Schistosoma mansoni: microfluorometric determination of circulating anodic antigen and antigen-antibody complexes in infected hamster serum

In this study the applicability of the defined antigen substrate spheres (DASS) system for the detection of circulating anodic antigen (CAA) in experimental Schistosoma mansoni infections in hamsters and mice is described. Using the DASS system as an antibody-antigen-labeled antibody technique, CAA could be detected both as a free circulating antigen and as the antigen-component of circulating antigen-antibody complexes.     

Mixotrophic growth of Thiobacillus A2 on acetate and thiosulfate as growth limiting substrates in the chemostat

During heterotrophic growth on acetate, in batch culture, the autotrophic growth potential of Thiobacillus A2, i.e. the capacity to oxidize thiosulfate and to fix carbon dioxide via the Calvin cycle, was completely repressed. The presence of thiosulfate in a batch culture with acetate as the organic substrate partly released the repression of the thiosulfate oxidizing system. Cultivation of the organism in continuous culture at a dilution rate of 0.05 h^-1 with different concentration ratios of thiosulfate and acetate in the reservoir medium led to mixotrophic growth under dual substrate limitation. Growth on the different mixtures of acetate and thiosulfate yielded upto 30% more cell dry weight than predicted from the growth yields on comparable amounts of these substrates separately. The extent to which the carbon dioxide fixation capacity and the maximum thiosulfate and acetate oxidation capacity are repressed appeared to be a function of the thiosulfate to acetate concentration ratio in the reservoir medium. The results of ¹⁴C-acetate assimilation experiments and of gas-analysis demonstrated that the extent to which acetate was assimilated depended also on the substrate ratio in the inflowing medium. Under the different growth conditions surprisingly little variation was found in some tri-carboxylic acid cycle enzyme activities. Cultivation of T. A2 at different growth rates with a fixed mixture of thiosulfate (18 mM) and acetate (11 mM) in the medium, showed that dual substrate limitation occured at dilution rates ranging from 0.03–0.20 h^-1.Abbreviations PPO 2,5-diphenoloxazol - RubPCase Ribulose-1,5-bisphophate carboxylase - Tris tris (hydroxymethyl) aminomethane - EDTA ethylenediaminetetra-acetic acid     

Observations on substrate preference of marine phytal harpacticoids (copepoda)

Field observations show that some marine macro-algae harbour large numbers of harpacticoid copepods while other species occupying similar zones of the shore harbour none or only small numbers. The possibility of active substrate selection of specific algae was investigated in Porcellidium dilatatum (Porcellidiidae) by providing it with a choice of chemical stimuli. P. dilatatum was significantly (P 0.05) attracted to Zonaria turneriana the alga it is primarily associated with in nature. The attractive property of the seaweed was reduced and destroyed by physico-chemical treatments. The nature of the attraction is presumed to lie in the interaction between the macrophytic substrate and the surface microbial film.     

Calling and Duetting Behavior in the Leafhopper <Emphasis Type="Italic">Balclutha incisa</Emphasis> (Hemiptera: Cicadellidae: Deltocephalinae): Opportunity for Female Choice?

Male Balclutha incisa (Cicadellidae: Deltocephalinae) produces substrate-borne signals as well as sounds produced by wing fluttering. Males produce 6 signal types that are associated with different behavioral activities: (i) cleaning, (ii) calling, (iii) precopulatory, (iv) copulation, (v) postcopulatory behavior, and (vi) aggression. Within each category signals have a high degree of stereotypy in regard to their temporal structure. Females produce two kinds of signals: (i) a duetting signal in response to the male, and (ii) an introductory or preparatory signal before the main signal. The sexes duet, with the female placing its signal immediately after the first two components of the males call, with an interval of some 136 ms. We discuss the role of the male calls in attracting females and the importance of the duet in maintaining a temporary pair-bond.     

