Living benthic foraminifera of the Okhotsk Sea: Faunal composition,standing stocks and microhabitats

Live (Rose Bengal stained) benthic foraminifera were investigated in surface sediment samples from the Okhotsk Sea to reveal the relationship between faunal characteristics and environmental parameters. Live benthic foraminifera were quantified in the size fraction > 125 µm in the upper 8 cm of replicate sediment cores, recovered with a multicorer at five stations along the Sakhalin margin, and at three stations on the southwestern Kamchatka slope. The stations are from water depths between 625 to 1752 m, located close or within the present Okhotsk Sea oxygen minimum zone, with oxygen levels between 0.3 and 1.5 ml l^- 1. At the high-productivity and ice-free Kamchatka stations, live benthic foraminifera are characterized by maximal standing stocks (about 1700-3700 individuals per 50 cm²), strong dominance of calcareous species (up to 87-91% of total live faunas), and maximal habitat depths (down to 5.2-6.7 cm depth). Vertical distributions of total faunal abundances exhibit a clear subsurface maximum in sediments. At the Sakhalin stations, which are seasonally ice-covered and less productive, live benthic foraminifera show lower standing stocks (about 200-1100 individuals per 50 cm²), lower abundance of calcareous species (10-64% of total live faunas), and shallower habitat depths (down to 2.5-5.4 cm depth). Faunal vertical distributions are characterized by maximum in the uppermost surface sediments. It is suggested that 1) lower and strongly seasonal organic matter flux, caused by the seasonal sea ice cover and seasonal upwelling, 2) lower bottom water oxygenation (0.3-1.1 ml l^- 1), and 3) more pronounced influence of carbonate undersaturated bottom water along the Sakhalin margin are the main factors responsible for the observed faunal differences. According to species downcore distributions and average living depths, common calcareous species were identified as preferentially shallow, intermediate and deep infaunal. Foraminiferal microhabitat occupation correlates with the organic matter flux and the depth of the oxygenated layer in sediments.     

Vegetation mapping and multivariate approach to indicator species of a forest ecosystem: A case study from the Thandiani sub Forests Division (TsFD) in the Western Himalayas

QuestionsDoes the plant species composition of Thandiani sub Forests Division (TsFD) correlate with edaphic, topographic and climatic variables? Is it possible to identify different plant communities in relation to environmental gradients with special emphasis on indicator species? Can this approach to vegetation classification support conservation planning?LocationThandiani sub Forests Division, Western Himalayas.MethodsQuantitative and qualitative characteristics of species along with environmental variables were measured using a randomly stratified design to identify the major plant communities and indicator species of the Thandiani sub Forests Division. Species composition was recorded in 10 × 2.5 × 2 and 0.5 × 0.5 m square plots for trees, shrubs and herbs, respectively. GPS, edaphic and topographic data were also recorded for each sample plot. A total of 1500 quadrats were established in 50 sampling stations along eight altitudinal transects encompassing eastern, western, northern and southern aspects (slopes). The altitudinal range of the study area was 1290 m to 2626 m above sea level using. The relationships between species composition and environmental variables were analyzed using Two Way Cluster Analysis (TWCA) and Indicator Species Analysis (ISA) via PCORD version 5.ResultsA total of 252 plant species belonging to 97 families were identified. TWCA and ISA recognized five plant communities. ISA additionally revealed that mountain slope aspect, soil pH and soil electrical conductivity were the strongest environmental factors (p ≤ 0.05) determining plant community composition and indicator species in each habitat. The results also show the strength of the environment-species relationship using Monte Carlo procedures.ConclusionsAn analysis of vegetation along an environmental gradient in the Thandiani sub Forests Division using the Braun-Blanquet approach confirmed by robust tools of multivariate statistics identified indicators of each sort of microclimatic zones/vegetation communities which could further be used in conservation planning and management not only in the area studied but in the adjacent regions exhibit similar sort of environmental conditions.     

