Will the Three Gorges Dam affect the underwater light climate of <Emphasis Type="Italic">Vallisneria spiralis</Emphasis> L. and food habitat of Siberian crane in Poyang Lake?

Almost 95% of the entire population of the Siberian crane (Grus leucogeranus) winter in Poyang Lake, China, where they forage on the tubers of the submerged aquatic macrophyte Vallisneria spiralis. The Three Gorges Dam on the Yangtze River may possibly affect this food source of the Siberian crane by affecting the light intensity reaching the top of the V. spiralis canopy. In this study, the photosynthetically active radiation at the top of the V. spiralis canopy (PAR_tc) in Lake Dahuchi was modeled from 1998 to 2006, and the potential impacts of changes in water level and turbidity on the underwater light climate of V. spiralis were analyzed. PAR_tc was calculated from incident irradiance while the losses due to reflection at the water surface, absorption, and scattering within the water column were taken into consideration. The results indicated significant differences in PAR_tc between years. Six years of water level and Secchi disk depth records revealed a seasonal switching of the lake from a turbid state at low water levels in autumn, winter, and spring to a clear state at high water levels during the monsoon in summer. The highest PAR_tc occurred at intermediate water levels, which were reached when the Yangtze River forces Lake Dahuchi out of its turbid state in early summer and the water becomes clear. The intended operation of the Three Gorges Dam, which will increase water levels in May and June, may advance the moment when Lake Dahuchi switches from turbid to clear. We suggest that this might increase production of V. spiralis and possibly improve the food habitat conditions for wintering Siberian crane in Poyang Lake.

Susceptibility to cell death induced by blockade of MAPK pathway in human colorectal cancer cells carrying Ras mutations is dependent on p53 status

Constitutive activation of mitogen-activated protein kinase (MAPK) pathway is implicated in a variety of human malignancies especially those that carry Ras mutations and is currently exploited as a cancer therapeutic target. The variability of response by cancer cells to the inhibition of the Ras/MAPK pathway both in vivo and in vitro, however, suggests that the genetic background of the tumor cell may modulate the effectiveness of this directed therapeutic. In a panel of colorectal cancer cell lines that carry Ras mutations and have constitutively active MEK/MAPK, we found that inhibition of the MAPK upstream kinase MEK by the small molecular MEK inhibitor U0126 induced cell death only in p53 wild-type cells. By contrast, p53-deficient cells were not affected by blocking the MEK/MAPK pathway. Using isogenic colon cancer cell lines and RNA interference, we show that loss of p53 significantly reduces MAPK phosphorylation and renders cells resistant to U0126 treatment. These findings reveal a critical role for p53 in MAPK-driven cell survival and place p53 upstream in the control cascade of MAPK activity. The therapeutic implication of these observations is that MAPK inhibitors will be most beneficial as a therapeutic agent in p53 normal colon cancers where constitutively active MAPK resulting from a Ras mutation is required for cell survival.     

Selection of monosomic addition plants in offspring families using repetitive DNA probes in Beta L.

The distribution of two repetitive DNA probes Sat-121 and PB6-4, specific for the section Procumbentes of the genus Beta, was tested in 16 B. patellaris monosomic addition families using a dot-blot hybridization procedure. All monosomic additions were accurately distinguished from diploid sib plants with both DNA probes. The probe PB6-4, with the strongest signal after hybridization, was selected for rapid screening of an extensive number of putative monosomic additions in B. patellaris or B. procumbens addition families using a squash-blot hybridization procedure. The probe PB6-4 detected 118 monosomic additions in 640 plants (18.4%) in eight different B. procumbens addition families. The addition family with chromosome 4 of B. procumbens was semi-lethal and could not be tested. The distribution of PB6-4 in B. patellaris addition families was confirmed in 63 addition families using the squash-blot procedure. In 4580 plants of these addition families, 628 individual monosomic additions (13.7%) were found. The relationship of the morphological characteristics of monosomic addition plants to the results of the squash-blot hybridization (plants with signal) using probe PB6-4 is quite rigorous but not complete. The correlation between plants with a signal and chromosome number (2n=19) is complete. These results indicate that sequences present on PB6-4 are probably present on all chromosomes of B. patellaris and B. procumbens. The possibility of utilizing the sequence information of Sat-121 for a PCR-based assay to screen for putative monosomic addition plants was also investigated as an alternative to chromosome counting. The DNA-amplification profiles using the primers REP and REP.INV clearly distinguished monosomic addition plants from their diploid sibs.     

