Gomphrena claussenii, a novel metal-hypertolerant bioindicator species, sequesters cadmium, but not zinc, in vacuolar oxalate crystals.

Gomphrena claussenii is a recently described zinc (Zn)- and cadmium (Cd)-hypertolerant Amaranthaceae species displaying a metal bioindicator Zn/Cd accumulation response. We investigated the Zn and Cd distribution in stem and leaf tissues of G. claussenii at the cellular level, and determined metabolite profiles to investigate metabolite involvement in Zn and Cd sequestration. Gomphrena claussenii plants exposed to high Zn and Cd supply were analysed by scanning electron microscopy with energy-dispersive X-ray (SEM-EDX) and micro-proton-induced X-ray emission (micro-PIXE). In addition, gas chromatography-time of flight-mass spectrometry (GC-TOF-MS) was used to determine metabolite profiles on high Zn and Cd exposure. Stem and leaf tissues of G. claussenii plants exposed to control and high Cd conditions showed the abundant presence of calcium oxalate (CaOx) crystals, but on high Zn exposure, their abundance was strongly reduced. Ca and Cd co-localized to the CaOx crystals in Cd-exposed plants. Citrate, malate and oxalate levels were all higher in shoot tissues of metal-exposed plants, with oxalate levels induced 2.6-fold on Zn exposure and 6.4-fold on Cd exposure. Sequestration of Cd in vacuolar CaOx crystals of G. claussenii is found to be a novel mechanism to deal with Cd accumulation and tolerance.

Physical gels based on charge-driven bridging of nanoparticles by triblock copolymers.

We have prepared an aqueous physical gel consisting of negatively charged silica nanoparticles bridged by ABA triblock copolymers, in which the A blocks are positively charged and the B block is neutral and water-soluble. Irreversible aggregation of the silica nanoparticles was prevented by precoating them with a neutral hydrophilic polymer. Both the elastic plateau modulus and the relaxation time increase slowly as the gel ages, indicating an increase both in the number of active bridges and in the strength with which the end blocks are adsorbed. The rate of this aging process can be increased significantly by applying a small shear stress to the sample. Our results indicate that charge-driven bridging of nanoparticles by triblock copolymers is a promising strategy for thickening of aqueous particle containing materials, such as water-based coatings.

Relation between gelation conditions and the physical properties of whey protein particles.

Whey protein particles have several applications in modulating food structure and for encapsulation, but there is a lack of methods to prepare particles with a very high internal protein content. In this study whey protein particles with high internal protein content were prepared through emulsification and heat gelation of 25% (w/w) whey protein isolate solution at different pH (6.8 or 5.5) and NaCl concentrations (50, 200, or 400 mM). Particles formed at pH 6.8 were spherical, whereas those formed at pH 5.5 were irregular and had a cauliflower-like appearance. Both particles had an average size of few micrometers, and the particles formed at pH 5.5 had higher protein content (∼39% w/v) than the particles formed at pH 6.8 (∼18% w/v). Similarly, particle morphology and protein density were also affected by initial NaCl concentration: particles formed at 50 mM NaCl (pH 6.8) were spherical, whereas particles formed at either 200 mM NaCl (pH 6.7) or 400 mM NaCl (pH 6.6) were irregular and protein density of the particles increased with increasing initial NaCl concentration. Whey protein particles formed at pH 5.5 showed an excellent heat stability: viscosity of the suspensions containing approximately 30% of protein particles formed at pH 5.5 did not show any change after heating at 90 °C for 30 min while the viscosity of suspensions containing protein particles prepared at other conditions increased after heating. In summary, whey protein particles with varying microstructure, shape, internal protein density, and heat stability can be formed by using heat-induced gelation of whey protein isolate at different gelling conditions.

Description of Meloidoderita salina sp. n. (Nematoda, Sphaeronematidae) from a micro-tidal salt marsh at Mont-Saint-Michel Bay in France.

Meloidoderita salinasp. n. is described and illustrated from the halophytic plant Atriplex portulacoides L. (sea purslane) growing in a micro-tidal salt marsh in the Mont-Saint-Michel Bay in France. This new species is the first member of Meloidoderita Poghossian, 1966 collected from a saline environment, and is characterized by the following features: sedentary mature females having a small swollen body with a clear posterior protuberance; slightly dorsally curved stylet, 19.9 µm long, with posteriorly sloping knobs; neck region irregular in shape and twisted; well developed secretory-excretory (S-E) pore, with markedly sclerotized S-E duct running posteriorly; prominent uterus bordered by a thick hyaline wall and filled with eggs. The adult female transforms into a cystoid. Eggs are deposited in both egg-mass and cystoid. Cystoids of Meloidoderita salinasp.n. display a unique sub-cuticular hexagonal beaded pattern. Male without stylet, pharyngeal region degenerated, S-E duct prominent, deirids small, developed testis 97.5 µm long, spicules 18.4 µm long, cloacal opening ventrally protruded, small phasmids posterior to cloaca opening and situated at 5.9 (3.2-7.7) µm from tail end, and conical tail ending in a rounded terminus marked with one (rarely two) ventrally positioned mucro. Additionally, some young malesof the new species were observed enveloped in the last J2 cuticle. Second-stage juvenile body 470 µm long, with a 16.4 µm long stylet, prominent rounded knobs set off from the shaft, hemizonid anterior and adjacent to S-E pore, small deirids located just above S-E pore level, genital primordium located at 68-77% of body length, phasmids small and located at about 19 µm from tail tip, and tail 38.7 µm long, tapering to finely pointed terminus with a finger-like projection. Phylogenetic analyses based on the nearly full length small subunit ribosomal DNA sequences of Meloidoderita salinasp. n. revealed a close relationship of the new species with Sphaeronema alni Turkina & Chizhov, 1986 and placed these two species sister to the rest of Criconematina.