Marine sediments from southeastern Brazilian continental shelf: A 1200 year record of upwelling productivity

A sediment core from the Cabo Frio coastal shelf (?23.19 S, ?41.8 W; 117 m depth), was analyzed for TOC, C/N ratio, organic petrography and planktonic foraminiferal content to evaluate variations in local productivity caused by changes in upwelling intensity and its relation to regional and global climatic variations during the last millennium. The Cabo Frio core recorded the last 1200 years of sedimentation, with rates varying from 0.11 to 0.32 mm yr^?1. Foraminiferal and organic geochemical analyses indicate the occurrence of three distinct periods of productivity. From 850 AD until 1070 AD, foraminifera fluxes consisting primarily of Turborotalita quinqueloba indicate stronger South Atlantic Central Water (SACW) transport onto the shelf, which induced high biological productivity that was also recorded by high TOC and marine palynomorphs content and a low C/N atomic ratio. This period coincided with a northward displacement of the atmospheric Intertropical Convergence Zone (ITCZ) and South Atlantic High (SAH) systems driven by positive temperature anomalies in the North Atlantic Ocean during the Medieval Climate Anomaly (MCA). From 1070 until 1500 AD, low TOC flux and planktonic foraminifera fluxes and high C/N atomic ratios suggest a reduction in marine productivity, probably driven by reduced transport of SACW associated with the southward displacement of the SAH and weakening of northeasterly winds. The period between 1500 and 1830 AD, which corresponds to the Little Ice Age, is marked by increased fluxes of planktonic foraminifera, principally of Globigerina bulloides and Globigerinita glutinata. These species mark an increase in productivity linked to SACW upwelling, supported by the enhancement of northeasterly winds and southward displacement of the ITCZ and SAH.     

Changes in the size distribution of goat steroidogenic luteal cells during pregnancy

Experiments were conducted to investigate the size distribution of goat steroidogenic luteal cells throughout pregnancy. Corpora lutea were collected from very early (<6 weeks), early (6–8 weeks), middle (9–14 weeks) or late (15–18 weeks) stages of pregnancy. Luteal tissue was dissociated into single-cell suspension by enzyme treatments. Cells were stained for 3β-hydroxysteroid dehydrogenase (3β-HSD) activity, a marker for steroidogenic cells. The steroidogenic cells covered a wide spectrum of size ranging from 5 to 45 μm in diameter. There was a significant increase in mean cell diameter (P>0.01) as pregnancy progressed. Mean diameter of 3β-HSD positive cells increased from 14.73±0.35 μm in the corpus luteum of very early pregnancy to 24.20±0.45 μm in the corpus luteum of late pregnancy. The ratio of large (>20 μm in diameter) to small (5–20 μm in diameter) luteal cells was 0.28:1.0 in very early pregnancy, with the 7.5–15 μm cell size class being dominant. However, the ratio of large-to-small luteal cells was increased to 1.77:1.0 μm as pregnancy advanced and 25–35 μm cell sizes became predominant. It is likely that small luteal cells could develop into large cells as pregnancy progresses. Development of pregnancy is also associated with an increase in size of steroidogenic luteal cells.     

Geographic position of sample grid and removal of uncommon species affect multivariate analyses of diverse assemblages: The case of oribatid mites (Acari: Oribatida)

In view of the rapid loss of biodiversity, large-scale environmental monitoring programs are urgently needed, over a range of local, regional and global scales. These programs can be made more efficient and cost-effective through shortcuts such as reduction of sampling effort and the use of low-cost surrogates. We revisited a large-scale dataset composed by 161 species recorded in 72 plots of 250 m, distributed over an 8 m × 8 m sampling grid in the tropical rainforest. Samples of litter and soil were collected and oribatid mites were extracted with a Berlese–Tullgren apparatus. Using a “moving window” procedure, we delimited smaller 5 km × 5 km grids in 16 possible positions within the larger grid. We first evaluated which fraction was more important to explain environmental and spatial patterns in the species composition: known environmental or spatial filters representing unknown causes of aggregation, and the confounded variance that might be associated with either or both. We used soil clay content, litter quantity, soil pH, number of trees, and distance to the nearest stream as environmental predictors. The spatial filters were generated using Moran Eigenvector mapping through the Principal Coordinates of Neighbor Matrices technique. To evaluate the influence of these fractions on the species composition, we used partial Redundancy Analysis. Using Principal Coordinates Analysis for abundance and presence/absence data, we evaluated if reduced matrices, discarding sets of less-frequent species, could identify the relationships captured with the complete dataset. All smaller grids contained more than 100 species. The effect of environmental variables on oribatid-mite community composition was always low, and each smaller grid position produced different results. Soil clay content and pH were the main factors associated with oribatid-mite distributions. The effects of unknown spatial patterns were greater than the environmental ones. Independently of the grid position, similar results were obtained for analyses with all oribatid-mite species, to the results obtained from analyses of only the most frequent species. Sets of more frequent and easily identifiable species proved to be a reliable surrogate for the complete assemblage. Omitting identifications of most species will improve the cost-effectiveness of monitoring programs. More emphasis should be placed on investigating the role of spatial heterogeneity and the effects of grid position in relation to patterns in species associations. Efficient biomonitoring could target surrogate species, to enable rapid tracking of environmental change while enlarging the sampling area to provide data for conservation strategies.     

