A Wnt1-regulated genetic network controls the identity and fate of midbrain-dopaminergic progenitors in vivo

Midbrain neurons synthesizing the neurotransmitter dopamine play a central role in the modulation of different brain functions and are associated with major neurological and psychiatric disorders. Despite the importance of these cells, the molecular mechanisms controlling their development are still poorly understood. The secreted glycoprotein Wnt1 is expressed in close vicinity to developing midbrain dopaminergic neurons. Here, we show that Wnt1 regulates the genetic network, including Otx2 and Nkx2-2, that is required for the establishment of the midbrain dopaminergic progenitor domain during embryonic development. In addition, Wnt1 is required for the terminal differentiation of midbrain dopaminergic neurons at later stages of embryogenesis. These results identify Wnt1 as a key molecule in the development of midbrain dopaminergic neurons in vivo. They also suggest the Wnt1-controlled signaling pathway as a promising target for new therapeutic strategies in the treatment of Parkinson's disease.     

Design and synthesis of 2,3,4,9-tetrahydro-1H-carbazole and 1,2,3,4-tetrahydro-cyclopenta[b]indole derivatives as non-nucleoside inhibitors of hepatitis C virus NS5B RNA-dependent RNA polymerase

A novel class of HCV NS5B RNA dependent RNA polymerase inhibitors containing 2,3,4,9-tetrahydro-1H-carbazole and 1,2,3,4-tetrahydro-cyclopenta[b]indole scaffolds were designed and synthesized. Optimization of the aromatic region showed preference for 5,8-disubstitution pattern in both the scaffolds examined while favoring the n-propyl moiety for the C-1 position. 1,2,3,4-tetrahydro-cyclopenta[b]indole scaffold was slightly more potent than the corresponding 2,3,4,9-tetrahydro-1H-carbazole and analogue 36 displayed an IC50 of 550 nM against HCV NS5B enzyme.     

MEL-28 is downstream of the Ran cycle and is required for nuclear-envelope function and chromatin maintenance

Early embryonic development depends on the faithful execution of basic cell biological processes whose coordination remains largely unknown. With a global network analysis, we found MEL-28 to be associated with two types of complexes, one implicated in nuclear-envelope function and the other in chromatin organization. Here, we show that MEL-28, a protein that shuttles between the nucleus and the kinetochore during the cell cycle, is required for the structural and functional integrity of the nuclear envelope. In addition, mel-28(RNAi) embryos exhibit defects in chromosome condensation, pronuclear migration, kinetochore assembly, and spindle assembly. This combination of mel-28(RNAi) phenotypes resemble those caused by depleting members of the Ran cycle in C. elegans, a conserved cellular signaling pathway that is required for mitotic spindle assembly, nuclear-envelope reformation after mitosis, and nucleocytoplasmic exchange (reviewed in). Although MEL-28 localization to the nuclear periphery is not dependent on nuclear pore components, it is dependent on RAN-1 and other key components of the Ran cycle. Thus, MEL-28 is downstream of the Ran cycle and is required for both proper nuclear-envelope function and chromatin maintenance.     

Bacterial community changes in TCE biodegradation detected in microcosm experiments

Laboratory microcosm experiments were set up to model biodegradation of trichloroethylene (TCE). Groundwater samples from two contaminated sites were taken, one of them with low (70 mg L⁻¹), the other with high sulfate (685 mg L⁻¹) concentration. In order to assess the biodegradative potential of natural microbiota, supplementary substrates (whey or molasses) were added to the bottles. At day 54, 98, 155, and 318, chemical and bacteriological parameters (i.e., Dehalococcoides test) were investigated. Terminal restriction fragment length polymorphism (T-RFLP) based diversity assessments were carried out to observe the bacterial community changes. Whey and molasses enhanced degradation at different rates. In the case of samples with high sulfate content and amended with whey, no ethylene, ethane, or methane was generated. Both ethylene and methane production was detected in samples of low sulfate content with added whey. The results of Dehalococcoides tests were positive for all control and amended samples. Based on T-RFLP analysis, the bacterial communities of high sulfate concentration groundwater microcosms amended with molasses or whey were very similar, while the communities of groundwater samples with low sulfate concentration were different when supplemented with whey or molasses. The rRNA and rDNA based investigations suggest that the proportions of the active microbes and the microbes present in the microcosms differ.     

