Role of Auxin in Maize Endosperm Development (Timing of Nuclear DNA Endoreduplication,Zein Expression,and Cytokinin)

The timing of developmental events and regulatory roles of auxin were examined in maize (Zea mays L.) endosperms. Zeatin, zeatin riboside, and indole-3-acetic acid (IAA) levels were determined by enzyme-linked immunosorbent (ELISA). Zeatin and zeatin riboside increased to maximal concentrations at an early stage (9 d after pollination [DAP]), corresponding to the stage when cell division rate was maximal. In contrast, IAA concentration was low at 9 DAP and abruptly increased from 9 to 11 DAP, thus creating a sharp decline in the cytokinin to auxin ratio. Coincident with the increase in IAA was an increase in DNA content per nucleus, attributed to postmitotic DNA replication via endoreduplication. Exogenous application of 2,4-dichlorophenoxyacetic acid (2,4-D) at 5 or 7 DAP hastened the time course of DNA accumulation per nucleus and increased the average nuclear diameter, whereas 2-(para-chlorophenoxy)isobutyric acid delayed such development. Exogenously applied 2,4-D hastened the accumulation of the zein polypeptides of apparent molecular masses of 12, 14, and 16 kD and the expression of mRNA hybridizing with a zein DNA probe. We conclude that an abrupt increase in auxin induces cellular differentiation events in endosperm, including endoredupliction and expression of particular zein storage proteins.     

Exploring the potential of biopharmaceutical production by Rigidoporus ulmarius: Cultivation,chemistry, and bioactivity studies

Rigidoporus ulmarius is used as a medicinal fungus in Asia. Three isolates (denoted #61, #62, and #63) of R. ulmarius were collected, and their biological activities were evaluated. Extracted polysaccharides from isolate #63 showed greater inhibition activity compared to isolates #61 and #62 in an in vitro endothelial cell tube formation assay, a standard evaluation of angiogenesis. The polysaccharides and ethanolic extract of isolate #63 dose-dependently suppressed the production of the interferon (IFN)-γ-induced inflammation marker, IP-10. Chemical analyses of the polysaccharides revealed that isolate #63 contained the highest value of fucose at a concentration of 59.1 ± 1.2 μmol/g polysaccharide. These results suggest that fucose-containing polysaccharides may play a role in the inhibitory effect. Isolate #63 showed the highest values of ADP among the three isolates in the ethanolic extract. These results suggest that different isolates from R. ulmarius exhibit different abilities to regulate antiangiogenic and anti-inflammatory processes.     

InsP₃receptors and Orai channels in pancreatic acinar cells: co-localization and its consequences

Orai1 proteins have been recently identified as subunits of SOCE (store-operated Ca2? entry) channels. In primary isolated PACs (pancreatic acinar cells), Orai1 showed remarkable co-localization and co-immunoprecipitation with all three subtypes of IP?Rs (InsP? receptors). The co-localization between Orai1 and IP?Rs was restricted to the apical part of PACs. Neither co-localization nor co-immunoprecipitation was affected by Ca2? store depletion. Importantly we also characterized Orai1 in basal and lateral membranes of PACs. The basal and lateral membranes of PACs have been shown previously to accumulate STIM1 (stromal interaction molecule 1) puncta as a result of Ca2? store depletion. We therefore conclude that these polarized secretory cells contain two pools of Orai1: an apical pool that interacts with IP?Rs and a basolateral pool that interacts with STIM1 following the Ca2? store depletion. Experiments on IP?R knockout animals demonstrated that the apical Orai1 localization does not require IP?Rs and that IP?Rs are not necessary for the activation of SOCE. However, the InsP?-releasing secretagogue ACh (acetylcholine) produced a negative modulatory effect on SOCE, suggesting that activated IP?Rs could have an inhibitory effect on this Ca2? entry mechanism.     

Characterization and biological functions of sulfated polysaccharides from sulfated-salt treatment of Antrodia cinnamomea

Sulfated polysaccharides (SPSs) of Antrodia cinnamomea were extracted by an exhaustive papain digestion and characterized, and their biological functions were evaluated. In this study, we demonstrated the existence of SPSs in the medicinal fungus, A. cinnamomea. Since no SPSs had previously been identified in any fungal organism, we attempted to characterize those from A. cinnamomea. SPSs from A. cinnamomea inhibited in vitro Matrigel tube formation, in an angiogenesis model, in a dose-dependent manner. Furthermore, using serum deprivation-induced apoptosis in neuronal-like PC12 cells as a stress model, the SPSs of A. cinnamomea were effective in preventing serum-deprived apoptosis. Compositional analysis revealed that myo-inositol, fucose, galactose, and glucose were the neutral sugars in SPSs of A. cinnamomea, and these SPSs had a high sulfate content. The sulfation degree paralleled their antiangiogenic and neuroprotective activities. In this work, we report novel data on the structure, antiangiogenic, and neuroprotective effects of these fungal SPSs.     