The threshold responses of phytoplankton community to nutrient gradient in a shallow eutrophic Chinese lake

Excessive nutrient loads resulted in cascading trophic effects and ecosystem responses. Aims of this study were to determine if the thresholds in nutrient gradient related to phytoplankton community composition could be identified in eutrophic lake, and further to analyze the change of phytoplankton assemblage along the nutrient concentration based on Threshold Indicator Taxa ANalysis (TITAN). The results presented the significant community thresholds estimate for negative taxa declining at 1.650 mg/L TN and 131.5 μg/L TP, as well as simultaneously increasing for positive taxa at 1.665 mg/L TN and 151.5 μg/L TP along nutrient enrichment gradient. However, there was unremarkable change point determined for TN:TP ratios in Lake Dianchi. Elevated TN and TP altered the phytoplankton assemblage, even may induce the fade of algal blooms across the threshold in the hypertrophic lake. The findings could provide implications for deeply deciphering abrupt transitions for phytoplankton assemblage and developing nutrient tactics to protect the lake ecosystems.     

Sources of variation contributing to production and quality attributes of Kyrgyz cashmere in Osh and Naryn provinces: Implications for industry development

We aimed to quantify the sources of variation contributing to the production and quality of cashmere produced in five districts in Osh and Naryn provinces of Kyrgyzstan. In early spring 2008 mid-side cashmere samples were taken from 719 cashmere adult females, and 41 cashmere adult males and castrates. Samples came from 53 villages and a total of 156 farmers’ flocks. For 91 goats from 33 farmers in 13 villages of two districts that had been sampled earlier, cashmere was combed from the goat at the time of a second visit (end of April 2008) when the cashmere would normally be harvested. Following standard cashmere objective measurement, data were examined using general linear modelling to quantify the effects of potential determinants. The mean fibre diameter (MFD) of cashmere differed between provinces (Osh 15.7 μm, Naryn 16.7 μm; P = 4.4 × 10⁻²⁰). About 42% of the cashmere was <16 μm, 48% was 16.0–18.0 μm and 9.5% was >18.0 μm. Most of the cashmere samples were coloured (81%), with 63% black and 19% white. The percentage of cashmere samples that were white declined as MFD increased (26% < 14 μm to 11% of >18 μm). The primary determinants of cashmere MFD of individual goats were age of goat (range 1.46 μm, P = 1.8 × 10⁻¹²) and farm (range 6.5 μm, P = 1.7 × 10⁻¹⁴). The lesser effects detected for sex (range 0.9 μm, P = 0.026) and colour of cashmere (range 1.8 μm, P = 0.023) were based on small sample sizes and are unreliable. Age of goat had important affects on fibre diameter variation (up to 1.7% in coefficient of variation, P = 5.8 × 10⁻⁶) and fibre curvature (2.5–5°/mm, P = 2.1 × 10⁻⁴). By far the greatest effect on fibre curvature was cashmere MFD (P = 3.0 × 10⁻¹⁰⁴) with a smaller effect of sex (about 5°/mm, P = 3.0 × 10⁻⁶). Village effects were detected on fibre diameter variability (range 4.5% in coefficient of variation, P = 0.027) and fibre curvature (range 15°/mm, P = 1.6 × 10⁻⁷). There was a strong negative association between increasing MFD and declining fibre curvature (−5.11 ± 0.181°/mm per 1 μm; P = 7.1 × 10⁻¹²¹; r² = 0.51). Average combed cashmere weight was 164 g, the clean cashmere content was 0.661 and median clean cashmere production was 110 g per goat (range 60–351 g). Combed cashmere production increased with altitude of the village, probably related to different moulting times as spring temperatures warmed up later in higher altitude villages up to 3200 masl. Measurements of combed cashmere MFD were coarser than the mid-side samples taken earlier in the year. There are farmers and cashmere goats in the sampled districts of Kyrgyzstan which produce the finest qualities of commercial cashmere as the vast majority of cashmere is fine, has low variation in fibre diameter and has fibre crimping (curvature) typical of Chinese and Mongolian cashmere. There is substantial scope to increase the production and commercial value of cashmere produced by Kyrgyz goats. In particular, some villages and farmers need to change their buck selection practices if they wish to produce acceptable cashmere. Farmers should separate their finer and white cashmere prior to sale.     

