Perchlorate Remediation by Electrokinetic Extraction and Electrokinetic Injection of Substrates

Perchlorate (ClO₄^-) contamination of groundwater has recently become a major concern across the nation. Electrokinetic (EK) extraction with the simultaneous EK injection of organic material to promote degradation could allow for the efficient removal of perchlorate while simultaneously promoting degradation of perchlorate. Column experiments were conducted to evaluate the technology. Lactate and glycine served as organic substrates to promote degradation after injection into the columns as well as maintaining the pH near neutral. Removal of perchlorate from contaminated materials kaolin, sand, and a natural soil historically contaminated by perchlorate was controlled by the ionic flux of perchlorate and not by transport from the osmotic flux which was only significant for kaolin experiments. Perchlorate was removed from contaminated sand and clay below our detection limits (5 ppb). Both lactic acid and glycine were successfully injected into clay and a sand matrix. Results from a contaminated site soil indicate that the Chemical Oxygen Demand was increased after electrokinetic injection of glycine and lactate. Experiments using soil from a contaminated site confirmed that EK can be used to both remove perchlorate and stimulate bioremediation by the injection of lactate or glycine. The use of EK technology to both remove and provide for continued source removal by bioremediation offers a potential new tool to treat low permeability systems.     

Use of otolith microchemistry to identify subbasin natal origin and use by invasive Lake Trout in Yellowstone Lake

Hydrobiologia - Nonindigenous lake trout (Salvelinus namaycush) expansion in Yellowstone Lake has led to a large decline in the native Yellowstone cutthroat trout (Oncorhynchus clarkii bouvieri)...     

Receptor-ligand requirements for increased NK cell polyfunctional potential in slow progressors infected with HIV-1 coexpressing KIR3DL1*h/*y and HLA-B*57

Carriage of the natural killer (NK) receptor genotype KIR3DL1*h/*y with its HLA-B*57 ligand (*h/*y+B*57) is associated with slow time to AIDS and low viral load (VL). To provide a functional basis for these epidemiological observations, we assessed whether HIV-1-infected slow progressors (SP) carrying the *h/*y+B*57 compound genotype would have increased NK cell polyfunctional potential in comparison to SP with other killer immunoglobulin-like receptor (KIR)/HLA compound genotypes and whether this enhanced polyfunctionality was dependent upon the coexpression of both KIR3DL1*h/*y and HLA-B*57. The functional potential of NK cells was investigated by stimulating peripheral blood mononuclear cells with HLA-devoid targets or single HLA transfectants. Multiparametric flow cytometry was used to detect NK cells with seven functional profiles representing all permutations of CD107a expression and gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) secretion. NK cells from individuals carrying KIR3DL1 receptor-HLA-Bw4 ligand pairs had greater trifunctional responses than those from KIR3DL1 homozygotes (hmz), who were Bw6 homozygotes. NK cells from subjects carrying the *h/*y+B*57 genotypes exhibited the highest trifunctional potential, and this was dependent on cocarriage of the NK receptor and its ligand. Trifunctional cells secreted more of each function tested on a per-cell basis than each corresponding monofunctional NK subset. Although VL influenced NK functionality, individuals with defined KIR/HLA genotypes exhibited differences in NK cell polyfunctionality that could not be accounted for by VL alone. The protective effect of HLA-B*57 on slow progression to AIDS and low VL may be mediated through its interaction with KIR3DL1 alleles to educate NK cells for potent activity upon stimulation.     

Homosexual behaviour in wild white-handed gibbons (Hylobates lar)

Homosexual mounting in apes and prosimians is rare. Male-male mounting was observed between an adult male white-handed gibbon and an adolescent male in the same group. The behaviour is discussed in terms of the gibbon group's social structure and the development of the adolescent. It is suggested that this behaviour provides reassurance for the adolescent gibbon.     

Acyl group specificity at the active site of tetrahydridipicolinate N-succinyltransferase

Tetrahydrodipicolinate N-succinyltransferase (DapD) catalyzes the succinyl-CoA-dependent acylation of L-2-amino-6-oxopimelate to 2-N-succinyl-6-oxopimelate as part of the succinylase branch of the meso-diaminopimelate/lysine biosynthetic pathway of bacteria, blue-green algae, and plants. This pathway provides meso-diaminopimelate as a building block for cell wall peptidoglycan in most bacteria, and is regarded as a target pathway for antibacterial agents. We have solved the X-ray crystal structures of DapD in ternary complexes with pimelate/succinyl-CoA and L-2-aminopimelate with the nonreactive cofactor analog, succinamide-CoA. These structures define the binding conformation of the cofactor succinyl group and its interactions with the enzyme and place its thioester carbonyl carbon in close proximity to the nucleophilic 2-amino group of the acceptor, in support of a direct attack ternary complex mechanism. The acyl group specificity differences between homologous tetrahydrodipicolinate N-acetyl- and N-succinyltransferases can be rationalized with reference to at least three amino acids that interact with or give accessible active site volume to the cofactor succinyl group. These residues account at least in part for the substrate specificity that commits metabolic intermediates to either the succinylase or acetylase branches of the meso-diaminopimelate/lysine biosynthetic pathway.     