Identification of IS elements in <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis> strains grown in a medium with ferrous iron or adapted to elemental sulfur

IS elements were identified in the genomes of five Acidithiobacillus ferrooxidans strains isolated from various media. IST2 elements were revealed in all the strains grown in a medium with ferrous iron, ISAfe1 elements were detected in four strains (TFBk, TFL-2, TFV-1 and TFO). Three strains (TFV-1, TFN-d and TFO) were found to contain IS elements, ~600 bp long. These were named preliminary as ISAfe600. Partial sequencing of the 5′- and 3′-terminal nucleotide stretches of an ISAfe1 element in TFBk and TFL-2 strains and complete sequencing of the ISAfe1 element in the TFBk strain has revealed nucleotide substitutions as compared to the prototype, i.e., the ISAfe1 element of an ATCC 19859 strain. Partial sequencing of the 5′- and 3′-terminal nucleotide stretches of the IST2 elements in TFO, TFBk and TFL-2 strains has shown numerous nucleotide substitutions when compared to the IST2 element of an ATCC 19859 strain. Complete sequencing of the IST2 element in the TFBk strain has revealed: the divergence between the IST2 elements in the TFBk strain and the prototype was 21.2%. Southern hybridization of EcoRI fragments of the chromosomal DNA from five A. ferrooxidans strains grown in a medium with ferrous iron using an internal region of ISAfe1, a full-length ISAfe1 or a full-length IST2 as probes has shown them to differ in the number of copies of IS elements and their localization on the chromosomes. Adaptation to elemental sulfur in A. ferrooxidans strains caused changes in the number, intensity and localization of hybridization bands. The authors discuss the role of IS elements in the adaptation of A. ferrooxidans to the new energy substrate. The nucleotide sequence data reported in this paper were deposited in GenBank under accession numbers: AY823401, the ISAfe1 from A. ferrooxidans TFBk; AY825254, the IST2 from TFBk; DQ002894, the 5′-terminal nucleotide sequence of ISAfe1 from TFL-2; DQ002895, the 3′-terminal nucleotide sequence of ISAfe1 from TFL-2; DQ005952, the 5′-terminal nucleotide sequence of IST2 from TFV-1; DQ005953, the 3′-terminal nucleotide sequence of IST2 from TFV-1.     

Effects of phorophyte determinants on lichen abundance in the <Emphasis Type="Italic">cerrado</Emphasis> of central Brazil

Previous studies have indicated that epiphytic lichens can be good bioindicators of fire history in tropical savannas. A Lichen Fire History (LFH) Key was developed to assess fire history in areas of cerrado (savanna) in central Brazil. However, the effectiveness and reliability of the LFH Key is much influenced by other lichen determinants. The aim of this study, therefore, was to investigate some of these factors in more detail, thereby allowing better estimates of fire history using the LFH Key. Fieldwork was carried out at the Reserva Ecológica do IBGE, 33 km outside Brasilia D.F., in plots of cerrado denso of varying fire history. Vegetation sampling took place in 20 × 20 m-quadrats within which measurements of the lichen abundance, scorch and phorophyte characteristics, including height, girth and tortuosity, were recorded for all the phorophytes encountered. Bark samples were collected from common cerrado phorophytes and tested for pH, conductivity, moisture content and absorbing capacity, texture and nutrient content. The results show that the greater the impact of fire, the lower the influence of other factors, such as bark characteristics, on the lichens. The strongest determinants of lichens in areas subjected to rare fires or protected from fire are bark aluminium content, bark pH, and microclimatic factors. Using the information gathered from the study, phorophyte species are grouped in terms of their reliability for use in the LFH Key. This study highlights the range of factors which can affect lichen abundance in the tropics, and the relationships between them.     

Tyrosine 769 of the keratinocyte growth factor receptor is required for receptor signaling but not endocytosis

Keratinocyte growth factor receptor (KGFR) is a receptor tyrosine kinase expressed on epithelial cells which belongs to the family of fibroblast growth factor receptors (FGFRs). Following ligand binding, KGFR is rapidly autophosphorylated on specific tyrosine residues in the intracellular domain, recruits substrate proteins, and is rapidly internalized by clathrin-mediated endocytosis. The role of different autophosphorylation sites in FGFRs, and in particular the role of the tyrosine 766 in FGFR1, first identified as PLCgamma binding site, has been extensively studied. We analyzed here the possible role of the tyrosine 769 in KGFR, corresponding to tyrosine 766 in FGFR1, in the regulation of KGFR signal transduction and MAPK activation as well as in the control of the endocytic process of KGFR. A mutant KGFR in which tyrosine 769 was substituted by phenylalanine was generated and transfected in NIH3T3 and HeLa cells. Our results indicate that tyrosine 769 is required for the binding to KGFR and tyrosine phosphorylation of PLCgamma as well as for the full activation of MAPKs and for cell proliferation through the regulation of FRS2 tyrosine phosphorylation, suggesting that this residue represents a key regulator of KGFR signal transduction. Our data also show that tyrosine 769 is not involved in the regulation of the endocytic process of KGFR.     