Proteomic analysis of the expression of proteins related to rice quality during caryopsis development and the effect of high temperature on expression

Proteins are essential to rice caryopsis development and quality formation. High temperature is an important environmental factor, which may decrease grain quality. In the present study rice caryopsis proteins were profiled by two-dimensional polyacrylamide gel electrophoresis, and differentially expressed proteins were analyzed by liquid chromatography/tandem mass spectrometry. Expressions of more than 400 polypeptide spots during caryopsis development, in response to temperature treatments or between varieties were monitored. Among them, more than 70 differentially expressed polypeptides were analyzed by liquid chromatography/tandem mass spectrometry. We identified 54 proteins with known functions. Of these, 21 were involved with carbohydrate metabolism, 14 with protein synthesis and sorting, and 9 with stress responses. Waxy (Wx) proteins and glutelins were the most significant spots, which increased significantly during development. Allergen-like proteins, PPDK and NADH-SDH, also were expressed during development, implying their physiological roles in caryopsis. Expression of large isoforms of Wx proteins was correlated with the amylose content of rice caryopses. One protein with high GC content in its DNA sequence was correlated with the chalky trait of kernels. High temperature (35/30 degrees C) decreased the expression of Wx proteins, allergen-like proteins, and elongation factor 1beta, but increased the expression of small heat shock proteins (sHSP), glyceraldehyde-3-phosphate dehydrogenase, and prolamin. sHSP was positively correlated with the appearance of chalky kernels. During development, glutelins were phosphorylated and glycosylated, indicating that these molecules were post-translationally modified. Possible functions of the expression of candidate proteins on the grain quality are discussed.     

Structural and Ultrastructural Alterations in BALB/c Mice: Effects of Penicillium Citrinum Metabolites

The aims of this work were to determine the effect of feeding BALB/c mice a diet containing culture materials of a citrinin producing strain of Penicillium citrinum (Thom). Changes in hematological parameters, serum chemistry and histological changes in liver, kidney and heart were determined. After 60 days, control treated (CT) mice appeared normal in all respects, whereas, the mice fed the feeds supplemented with Penicillium (CMT) showed decreased weight gain, lower hematocrits, increased serum alanine aminotransferase (ALT) and clear signs of renal and hepatotoxicity based on histological changes. Changes observed in the liver of CMT mice included portal and lobular infiltration of polymorphonuclear cells, with concomitant hepatocellular necrosis, hepatic steatosis, prominent Kupffer's cells, hemosiderin granules in the cytoplasm of periportal hepatocytes and other lipid inclusions in the surrounding mitochondria were also observed. Our findings suggest that in vivo, P. citrinum Thom metabolites, which contain citrinin, could cause illnesses such as toxic hepatitis or intravascular hemolysis.     

Impact of metal pollution and Thlaspi caerulescens growth on soil microbial communities

Soil microorganisms drive critical functions in plant-soil systems. As such, various microbial properties have been proposed as indicators of soil functioning, making them potentially useful in evaluating the recovery of polluted soils via phytoremediation strategies. To evaluate microbial responses to metal phytoextraction using hyperaccumulators, a microcosm experiment was carried out to study the impacts of Zn and/or Cd pollution and Thlaspi caerulescens growth on key soil microbial properties: basal respiration; substrate-induced respiration (SIR); bacterial community structure as assessed by PCR-denaturing gradient gel electrophoresis (DGGE); community sizes of total bacteria, ammonia-oxidizing bacteria, and chitin-degrading bacteria as assessed by quantitative PCR (Q-PCR); and functional gene distributions as determined by functional gene arrays (GeoChip). T. caerulescens proved to be suitable for Zn and Cd phytoextraction: shoots accumulated up to 8,211 and 1,763 mg kg(-1) (dry weight [DW]) of Zn and Cd, respectively. In general, Zn pollution led to decreased levels of basal respiration and ammonia-oxidizing bacteria, while T. caerulescens growth increased the values of substrate-induced respiration (SIR) and total bacteria. In soils polluted with 1,000 mg Zn kg(-1) and 250 mg Cd kg(-1) (DW), soil bacterial community profiles and the distribution of microbial functional genes were most affected by the presence of metals. Metal-polluted and planted soils had the highest percentage of unique genes detected via the GeoChip (35%). It was possible to track microbial responses to planting with T. caerulescens and to gain insight into the effects of metal pollution on soilborne microbial communities.     

Cellular geography of IP3 receptors, STIM and Orai: a lesson from secretory epithelial cells

Pancreatic acinar cells exhibit a remarkable polarization of Ca2+ release and Ca2+ influx mechanisms. In the present brief review, we discuss the localization of channels responsible for Ca2+ release [mainly IP3 (inositol 1,4,5-trisphosphate) receptors] and proteins responsible for SOCE (store-operated Ca2+ entry). We also place these Ca2+-transporting mechanisms on the map of cellular organelles in pancreatic acinar cells, and discuss the physiological implications of the cellular geography of Ca2+ signalling. Finally, we highlight some unresolved questions stemming from recent observations of co-localization and co-immunoprecipitation of IP3 receptors with Orai channels in the apical (secretory) region of pancreatic acinar cells.