d-Tagatose production in the presence of borate by resting Lactococcus lactis cells harboring Bifidobacterium longum l-arabinose isomerase

Bifidobacterium longum NRRL B-41409 l-arabinose isomerase (l-AI) was overexpressed in Lactococcus lactis using a phosphate depletion inducible expression system. The resting L. lactis cells harboring the B. longum l-AI were used for production of d-tagatose from d-galactose in the presence of borate buffer. Multivariable analysis suggested that high pH, temperature and borate concentration favoured the conversion of d-galactose to d-tagatose. Almost quantitative conversion (92 %) was achieved at 20 g L^?1 substrate and at 37.5 °C after 5 days. The d-tagatose production rate of 185 g L^?1 day^?1 was obtained at 300 g L^?1 galactose, at 1.15 M borate, and at 41 °C during 10 days when the production medium was changed every 24 h. There was no significant loss in productivity during ten sequential 24 h batches. The initial d-tagatose production rate was 290 g L^?1 day^?1 under these conditions.     

Biodistribution of the chimeric monoclonal antibody U36 radioiodinated with a closo-dodecaborate-containing linker. Comparison with other radioiodination methods

We have evaluated the applicability of the [(4-isothiocyanatobenzylammonio)undecahydro-closo-dodecaborate (1-)] (DABI) linker molecule for antibody radiohalogenation and compared it to radiohalogenation using the linker N-succinimidyl 4-iodobenzoate (PIB) and to direct radiohalogenation using Chloramine T. These studies were performed to assess the potential of DABI conjugates and to optimize the biological properties of halogen-labeled cMAb U36. The three conjugates were evaluated in vitro for their specificity and affinity and in vivo for their biodistribution patterns in normal mice at 1.5, 6, 24, and 96 h pi. Labeling efficiencies of direct CAT labeling, indirect PIB labeling, and indirect DABI labeling were 90-95%, 60%, and 68%, respectively. This resulted in a PIB:cMAb U36 molar ratio of 1.8-2.5 and a DABI:cMAb U36 molar ratio of 4.1. The in vitro data demonstrated specific binding for all conjugates and similar affinities with values around 1 x 10(8) M(-)(1). However, the in vivo data revealed accumulation of the radioiodine uptake in thyroid for the directly labeled conjugate, with a value 10 times higher than the indirectly labeled conjugates 96 h pi. Both the (125)I-PIB-cMAb U36 and (125)I-DABI-cMAb U36 conjugates yielded a low thyroid uptake with no accumulation, indicating different catabolites for these conjugates. This may favor the use of the indirectly labeled conjugates for future studies. Apart from the specific results obtained, these findings also demonstrate how the right linker molecule will provide additional opportunities to further improve the properties of an antibody-radionuclide conjugate.     

Growth of pike larvae: effects of prey, turbidity and food quality

We studied experimentally the effects of turbidity and prey composition on pike larval growth and hypothesized that pike larval growth varies with turbidity and food quality. We reared the first-feeding pike larvae (Esox lucius) in laboratory tanks with (1) clear or (2) turbid water provided with zooplankton rations from (3) an inner and (4) an outer archipelago site. The sites differ in physical features, salinity, eutrophication status, zooplankton community structure and density. Pike larvae showed the highest weight increase in clear water with zooplankton from the outer site and the poorest weight increase in turbid water with zooplankton as prey from the inner site. Our fatty acid analysis revealed that unsaturated fatty acid levels were highest in the outer site. The relative percentage of copepods was also higher in the outer site. This study supports the hypothesis that turbidity weakens the ability of pike larvae to capture certain prey. Further, zooplankton community composition matters in turbid water, but is not a primary factor in clear water.     

Electrophoretic and histochemical studies of carbonic anhydrase activity

Summary A simple method for separation of carbonic anhydrase activity into components by electrophoresis on cellulose acetate strips is described. With this method, using barbiturate buffer systems at various pH values, two main components of CAH in rat erythrocytes, and the splitting of each of these into two minor components were revealed. Two components were also observed in the CAH activity in kidney and lens homogenates, and one component in brain homogenate. A modification of Häusler's histochemical method for CAH was adapted for visualization of the electrophoretically separated bands. This rendered the evalution of the results easier than with the quantitative measurements alone. The quantitative measurement of CAH activity in electrophoretic strips corresponded with the degree of staining by the histochemical method. This among other facts supports the view of the specificity of the histochemical method used. Some examples of the histochemical staining pattern of the CAH activity in rat tissues are given.     

Effects of intensity and duration of infection by a hemiparasitic plant, Rhinanthus serotinus, on growth and reproduction of a perennial grass, Agrostis capillaris

Arising from annual variation in parasitic plant population densities, substantial yearly changes may occur in the parasitic load of an individual perennial host. We conducted two two-year greenhouse pot experiments to examine the effects of varying intensities and duration of infection by an annual root hemiparasitic plant. Rhinanthus serotinus, on the growth and reproduction of its perennial host grass. Agrostis capillaris. In the first experiment, one host plant was growing either alone or under a load of 1 or 3 root hemiparasitic plants for one growing season, and during the next season all hosts continued their life free of hemiparasites. In the second experiment, the host plants either grew alone or were parasitised by 1 or 2 root hemiparasitic plants either during the first growing season only or during two successive seasons (the parasitic load being the same in the two seasons). In both experiments, the root hemiparasites markedly reduced the growth and reproduction of their perennial hosts. In the first experiment, the negative effects of parasites on host performance increased with the increase in intensity of parasitic infection from one to three parasites. The harmful effects of hemiparasitim were carried over to the following season; hosts parasitised during the previous season with one or three parasites produced significantly less biomass than those without parasites. In addition, hosts parasitised by three parasites during the first season produced significantly less panicles in the second season than those parasitised by one parasite and those without parasites. The second experiment showed that the production of biomass of A. capillaris during the second season was, but the production of panicles was not affected by the duration of parasitic infection. In addition, in this experiment, the second season biomass of A. capillaris depended on the intensity of infection (1 vs 2 parasites), but the production of panicles was unaffected by the number of parasites.     

