Isolation and structural identification of the trihydroxamate siderophore vicibactin and its degradative products from Rhizobium leguminosarum ATCC 14479 bv. trifolii

The Rhizobia are a group of free-living soil bacteria known for their ability to symbiotically infect the roots of specific host plants as well as to produce siderophores in order to compete with other microorganisms for the limited availability of iron in the rhizosphere. In this study, Rhizobium leguminosarum ATCC 14479, which preferentially infects the red clover Trifolium pratense, was found to produce the trihydroxamate siderophore vicibactin (C₃₃H₅₅N₆O₁₅) under iron restricted conditions. In addition, two other iron-binding, siderophore-like compounds: C₂₀H₃₆N₄O₁₀, C₃₁H₅₅N₆O₁₅, were isolated and purified from the culture media. Due to the structural similarity of the latter compounds to vicibactin based on electrospray-mass spectrometry and nuclear magnetic resonance data, these heretofore unreported molecules are thought to be either modified or degraded products of vicibactin. Although vicibactin has previously been found to be commonly produced by other rhizobial strains, this is the first time it has been chemically characterized from a clover infecting strain of R. leguminosarum.     

Linking Restoration and Landscape Ecology

Landscape ecology focuses on questions typically addressed over broad spatial scales. A landscape approach embraces spatial heterogeneity, consisting of a number of ecosystems and/or landscape structures of different types, as a central theme. Such studies may aid restoration efforts in a variety of ways, including (1) provision of better guidance for selecting reference sites and establishing project goals and (2) suggestions for appropriate spatial configurations of restored elements to facilitate recruitment of flora/fauna. Likewise, restoration efforts may assist landscape–level studies, given that restored habitats, possessing various patch arrangements or being established among landscapes of varying diversity and conditions of human alteration, can provide extraordinary opportunities for experimentation over a large spatial scale. Restoration studies can facilitate the rate of information gathering for expected changes in natural landscapes for which introduction of landscape elements may be relatively slow. Moreover, data collected from restoration studies can assist in validation of dynamic models of current interest in landscape ecology. We suggest that restoration and landscape ecology have an unexplored mutualistic relationship that could enhance research and application of both disciplines.     

Twisted gastrulation promotes BMP signaling in zebrafish dorsal-ventral axial patterning

In vertebrates and invertebrates, the bone morphogenetic protein (BMP) signaling pathway patterns cell fates along the dorsoventral (DV) axis. In vertebrates, BMP signaling specifies ventral cell fates, whereas restriction of BMP signaling by extracellular antagonists allows specification of dorsal fates. In misexpression assays, the conserved extracellular factor Twisted gastrulation (Tsg) is reported to both promote and antagonize BMP signaling in DV patterning. To investigate the role of endogenous Tsg in early DV patterning, we performed morpholino (MO)-based knockdown studies of Tsg1 in zebrafish. We found that loss of tsg1 results in a moderately strong dorsalization of the embryonic axis, suggesting that Tsg1 promotes ventral fates. Knockdown of tsg1 combined with loss of function of the BMP agonist tolloid (mini fin) or heterozygosity for the ligand bmp2b (swirl) enhanced dorsalization, supporting a role for Tsg1 in specifying ventral cell fates as a BMP signaling agonist. Moreover, loss of tsg1 partially suppressed the ventralized phenotypes of mutants of the BMP antagonists Chordin or Sizzled (Ogon). Our results support a model in which zebrafish Tsg1 promotes BMP signaling, and thus ventral cell fates, during DV axial patterning.     

Recombinant human monoclonal antibodies

To produce human monoclonal antibodies in bacteria, a gene repertoire of IgM variable regions was isolated from human peripheral B lymphocytes by the polymerase chain reaction. Alternatively, synthetic antibody genes with random hypervariable regions are being generated that may provide libraries of even higher complexity. For the selection of specific monoclonal antibodies from these libraries, we have developed twoE. coli vector systems that facilitate the surface display of an antibody physically linked to its own gene. The phagemid pSEX encodes a fusion protein of an antigen binding domain (Fv-antibody) with the docking protein (pIII) of filamentous phages. Specific antibody genes can therefore be enriched by antigen affinity chromatography. The plasmid pAP1 encodes a fusion protein of an Fv-antibody with a bacterial cell-wall protein. Bacteria carrying this plasmid express functional Fv-antibodies tightly bound to their surface. This should enable the selection of single cells with a fluorescence-assisted cell sorter (FACS) using labeled antigen or by adsorption to immobilized antigen. These vectors permit three major principles of the antibody response to be mimicked inE. coli:

1. Generation of a highly complex antibody repertoire;
2. Clonal selection procedures for library screening; and
3. The possibility of increasing a given affinity by repeated rounds of mutation and selection.

