Elimination of heparan sulfate by heparitinases induces abnormal mesodermal and neural formation in Xenopus embryos

The expression of heparan sulfate glycosaminoglycan (HS-GAG) was examined in Xenopus embryos during the developmental stages. Chemical analysis showed the existence of HS-GAG in the ³⁵S-labeled embryos. By western blot analysis using a specific anti-HS monoclonal antibody, HS-GAG related epitope was found after the neurulation on two protein bands, whose molecular weights were approximately 90 kDa and 100 kDa, respectively. Immunohistochemistry revealed that HS-GAG occurred exclusively in the animal hemisphere in early gastrulae, and then appeared predominantly on the sheath of the neural tube, the notochord and epithelium. To address whether HS-GAG chains contribute to Xenopus embryonic development, we eliminated the embryonic HS-GAG by injecting purified Flavobacterium heparitinases (HSase) into their blastocoels. Most of the injected embryos were aberrant in mesodermal and neural formation, and became acephalic. Histological examination showed that these embryos were completely devoid of the central nervous system and the mesodermal tissues. Neither heat-inactivated heparitinase nor chondroitinase showed such abnormality. The HS-GAG-eliminated embryos showed decreased expression of both muscular and neural-specific markers. These results suggest that HS-GAG plays an indispensable role in establishing the fundamental body plan during early Xenopus development.     

Spatial and Temporal variations of mangrove fish assemblages in Martinique (French West Indies)

A study of the mangrove fish fauna in a bay of Martinique Island (French West Indies) was carried out at different seasons during two consecutive years. fishes were sampled with specific hoop-nets in the coastal areas at 8 stations.A total of 87 species was collected in the bay. Most individuals were represented by small-size specimens and juveniles. The overall species richness varied according to the stations and the sampling periods. The biomass and number of individuals were variable according to the location but remained stable in time. A factor correspondence analysis and a hierarchical clustering with median links were used to follow the evolution of the stations in space and time. Two types of stations were differentiated: the stations characterized by the mangrove and those under the influence of seagrass beds. A seasonal cycle, opposing the dry periods to the others, was observed.Thus, it seems that the use of the mangrove habitat by the fishes is optimized through a complete reorganization of communities in terms of species composition whereas the overall number and biomass remain stable. This model remains valid even for the most constraining biota of the mangrove ecosystem inhabited by a small number of well adapted species.     

Dynamics and distribution of microphytobenthic chlorophyll-a in the Western Scheldt estuary (SW Netherlands)

The temporal dynamics and spatial distribution of microphytobenthic chlorophyll-a in the layer 0–1 cm were determined in the Western Scheldt estuary over the period 1991–1992. Connections between the annually averaged benthic chlorophyll-a and station elevation and sediment composition (as a measure of the hydrodynamic energy caused by currents and waves) were also examined.Microphytobenthic chlorophyll-a showed one main peak in early summer and a smaller peak in autumn. The mean chlorophyll-a concentration of 113 mg Chl-a m^–2 in the upper centimeter is of the same order of magnitude as in other estuarine areas. The average annual primary production of the microphytobenthos has been estimated at 136 g C m^–2 y^–1 The primary production of sediment inhabiting microalgae is at least 17% of the total primary production in the estuary.Considerable differences in annually averaged chlorophyll-a emerges between the stations. These differences are related mainly to the interaction between station elevation and clay content of the sediment.     

Zooplankton as a compound mineralising and synthesizing system: phosphorus excretion

Data on phosphate excretion rates of zooplankton are based on measurements using the pelagic crustacean zooplankton of Lake Vechten and laboratory-cultured Daphnia galeata. In case of Daphnia sp we measured the effects of feeding on P-rich algae and P-poor algae (Scenedesmus) as food on the P-excretion rates at 20°C. The excretion rates of the natural zooplankton community, irrespective of the influence of the factors mentioned, varied by an order of magnitude: 0.025–0.275µg PO₄-Pmg^–1C in zooplankton (C _zp ) h^–1. The temperature accounted for about half the observed variation in excretion rates. The mean excretion rates in the lake, computed for 20°C, varied between 0.141 and 0.260 µg Pmg^–1C _zp h^–1. Based on data of zooplankton biomass in the lake the P-regeneration rates by zooplankton covered between 22 and 239% of the P-demand of phytoplankton during the different months of the study period.In D. galeata, whereas the C/P ratios of the Scenedesmus used as food differed by a factor 5 in the experiments, the excretion rates differed by factor 3 only. Despite the higher P-excretion rates (0.258± 0.022 µg PO₄-P mg^–1 C h^–1) of the daphnids fed with P-rich food than those fed with P-poor food (0.105 ± 0.047 µg PO₄-P mg^–1 C h^p–1), both the categories of the animals were apparently conserving P. A survey of the literature on zooplankton excretion shows that in Daphnia the excretion rates vary by a factor 30, irrespective of the species and size of animals and method of estimation and temperature used.About two-thirds of this variation can be explained by size and temperature. A major problem of comparability of studies on P-regeneration by zooplankton relates to the existing techniques of P determination, which necessitates concentrating the animals several times above the in situ concentration (crowding) and prolonged experimental duration (starving), both of which manifest in marked changes that probably lead to underestimation of the real rates.     