Protection of oxidative stress induced apoptosis in osteosarcoma cells by dihydromyricetin through down-regulation of caspase activation and up-regulation of BcL-2

Current study was aimed to investigate the effect of dihydromyricetin on hydrogen peroxide induced oxidative stress in the osteosarcoma cells. MTT assay showed that hydrogen peroxide treatment at a concentration of 100 μM caused a significant (p < 0.005) reduction in the viability of MG63 cells. However, reduction in cell viability caused by 100 μM concentration of hydrogen peroxide was completely prevented on incubation with 30 μM dose of dihydromyricetin. Treatment with 100 μM concentration of hydrogen peroxide for 24 h led to condensation of chromatin material, rounding of cell shape and detachment of cells. The results from flow cytometry using annexin V-FITC and PI double staining showed apoptosis induction in 47.84 ± 5.21% cells on treatment with 100 μM concentration of hydrogen peroxide compared to 2.32 ± 0.54% in controlcells. The apoptotic alterations in MG63 cell morphology were prevented significantly on pre-treatment with 30 μM doses of dihydromyricetin for 48 h. Annexin V-FITC and PI staining showed reduction of hydrogen peroxide induced apoptotic cell percentage to 3.07 ± 0.86% on pre-treatment of MG63 cells with 30 μM dose of dihydromyricetin. Western blot analysis showed a significant increase in the activation of caspase-3 and -9 on treatment of MG63 cells for 24 h with 100 μM concentration of hydrogen peroxide. The expression level of Bcl-2 was decreased significantly by 100 μM concentration of hydrogen peroxide in MG63 cells. However, pre-treatment of MG63 cells with 30 μM dose of dihydromyricetin for 48 h significantly prevented hydrogen peroxide induced increase in caspase-3 and -9 levels and reduction in Bcl-2 level. Thus dihydromyricetin prevents hydrogen peroxide induced reduction in viability and induction of apoptosis in MG63 cells through down-regulation of caspase activation and up-regulation of Bcl-2 levels.     

Characterization of azaspiracids in plankton size-fractions and isolation of an azaspiracid-producing dinoflagellate from the North Sea

Azaspiracids (AZAs) are a group of lipophilic polyether toxins implicated in incidents of shellfish poisoning in humans, particularly in northern Europe. In an attempt to establish the biogeographical distribution of AZA toxins, their association with plankton size-fractions, and to confirm the identity of the causative species responsible for human poisoning, a month-long oceanographic study was undertaken in coastal North Sea waters. The occurrence and abundance of AZA analogues was measured by on board triple quadrupole mass spectrometry coupled to liquid chromatography (LC-MS/MS). In size-fractionated plankton samples collected by net tows (20 μm mesh-size), by pumping from discrete depths and from Niskin entrapment bottle casts to fixed depths, AZA-1 was consistently the major azaspiracid component. In eastern Scottish coastal waters, the highest amounts of AZA-1 in net tow samples were in the 50–200 μm fractions, with lesser amounts detected in the >200 μm and 50–20 μm fractions. At these stations, the 50–200 μm fractions were rich in the ciliate Favella ehrenbergii. Cells of F. ehrenbergii isolated by microcapillary indeed contained AZA-1, but isolated cells grown and fed the non-toxic dinoflagellate Scrippsiella trochoidea for one week failed to contain any detectable AZA-1—evidence that F. ehrenbergii is merely a vector for AZA. Detailed analysis of plankton from Niskin bottle samples from around the North Sea typically showed highest amounts of AZA in the 3–20 μm fraction. From this fraction, a large number of crude cultures were established and subsequently screened for the presence of AZAs. A small photosynthetic thecate dinoflagellate, provisionally designated as strain 3D9, was isolated by microcapillary and brought into pure culture. This dinoflagellate strain was found to produce AZA-1, AZA-2 and an isomer of AZA-2. Sequence comparisons by molecular genetic techniques also indicated that this genotype was present in field samples rich in AZA. This discovery of a novel causative dinoflagellate for AZA toxicity essentially explains the lack of correlation of AZA with the abundance and distribution of the previously postulated culprit species Protoperidinium crassipes. We instead propose that such large phagotrophic dinoflagellates can act as an AZA vector following grazing upon a proximal source, such as the dinoflagellate 3D9 strain.     

