Hairless regulates p53 target genes to exert tumor suppressive functions in glioblastoma

Glioblastoma (GBM) is the most common malignant brain tumor and is associated with a poor prognosis, with most patients living less than a year after diagnosis. Given that GBM nearly always recurs after conventional treatments, there is an urgent need to identify novel molecular targets. Hairless (HR) is a nuclear factor enriched in the skin and has been previously implicated in hair cycling. HR is also highly expressed in the brain, but its significance is unknown. We found that human hairless gene (HR) expression is significantly decreased in all GBM subtypes compared with normal brain tissue and is predictive of prognosis, which suggests that loss of HR expression can contribute to GBM pathogenesis. HR was recently discovered to bind to and regulate p53 responsive elements, and thus we hypothesized that HR may have a tumor suppressive function in GBM by modulating p53 target gene expression. We found that HR indeed regulates p53 target genes, including those implicated in cell cycle progression and apoptosis in the GBM-derived U87 cell line, and restoring HR expression triggered G2/M arrest and apoptosis. An analysis of sequenced genomes from patients with GBM revealed 10 HR somatic mutations in patients with glioma, two of which are located in the histone demethylase domain of HR. These two mutations, P996S and K1004N, were reconstructed and found to have impaired p53 transactivating properties. Collectively, the results of our study suggest that HR has tumor suppressive functions in GBM, which may be clinically relevant and a potential avenue for therapeutic intervention.     

Influence of die-back syndrome on reproductive strategies within Phragmites australis populations

Sexual plant reproduction is a strategy that allows plant populations to increase genetic variability, and consequently to be more efficient in adapting to new environments and to overcome stress conditions. Here, we focus on the reproductive mode of Phragmites australis, an important sub-cosmopolite species that can spread both by clonal propagation and sexual reproduction. In Europe, P. australis is affected by severe decline (known as Reed Die-Back Syndrome or RDBS); this study aims to understand if in RDBS condition the reproductive strategy of P. australis was altered, based on the hypothesis that if stress occurs, plants are likely to use their energy to increase genetic variability to overcome the stress. Inflorescences in five Italian wetlands were sampled from healthy and RDBS-affected stands of P. australis. Seed production, seed viability and seed germination were evaluated, with germination monitored under two different water conditions: moist and submerged. Stress induced by RDBS appears to influence the reproductive strategy of P. australis. In RDBS-affected stands, seed production was significantly higher than healthy stands for four of five sites. Seed viability and germination were also higher in RDBS-affected stands, but these differences were lower and site dependent.     

Analysis of genetic structure of a Suillus grevillei population in a Larix kaempferi stand by polymorphism of inter-simple sequence repeat (ISSR)

Clones of ectomycorrhizal fungi can colonize new areas through production of vegetative mycelium or spore dispersal, but the relative importance of these processes in nature is not known. In this study, sporocarps of an ectomycorrhizal fungus, Suillus grevillei , were mapped and sampled from a Larix kaempferi stand at the foot of Mt Fuji. DNA was extracted directly from each sporocarp, and DNA polymorphism was analysed by polymerase chain reaction (PCR) amplification of inter-simple sequence repeat (ISSR) regions primed by (GTG)₅, (GCC)₅ and (GACA)₄. Different sensitivities to detect polymorphism were found among the three primers, with (GACA)₄ showing the highest sensitivity. Forty seven sporocarps were analysed by the three ISSR primers and divided into 34 genets based on combination of PCR fingerprints. In the population 28 genets were represented by individual sporocarps. In most cases, sporocarps grown in aggregation (within a circle of 50 cm diameter) showed some different ISSR band patterns. These results suggest that genets of S. grevillei at the test site are relatively small. The genetic similarities between the 34 genets were also calculated and similarity groups were determined by the criterion that all similarity F values of genets within a group were not <80%. In general, the genets within a similarity group located close to each other. The results of multiple different but highly related genets in a small area suggest that the population of S. grevillei in this stand is not spread and maintained by clonal mycelium extension but is reproduced by spore dispersal.     

Procurement of spore‐free Bacillus anthracis for molecular typing outside of BSL3 environment

Aims: To (i) develop a protocol that would eliminate or greatly reduce sporulation within Bacillus anthracis vegetative cells, and (ii) harvest an adequate number of cells and sufficient DNA suitable for molecular methods including Riboprint^® analysis and pulse field gel electrophoresis (PFGE). Methods and Results: Seven strains of B. anthracis (Ames, French B2, Heluky, Kruger, Pasteur, Sterne, and Vollum) were grown at 37, 42 and 45°C under normal air, enhanced CO₂, microaerophilic, and anaerobic conditions on solid media and subcultured in two broths with and without supplements. The bacterial cells were centrifuged and washed. Slides made from the cell pellets were stained with Malachite Green and observed for the presence of spores. Cell preparations were subjected to 80°C for 30 min and processed for and analysed by either Riboprinter^® or PFGE. Multiple pellets of each strain were processed, stained, placed onto solid culture media, incubated for 7 days and observed for growth. The cell preparations yielded clear and reproducible results with both molecular methods. None of the cell preparations yielded growth on the culture media. Conclusions: This method eliminated viable spores in cell preparations of B. anthracis, yet still allowed the growth of vegetative cells to provide sufficient DNA suitable for analysis by Riboprinter^® and PFGE. Significance and Impact of the Study: This method will provide safe cell preparations, prevent instrument contamination, and may be useful for other aerobic and anaerobic spore‐formers.     