Surfactant aggregates (solloids) adsorbed on silica as stationary chromatographic phases: structures and properties

The structure and physical properties of solloids (surfactant aggregates adsorbed on surfaces) adsorbed on particles are of general interest. The relationship between solloid structure and properties of hexadecyltrimethylammonium bromide (HTAB), cetylpyridinium chloride (CPC) and cetylpyridinium salicylate (CPS) adsorbed on silica particles was studied by electron paramagnetic resonance (EPR) spectroscopy using the spin-probes peroxylaminedisulfonate (PADS) and 4-[N,N-dimethyl-N-(n-hexadecyl)ammonium]-2,2,6,6-tetramethylpiperidinyl-N-oxy bromide (HTAB*). Using HTAB* incorporated in HTAB, CPC and CPC solloids and comparing the results to those in micelles, it was determined that for silica around pH 4 the solloids are very similar in properties to the micelles. This is consistent with a linear solvation–energy relationship (LSER) analysis of solute equilibration data which indicates that at pH 5 HTAB solloids have similar properties to HTAB micelles. The PADS spin-probe appears to be more sensitive to changes in the properties of the double layer, and substantial differences were observed between HTAB, CPC and CPS and as a function of HTAB concentration for HTAB solloids on silica.     

A Single Amino Acid Substitution Prevents Recognition of a Dominant Human Aquaporin-4 Determinant in the Context of HLA-DRB1*03:01 by a Murine TCR

BackgroundAquaporin 4 (AQP4) is considered a putative autoantigen in patients with Neuromyelitis optica (NMO), an autoinflammatory disorder of the central nervous system (CNS). HLA haplotype analyses of patients with NMO suggest a positive association with HLA-DRB1* 03:01. We previously showed that the human (h) AQP4 peptide 281–300 is the dominant immunogenic determinant of hAQP4 in the context of HLA-DRB1*03:01. This immunogenic peptide stimulates a strong Th₁ and Th₁₇ immune response. AQP4_281-300-specific encephalitogenic CD4⁺ T cells should initiate CNS inflammation that results in a clinical phenotype in HLA-DRB1*03:01 transgenic mice.MethodsControlled study with humanized experimental animals. HLA-DRB1*03:01 transgenic mice were immunized with hAQP4_281-300, or whole-length hAQP4 protein emulsified in complete Freund’s adjuvant. Humoral immune responses to both antigens were assessed longitudinally. In vivo T cell frequencies were assessed by tetramer staining. Mice were followed clinically, and the anterior visual pathway was tested by pupillometry. CNS tissue was examined histologically post-mortem. Flow cytometry was utilized for MHC binding assays and to immunophenotype T cells, and T cell frequencies were determined by ELISpot assay.ResultsImmunization with hAQP4_281-300 resulted in an in vivo expansion of antigen-specific CD4⁺ T cells, and an immunoglobulin isotype switch. HLA-DRB1*03:01 TG mice actively immunized with hAQP4_281-300, or with whole-length hAQP4 protein were resistant to developing a neurological disease that resembles NMO. Experimental mice show no histological evidence of CNS inflammation, nor change in pupillary responses. Subsequent analysis reveals that a single amino acid substitution from aspartic acid in hAQP4 to glutamic acid in murine (m)AQP4 at position 290 prevents the recognition of hAQP4_281-300 by the murine T cell receptor (TCR).ConclusionInduction of a CNS inflammatory autoimmune disorder by active immunization of HLA-DRB1*03:01 TG mice with human hAQP4_281-300 will be complex due to a single amino acid substitution. The pathogenic role of T cells in this disorder remains critical despite these observations.     

Amyloplast sedimentation dynamics in maize columella cells support a new model for the gravity-sensing apparatus of roots

Quantitative analysis of statolith sedimentation behavior was accomplished using videomicroscopy of living columella cells of corn (Zea mays) roots, which displayed no systematic cytoplasmic streaming. Following 90 degrees rotation of the root, the statoliths moved downward along the distal wall and then spread out along the bottom with an average velocity of 1.7 microm min(-1). When statolith trajectories traversed the complete width or length of the cell, they initially moved horizontally toward channel-initiation sites and then moved vertically through the channels to the lower side of the reoriented cell where they again dispersed. These statoliths exhibited a significantly lower average velocity than those sedimenting on distal-to-side trajectories. In addition, although statoliths undergoing distal-to-side sedimentation began at their highest velocity and slowed monotonically as they approached the lower cell membrane, statoliths crossing the cell's central region remained slow initially and accelerated to maximum speed once they reached a channel. The statoliths accelerated sooner, and the channeling effect was less pronounced in roots treated with cytochalasin D. Parallel ultrastructural studies of high-pressure frozen-freeze-substituted columella cells suggest that the low-resistance statolith pathway in the cell periphery corresponds to the sharp interface between the endoplasmic reticulum (ER)-rich cortical and the ER-devoid central region of these cells. The central region is also shown to contain an actin-based cytoskeletal network in which the individual, straight, actin-like filaments are randomly distributed. To explain these findings as well as the results of physical simulation experiments, we have formulated a new, tensegrity-based model of gravity sensing in columella cells. This model envisages the cytoplasm as pervaded by an actin-based cytoskeletal network that is denser in the ER-devoid central region than in the ER-rich cell cortex and is linked to stretch receptors in the plasma membrane. Sedimenting statoliths are postulated to produce a directional signal by locally disrupting the network and thereby altering the balance of forces acting on the receptors in different plasma membrane regions.