A revision of Microbothriorhynchus Yamaguti, 1952 (Cestoda: Trypanorhyncha), with the redescription of M coelorhynchi Yamaguti, 1952 and the description of M reimeri n sp

The genus Microbothriorhynchus Yamaguti, 1952 is revised. Plerocerci of M. coelorhynchi Yamaguti, 1952 from the body-cavity of Brotula barbata (Bloch & Schneider) off the Angolan coast are re-described, and M. reimeri n. sp. from the body-cavity of Helicolenus maculatus (Cuvier) and Caelorinchus parallelus (Günther) from coastal waters off Mozambique is added to the genus. Both species are characterised in having an elongated, slightly craspedote scolex with small, collar-like bothridia, an elongate pars vaginalis and very long bulbs. The tentacular armature is heteroacanthous atypica, with eight principal hooks and intercalary hooks merging with a band of spiniform hooks on the external tentacle surface. The adult is unknown. Microbothriorhynchus is allocated to the Lacistorhynchidae Guiart, 1927, in having a blastocyst, two bothridia and a heteroacanthous atypica armature. Dasyrhynchus Pintner, 1928 and Pseudogrillotia Dollfus, 1969 are considered the most closely related genera, sharing characters such as the craspedote scolex, similar scolex proportions and hook patterns. Microbothriorhynchus appears to link the heteroacanthous atypica genera Grillotia Guiart, 1927 and Pseudogrillotia with the poeciloacanthous genus Dasyrhynchus.     

Mutational spectrum in the cardioauditory syndrome of Jervell and Lange-Nielsen

Jervell and Lange-Nielsen syndrome (JLNS) is an autosomal recessive syndrome characterised by profound congenital sensorineural deafness and prolongation of the QT interval on the electrocardiogram, representing abnormal ventricular repolarisation. In a study of ten British and Norwegian families with JLNS, we have identified all of the mutations in the KCNQ1 gene, including two that are novel. Of the nine mutations identified in this group of 10 families, five are nonsense or frameshift mutations. Truncation of the protein proximal to the recently identified C-terminal assembly domain is expected to preclude assembly of KCNQ1 monomers into tetramers and explains the recessive inheritance of JLNS. However, study of a frameshift mutation, with a dominant effect phenotypically, suggests the presence of another assembly domain nearer to the N-terminus.     

Crystal structure of RNA 3'-terminal phosphate cyclase, a ubiquitous enzyme with unusual topology

BACKGROUND: RNA cyclases are a family of RNA-modifying enzymes that are conserved in eucarya, bacteria and archaea. They catalyze the ATP-dependent conversion of the 3'-phosphate to the 2',3'-cyclic phosphodiester at the end of RNA, in a reaction involving formation of the covalent AMP-cyclase intermediate. These enzymes might be responsible for production of the cyclic phosphate RNA ends that are known to be required by many RNA ligases in both prokaryotes and eukaryotes. RESULTS: The high-resolution structure of the Escherichia coli RNA 3'-terminal phosphate cyclase was determined using multiwavelength anomalous diffraction. Two orthorhombic crystal forms of E. coli cyclase (space group P2(1)2(1)2(1) and P2(1)2(1)2) were used to solve and refine the structure to 2.1 A resolution (R factor 20.4%; R(free) 27.6%). Each molecule of RNA cyclase consists of two domains. The larger domain contains three repeats of a folding unit comprising two parallel alpha helices and a four-stranded beta sheet; this fold was previously identified in translation initiation factor 3 (IF3). The large domain is similar to one of the two domains of 5-enolpyruvylshikimate-3-phosphate synthase and UDP-N-acetylglucosamine enolpyruvyl transferase. The smaller domain uses a similar secondary structure element with different topology, observed in many other proteins such as thioredoxin. CONCLUSIONS: The fold of RNA cyclase consists of known elements connected in a new and unique manner. Although the active site of this enzyme could not be unambiguously assigned, it can be mapped to a region surrounding His309, an adenylate acceptor, in which a number of amino acids are highly conserved in the enzyme from different sources. The structure of E. coli cyclase will be useful for interpretation of structural and mechanistic features of this and other related enzymes.     