Morphometric and phylogenetic analysis of theDaviesia ulicifolia complex (Fabaceae, Mirbelieae)

Daviesia ulicifolia is a widespread species that exhibits complex variation throughout its range. Using ordination and cluster analysis of morphometric characters we resolved ten terminal taxa for phylogenetic analysis. A data set including these and five closely related species was coded for a combination of morphometric and qualitative characters and analysed using parsimony. This revealed thatD. ulicifolia is paraphyletic by inclusion ofD. acicularis, D. arenaria andD. microcarpa. One terminal cluster is more similar to an outgroup species (D. arthropoda) than toD. ulicifolia and should be treated as a new species. Given recent theoretical and empirical studies showing paraphyly to be both expected and observed at species level, we propose that all existing species in this group continue to be recognised taxonomically. We suggest subdividingD. ulicifolia into several subspecies.     

Costs of jasmonate-induced responses in plants competing for limited resources

We present a design to quantify fitness consequences of jasmonate-induced responses in plants that are competing for limited resources with a conspecific. Under both high and low nitrogen supply rates, uninduced (control) Nicotiana attenuata plants growing next to a plant induced with 250 μg methyl jasmonate (MJ) yielded more seed capsules than control plants competing with another control plant. We conclude that there is a opportunity benefit for control plants growing next to an induced plant. Initially, MJ-induced plants grew more slowly, but by senescence they had produced the same number of seed capsules as control plants that had competed with another control plant. Replacement series showed that the fitness of MJ-induced plants is not influenced by the competitive status of their neighbour plant. We argue that competitive designs are useful tools for evaluating the phenotypic costs of ecologically important traits.     

A light and electron microscopic study of the pineal gland of the ground squirrel, Citellus tridecemlineatus.

The pineal gland of the 13-lined ground squirrel (Citellus tridecemlineatus) has been examined at the light and electron microscopic level. This gland is composed of low-density parenchymal cells interspersed among which are occasional glial, vascular and neural elements. Punctuating the glandular parenchymal mass are prominent perivascular and intercellular spaces. The parenchymal cells possess numerous mitochondria and less prominent profiles of rough and smooth-surfaced endoplasmic reticulum. Golgi apparatus, microtubules and lipid droplets of varying size and electron density constitute regular cytoplasmic features, with dense-core vesicles being present occasionally. The parenchymal cells have numberous processes. One among these in each cell extends for several micra to terminate in a bulbous expansion containing both clear and dense-core vesicles and occasional electron-dense inclusions. These bulbous terminals are found within the perivascular and intercellular spaces where they course in close proximity to both other parenchymal elements and axon terminals. Glial cells and their processes invest the pineal periphery and incompletely separate the parenchymal cells.     

Nuclear factor TDP-43 and SR proteins promote in vitro and in vivo CFTR exon 9 skipping

Alternative splicing of human cystic fibrosis transmembrane conductance regulator (CFTR) exon 9 is regulated by a combination of cis-acting elements distributed through the exon and both flanking introns (IVS8 and IVS9). Several studies have identified in the IVS8 intron 3' splice site a regulatory element that is composed of a polymorphic (TG)m(T)n repeated sequence. At present, no cellular factors have been identified that recognize this element. We have identified TDP-43, a nuclear protein not previously described to bind RNA, as the factor binding specifically to the (TG)m sequence. Transient TDP-43 overexpression in Hep3B cells results in an increase in exon 9 skipping. This effect is more pronounced with concomitant overexpression of SR proteins. Antisense inhibition of endogenous TDP-43 expression results in increased inclusion of exon 9, providing a new therapeutic target to correct aberrant splicing of exon 9 in CF patients. The clinical and biological relevance of this finding in vivo is demonstrated by our characterization of a CF patient carrying a TG10T9(DeltaF508)/TG13T3(wt) genotype leading to a disease-causing high proportion of exon 9 skipping.     

Development of non-phosphorylated cyclic thioether peptide binding to the Grb2-SH2 domain

Summary One of the critical intracellular signaling pathways involves specific interactions between growth factor receptors and the adaptor protein Grb2. These interactions normally involve specific tyrosine phosphorylated regions in receptors and other cognate proteins. Following the lead of our recent findings that a phage library based non-phosphorylated disulfide linked 11-mer peptide inhibited such interactions, we report here the synthesis of novel redox-stable cyclic peptide analogs. These include thioether cyclized and backbone cyclized structures. The thioether analog was prepared under mild conditions from an N-terminally chloroacetylated and C-terminally cysteine extended peptide precursor. The thioether peptide showed equipotent binding affinity for the Grb2-SH2 domain (IC₅₀=10–15 μM) when compared to the disulfide cyclized lead-peptide. The bioactive thioether linked peptide was demonstrated to offer advantages to the disulfide cyclized peptides under physiological conditions.