Molecular markers of nuclear restoration gene Rf1 in sunflower using bulked segregant analysis-RAPD

Restoration of cytoplasmic male sterility (CMS) in sunflower was demonstrated to be controlled by polygenes by analysing 982 effective crosses among 109 self-crossed lines and 16 CMS lines. Two self-crossed lines and one CMS line with distinct genotypes were applied to creation of segregating populations for DNA bulks of the target gene Rfl. Bulked DNA was prepared in order to investigate single gene Rfl and its gene marker among polygenic characters at the same genetic background. Using 80 10-mer operon primers, 620 RAPD reactions were carried out between fertile and sterile DNA bulks. In about 800 loci, primary results showed that 8 were related to the restoration genes. Furthermore. 2 were confirmed as RAPD markers for gene Rfl by examining 9 maintenance and 7 restoration lines. This method is the improvement for bulked segregant analysis[1] with which markers of single gene of target can be identified rapidly among polygenic characters.     

Bacterial tail anchors can target to the mitochondrial outer membrane

Background

During the generation and evolution of the eukaryotic cell, a proteobacterial endosymbiont was re-fashioned into the mitochondrion, an organelle that appears to have been present in the ancestor of all present-day eukaryotes. Mitochondria harbor proteomes derived from coding information located both inside and outside the organelle, and the rate-limiting step toward the formation of eukaryotic cells may have been development of an import apparatus allowing protein entry to mitochondria. Currently, a widely conserved translocon allows proteins to pass from the cytosol into mitochondria, but how proteins encoded outside of mitochondria were first directed to these organelles at the dawn of eukaryogenesis is not clear. Because several proteins targeted by a carboxyl-terminal tail anchor (TA) appear to have the ability to insert spontaneously into the mitochondrial outer membrane (OM), it is possible that self-inserting, tail-anchored polypeptides obtained from bacteria might have formed the first gate allowing proteins to access mitochondria from the cytosol.

Results

Here, we tested whether bacterial TAs are capable of targeting to mitochondria. In a survey of proteins encoded by the proteobacterium Escherichia coli, predicted TA sequences were directed to specific subcellular locations within the yeast Saccharomyces cerevisiae. Importantly, TAs obtained from DUF883 family members ElaB and YqjD were abundantly localized to and inserted at the mitochondrial OM.

Conclusions

Our results support the notion that eukaryotic cells are able to utilize membrane-targeting signals present in bacterial proteins obtained by lateral gene transfer, and our findings make plausible a model in which mitochondrial protein translocation was first driven by tail-anchored proteins.

Reviewers

This article was reviewed by Michael Ryan and Thomas Simmen.

     

The Interplay of Proton,Electron, and Metabolite Supply for Photosynthetic H2 Production in Chlamydomonas reinhardtii

To obtain a detailed picture of sulfur deprivation-induced H₂ production in microalgae, metabolome analyses were performed during key time points of the anaerobic H₂ production process of Chlamydomonas reinhardtii. Analyses were performed using gas chromatography coupled to mass spectrometry (GC/MS), two-dimensional gas chromatography combined with time-of-flight mass spectrometry (GCxGC-TOFMS), lipid and starch analysis, and enzymatic determination of fermentative products. The studies were designed to provide a detailed metabolite profile of the solar Bio-H₂ production process. This work reports on the differential analysis of metabolic profiles of the high H₂-producing strain Stm6Glc4 and the wild-type cc406 (WT) before and during the H₂ production phase. Using GCxGC-TOFMS analysis the number of detected peaks increased from 128 peaks, previously detected by GC/MS techniques, to ∼1168. More detailed analysis of the anaerobic H₂ production phase revealed remarkable differences between wild-type and mutant cells in a number of metabolic pathways. Under these physiological conditions the WT produced up to 2.6 times more fatty acids, 2.2 times more neutral lipids, and up to 4 times more fermentation products compared with Stm6Glc4. Based on these results, specific metabolic pathways involving the synthesis of fatty acids, neutral lipids, and fermentation products during anaerobiosis in C. reinhardtii have been identified as potential targets for metabolic engineering to further enhance substrate supply for the hydrogenase(s) in the chloroplast.     

