Effects of nutrient and soil moisture on competition between shape Carex stricta,shape Phalaris arundinacea,and shape Typha latifolia

We investigated the importance of nutrients, soil moisture, arbuscular mycorrhizal fungi (AMF), and interspecific competition levels on the biomass allocation patterns of three wetland perennial plant species, Carex stricta Lam., Phalaris arundinacea L., and Typha latifolia L. A factorial experiment was conducted with high-low nutrient levels, high-low soil moisture levels, and with and without AMF inoculation. Under the experimental conditions, plant inoculation by AMF was too low to create a treatment and the AMF treatment was dropped from the total analysis. P. arundinacea and T. latifolia biomass were 73% and 77% higher, respectively, in the high nutrient treatment compared to the low nutrient treatment. Biomass allocation between shoots and roots remained relatively constant between environmental treatments, although shoot:root ratios of P. arundinacea declined in the low nutrient treatment. For C. stricta, the high nutrient and soil moisture treatments resulted in an increase in biomass of 50% and 15%, respectively. Shoot:root ratios were nearly constant among all environmental conditions. Biomass of T. latifolia and C. stricta was greatly decreased when grown with P. arundinacea. The rapid, initial height growth of P. arundinacea produced a spreading, horizontal canopy that overshadowed the vertical leaves of T. latifolia and C. stricta throughout the study. This pattern was repeated in both high and low nutrient and soil moisture treatments. When grown with P. arundinacea, C. stricta and T. latifolia significantly increased their mean shoot height, regardless of the nutrient or soil moisture level. The results of this experiment suggest that C. stricta and T. latifolia were light limited when growing with P. arundinacea and that canopy architecture is more important for biomass allocation than the other environmental conditions tested. The results also suggest that Phalaris arundinacea is an inherently better competitor (sensu Grime 1979) than C. stricta or T. latifolia.     

Hepatopancreas but not ovary is the site of vitellogenin synthesis in female fresh water crab, Oziothelphusa senex senex

The objective of the present study was to explore the site of synthesis of vitellogenin (Vtg) in fresh water edible crab, Oziothelphusa senex senex. Vtg cDNA fragments were isolated from the hepatopancreas of female crabs using RT-PCR method, and the deduced amino acid sequence of O. senex senex showed more than 60% identity with other brachyuran Vtg sequences. RT-PCR analysis showed that Vtg mRNA can be detected only in hepatopancreas of female Oziothelphusa but not in other tissues including eyestalks, Y-organs, mandibular organs, thoracic ganglion, hypodermis and ovary. Antibodies were raised against vitellin purified from the ovary of O. senex senex. Immunoprecipitation analysis revealed the presence of Vtg in the hepatopancreas of vitellogenic stage I females and in the hemolymph, hepatopancreas and ovary extracts from vitellogenic stage II females but absent in hemolymph and hepatopancreas extract of males. These results suggest that Vtg is synthesized only in hepatopancreas but not in the ovaries of O. senex senex. In addition, Vtg synthesized in hepatopancreas is transported to ovary through hemolymph.     

Polymorphism in the intermediates and products of amyloid assembly

Amyloid formation reactions exhibit two classes of polymorphisms: the metastable intermediates commonly observed during amyloid formation and the range of conformationally distinct mature fibrils often seen at the reaction endpoint. Although recent data suggest that spherical oligomers and protofibrils in most cases are not obligate intermediates of amyloid assembly, oligomeric states might sometimes serve as on-pathway intermediates. Mature amyloid polymorphs self-propagate as a result of the normally very high fidelity of amyloid elongation, giving rise to strain behavior and species barriers in prion phenomena. Oligomers, protofibrils and various polymorphic forms of mature amyloid fibrils seem to be distinguished by differences in atomic structure that give rise to differences in observed morphologies.     