Bacterioplankton in a small polyhumic lake with an anoxic hypolimnion

Bacterioplankton biomass and dark fixation of inorganic carbon were measured in the highly humic (water colour up to 550 mg Pt l^?1) and acidic lake, Mekkojärvi. Strong thermal and chemical stratification developed in the water column early in spring and led rapidly to anoxia in the hypolimnion, which extended to less than 1.0 m from the surface. In the epilimnion only small bacteria were abundant. In the anoxic zone both the abundance and the mean size of bacteria were considerably higher than in the epilimnion. These differences are thought to be the result of different grazing pressure from zooplankton in the two zones. In late summer a high concentration of bacteriochlorophyll d in the upper hypolimnion indicated a high density of photosynthetic bacteria. Bacterial biomass was similar to that of phytoplankton in the epilimnion, but 23 times higher in the whole water column. In August, dark fixation of inorganic radiocarbon in the anaerobic zone was 51% of the total ¹⁴C-incorporation and the contribution of light fixation was only 5.4%. In the polyhumic Mekkojarvi, bacterioplankton was evidently a potentially significant carbon source for higher trophic levels, but bacterioplankton production could not be supported by phytoplankton alone. Allochthonous inputs of dissolved organic matter probably support most of the bacterial production.     

Classifying clonal growth forms based on vegetative mobility and ramet longevity: a whole community analysis

We measured rhizome branching, clonal mobility, and ramet longevity of 98 meadow plant species. A cluster analysis applied to this dataset revealed nine clonal growth types that differ mainly by the ramet lifespan and vegetative mobility. Then we compared the abundance of these groups of clonal species between the three following plant communities: (1) open, (2) restored and (3) overgrown wooded meadows in the Laelatu-Nehatu-Puhtu Nature Reserve, Estonia. This is the first study where the quantitative values of belowground clonal traits have been measured for all species of a species-rich community. We show that species with annual ramets and with a low vegetative mobility were most abundant in open grasslands. The relative abundance of perennial species with annual ramets was positively correlated with shoot density and species diversity, indicating that high ramet turnover rates combined with a high genet longevity can positively affect species coexistence in meadow communities. Hence, this study provides evidence for the fact that the average values of clonal life-history parameters differ between these communities. Herb communities under forest canopy consist, in average, of species with ramets that live longer and are clonally more mobile than in the communities of open sites.     

Infection of human peripheral blood mononuclear cells with a temperature-sensitive mutant of measles virus.

A stable temperature-sensitive mutant of measles virus (MV ts38) was used to study the mechanism of virus-mediated immune suppression of peripheral blood mononuclear cells in vitro. Both unstimulated and phytohemagglutinin-stimulated cultures released infectious virus at 32 degrees C, whereas no virus was released at 37 degrees C, although both viral RNA and viral proteins were synthesized. However, the response of the lymphoid cells to phytohemagglutinin, concanavalin A, and herpes simplex virus antigen was decreased in the presence of MV ts38 at 37 degrees C. The viability of infected cells was not diminished, therefore excluding cell death as a reason for immunosuppression. Interleukin 2 did not play a role in the inhibitory effect of MV ts38. Antibodies to alpha interferon partially reversed the inhibitory effect of the virus infection on lymphocyte mitogenesis, thus implying that alpha interferon plays a role in the immunosuppression. Depletion experiments indicated that adherent cells play a greater role in the measles virus-induced immunosuppression than nonadherent cells. However, monocyte maturation to macrophages had no effect on the degree of immunosuppression.     

Fossil remains of an unknown Alona species (Chydoridae, Aloninae) from a high arctic lake in Nordaustlandet (Svalbard) in relation to glaciation and Holocene environmental history

We analyzed a lacustrine sediment core covering the Holocene from Lake Einstaken, Nordaustlandet, for its fossil Cladocera (Crustacea) with an aim to reconstruct past aquatic communities in this environmentally extreme and unexplored region. In the analysis, we encountered remains (carapaces, ephippia, headshields, and postabdomens) of an unknown chydorid (Chydoridae, Aloninae) species during two separate periods in the early Holocene. The remains had some comparable morphological characters with the European Alona guttata s.str. Sars, 1862 and with the glacial relict Alona werestschagini Sinev, 1999, but they differed clearly from the previous species; the headshield had broadly rounded rostrum and narrow fornices, the ephippium was heavily pigmented and reticulated, and the postabdomen had convex dorsal and ventral margins. The postabdomen had evidently similar morphology with Alona bergi R?en, 1992, which has been described, although inadequately, from arctic Canada and northern Greenland. We conclude, based on the morphology of the postabdomen, that the unknown remains belong to species closely resembling A. bergi, named here Alona cf. bergi, and assume that the species, whether the true A. bergi or some other cryptic species of the A. guttata group, is a postglacial relict of the high arctic adapted to cold climate. Herewith, we emphasize the need for extensive biogeographical investigations into both fossil and intact specimens of chydorids in the arctic.