Human Impacts on the Fire Regime of Interior Alaska: Interactions among Fuels, Ignition Sources, and Fire Suppression

Wildfire is the major natural agent of disturbance in interior Alaska. We examined the magnitude of human impact on fire by comparing fire regime between individual 1-km² grid cells designated for fire suppression with lands where fires are allowed to burn naturally. Two-thirds of interior Alaska has an essentially natural fire regime, with few human ignitions, negligible suppression activity, and many large lightning-caused fires. In the 17% of land that is designated for fire suppression due to its proximity to communities and roads, there was a 50% reduction in the proportion of area burned from 1992–2001, relative to areas without suppression. The remaining 16% of land serves as a buffer, receives some suppression, and has an intermediate fire regime. Even though there were 50 times more fires and the fire season began two months earlier in lands designated for suppression, most of these fires were lit by people and remained small because fires tended to occur at times and places less favorable for fire spread and were more accessible to fire fighters compared to lands not designated for suppression. Even in the absence of fire suppression, human-caused fires were less likely to exceed 400 ha compared to lightning-caused fires. Fire suppression reduced area burned in all fuel types but was somewhat more effective in less flammable (non-forest) vegetation. Alaska’s fire policy of focusing suppression efforts on a small proportion of the fire-prone region maximizes the ecological and social benefits associated with fire-dependent ecosystem services, while minimizing the social and ecological costs of suppression. Application of this policy to other areas would require well-informed managers and stakeholders to make difficult decisions about the relative costs and benefits of fire across ecologically and culturally variable landscapes.     

First-In-Human,Double-Blind,Placebo-Controlled,Randomized, Dose-Escalation Study of BG00010, a Glial Cell Line-Derived Neurotrophic Factor Family Member,in Subjects with Unilateral Sciatica

ObjectiveTo evaluate the safety, tolerability, and pharmacokinetics of single doses of BG00010 (neublastin, artemin, enovin) in subjects with unilateral sciatica.MethodsThis was a single-center, blinded, placebo-controlled, randomized Phase 1 sequential-cohort, dose-escalation study (ClinicalTrials.gov identifier {"type":"clinical-trial","attrs":{"text":"NCT00961766","term_id":"NCT00961766"}}NCT00961766; funded by Biogen Idec). Adults with unilateral sciatica were enrolled at The Royal Adelaide Hospital, Australia. Four subjects were assigned to each of eleven cohorts (intravenous BG00010 0.3, 1, 3, 10, 25, 50, 100, 200, 400, or 800 μg/kg, or subcutaneous BG00010 50 μg/kg) and were randomized 3:1 to receive a single dose of BG00010 or placebo. The primary safety and tolerability assessments were: adverse events; clinical laboratory parameters and vital signs; pain as measured by a Likert rating scale; intra-epidermal nerve fiber density; and longitudinal assessment of quantitative sensory test parameters. Blood, serum, and plasma samples were collected for pharmacokinetic and pharmacodynamic assessments. Subjects were blinded to treatment assignment throughout the study. The investigator was blinded to treatment assignment until the Data Safety Review Committee review of unblinded data, which occurred after day 28.ResultsBeyond the planned enrollment of 44 subjects, four additional subjects were enrolled into to the intravenous BG00010 200 μg/kg cohort after one original subject experienced mild generalized pruritus. Therefore, a total of 48 subjects were enrolled between August 2009 and December 2011; all were included in the safety analyses. BG00010 was generally well tolerated: in primary analyses, the most common treatment-emergent adverse events were changes in temperature perception, pruritus, rash, or headache; no trends were observed in clinical laboratory parameters, vital signs, intra-epidermal nerve fiber density, or quantitative sensory testing. BG00010 was not associated with any clear, dose-dependent trends in Likert pain scores. BG00010 was rapidly distributed, with a prolonged terminal elimination phase.ConclusionsThese data support the development of BG00010 for the treatment of neuropathic pain.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00961766","term_id":"NCT00961766"}}NCT00961766     

甘薯品种营养成份与抗小象虫的相关性研究

通过田间和网室测定结果,抗小象虫较好的甘薯品种有抗虫1号、台农26、Tis2534、Ricin和鸡蛋黄;抗小象虫较弱的品种有新种花、惠红早、“329”、广薯15等。甘薯品种的营养成份与抗小象虫相关性分析结果表明,抗、感品种与粗纤维、粗脂肪含量无明显相关,与粗淀粉有显负相关,R1=-0．9935,而与粗蛋白和18种氨基酸总量有显正相关,R2=0．9741,R3=0．9621。表现粗淀粉含量高的品种,其虫害指敦较低,抗虫性强;而粗蛋白含量和18种氨基酸总量高的品种,其虫害指数较高,抗虫性则表现较弱。测定分析说明了甘薯品种营养成份与抗虫性存在一定的相关性。     

Trichobilharzia regenti: host immune response in the pathogenesis of neuroinfection in mice

