Dissection of C. elegans behavioral genetics in 3-D environments

The nematode Caenorhabditis elegans is a widely used model for genetic dissection of animal behaviors. Despite extensive technical advances in imaging methods, it remains challenging to visualize and quantify C. elegans behaviors in three-dimensional (3-D) natural environments. Here we developed an innovative 3-D imaging method that enables quantification of C. elegans behavior in 3-D environments. Furthermore, for the first time, we characterized 3-D-specific behavioral phenotypes of mutant worms that have defects in head movement or mechanosensation. This approach allowed us to reveal previously unknown functions of genes in behavioral regulation. We expect that our 3-D imaging method will facilitate new investigations into genetic basis of animal behaviors in natural 3-D environments

An improved baculovirus insecticide producing occlusion bodies that contain Bacillus thuringiensis insect toxin

Baculovirus occlusion bodies, large proteinaceous structures which contain virions, have recently been engineered to incorporate foreign proteins. The major constituent protein of occlusion bodies from the baculovirus Autographa californica nucleopolyhedrovirus is polyhedrin, and assembly of recombinant occlusion bodies which incorporate a foreign protein depends on an interaction between native polyhedrin and a polyhedrin–foreign protein fusion. This technology has now been applied to the generation of a recombinant baculovirus (ColorBtrus) that produces occlusion bodies incorporating the Bacillus thuringiensis (Bt) insecticidal Cry1Ac toxin protein. ColorBtrus coexpresses native polyhedrin and a fusion protein in which polyhedrin is fused to the Bt toxin, which is in turn fused to green fluorescent protein (GFP). Analysis of ColorBtrus occlusion bodies confirmed that they include both Bt toxin and GFP, yet still incorporate virions. Bioassay of ColorBtrus demonstrated that its speed of action and pathogenicity are strikingly enhanced compared to wild-type virus. ColorBtrus represents a novel, powerful biological insecticide that combines positive attributes of both Bt toxin and baculovirus based systems

Upregulation of smpd3 via BMP2 stimulation and Runx2.

Deletion of smpd3 induces osteogenesis and dentinogenesis imperfecta in mice. smpd3 is highly elevated in the parietal bones of developing mouse calvaria, but not in sutural mesenchymes. Here, we examine the mechanism of smpd3 regulation, which involves BMP2 stimulation of Runx2. smpd3 mRNA expression increased in response to BMP2 treatment and Runx2 transfection in C2C12 cells. The Runx2-responsive element (RRE) encoded within the -562 to -557 region is important for activation of the smpd3 promoter by Runx2. Electrophoretic mobility shift assays revealed that Runx2 binds strongly to the -355 to -350 RRE and less strongly to the -562 to -557 site. Thus, the smpd3 promoter is activated by BMP2 and is directly regulated by the Runx2 transcription factor. This novel description of smpd3 regulation will aid further studies of bone development and osteogenesis. [BMB reports 2009; 42(2): 86-90

Protective Effects of Emodin and Chrysophanol Isolated from Marine Fungus Aspergillus sp. on Ethanol-Induced Toxicity in HepG2/CYP2E1 Cells

Alcohol-induced liver injury progresses from fatty infiltration followed by a harmful cause of inflammation leading to an irreversible damage. In this study, two compounds (emodin and chrysophanol) isolated from marine fungus Aspergillus sp. were examined for their protective effects against ethanol-induced toxicity in vitro. Ethanol-induced HepG2/CYP2E1 cells were treated with the compounds at various concentrations, and the results showed that there was a dose-dependent decrease of gamma-glutamyl transpeptidase (GGT) activity and increase of glutathione (GSH) in the culture media with an increase in cell viability. Furthermore, the protective effects of the compounds were evaluated by protein expression levels of GGT, GSH, and CYP2E1 using Western blot. Among the compounds, emodin addressed to the ethanol-induced cytotoxicity more effectively compared to the chrysophanol. It could be suggested that emodin isolated from this genus would be a potential candidate for attenuating ethanol induced liver damage for further industrial applications such as functional food and pharmaceutical developments

Full-length cDNA sequences from Rhesus monkey placenta tissue: analysis and utility for comparative mapping

<p>Abstract</p> <p>Background</p> <p>Rhesus monkeys (<it>Macaca mulatta</it>) are widely-used as experimental animals in biomedical research and are closely related to other laboratory macaques, such as cynomolgus monkeys (<it>Macaca </it><it>fascicularis</it>), and to humans, sharing a last common ancestor from about 25 million years ago. Although rhesus monkeys have been studied extensively under field and laboratory conditions, research has been limited by the lack of genetic resources. The present study generated placenta full-length cDNA libraries, characterized the resulting expressed sequence tags, and described their utility for comparative mapping with human RefSeq mRNA transcripts.</p> <p>Results</p> <p>From rhesus monkey placenta full-length cDNA libraries, 2000 full-length cDNA sequences were determined and 1835 rhesus placenta cDNA sequences longer than 100 bp were collected. These sequences were annotated based on homology to human genes. Homology search against human RefSeq mRNAs revealed that our collection included the sequences of 1462 putative rhesus monkey genes. Moreover, we identified 207 genes containing exon alterations in the coding region and the untranslated region of rhesus monkey transcripts, despite the highly conserved structure of the coding regions. Approximately 10% (187) of all full-length cDNA sequences did not represent any public human RefSeq mRNAs. Intriguingly, two rhesus monkey specific exons derived from the transposable elements of AluYRa2 (SINE family) and MER11B (LTR family) were also identified.</p> <p>Conclusion</p> <p>The 1835 rhesus monkey placenta full-length cDNA sequences described here could expand genomic resources and information of rhesus monkeys. This increased genomic information will greatly contribute to the development of evolutionary biology and biomedical research.</p

Relationship between the Retinal Thickness Analyzer and the GDx VCC Scanning Laser Polarimeter, Stratus OCT Optical Coherence Tomograph, and Heidelberg Retina Tomograph II Confocal Scanning Laser Ophthalmoscopy

PURPOSE: To assess the relationship between the retinal thickness analyzer (RTA) parameters, and those of the GDx VCC scanning laser polarimeter (GDx VCC), Stratus OCT optical coherence tomography (Stratus OCT), and Heidelberg retinal tomograph II confocal scanning laser ophthalmoscopy (HRT II). METHODS: Twenty-nine primary open-angle glaucoma patients were retrospectively included in this study. Measurements were obtained using the RTA, GDx VCC, Stratus OCT, and HRT II. We calculated the correlation coefficients between the parameters of RTA and those of the other studies. RESULTS: Among the optic disc parameters of RTA, the cup volume was best correlated with Stratus OCT (R=0.780, p<0.001) and HRT II (R=0.896, p<0.001). Among the posterior pole retinal thickness parameters, the posterior pole abnormally thin area (PPAT) of the RTA and the inferior average of the GDx VCC were best correlated (R=-0.596, p=0.001). The PPAT of the RTA and the inferior maximum of the Stratus OCT were best correlated (R=-0.489, p=0.006). The perifoveal minimum thickness (PFMT) of the RTA and the cup shape measurement of the HRT II were best correlated (R=-0.565, p=0.004). CONCLUSIONS: Many RTA optic disc parameters were significantly correlated with those of the Stratus OCT and HRT II. The RTA posterior pole retinal thickness parameters were significantly correlated with those of the GDx VCC, Stratus OCT and HRT II. The RTA optic disc and posterior pole retinal thickness parameters may be valuable in the diagnosis of glaucomaope