Canonical variables of aquatic bryophyte combinations for predicting water trophic level

Concentrations of soluble reactive phosphorus (SRP), nitrate, and soluble reactive silicon (SRSi) were monitored in 12 streams draining small catchments (<10 km2) in the English Lake District. The catchments varied with respect to underlying geology, soil type and land cover. Average concentrations of SRP were in the range 0.5–11.2 μg P l-1, and estimated loads ranged from 0.01 to 0.14 kg P ha-1 a-1. The higher concentrations and loads were associated with catchments containing improved pasture. Mean streamwater concentrations of nitrate varied from 55 to 660 μg N l-1, while loads were in the range 0.8–9.6 kg N ha-1 a-1; no general dependence on catchment properties was discerned. Concentrations of SRSi were similar in all the streams (0.8–2 mg Si l-1), and annual loads were in the range 10–26 kg Si ha-1 a-1. Loads of all three nutrients were greatest during the winter, because of higher discharges, but in some catchments containing improved pasture, considerable transport of P also took place during the summer. Concentrations of nitrate in streams draining unimproved moorland catchments are approximately twice those reported for samples taken from similar streams in 1973 and 1974, possibly because of increased atmospheric deposition of inorganic nitrogen (ammonium and nitrate). Concentrations of SRP in such streams were similar to those reported for the earlier samples. Comparisons of stream loads of SRP and nitrate with estimated inputs suggest that catchment soils retain substantial amounts of these nutrients. Implications for surface water eutrophication of changes in P retention by soils are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Soil phosphorus fractions and adsorption as affected by organic and inorganic sources

The effect of organic and inorganic sources of phosphorus (P) on soil P fractions and P adsorption was studied in a field without plant growth on a Kandiudalf in western Kenya. A high-quality organic source, Tithonia diversifolia (Hemsley) A. Gray leaves, and a low-quality source, maize (Zea mays L.) stover, were applied alone or in combination with triple superphosphate (TSP). The P rate was kept constant at 15 kg P ha^-1. Soil extractable P (resin, bicarbonate and sodium hydroxide), microbial biomass P and C and P adsorption isotherms were determined during 16 weeks after application of treatments. Application of tithonia either alone or with TSP increased resin P, bicarbonate P, microbial P, and sodium hydroxide inorganic P. Tithonia alone reduced P adsorption at 2–16 weeks. Maize stover had no effect on any of the P fractions or P adsorption. At 8 weeks, the application of tithonia reduced microbial C-to-P ratio (20) as compared to maize stover, TSP and the control (31–34). The reduction in P adsorption by tithonia was accompanied by increases in all measured P fractions, the sum of P in those fractions (resin, bicarbonate and sodium hydroxide) being larger than the P added. The reduction in P adsorption apparently resulted from competition for adsorption sites, probably by organic anions produced during decomposition of the high quality tithonia. Integration of inorganic P (TSP) with organic materials had little added benefit compared to sole application of TSP, except that combination of tithonia with TSP increased microbial biomass. The results indicate that a high quality organic input can be comparable to or more effective than inorganic P in increasing P availability in the soil.     

The role of the rock cod Notothenia coriiceps Richardson, 1844 in the life-cycle of Antarctic parasites

Fifty specimens of Notothenia coriiceps caught in Potter Cove, King George Island, were examined for ecto- and endoparasites. Of the 22 parasite species found, 18 were helminths, 2 were hirudineans and 2 were crustaceans. The isopod Aega antarctica and an unidentified hirudinean are reported for the first time from this fish host. Dominant parasites were the adults of Aspersentis megarhynchus, the invasive stage of Corynosoma spp. (cystacanth) and the adults of Macvicaria pennelli, with respective prevalences of infestation of 94, 76 and 74%. The preferred sites of infestation were the pylorus and intestine, where five different larval (nematodes and cestodes) and eight adult (digeneans and acanthocephalans) parasite species were found. No adult nematodes and cestodes were found and no parasites could be isolated from the musculature. The results of the present study are related to previous findings on the parasite fauna of N. coriiceps. The comparison implies a high parasite diversity in this benthic Antarctic fish species. Most parasites found appear to have a wide range of distribution within Antarctic waters together with a low host specificity. Besides its role as final host for several species of trematodes and acanthocephalans, N. coriiceps serves as transmitter of parasite larvae to piscivorous birds and seals. It is concluded that the parasite fauna in Antarctic fish species provides important insights into the different habitat use and trophic relationship of their fish hosts. Received: 11 September 1997 / Accepted: 12 January 1998     