Bioaugmentation of UASB reactors with immobilized Sulfurospirillum barnesii for simultaneous selenate and nitrate removal.

Whole-cell immobilization of selenate-respiring Sulfurospirillum barnesii in polyacrylamide gels was investigated to allow the treatment of selenate contaminated (790 microg Se x L(-1)) synthetic wastewater with a high molar excess of nitrate (1,500 times) and sulfate (200 times). Gel-immobilized S. barnesii cells were used to inoculate a mesophilic (30 degrees C) bioreactor fed with lactate as electron donor at an organic loading rate of 5 g chemical oxygen demand (COD) x L(-1) day(-1). Selenate was reduced efficiently (>97%) in the nitrate and sulfate fed bioreactor, and a minimal effluent concentration of 39 microg Se x L(-1) was obtained. Scanning electron microscopy with energy dispersive X-ray (SEM-EDX) analysis revealed spherical bioprecipitates of

Biological alkylation and colloid formation of selenium in methanogenic UASB reactors.

Bioalkylation and colloid formation of selenium during selenate removal in upflow anaerobic sludge bed (UASB) bioreactors was investigated. The mesophilic (30 degrees C) UASB reactor (pH = 7.0) was operated for 175 d with lactate as electron donor at an organic loading rate of 2 g COD L(-1) d(-1) and a selenium loading rate of 3.16 mg Se L(-1) d(-1). Combining sequential filtration with ion chromatographic analysis for selenium oxyanions and solid phase micro extraction gas chromatography mass spectrometry (SPME-GC-MS) for alkylated selenium compounds allowed to entirely close the selenium mass balance in the liquid phase for most of the UASB operational runtime. Although selenate was removed to more than 98.6% from the liquid phase, a less efficient removal of dissolved selenium was observed due to the presence of dissolved alkylated selenium species (dimethylselenide and dimethyldiselenide) and colloidal selenium particles in the effluent. The alkylated and the colloidal fractions contributed up to 15 and 31%, respectively, to the dissolved selenium concentration. The size fractions of the colloidal dispersion were: 4 to 0.45 mum: up to 21%, 0.45 to 0.2 mum: up to 11%, and particles smaller than 0.2 mum: up to 8%. Particles of 4 to 0.45 mum were formed in the external settler, but did not settle. SEM-EDX analysis showed that microorganisms form these selenium containing colloidal particles extracellularly on their surface. Lowering the temperature by 10 degrees C for 6 h resulted in drastically reduced selenate removal efficiencies (after a delay of 1.5 d), accompanied by the temporary formation of an unknown, soluble, organic selenium species. This study shows that a careful process control is a prerequisite for selenium treatment in UASB bioreactors, as disturbances in the operational conditions induce elevated selenium effluent concentrations by alkylation and colloid formation.

Lipophilic and hydrophilic moisturizers show different actions on human skin as revealed by cryo scanning electron microscopy.

To study the mode of action of moisturizers on human skin, hydrophilic moisturizers in water and neat lipophilic moisturizers were applied on excised skin for 24 h at 32 degrees C. Samples of the treated skin were subsequently visualized in a cryoscanning electron microscope. The stratum corneum (SC) appeared as a region of swollen corneocytes (the swollen region) sandwiched between two layers of relatively dry corneocytes (the upper and lower non-swelling regions respectively). Lipophilic moisturizers increased the water content of the SC, whereas hydrophilic moisturizers can also reduce the water content of the SC. When focusing on the effect of the moisturizers on the three different regions, it was observed that cells in the swelling region are most sensitive to the application of the moisturizers and that the change in SC thickness is most influenced by the change in the thickness of the swelling region. Summarizing, SC cells are not equally sensitive to moisturizer application: centrally located corneocytes are more sensitive than corneocytes in the upper and the lowest regions of the SC.

Abscisic acid controls embryo growth potential and endosperm cap weakening during coffee (Coffea arabica cv. Rubi) seed germination.