Coralline algae as depth indicators in the Miocene carbonates of the Eratosthenes Seamount (ODP Leg 160, Hole 966F)

The abundance of the major coralline algal groups has been investigated and quantified in the coralline-rich facies of the Miocene shallow-water carbonates of the Eratosthenes Seamount (eastern Mediterranean, off-shore Cyprus). The analysis is based on the quantification of the most easily-recognizable groups of coralline algae in order to provide a user-friendly approach for palaeobathymetric reconstructions. Coralline algal distribution through the core suggests water depth estimates generally similar to those based on the composition of the skeletal assemblage and the benthic foraminiferal association in particular. The only noticeable difference occurs in the rhodolith and coral facies, where algal distribution suggests deeper waters than those indicated by benthic foraminifera. The distribution pattern of the major groups suggests that the ratio between Hapalidiales and Corallinales is the most reliable indicator of water-depth. The comparison with other models available in literature highlights a general zonation useful for the study of tropical, middle to late Miocene oligotrophic carbonates. Very shallow settings (0–20 m) are overwhelmingly dominated by Corallinales; in slightly deeper settings (20–40 m) Hapalidiales are more abundant, especially if the sea-floor is shaded (for example by a macrophyte canopy). Between 40 and 60 m, Hapalidiales dominate but Corallinales are still common, while below 60 m Corallinales are very rare. In non-oligotrophic environments this zonation is not reliable and, due to the reduced water clarity related to the high primary productivity, Hapalidiales clearly dominate even in very shallow settings.     

Magnesium and strontium compositions of recent symbiont-bearing benthic foraminifera

Minor and trace elements in foraminiferal carbonates are potential paleo-proxies of climate, nutrient and seawater composition. There are very few reports of trace element composition of symbiont-bearing, larger foraminifera that are known to be important constituents of shallow-marine, modern and ancient carbonates. In this paper we examine the range of variation in Mg and Sr content of Recent species of these foraminifera from a lagoon of Lakshadweep Atoll (Indian Ocean) and Akajima Islands, Japan. Two hyaline species, Amphistegina lessonii and Neorotalia calcar,and two porcellaneous species, Amphisorus hemprichii and Marginopora vertebralis were collected live from Lakshadweep islands. Mg / Ca in these foraminifera is of an order of magnitude higher than the values reported for planktonic and symbiont-free benthic foraminifera. The Sr / Ca values are, however, comparable with the reported values in other foraminiferal taxa and they are found to vary within a narrow range. Electron-probe micro-analysis of three symbiont-bearing benthic species indicates spatial heterogeneity of high orders in Mg / Ca composition in all the species. The annual variation in temperature and pH of the lagoon water cannot explain the observed amplitude of the compositional variation. The photosynthesis and respiration of the symbionts and host foraminifera are possibly the major cause of compositional heterogeneity in individual tests, as has also been recently postulated for symbiont-bearing planktonic foraminiferal species. It highlights the need to isolate biological factors and necessitates species-specific paleotemperature scale in paleoclimatic analysis. We also analyzed δ¹⁸O, δ¹³C, Ca, Mg and Sr in carefully dissected chambers of a reef-dwelling, porcellaneous benthic foraminifer, Marginopora kudakajimaensis, collected live in four seasons. A moderate positive correlation is observed between Mg / Ca and temperature. However, large inter- and intra-test variation in Mg limits the precision of Mg / Ca as palaeotemperature proxy. The Sr / Ca of the test calcite is unrelated to temperature of the sea water. The δ¹³C of M. kudakajimaensis does not correlate with δ¹⁸O, Mg / Ca or Sr / Ca.     