Binding Affinity,Specificity and Comparative Biodistribution of the Parental Murine Monoclonal Antibody MX35 (Anti-NaPi2b) and Its Humanized Version Rebmab200

The aim of this preclinical study was to evaluate the characteristics of the monoclonal antibody Rebmab200, which is a humanized version of the ovarian-specific murine antibody MX35. This investigation contributes to the foundation for future clinical α-radioimmunotherapy of minimal residual ovarian cancer with ²¹¹At-Rebmab200. Here, the biodistribution of ²¹¹At-Rebmab200 was evaluated, as was the utility of ^99mTc-Rebmab200 for bioimaging. Rebmab200 was directly compared with its murine counterpart MX35 in terms of its in-vitro capacity for binding the immobilized NaPi2B epitope and live cells; we also assessed its biodistribution in nude mice carrying subcutaneous OVCAR-3 tumors. Tumor antigen and cell binding were similar between Rebmab200 and murine MX35, as was biodistribution, including normal tissue uptake and in-vivo tumor binding. We also demonstrated that ^99mTc-Rebmab200 can be used for single-photon emission computed tomography of subcutaneous ovarian carcinomas in tumor-bearing mice. Taken together, our data support the further development of Rebmab200 for radioimmunotherapy and diagnostics.     

Short-term adaptation during propagation improves the performance of xylose-fermenting <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> in simultaneous saccharification and co-fermentation

Background

Inhibitors that are generated during thermochemical pretreatment and hydrolysis impair the performance of microorganisms during fermentation of lignocellulosic hydrolysates. In omitting costly detoxification steps, the fermentation process relies extensively on the performance of the fermenting microorganism. One attractive option of improving its performance and tolerance to microbial inhibitors is short-term adaptation during propagation. This study determined the influence of short-term adaptation on the performance of recombinant Saccharomyces cerevisiae in simultaneous saccharification and co-fermentation (SSCF). The aim was to understand how short-term adaptation with lignocellulosic hydrolysate affects the cell mass yield of propagated yeast and performance in subsequent fermentation steps. The physiology of propagated yeast was examined with regard to viability, vitality, stress responses, and upregulation of relevant genes to identify any links between the beneficial traits that are promoted during adaptation and overall ethanol yields in co-fermentation.

Results

The presence of inhibitors during propagation significantly improved fermentation but lowered cell mass yield during propagation. Xylose utilization of adapted cultures was enhanced by increasing amounts of hydrolysate in the propagation. Ethanol yields improved by over 30 % with inhibitor concentrations that corresponded to ≥2.5 % water-insoluble solids (WIS) load during the propagation compared with the unadapted culture. Adaptation improved cell viability by >10 % and increased vitality by >20 %. Genes that conferred resistance against inhibitors were upregulated with increasing amounts of inhibitors during the propagation, but the adaptive response was not associated with improved ethanol yields in SSCF. The positive effects in SSCF were observed even with adaptation at inhibitor concentrations that corresponded to 2.5 % WIS. Higher amounts of hydrolysate in the propagation feed further improved the fermentation but increased the variability in fermentation outcomes and resulted in up to 20 % loss of cell mass yield.

Conclusions

Short-term adaptation during propagation improves the tolerance of inhibitor-resistant yeast strains to inhibitors in lignocellulosic hydrolysates and improves their ethanol yield in fermentation and xylose-fermenting capacity. A low amount of hydrolysate (corresponding to 2.5 % WIS) is optimal, whereas higher amounts decrease cell mass yield during propagation.

     

Structure of the Ni(II) complex of Escherichia coli peptide deformylase and suggestions on deformylase activities depending on different metal(II) centres

Crystal structures of polypeptide deformylase (PDF) of Escherichia coli with nickel(II) replacing the native iron(II) have been solved with chloride and formate as metal ligands. The chloro complex is a model for the correct protonation state of the hydrolytic hydroxo ligand and the protonated status of the Glu133 side chain as part of the hydrolytic mechanism. The ambiguity that recently some PDFs have been identified with Zn²⁺ ion as the active-site centre whereas others are only active with Fe²⁺ (or Co²⁺, Ni²⁺) is discussed with respect to Lewis acid criteria of the metal ion and substrate activation by the CD loop.