Does universal temperature dependence apply to communities? An experimental test using natural marine plankton assemblages

The metabolic theory of ecology (MTE) is an intriguing but controversial theory that tries to explain ecological patterns at all scales on the basis of first principles. Temperature plays a pivotal role in this theory. According to MTE, the Arrhenius relationship that describes the effect of temperature on biochemical reactions extends to a 'universal temperature dependence' that encompasses all kinds of processes and scales up to the cellular, the organismal, and the community level. In this study we test the prediction that community growth rate is temperature dependent in an Arrhenius-like way. First, we performed a literature review of the scanty data on the temperature dependence of the rates of metabolism, photosynthesis and growth of communities. In contrast to the predictions of MTE, the community activation energies did not cluster around 0.32 eV, the activation energy of photosynthesis and primary production or around 0.65 eV, the activation energy of metabolism. However, in none of the published studies the conditions were sufficiently controlled to allow firm conclusions. We therefore also performed replicated and controlled experiments using natural assemblages of marine plankton. As predicted by MTE, the maximal growth rates of community biomass increased linearly in an Arrhenius plot, with a slope close to 0.32 eV. However, a diversity of other models for the temperature dependence of community growth rates fit our data equally well. Hence, our results are at best a weak confirmation of MTE.     

Low crop plant population densities promote pollen-mediated gene flow in spring wheat (Triticum aestivum L.)

Transgenic wheat is currently being field tested with the intent of eventual commercialization. The development of wheat genotypes with novel traits has raised concerns regarding the presence of volunteer wheat populations and the role they may play in facilitating transgene movement. Here, we report the results of a field experiment that investigated the potential of spring wheat plant population density and crop height to minimize gene flow from a herbicide-resistant (HR) volunteer population to a non-HR crop. Pollen-mediated gene flow (PMGF) between the HR volunteer wheat population and four conventional spring wheat genotypes varying in height was assessed over a range of plant population densities. Natural hybridization events between the two cultivars were detected by phenotypically scoring plants in F₁ populations followed by verification with Mendelian segregation ratios in the F_1:2 families. PMGF was strongly associated with crop yield components, but showed no association with flowering synchrony. Maximum observed PMGF was always less than 0.6%, regardless of crop height and density. The frequency of PMGF in spring wheat decreased exponentially with increasing plant population density, but showed no dependence on either crop genotype or height. However, increasing plant densities beyond the recommended planting rate of 300 cropped wheat plants m⁻² provided no obvious benefit to reducing PMGF. Nevertheless, our results demonstrate a critical plant density of 175–200 cropped wheat plants m⁻² below which PMGF frequencies rise exponentially with decreasing plant density. These results will be useful in the development of mechanistic models and best management practices that collectively facilitate the coexistence of transgenic and nontransgenic wheat crops.     

Chemical characterization of a marine conditioning film

Conditioning film grown on glass panels was analyzed for organic carbon and nitrogen, total carbohydrates, aldoses, uronic acids, and proteins. These compounds showed non-linear increase over the period of immersion. Total carbohydrate-C, aldose-C, uronic acid-C, and protein-C accounted for 5.59–23.50%, 0.6–5%, 1–5%, and 4–14% of organic carbon, respectively, while aldose-C accounted for 10–72% of the total carbohydrate-C. Aldoses such as rhamnose, arabinose, xylose, mannose, and glucose were generally present in the conditioning film. A principal component analysis based on weight percentages of aldoses established three factors that contributed ～77% of the total variance. The first factor was influenced by the abundance of rhamnose and accounted for 41% of the total variance. The second factor was influenced by the concentrations of arabinose and xylose, whereas the third factor was controlled by ribose and galactose, thereby implicating the role of terrestrial and microbial input to the conditioning film. These changes in the chemistry of the conditioning film may play some role in influencing microbial adhesion to surfaces.