Evaluation of stream ecosystem health and species association based on multi-taxa (benthic macroinvertebrates,algae, and microorganisms) patterning with different levels of pollution

Benthic communities of macroinvertebrates, algae, and microorganisms were concurrently collected using a Surber sampler (30 × 30 m²; 300 μm mesh), brush (5 × 5 cm²), and syringe (100 mL; Denaturing Gradient Gel Electrophoresis), respectively, to determine the ecological integrity of streams with different levels of pollution. Macroinvertebrates provided a clearer representation of the gradient of pollution, while a broader scope of species distribution was observed for algae and microorganisms, including sites severely polluted with heavy metals. Species associations among different taxa were presented on the Self-Organizing Map (SOM) and Nonmetric Multidimensional Scaling (NMDS) based on environmental factors. After screening, indicator species visualized on the SOM represented a wider range of environmental impacts and were more illustrative with benthic macroinvertebrates in least polluted sites. In contrast NMDS presented species more closely associated with overall variance of communities with severe pollution, mainly in microorganisms and algae. Multi-taxa community analysis using SOM and NMDS in combination would provide a comprehensive assessment for addressing ecological integrity in streams.     

Paleoceanographic changes during the past 1.9 Myr at DSDP Site 238, Central Indian Ocean Basin: Benthic foraminiferal proxies

Deep-sea benthic foraminifera have been quantitatively analyzed in samples (> 125 μm size fraction) from Ocean Drilling Program (ODP) Site 238, to understand paleoceanographic changes in the Central Indian Basin over the past 1.9 Myr. Factor and cluster analyses of the 25 highest-ranked species made it possible to identify five biofacies, characterizing distinct deep-sea environmental settings. The environmental interpretation of each biofacies is based on the ecology of recent deep-sea benthic foraminifera. The benthic faunal record indicates fluctuating deep-sea conditions in environmental parameters including oxygenation, surface productivity and organic food supply. These changes appear to be linked to Indian summer monsoon variability, the main climatic feature of the Indian Ocean region. The benthic assemblages show a major shift at ∼ 0.7 to 0.6 Ma, marked by major turnovers in the relative abundances of species, coinciding with an increased amplitude of glacial cycles. These cycles appear to have influenced low latitude monsoonal climate as well as deep-sea conditions in the Central Indian Ocean Basin.     

Design,synthesis and biological evaluation of 3-benzyloxy-linked pyrimidinylphenylamine derivatives as potent HIV-1 NNRTIs

A novel series of 3-benzyloxy-linked pyrimidinylphenylamine derivatives (8a–8s) was designed, synthesized and evaluated for their in vitro anti-HIV activity in MT-4 cell cultures. Most of the compounds inhibited wild-type (wt) HIV-1 replication in the lower micromolar concentration range (EC₅₀ = 0.05–35 μM) with high selectivity index (SI) values (ranged from 10 to >4870). In particular, 8h and 8g displayed excellent antiretroviral activity against wt HIV-1 with low cytotoxicity (EC₅₀ = 0.07 μM, CC₅₀ >347 μM, SI >4870; EC₅₀ = 0.05 μM, CC₅₀ = 42 μM, SI = 777, respectively), comparable to that of the marked drug nevirapine (EC₅₀ = 0.113 μM, CC₅₀ >15 μM, SI >133). In order to confirm the binding target, 8h was selected to perform the anti-HIV-1 RT assay. Additionally, preliminary structure activity relationship (SAR) analysis and molecular docking studies of newly synthesized compounds were also discussed, as well as the predicted physicochemical properties.     

Regulation of plasma membrane Ca2 +-ATPase activity by acetylated tubulin: Influence of the lipid environment

We demonstrated previously that acetylated tubulin inhibits plasma membrane Ca^2 +-ATPase (PMCA) activity in plasma membrane vesicles (PMVs) of rat brain through a reversible interaction. Dissociation of the PMCA/tubulin complex leads to restoration of ATPase activity. We now report that, when the enzyme is reconstituted in phosphatidylcholine vesicles containing acidic or neutral lipids, tubulin not only loses its inhibitory effect but is also capable of activating PMCA. This alteration of the PMCA-inhibitory effect of tubulin was dependent on concentrations of both lipids and tubulin. Tubulin (300 μg/ml) in combination with acidic lipids at concentrations > 10%, increased PMCA activity up to 27-fold. The neutral lipid diacylglycerol (DAG), in combination with 50 μg/ml tubulin, increased PMCA activity > 12-fold, whereas tubulin alone at high concentration (≥ 300 μg/ml) produced only 80% increase. When DAG was generated in situ by phospholipase C incubation of PMVs pre-treated with exogenous tubulin, the inhibitory effect of tubulin on PMCA activity (ATP hydrolysis, and Ca^2 + transport within vesicles) was reversed. These findings indicate that PMCA is activated independently of surrounding lipid composition at low tubulin concentrations (< 50 μg/ml), whereas PMCA is activated mainly by reconstitution in acidic lipids at high tubulin concentrations. Regulation of PMCA activity by tubulin is thus dependent on both membrane lipid composition and tubulin concentration.     