Predicted population trends for Cozumel Curassows (Crax rubra griscomi): empirical evidence and predictive models in the face of climate change

ABSTRACT We assessed the current conservation status and population trends of Cozumel Curassows (Crax rubra griscomi), a critically endangered and endemic cracid of Cozumel Island, Mexico. Distance sampling was used to estimate the population size in 2005 and compare it with a previous estimate in 1995. Population viability analysis (PVA) was used to model population trends, considering ecological and environmental conditions prevailing in 1995 and 2005. Additional PVA models were constructed using different hypothetical scenarios to assess the effects of particular factors and management strategies on population trends. We estimated a population size of 372 ± 155 curassows in the island's tropical semideciduous forest, before two hurricanes hit Cozumel Island in 2005. PVA modeling predicted a steep decline in population size in the ensuing decades as a result of an increased frequency and intensity of hurricanes related to global climate change, and an observed female bias in the sex ratio likely caused by a higher mortality rate for adult males. We recommend urgent management actions for the long‐term conservation of Cozumel Curassows, including a ban on hunting, eradication of the feral fauna, particularly dogs, and implementation of a captive‐breeding program to supplement the wild population.     

3种农业措施对柑橘丛枝菌根发育的影响

研究了土施N肥(2L 0.5%尿素)、覆草(5~6cm厚杂草)和翻耕(20cm表层土翻耕2次)三种措施,对田间条件下的国庆1号/枳和国庆4号/枳的菌根侵染率和孢子密度的影响。结果表明,覆草极显著地提高了2种柑橘根系的孢子密度,而对菌根侵染率无显著影响;施N肥极显著降低2种柑橘的孢子密度;翻耕极显著抑制了2种柑橘的菌根侵染率。2种柑橘的根系菌根侵染率和孢子密度间都存在显著或极显著正相关。     

水分胁迫对澳洲坚果花粉生活力和贮藏性的影响

采集经不同水分胁迫处理的澳洲坚果植株的花粉进行生活力测定和贮藏性研究,结果表明,新鲜花粉生活力随贮藏时间增加而下降。常温下正常发育花粉可保存6d左右,此后萌发率逐渐下降;而处于水分胁迫下的花粉,6d后生活力迅速降低;水分胁迫程度越严重,花粉生活力下降越快。几种处理的花粉有效贮藏期分别是:饱和灌溉处理6d,正常灌溉处理30d,水分亏缺处理10d,严重干旱处理(CK)4d。研究还表明,品种间花粉贮藏性表现为Kau >Pahala >O.C.。澳洲坚果花粉生活力的最佳染色鉴定方法为联苯胺染色法。     

阔鳞瘤蕨(水龙骨科)配子体形态发育的研究

利用光学显微镜详细观察记录了阔鳞瘤蕨孢子萌发、配子体发育及幼孢子体形成的整个形态发育过程。结果表明:成熟孢子黄色,极面观椭圆形,赤道面观肾形,近极单裂缝,表面具大小不一的球形颗粒。接种后6~8 d孢子萌发,萌发类型为书带蕨型,形成3~7个细胞的丝状体。原叶体发育为槲蕨型,接种后30 d左右进入片状体阶段,毛状体出现在片状体形成之后,幼原叶体短舌状,成熟原叶体心脏形。精子器在接种后60 d左右开始产生,颈卵器在精子器发生后10 d左右形成,每个原叶体上只能形成一个受精卵。培养基培养的片状体常在基部进行营养繁殖。     

Magnetic sorting of leukocytes

Magnetic filtration of surface-labeled cells has been applied to the fractionation of leukocytes in a model system, using a colloidal magnetite reagent to label mouse spleen cells. This reagent was completely free from problems of aggregation or settling. Since the individual submicron particles were invisible under the microscope, cells were not visibly altered by labeling. Viability also was unaffected by either labeling or magnetic filtration. Using a 10-kG magnet and a 5-mL filter column, 50 million cells were fractionated in less than 10 min, with 99% removal of labeled T lymphocytes. The efficiency of the magnetic method is limited at present by the fact that cells that do not have the surface target antigen of interest, and so are not antibody coated, may adsorb a small amount of label nonspecifically. These then have a nonzero chance of being captured in the filter along with the labeled cells.