Besides their natural bird hosts, Trichobilharzia regenti cercariae are able to penetrate skin of mammals, including humans. Experimental infections of mice showed that schistosomula of this species are able to avoid the immune response in skin of their non-specific mammalian host and escape the skin to migrate to the CNS. Schistosomula do not mature in mammals, but can survive in nervous tissue for several days post infection. Neuroinfections of specific bird hosts as well as accidental mammalian hosts can lead to neuromotor effects, for example, leg paralysis and thus this parasite serves as a model of parasite invasion of the CNS.Here, we show by histological and immunohistochemical investigation of CNS invasion of immunocompetent (BALB/c) and immunodeficient (SCID) mice by T. regenti schistosomula that the presence of parasites in the nervous tissue initiated an influx of immune cells, activation of microglia, astrocytes and development of inflammatory lesions. Schistosomula elimination in the tissue depended on the host immune status. In the absence of CD3+ T-cells in immunodeficient SCID mice, parasite destruction was slower than that in immunocompetent BALB/c mice. Axon injury and subsequent secondary demyelination in the CNS were associated with mechanical damage due to migration of schistosomula through the nervous tissue, and not by host immune processes. Immunoreactivity of the parasite intestinal content for specific antigens of oligodendrocytes/myelin and neurofilaments showed for the first time that schistosomula ingest the nervous tissue components during their migration.     

Effect of density on age-specific mortality in Drosophila: a density supplementation experiment

Age-specific mortality rates were studied at two adult density levels in four inbred lines of Drosophila melanogaster. In experimental populations, adult densities were maintained at constant levels throughout the experiment by replacing dead flies with live, marked mutants. In control populations, densities declined naturally as the cohorts aged. For all experimental populations the best mortality model is the two-stage Gompertz model, with slower mortality acceleration at older ages. Flies in the experimental populations generally lived longer than flies in control populations, regardless of sex, genotype, or initial density level. The data demonstrate that deceleration of age-specific mortality rates at older ages is not caused by declining cohort densities. Mortality deceleration is a real phenomenon that raises serious questions about the evolution of senescence.     

Effect of selenium supplementation in asthmatic subjects on the expression of endothelial cell adhesion molecules in culture

Endothelial cells play a major role in immunologic reactions, in which cellular adhesion molecules P-selectin, ICAM-1, VCAM-1, and ELAM-1 are important mediators in the recruitment of leukocytes in pulmonary inflammation. Selenium (Se) is known to modulate immunological mechanisms of asthma. The aim of our investigation was to examine whether Se supplementation in cortico-dependent asthmatic patients may modulate adhesion molecule expression in cultured endothelium. Our findings indicated that P-selectin, VCAM-1, and ELAM-1 expression on human umbilical vein endothelial cells stimulated with peripheral blood mononuclear cells obtained from asthmatics before supplementation with Se was significantly higher than from healthy donors (p < 0.05). The production of ICAM-1 showed only slight augmentation. The levels of VCAM-1 and ELAM-1 expression were significantly decreased after 3 mo of Se supplementation (p < 0.05). After 6 mo of intervention period the intensity of P-selectin and ICAM-1 expression was also significantly reduced (p < 0.05 andp < 0.01, respectively). The inhibitory effect of Se on the adhesion molecule expression was studied in cultured endothelial cells after interferon-γ stimulation. Our data suggest that Se affects the expression of P-selectin, ICAM-1, VCAM-1, and ELAM-1 in a dosedependent manner and the half-maximal inhibitory concentrations were 3.4, 0.5, 4, and 3.8 μg/mL, respectively. The maximal inhibitions (greater than 80%) were observed in vitro with 10 μg/mL Se (p < 0.01). Regulation of adhesion molecule expression may be an important mechanism through which the inflammation may be controlled.     

1H, 13C and 15N assignments of a camelid nanobody directed against human α-synuclein

Nanobodies are single chain antibodies that are uniquely produced in Camelidae, e.g. camels and llamas. They have the desirable features of small sizes (Mw < 14 kDa) and high affinities against antigens (Kd ~ nM), making them ideal as structural probes for biomedically relevant motifs both in vitro and in vivo. We have previously shown that nanobody binding to amyloidogenic human lysozyme variants can effectively inhibit their aggregation, the process that is at the origin of systemic amyloid disease. Here we report the NMR assignments of a new nanobody, termed NbSyn2, which recognises the C-terminus of the intrinsically disordered protein, human α-synuclein (aS), whose aberrant self-association is implicated in Parkinson’s disease.