Vegetation change from chronic stress events: Detection of the effects of tide gate removal and long‐term drought on a tidal marsh

Question: Chronic stress events are defined as disturbance events that exceed the lifespan of the dominant plant species, fluctuate in intensity and lack abruptness or physical destruction of biomass. Can the effects of chronic stress events be measured on vegetation communities? Did two chronic stress events, the removal of a tide gate and a four year drought, cause a temporary or permanent shift in the vegetation communities of a tidal marsh? Location: Tidal marsh in southeastern United States. Methods: Change in species composition and dominance and community change on a landscape level salinity gradient were measured between time periods ranging from four months to seven years to construct a statistical baseline reference community at freshwater, oligohaline, and mesohaline sections of a tidal marsh. Statistical shifts in the plant community were defined as changes in the plant community that fell outside of the defined baseline reference community. Results: Plant community changes outside of the reference community occurred in 13 out of 378 community comparisons. Removal of the tide gate had a greater effect on interstitial salinity levels than the drought and was most intense in the oligohaline marsh, where between 20 to 45% of the freshwa‐ter/oligohaline community types permanently converted to oligohaline community types. However, community shifts in the freshwater and oligohaline marsh induced by the drought were temporary, lasting from 1 to 3+ years. Neither chronic stress event permanently altered the mesohaline plant communities. Conclusion: The effects of chronic stress events could be detected; an extended historical record of vegetation change (18 years) was necessary to identify community shifts outside of a reference condition of the community and to determine if those shifts were permanent or temporary.     

Identification of novel allosteric modulators for the G-protein coupled US28 receptor of human cytomegalovirus

The highly constitutively active G-protein coupled receptor US28 of human cytomegalovirus (HCMV) is an interesting pharmacological target because of its implication on viral dissemination, cardiovascular diseases and tumorigenesis. We found that dihydroisoquinolinone and tetrahydroisoquinoline scaffolds may be promising lead structures for novel US28 allosteric inverse agonists. These scaffolds were rapidly synthesized by radical carboamination reactions followed by non-radical transformations. Our novel US28 allosteric modulators provide valuable scaffolds for further ligand optimization and may be helpful chemical tools to investigate molecular mechanisms of US28 constitutive signaling and its role in pathogenesis.     

Na,K-ATPase from mice lacking the gamma subunit (FXYD2) exhibits altered Na+ affinity and decreased thermal stability

The gamma subunit of the Na,K-ATPase, a 7-kDa single-span membrane protein, is a member of the FXYD gene family. Several FXYD proteins have been shown to bind to Na,K-ATPase and modulate its properties, and each FXYD protein appears to alter enzyme kinetics differently. Different results have sometimes been obtained with different experimental systems, however. To test for effects of gamma in a native tissue environment, mice lacking a functional gamma subunit gene (Fxyd2) were generated. These mice were viable and without observable pathology. Prior work in the mouse embryo showed that gamma is expressed at the blastocyst stage. However, there was no delay in blastocele formation, and the expected Mendelian ratios of offspring were obtained even with Fxyd2-/- dams. In adult Fxyd2-/- mouse kidney, splice variants of gamma that have different nephron segment-specific expression patterns were absent. Purified gamma-deficient renal Na,K-ATPase displayed higher apparent affinity for Na+ without significant change in apparent affinity for K+. Affinity for ATP, which was expected to be decreased, was instead slightly increased. The results suggest that regulation of Na+ sensitivity is a major functional role for this protein, whereas regulation of ATP affinity may be context-specific. Most importantly, this implies that gamma and other FXYD proteins have their effects by local and not global conformation change. Na,K-ATPase lacking the gamma subunit had increased thermal lability. Combined with other evidence that gamma participates in an early step of thermal denaturation, this indicates that FXYD proteins may play an important structural role in the enzyme complex.     