     

Toxicity and mutual interactions of cadmium and zinc ions in normal and carcinogen-transformed mouse cells

The toxicity of chloride salts of physiological (zinc, manganese, nickel) and non physiological (cadmium) bivalent metal ions was studied in normal or carcinogen-transformed mouse embryo fibroblast cells. The dose response curves for toxicity to both types of cells exhibited similar shapes. The transformed cells, however, were about twice as sensitive to zinc toxicity as normal cells. When normal and transformed cells were grown together and incubated for several hours with an appropriate concentration of zinc, the malignant cells were selectively killed. Cadmium was much more toxic than the three other metal ions in both types of cells. Its toxic effect was reversed by simultaneous addition of zinc at nontoxic concentrations.Abbrevications CFA colony forming ability - MCA 3-methylcholanthrene     

Has treatment for childhood gastroenteritis changed?

Because so many children with gastroenteritis in our area were being treated with drugs, which are potentially harmful, we assessed the extent of treatment before admission to hospital of 288 children. Sixty four had been treated: 45 with antibiotic, antidiarrhoeal, or antiemetic drugs and 34 had been given glucose-electrolyte solution, 15 of those had also been given drugs; 119 had had no treatment. Since 1979 there has been a decrease in the use of drugs for gastroenteritis, but glucose-electrolyte mixtures are still underused.     

Splenic hypertrophy and extramedullary hematopoiesis induced in male Syrian hamsters by short photoperiod or melatonin injections and reversed by melatonin pellets or pinealectomy

Adult male Syrian hamsters either placed in a short photoperiod alone or kept in a long photoperiod and given daily afternoon injections of the pineal indole melatonin (25 micrograms) exhibited splenic hypertrophy and extramedullary hematopoiesis in addition to a marked regression in testicular weight. The testicular regression as well as the changes in spleen weight and histology could be prevented if the animals in short photoperiod were either pinealectomized or implanted subcutaneously with a pellet containing 1 mg melatonin. Female Syrian hamsters given afternoon injections of melatonin for 7 or 12 weeks had ovaries devoid of corpora lutea; additionally, these animals had reduced relative spleen weights compared to the control animals. In conclusion, it is apparent that spleen weight varies with the functional status of the gonads. Splenic hypertrophy accompanied by pineal-induced testicular regression in males may be related to splenic extramedullary hematopoiesis.     

Immune activation and CD8+ T-cell differentiation towards senescence in HIV-1 infection

Progress in the fight against the HIV/AIDS epidemic is hindered by our failure to elucidate the precise reasons for the onset of immunodeficiency in HIV-1 infection. Increasing evidence suggests that elevated immune activation is associated with poor outcome in HIV-1 pathogenesis. However, the basis of this association remains unclear. Through ex vivo analysis of virus-specific CD8⁺ T-cells and the use of an in vitro model of naïve CD8⁺ T-cell priming, we show that the activation level and the differentiation state of T-cells are closely related. Acute HIV-1 infection induces massive activation of CD8⁺ T-cells, affecting many cell populations, not only those specific for HIV-1, which results in further differentiation of these cells. HIV disease progression correlates with increased proportions of highly differentiated CD8⁺ T-cells, which exhibit characteristics of replicative senescence and probably indicate a decline in T-cell competence of the infected person. The differentiation of CD8⁺ and CD4⁺ T-cells towards a state of replicative senescence is a natural process. It can be driven by excessive levels of immune stimulation. This may be part of the mechanism through which HIV-1-mediated immune activation exhausts the capacity of the immune system.     

Duabanga-like leaves from the Middle Eocene Princeton chert and comparative leaf histology of Lythraceae sensu lato

A permineralized lythraceous leaf type found in close association with fruits, stems, and roots of Decodon allenbyensis Cevallos-Ferriz et Stockey in the Middle Eocene Princeton chert of British Columbia, Canada, is described. Midribs have a prominent C-shaped midvein surrounded by sclerenchyma, with an adaxial epidermis of rectangular to rounded cells lacking enlarged mucilage cells. Leaves are dorsiventral, 180-270 μm thick at the lamina, with a double palisade layer. Abaxial epidermal cells have prominent papillae, and these epidermal cells can be infected by fungi, forming dark sterile stromata. Fossil leaves are similar to those of Myrtales and are compared to those of Lythraceae sensu lato. Although these leaves are thought to belong to the previously described Decodon allenbyensis found in the same chert layer, they lack the diagnostic features of extant Decodon leaves. Instead they share most anatomical similarities with Duabanga grandiflora Roxburgh ex DC Walpers (Lythraceae, subfamily Duabangoideae) including vascular tissues, palisade and spongy mesophyll, bundle fibers, and abaxial epidermal papillae. Duabanga grandiflora differs from the fossil in having mucilaginous cells and a consistently V-shaped abaxial midrib. Although anatomically similar to Duabanga, the fossil leaves are considered those of D. allenbyensis, based on association and the depositional environment prior to preservation. Recent phylogenetic analyses place Duabanga and Decodon in separate clades within Lythraceae, but relationships between these clades are not well supported, indicating that fossil leaves should provide useful anatomical characters for elucidating relationships within Lythraceae.