Metabolism of exogenous auxin by Arabidopsis thaliana: Identification of the conjugate Nα-(indol-3-ylacetyl)-glutamine and initiation of a mutant screen

The accumulation of conjugates of indole-3-acetic acid (IAA) in Arabidopsis thaliana was studied by incubating tissues with high concentrations of exogenous IAA, followed by reverse phase HPLC analysis of the extracts. Using fluorescence detection, indole-3-acetyl-aspartate, indole-3-acetyl-glutamate, and indole-3-acetyl-glucose were observed and quantitated in extracts of tissue after 24 h incubation with 500 μ M IAA. In addition, a new metabolite was detected and positively identified as indole-3-acetyl-glutamine by fast atom bombardment mass spectrometry, exact mass measurement, and tandem mass spectrometry in comparison with a synthetic standard. The amounts of individual conjugates formed differed between leaves, shoot axes and roots. In all three tissues, indole-3-acetyl-aspartate was the most abundant conjugate, the highest level being observed in roots. Highest levels of indole-3-acetyl-glutamine were observed in leaves, where it was the second most abundant conjugate and comprised approximately 12% of the fluorescent metabolites. Accumulation of the three amide conjugates was dramatically inhibited by cycloheximide, whereas accumulation of indole-3-acetyl-glucose was little affected. Based on these data, a screen for Arabidopsis mutants altered in the IAA-inducible system for auxin conjugate formation was initiated. The first mutant to be isolated and characterized produces more indole-3-acetyl-glutamine and less indole-3-acetyl-aspartate than wild-type, and is allelic to an existing class of photorespiration mutants ( gluS ) deficient in chloroplastic glutamate synthase.     

Clonal integration improves compensatory growth in heavily grazed ramet populations of two inland-dune grasses

Herbaceous species possess several mechanisms to compensate for tissue loss. For clonal herbaceous species, clonal integration may be an additional mechanism. This may especially hold true when tissue loss is very high, because other compensatory mechanisms may be insufficient. On inland dunes in northern China, we subjected Bromus ircutensis and Psammochloa villosa ramets within 0.5 m×0.5 m plots to three clipping treatments, i.e., no clipping, moderate (50% shoot removal) and heavy clipping (90% shoot removal), and kept rhizomes at the plot edges connected or disconnected. Moderate clipping did not reduce ramet, leaf or biomass density of either species. Under moderate clipping, rhizome connection significantly improved the performance of Psammochloa, but not that of Bromus. Heavy clipping reduced ramet, leaf and biomass density in the disconnected plots of both species, but such negative effects were negated or greatly ameliorated when the rhizomes were connected. Therefore, clonal integration contributed greatly to the compensatory growth of both species. The results suggest that clonal integration is an additional compensatory mechanism for clonal plants and may be important for their long-term persistence in the heavily grazed regions in northern China.     

Nanostructure and nanomechanics analysis of lymphocyte using AFM: From resting,activated to apoptosis

The ultrastructural and mechanical properties of single resting, activated and apoptosis lymphocyte have been investigated by atomic force microscopy (AFM). Using topographic imaging, we showed that the surface of the resting lymphocyte is smooth, while lymphocyte activation and apoptosis are often accompanied by changes in cell morphology. The apoptosis lymphocyte is rougher than those of the two other morphotypes, and coated with many big particles. Using spatially resolved force–distance curves, we found that the valve of the activated lymphocyte is about two to three times stiffer (Young's modulus of ～20 kPa) than those of the two other morphotypes (5–11 kPa). These results can improve our understanding of the mechanical properties of cells during growth and differentiation.     

Regulation of the G2/M phase of the cell cycle by sperm associated antigen 8 (SPAG8) protein

Sperm associated antigen 8 (SPAG8), a testis‐specific protein produced during male germ cell differentiation, was isolated from a human testis expression library using antibodies found in the serum obtained from an infertile woman. It was found to have a close functional relationship with microtubules. In this study, we generated a stably expressing SPAG8 CHO‐K1 cell line. Immunofluorescence confocal microscopy showed that SPAG8 was concentrated at the microtubule‐organizing center (MTOC) during prophase. As the cells progressed into metaphase, it co‐localized with α‐tubulin on the spindle. In anaphase, it was detected on both astral microtubules and mid‐zone. Following cytokinesis, SPAG8 resumed its localization on the MTOC. Meanwhile, flow cytometry analysis found that SPAG8 prolonged the G2/M phase of CHO‐K1 cells stably expressing SPAG8. Furthermore, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay showed that SPAG8 inhibited the proliferation of the stable cells. SPAG8 might be involved in the regulation of cell cycle by changing the phosphorylation level of Tyr15 on cdc2. These results suggest that SPAG8 might play a role in cell division during spermatogenesis. Copyright © 2009 John Wiley & Sons, Ltd.     