A novel type of detergent-resistant membranes may contribute to an early protein sorting event in epithelial cells

One sorting mechanism of apical and basolateral proteins in epithelial cells is based on their solubility profiles with Triton X-100. Nevertheless, apical proteins themselves are also segregated beyond the trans-Golgi network by virtue of their association or nonassociation with cholesterol/sphingolipid-rich microdomains (Jacob, R., and Naim, H. Y. (2001) Curr. Biol. 11, 1444-1450). Therefore, extractability with Triton X-100 does not constitute an absolute criterion of protein sorting. Here, we investigate the solubility patterns of apical and basolateral proteins with other detergents and demonstrate that the mild detergent Tween 20 is adequate to discriminate between apical and basolateral proteins during early stages in their biosynthesis. Although the mannose-rich forms of the apical proteins sucrase-isomaltase, lactase-phlorizin hydrolase, aminopeptidase N, and dipeptidylpeptidase IV reveal similar solubility profiles comprising soluble and nonsoluble fractions, the basolateral proteins, vesicular stomatitis virus G protein, major histocompatibility complex class I, and CD46 are entirely soluble with this detergent. The insoluble Tween 20 membranes are enriched in phosphatidylinositol and phosphatidylglycerol compatible with their synthesis in the endoplasmic reticulum and the existence of a novel class of detergent-resistant membranes. The association of the mannose-rich biosynthetic forms of the apical proteins, sucraseisomaltase, lactase-phlorizin hydrolase, aminopeptidase N, and dipeptidylpeptidase IV with the Tween 20-resistant membranes suggests an early polarized sorting mechanism prior to maturation in the Golgi apparatus.     

Hydrogen-deuterium (H/D) exchange mapping of Abeta 1-40 amyloid fibril secondary structure using nuclear magnetic resonance spectroscopy

We describe here details of the hydrogen-deuterium (H/D) exchange behavior of the Alzheimer's peptide Abeta(1)(-)(40), while it is a resident in the amyloid fibril, as determined by high-resolution solution NMR. Kinetics of H/D exchange in Abeta(1)(-)(40) fibrils show that about half the backbone amide protons exchange during the first 25 h, while the other half remain unexchanged because of solvent inaccessibility and/or hydrogen-bonded structure. After such a treatment for 25 h with D(2)O, fibrils of (15)N-enriched Abeta were dissolved in a mixture of 95% dimethyl sulfoxide (DMSO) and 5% dichloroacetic acid (DCA) and successive heteronuclear (1)H-(15)N HSQC spectra were collected to identify the backbone amides that did not exchange in the fibril. These studies showed that the N and C termini of the peptide are accessible to the solvent in the fibril state and the backbone amides of these residues are readily exchanged with bulk deuterium. In contrast, the residues in the middle of the peptide (residues 16-36) are mostly protected, suggesting that that many of the residues in this segment of the peptide are involved in a beta structure in the fibril. Two residues, G25 and S26, exhibit readily exchangeable backbone amide protons and therefore may be located on a turn or a flexible part of the peptide. Overall, the data substantially supports current models for how the Abeta peptide folds when it engages in the amyloid fibril structure, while also addressing some discrepancies between models.     

Functional complementation between the PDX1 vitamin B6 biosynthetic gene of Cercospora nicotianae and pdxJ of Escherichia coli

The pathway for de novo vitamin B(6) biosynthesis has been characterized in Escherichia coli, however plants, fungi, archaebacteria, and most bacteria utilize an alternative pathway. Two unique genes of the alternative pathway, PDX1 and PDX2, have been described. PDX2 encodes a glutaminase, however the enzymatic function of the product encoded by PDX1 is not known. We conducted reciprocal transformation experiments to determine if there was functional homology between the E. coli pdxA and pdxJ genes and PDX1 of Cercospora nicotianae. Although expression of pdxJ and pdxA in C. nicotianae pdx1 mutants, either separately or together, failed to complement the pyridoxine mutation in this fungus, expression of PDX1 restored pyridoxine prototrophy to the E. coli pdxJ mutant. Expression of PDX1 in the E. coli pdxA mutant restored very limited ability to grow on medium lacking pyridoxine. We conclude that the PDX1 gene of the alternative B(6) pathway encodes a protein responsible for synthesis of the pyridoxine ring.