Effect of salinity and seawater calcite saturation state on Mg and Sr incorporation in cultured planktonic foraminifera

Trace elements incorporated in planktonic foraminiferal test carbonate are commonly used as paleoproxies. For instance, Mg/Ca ratios are frequently used for reconstructing sea surface temperature and, together with the foraminiferal stable oxygen isotope ratios, are also used as paleosalinity proxy. Foraminiferal Sr/Ca ratios constitute another example of the application of trace elements in paleostudies since they may reflect the Sr/Ca values of seawater. However, over the past few decades it has been proven that the incorporation of trace elements in foraminiferal calcite is controlled by more than one environmental parameter. To quantify the effect of salinity on Mg and Sr incorporation planktonic foraminifera Globigerinoides sacculifer (sensu stricto) were grown in the laboratory under different environmental conditions. Laboratory experiments allowed us to separate a direct salinity effect from a possible independent impact through differences in the calcite saturation state of the seawater (Ω). Although the temperature effect is more important than the salinity effect, a change of 4 salinity units is equivalent to a 1 °C bias on Mg/Ca-based temperatures. This effect of salinity on Mg incorporation is minor. However, when using Mg/Ca-based temperatures in combination with foraminiferal δ¹⁸O to calculate salinity, it cannot be neglected. The present study shows salinity as the overriding control on Mg incorporation within the range of Ω studied (Ω between 5.25 and 6.50; [CO₃²⁻] between 218 and 270 μmol/kg) at a constant temperature of 26 °C. In contrast, Ω appears to be the main control on foraminiferal Sr incorporation (0.10 mmol/mol per 100 µmol/kg rise in [CO₃²⁻]), whereas salinity has a non significant influence on Sr/Ca.     

Isolation of antioxidant constituents from Combretum apiculatum subsp. apiculatum

Species of the family Combretaceae are used extensively in traditional medicine against inflammation and infections, and although antibacterial activity has been reported in non-polar extracts, further rationale for the widespread use of the Combretaceae is expected to exist. Methanol extracts of leaves of ten different Combretum species were evaluated for antioxidant activity by spraying TLC chromatograms of each leaf extract with 2,2-diphenyl-1-picrylhydrazyl (DPPH). Compounds with antioxidant activity were detected by bleaching of the purple DPPH colour. Leaf extracts of Combretum apiculatum subsp. apiculatum had the most antioxidant compounds. This species was consequently selected for phytochemical investigation. A DPPH assay-directed fractionation of the leaf extracts of C. apiculatum led to the isolation of four antioxidant compounds from the ethyl acetate and butanol soluble fractions. The structures of the compounds were determined by spectroscopic analyses (¹H-NMR, ¹³C-NMR and MS) and identified as: cardamonin (1), pinocembrin (2), quercetrin (3) and kaempferol (4). In a quantitative antioxidant assay, the more polar fractions (ethyl acetate and butanol) obtained by solvent–solvent fractionation had the highest antioxidant activity among the solvent fractions obtained from C. apiculatum, with EC₅₀ values of 3.91 ± 0.02 and 2.44 ± 0.02 μg/ml respectively. Of the four isolated compounds, quercetrin (4) and kaempferol (3) had the strongest antioxidant activity, with EC₅₀ values of 11.81 ± 85 and 47.36 ± 0.03 μM respectively. Cardamonin (1) and pinocembrin (2) did not demonstrate strong activity. L-ascorbic acid was used as standard antioxidant agent (EC₅₀ = 13.37 ± 0.20 μM or 2.35 μg/ml). The cytotoxicity of cardamonin and pinocembrin was evaluated on Vero kidney cells using the MTT (3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide) assay with berberine as positive control. At concentrations higher than 50 μg/ml of cardamonin or pinocembrin, the cells were not viable. Cardamonin was more toxic (LC₅₀ = 1.97 μg/ml) than pinocembrin (LC₅₀ = 29.47 μg/ml) and even the positive control, berberine (LC₅₀ = 12.35 μg/ml).     