Using embryonic stem cells to form a biological pacemaker via tissue engineering technology

Biological pacemakers can be achieved by various gene‐based and cell‐based approaches. Embryonic stem cells (ESCs)‐derived pacemaker cells might be the most promising way to form biological pacemakers, but there are challenges as to how to control the differentiation of ESCs and to overcome the neoplasia, proarrhythmia, or immunogenicity resulting from the use of ESCs. As a potential approach to solve these difficult problems, tissue‐engineering techniques may provide a precise control on the different cell components of multicellular aggregates and the forming of a construct with‐defined architectures and functional properties. The combined interactions between ESC‐derived pacemaker cells, supporting cells, and matrices may completely reproduce pacemaker properties and result in a steady functional unit to induce rhythmic electrical and contractile activities. As ESCs have a high capability for self‐renewal, proliferation, and potential differentiation, we hypothesize that ESCs can be used as a source of pacemaker cells for tissue‐engineering applications and the ambitious goal of biological cardiac pacemakers may ultimately be achieved with ESCs via tissue‐engineering technology.     

Bioactive compounds from Rumex plants

Two new naphthalene acylglucosides, rumexneposides A (1) and B (2), together with 12 known compounds (3-14), were isolated from the roots of Rumex nepalensis. Their structures were established by chemical and spectroscopic methods. The biological activities of compounds 1-14 as well as an additional 11 compounds previously isolated from R. nepalensis and Rumex hastatus (15–25) were evaluated against Mycobacterium tuberculosis, para-aminobenzoic acid (pAba) pathway, and a panel of human cancer cell lines. The results showed that compound 15 was the most active against M. tuberculosis with an MIC value of 2.85 μM similar to that of isoniazid. Compound 5 could inhibit pAba synthetic pathway with an MIC value of 12.6 μM, comparable to that of positive control abyssomicin C, representing a new example of the rare pAba pathway inhibitors.     

Marine fouling on floating installations west of Dongsha Islands,the northern South China Sea

Marine fouling on floating installations west of Dongsha Islands, the northern South China Sea, was investigated from August 1987 to October 1990. Two buoy stations were deployed at approximately 110 km and 115 km west of Dongshadao, Dongsha Islands, anchored in water at 325 and 345 m depths, and exposed for 8 and 12 months. The experimental panels were fixed on iron structure frames which were connected to the buoy and its mooring system immersed in water at different depths, and retrieved after 4, 8 and 12 months of exposure. Fouling organisms colonizing the buoys and their mooring systems were sampled after 8 and 12 months of deployment. A total of 86 species was collected and identified at these two stations and the fouling community showed a typical oceanic characteristics. Most of the species were mainly found in the top 50 m depth and a striking vertical zonation of species was also observed with depth. The distance from offshore is a major factor in determining the fouling community composition. In terms of biomass, the fouling community was dominated by pedunculate barnacles, hydroids, and algae, followed by common oysters, pearl oysters and acorn barnacles. Biomass of hard fouling organisms increased over time. Under the influence of hydrological currents propagules of communities around the Dongsha Islands may also affect the development of the fouling communities in the offshore waters.     

Recycling wasted biomaterial,crab shells,as an adsorbent for the removal of high concentration of phosphate

In this study, crab shells were recycled as an adsorbent for the removal of phosphate. The effects of shell particle size, temperature, pH and phosphate concentration on phosphate removal were investigated. Shell particles less than 1000 μm in diameter removed more than 85% of 500 mg/L phosphate in 24 h while particles 3350 μm in diameter exhibited only 50% removal efficiency. Temperature showed negligible effect on phosphate removal in the range of 15–45 °C. Although removal efficiency was highest at pH 2.0, the efficiency remained 50–60% at pH of 4.0–10.0. The maximum removal capacity was calculated as 108.9 mg/g through Langmuir isotherm plotting, which was 17.0 and 4.7 times higher than those of coal fly ash and scallop shells, respectively. Although calcium carbonate played an active role in the removal of phosphate, both proteins composing 12.5% of crab shells and cellulose-like backbone of the crab shells also played an important role in phosphate removal.