The substrate specificity of SARS coronavirus 3C-like proteinase

The 3C-like proteinase of severe acute respiratory syndrome coronavirus (SARS) has been proposed to be a key target for structural based drug design against SARS. We have designed and synthesized 34 peptide substrates and determined their hydrolysis activities. The conserved core sequence of the native cleavage site is optimized for high hydrolysis activity. Residues at position P4, P3, and P3' are critical for substrate recognition and binding, and increment of beta-sheet conformation tendency is also helpful. A comparative molecular field analysis (CoMFA) model was constructed. Based on the mutation data and CoMFA model, a multiply mutated octapeptide S24 was designed for higher activity. The experimentally determined hydrolysis activity of S24 is the highest in all designed substrates and is close to that predicted by CoMFA. These results offer helpful information for the research on the mechanism of substrate recognition of coronavirus 3C-like proteinase.     

Substrate uptake tests and quantitative FISH show differences in kinetic growth of bulking and non-bulking activated sludge

The competition between filaments and floc formers in activated sludge has been historically described using kinetic selection. However, recent studies have suggested that bacterial storage may also be an important factor in microbial selection, since the dynamic nature of substrate flows into wastewater treatment plants elicit transient responses from microorganisms. Respirometry-based kinetic selection should thus be reevaluated by considering cell storage, and a more reliable method should be developed to include bacterial storage in the analysis of growth of filaments and floc formers in activated sludge. In this study, we applied substrate uptake tests combined with metabolic modeling to determine the growth rates, yields and maintenance coefficients of bulking and non-bulking activated sludge developed in lab scale reactors under feast and famine conditions. The results of quantitative fluorescence in situ hybridization (FISH) showed that the filaments Eikelboom Type 1851, Type 021N, and Thiothrix nivea were dominant in bulking sludge, comprising 42.0 % of mixed liquor volatile suspended solids (MLVSS), with 61.6% of the total filament length extending from flocs into bulk solution. Only low levels of Type 1851 filament length (4.9% of MLVSS) occurred in non-bulking sludge, 83.0% of which grew inside the flocs. The kinetic parameters determined from the substrate uptake tests were consistent with those from respirometry and showed that filamentous bulking sludge had lower growth rates and maintenance coefficients than non-bulking sludge. These results provide support for growth kinetic differences in explaining the competitive strategy of filamentous bacteria.     

Species composition and assemblage structure of chironomid larvae (Diptera: Chironomidae) attaching to the artificial substrates in a Japanese temperate basin,in relation to the longitudinal gradient

The relative importance of natural and anthropogenic factors, especially topographic type, riparian canopy, altitude, temperature and bank protection, on larval chironomid assemblage was investigated in a Japanese basin. To focus on the macro-scale factors, a concrete block, as an artificial substrate, was used for chironomid collection so that sampling regime may be identical among the sites. Partial CCA using sampling month as a covariable revealed that topographic type, riparian canopy coverage, water temperature and altitude were the main factors influencing species distribution. Stempellinella tamaseptima, Polypedilum tamanigrum and five Rheotanytarsus species showed positive, whereas five Cricotopus species showed negative associations with canopy coverage. Some traditional longitudinal zonations of species were still shown. Chironomus flaviplumus and Chironomus yoshimatsui were merely associated with lower reaches. Stepwise multiple regressions of the assemblage indices on the environmental variables were applied. Bank protection and depth showed negative correlations with Shannon diversity H′. Both topographic type and depth showed negative correlations with Pielou equitability J. Topographic type (lower reach) and specific conductance showed positive, while bank protection showed a negative correlation with abundance. Species richness was not explained by any variables. As a whole, topographic type was the most directly related factor to chironomid assemblages.