Role and characteristics of problematic biofilms within the removal and mobility of trace metals in a pilot-scale membrane bioreactor

The characteristics of problematic biofilms (i.e., fouling and clogging layers) were studied with regards to the removal and fate of trace metals (contents well under 100 μg/L) during the long-term operation of a pilot-scale membrane bioreactor for the treatment of real wastewaters from a large industrial area.Results showed that clogging layer was more effective than suspended activated sludge in the biosorption of As > Zn > Ni > Cd > Sb > Fe > Se due to the synergic effects of extracellular polymeric compounds and metal-resistant bacteria. In fact the selective microbial speciation of the phylum of Bacteroidetes, which is highly resistant to heavy metals, was observed in the clogging sludge in spite of the very low concentration of dissolved metals in the bioreactor.Compared to the suspended activated sludge, the clogging layer enhanced the biosorption of very toxic substances such as As, Cd and Ni. In fact, the metal contents were respectively: 7.9–7.4 vs. 690–840 μgAs/kgTS; 1.5–2.2 vs. 149–219 μgCd/kgTS; 58.8–71.7 vs. 227–298 μgNi/kgTS. Then, the potential desorption of metals during the membrane acid cleanings was estimated as relevant as 10–15% of the metals associated to the clogging sludge. The combined effects of pH and the selected microbial community, and the minor effect of the redox potential, let us conclude on the major importance of bio-sorption/desorption mechanisms with respect to bio-precipitation/dissolution.     

Enzymatic decolorization of sulfonphthalein dyes

The white rot fungus (WRF) Pleurotus ostreatus produced manganese peroxidase (MnP) and manganese-independent peroxidase (MIP) activities during solid state fermentation of wheat straw, a natural lignocellulosic substrate. Most of the sulfonphthalein (SP) dyes were decolorized by MnP at pH 4.0. The higher K_m for meta-cresol purple (40 μM) and lower K_m for ortho-cresol red (26 μM) for MnP activities explained the preference for the position of methyl group at ortho than at meta on chromophore. Bromophenol blue decolorizing activity was higher at pH 3.5 and decreased as the concentration of Mn^II was increased. SP-decolorizing activity was associated not only with MnP but also with MIP. Additional bromine group along with the methyl group on SP chromophores decreases the rate of decolorization. Bromination of sulfonphthalein chromophore makes them the poorer substrate for MnP. This is evident from the higher K_m for bromocresol green (117 μM) when compared to bromocresol purple (36 μM) and bromophenol blue (78 μM). The order of preference for the SP dyes as substrate for the MnP-catalyzed decolorizing activity is phenol red > ortho-cresol red > meta-cresol purple > bromophenol red > bromocresol purple > bromophenol blue > bromocresol green and the order of preference for the SP dyes as substrate for the MIP-catalyzed decolorizing activity is bromocresol green > bromophenol blue > bromocresol purple > bromophenol red > meta-cresol purple > ortho-cresol red > phenol red. Inhibition of PR decolorizing activity by NaN₃ provided the evidence of decolorizing activity as an oxidative process.     

Morphological characteristics of phytoliths from representative conifers in China

Coniferous phytoliths in sediments are an effective tool for detecting the historical appearance of conifers. However, at the timberline in mountainous areas, such coniferous phytoliths are easily confused with grass phytoliths. This study analyses modern phytoliths from 17 conifer plants. Six common types and six rare types were identified. The conifers studied produce abundant blocky polyhedral and cubic (in the average 30–40 μm size range), blocky scrobiculate (average 30–40 μm), tabular elongate unsculpted (length 50–100 μm, width 10–20 μm), tabular elongate cavate (length 50–150 μm, width ∼10 μm), tabular elongate dendritic (50–100 μm × 10–20 μm), and irregular oblong (20–40 μm) phytoliths. This paper aims to show morphological characteristics of coniferous phytoliths in China, and to show how the common coniferous phytoliths differ from similar grass phytolith types, such as blocky polyhedral coniferous phytoliths from silicified parallelepipedal bulliform cells produced by grass. Blocky polyhedral and cubic phytoliths are the commonest coniferous phytoliths found in the sediments, but need to be carefully distinguished from grass parallelepipedal bulliform cells. This study indicates that clearly protruding ridges and irregular inward edges are essential features of cubic and polyhedral morphotypes produced by conifers. Results of this paper might provide important material for the study of paleovegetation and paleoecology of mountainous areas, especially at the timberline.