress nonsense mutations located in the defective dystophin gene in mdx mice, suggesting a possible treatment for Duchenne muscular dystrophy (DMD). However, it is very difficult to find patients that are applicable for this therapy, because: (1) only 5-13% of DMD patients have nonsense mutations in the dystrophin gene, (2) it is challenging to find nonsense mutations in the gene because dystrophin cDNA is very long (14 kb), and (3) the efficiency of aminoglycoside-induced read-through is dependent on the kind of nonsense mutation. In order to develop a system for identifying candidates that qualify for aminoglycoside therapy, fibroblasts from nine DMD patients with nonsense mutation of dystrophin gene were isolated, induced to differentiate to myogenic lineage by AdMyoD, and exposed with gentamicin. The dystrophin expression in gentamicin-exposed myotubes was monitored by in vitro dystrophin staining and western blotting analysis. The results showed that gentamicin was able to induce dystrophin expression in the differentiated myotubes by the read-through of the nonsense mutation TGA in the gene; a read-through of the nonsense mutations TAA and TAG did not occur and consequently did not lead to dystrophin expression. Therefore, it is speculated that the aminoglycoside treatment is far more effective for DMD patients that have nonsense mutation TGA than for patients that have nonsense mutation TAA and TAG. In this study, we introduce an easy system to identify patients for this therapy and report for the first time, that dystrophin expression was detected in myotubes of DMD patients using gentamicin.

Levels and distribution of polybrominated diphenyl ethers and organochlorine compounds in sea turtles from Japan.

Three species of sea turtles (green, hawksbill and loggerhead turtles) stranded along the coasts or caught (by-catch) around Ishigaki Island and Kochi, Japan were collected between 1998 and 2006 and analyzed for six organohalogen compounds viz., PBDEs, PCBs, DDTs, CHLs, HCHs and HCB. The present study is the first and foremost to report the occurrence of organohalogen compounds in the sea turtles from Japan. Among the compounds analyzed, concentrations of PCBs, DDTs and CHLs were the highest in all the turtle samples. PBDEs were ubiquitously present in all the turtle species. Comparing with the other two species, concentrations of organohalogens in green turtle were relatively low and decreasing trend in the concentrations were noted with increasing carapace length. Concentrations of OCs in sea turtles from the coasts of Ishigaki Island and Kochi were relatively low as compared to those from other locations in the world.

Contamination status and spatial distribution of organochlorine compounds in fishes from Nansei Islands, Japan.

Two species of fishes (n=52; tilapia and mullet) from industrialized and urbanized areas of Okinawa Island (Manko-Noha river, Hija river and Shikaza river) and from a remote area of Ishigaki Island (Anparu mudflat), Japan were collected between August 2005 and July 2006, and analyzed for five organochlorine compounds (OCs), viz., DDTs, PCBs, CHLs, HCHs and HCB. Concentrations and the contamination patterns of OCs in fishes varied between locations. Considerable residue levels of OCs, especially CHLs and DDTs were found in both fishes from the main Okinawa Island. These levels were relatively higher than the reported values for seafood from Japanese coasts, indicating that even now pollution sources of these contaminants still exist in this region. On the other hand, lower concentrations of OCs were detected in fishes from Ishigaki Island waters than those of other Japanese coastal waters, suggesting that this region is less contaminated by OC contaminants.

Absence of Cajal-Retzius cells and subplate neurons associated with defects of tangential cell migration from ganglionic eminence in Emx1/2 double mutant cerebral cortex.

Emx1 and Emx2, mouse orthologs of the Drosophila head gap gene, ems, are expressed during corticogenesis. Emx2 null mutants exhibit mild defects in cortical lamination. Segregation of differentiating neurons from proliferative cells is normal for the most part, however, reelin-positive Cajal-Retzius cells are lost by the late embryonic period. Additionally, late-born cortical plate neurons display abnormal position. These types of lamination defects are subtle in the Emx1 mutant cortex. In the present study we show that Emx1 and Emx2 double mutant neocortex is much more severely affected. Thickness of the cerebral wall was diminished with the decrease in cell number. Bromodeoxyuridine uptake in the germinal zone was nearly normal; moreover, no apparent increase in cell death or tetraploid cell number was observed. However, tangential migration of cells from the ganglionic eminence into the neocortex was greatly inhibited. The wild-type ganglionic eminence cells transplanted into Emx1/2-double mutant telencephalon did not move to the cortex. MAP2-positive neuronal bodies and RC2-positive radial glial cells emerged normally, but the laminar structure subsequently formed was completely abnormal. Furthermore, both corticofugal and corticopetal fibers were predominantly absent in the cortex. Most importantly, neither Cajal-Retzius cells nor subplate neurons were found throughout E11.5-E18.5. Thus, this investigation suggests that laminar organization in the cortex or the production of Cajal-Retzius cells and subplate neurons is interrelated to the tangential movement of cells from the ganglionic eminence under the control of Emx1 and Emx2.