Distribution and production ofChironomus salinarius (Diptera: Chironomidae) in a shallow coastal lagoon in the Bay of Cádiz

The abundance, generation time and production ofChironomus salinarius larvae in a lagoon fish-pond system in the Bay of Cádiz were studied by taking monthly samples at 3 sites during 1991 and 1992. Numerical abundance and biomass of larvae showed considerable spatial, seasonal and interannual variation (ANCOVAs,P<0.001). The maximum mean annual density was 7048 larvae m^–2, and corresponded to a biomass of 3.08 g dry weight (DW) m^–2. It was recorded at the site with the lowest rate of water renewal. Seasonal patterns were similar at all sites, with main annual peaks of abundance and biomass in autumn-early winter. Chironomid density was positively related to the biomass of benthic macroalgae (P<0.001). The population studied was multivoltine with a probable average of five generations per year, with overlapping cohorts and a predominance of third- and fourth-instar larvae. Estimates of annual production ranged between 72.2 g DW m^–2 yr^–1 at the site with the lowest rate of water renewal in 1991 and 0.1 g DW m^–2 yr^–1 at the site with the highest rate of water renewal in 1992. Mean annual production and the production/biomass ratio for the system was estimated to be 16.8 g DW m^–2 yr^–1 and 12.7, respectively. Possible factors leading to the observed density fluctuations are discussed, as well as possible sources of error in production estimates.     

Environmental and macroinvertebrate dynamics in the Lower Rhône River and a lateral dike field: a study matching two functioning descriptors

Environmental variables and macroinvertebrate communities are studied in two sites of the Lower Rhône River, the main channel and a lateral, occasionally connected dike field. Environmental variables and faunistic communities allow discrimination the two compartments. The environmental and faunistic differences the two sites change over time. The physical and chemical differences are significantly correlated with water discharge of the main channel. The faunistic ones are significantly correlated with the temperature of the dike field water. The connections between the main channel and the dike field could be very important to maintain a high heterogeneity of the habitat, and for recolonization of the main channel after a perturbation.     

Establishing apoptosis resistant cell lines for improving protein productivity of cell culture

The authors established apoptosis resistant COS–1, myeloma, hybridoma, and Friend leukemia cell lines by genetically engineering cells, aiming at more efficient protein production by cell culture. COS–1 cells, which are most widely used for eukariotic gene expression, were transfected with human bcl–2 gene. Both bcl–2 and mock transfected COS–1 cells were cultured at low (0.2%) serum concentration for 9 days. The final viable cell number of the bcl–2 transfected cells was ninefold of that of the mock transfectants. Both bcl–2 and mock transfectants were further transfected with the vector pcDNA- containing SV40 ori and immunoglobulin gene for transiently expressing protein. The bcl–2 expressing COS–1 cells produced more protein than the mock transfected COS–1 cells after 4 days posttransfection.Mouse myeloma p3-X63-Ag.8.653 cells, which are widely used as the partner for preparing hybridoma, and hybridoma 2E3 cells were transfected with human bcl–2 gene. Both bcl–2 transfected myeloma and hybridoma survived longer than the corresponding original cells in batch culture. The bcl–2 transfected 2E3 cells survived 2 to 4 four days longer in culture, producing 1.5- to 4-fold amount of antibody in comparison with the mock transfectants.Coexpression of bag–1 with bcl–2 improved survival of hybridoma 2E3 cells more than bcl–2 expression alone. The bag–1 and bcl–2 coexpressing cells produced more IgG than the the cells expressing bcl–2 alone.Apoptosis of Friend murine erythroleukemia(F-MEL) cells was suppressed with antisense c-jun expression. The antisense c-jun expressing cells survived 16 days at non-growth state.     

On the Modeling of Some Anti-inflammatory and Anti-thrombotic Drugs by AM1

Two families of autacoids from cell membrane phospholipids have been identified. The first, the icosanoids, which are formed from arachidonic acid, include prostaglandins and leukotrienes. The other includes modified phospholipids, as the platelet aggregating factor (PAF). These compounds are related to inflammatory and cardiovascular diseases. We review in this paper some of the work that has been done in our laboratories in the last few years relating to the modeling of new potential thromboxane synthase (TXS) and 5-lipoxygenase (5-LO) and cyclooxygenase (COX) inhibitors, and TXA₂ receptor antagonists derived from nitrogenated heterocycles. We include the results of the modeling of a group of proposed PAF antagonists, and compare their structures with PAF itself and with a recently proposed PAF antagonist model. This revised version was published online in June 2006 with corrections to the Cover Date.     

Rehabilitating degraded forests in Central Europe into self–sustaining forest ecosystems

There is a need for a consistent forest restoration strategy for the `Black Triangle', Central Europe. In the past 50 years, forests in this area have been heavily affected by industrial pollution. Recently, the amount of pollutants has decreased. This means that effective forest restoration programmes can be started. Forest decline must be seen as an ecological disturbance which cannot be solved by applying technical measures only. An ecosystem approach to forest restoration must be introduced into restoration policy and management. Basic principles and working methods of such an approach are briefly described and proposals are made for forest restoration policy, management and research in the area.