The mechanism and regulation of coffee seed germination were studied in Coffea arabica L. cv. Rubi. The coffee embryo grew inside the endosperm prior to radicle protrusion and abscisic acid (ABA) inhibited the increase in its pressure potential. There were two steps of endosperm cap weakening. An increase in cellulase activity coincided with the first step and an increase in endo-beta-mannanase (EBM) activity with the second step. ABA inhibited the second step of endosperm cap weakening, presumably by inhibiting the activities of at least two EBM isoforms and/or, indirectly, by inhibiting the pressure force of the radicle. The increase in the activities of EBM and cellulase coincided with the decrease in the force required to puncture the endosperm and with the appearance of porosity in the cell walls as observed by low-temperature scanning electronic microscopy. Tissue printing showed that EBM activity was spatially regulated in the endosperm. Activity was initiated in the endosperm cap whereas later during germination it could also be detected in the remainder of the endosperm. Tissue printing revealed that ABA inhibited most of the EBM activity in the endosperm cap, but not in the remainder of the endosperm. ABA did not inhibit cellulase activity. There was a transient rise in ABA content in the embryo during imbibition, which was likely to be responsible for slow germination, suggesting that endogenous ABA also may control embryo growth potential and the second step of endosperm cap weakening during coffee seed germination.

Antennal sensilla of two parasitoid wasps: a comparative scanning electron microscopy study.

Two closely related parasitoid wasp species, Cotesia glomerata (L.) and Cotesia rubecula (Marshall) (Hymenoptera:Braconidae), are different in their associative learning of plant odors. To provide a solid basis for our research on the mechanisms that underlie this difference, we described the morphology of the antennal sensilla of these two species using scanning electron microscopy complemented with transmission electron microscopy. Female and male antennae of both species have the same six types of sensilla. We classified these sensilla as sensilla trichodea without pores, sensilla trichodea with a tip pore, sensilla trichodea with wall pores, sensilla coeloconica type I, sensilla coeloconica type II, and sensilla placodea. We conclude that the morphology, numbers, and distribution of the sensory receptors are highly similar in these two closely related wasp species. Differences between species and sexes occurred only in sensilla placodea numbers. C. rubecula has more sensilla placodea than C. glomerata and males of both species have a larger number and a higher density of sensilla placodea compared to females of the same species.

Diffusional properties of methanogenic granular sludge: 1H NMR characterization.

The diffusive properties of anaerobic methanogenic and sulfidogenic aggregates present in wastewater treatment bioreactors were studied using diffusion analysis by relaxation time-separated pulsed-field gradient nuclear magnetic resonance (NMR) spectroscopy and NMR imaging. NMR spectroscopy measurements were performed at 22 degrees C with 10 ml of granular sludge at a magnetic field strength of 0.5 T (20 MHz resonance frequency for protons). Self-diffusion coefficients of H(2)O in the investigated series of mesophilic aggregates were found to be 51 to 78% lower than the self-diffusion coefficient of free water. Interestingly, self-diffusion coefficients of H(2)O were independent of the aggregate size for the size fractions investigated. Diffusional transport occurred faster in aggregates growing under nutrient-rich conditions (e.g., the bottom of a reactor) or at high (55 degrees C) temperatures than in aggregates cultivated in nutrient-poor conditions or at low (10 degrees C) temperatures. Exposure of aggregates to 2.5% glutaraldehyde or heat (70 or 90 degrees C for 30 min) modified the diffusional transport up to 20%. In contrast, deactivation of aggregates by HgCl(2) did not affect the H(2)O self-diffusion coefficient in aggregates. Analysis of NMR images of a single aggregate shows that methanogenic aggregates possess a spin-spin relaxation time and self-diffusion coefficient distribution, which are due to both physical (porosity) and chemical (metal sulfide precipitates) factors.

Structure of the developing pea seed coat and the post-phloem transport pathway of nutrients.

An important function of the seed coat is to deliver nutrients to the embryo. To relate this function to anatomical characteristics, the developing seed coat of pea (Pisum sativum L.) was examined by light- and cryo-scanning electron microscopy (cryo-SEM) from the late pre-storage phase until the end of seed filling. During this time the apparently undifferentiated seed coat tissues evolve into the epidermal macrosclereids, the hypodermal hourglass cells, chlorenchyma, ground parenchyma and branched parenchyma. Using the fluorescent symplast tracer 8-hydroxypyrene-1,3,6-trisulfonic acid, it could be demonstrated that solutes imported by the phloem move into the chlorenchyma and ground parenchyma, but not into the branched parenchyma. From a comparison with literature data of common bean (Phaseolus vulgaris L.) and broad bean (Vicia faba L.), it is concluded that in the three species different parenchyma layers, but not the branched parenchyma, may be involved in the post-phloem symplasmic transport of nutrients in the seed coat. In pea, the branched parenchyma dies during the storage phase, and its cell wall remnants then form the boundary layer between the living seed coat parenchyma cells and the cotyledons. Using cryo-SEM, clear images were obtained of this boundary layer which showed that many intracellular spaces in the seed coat parenchyma are filled with an aqueous solution. This is suggested to facilitate the diffusion of nutrients from the site of unloading towards the cotyledons.

Water distribution and related morphology in human stratum corneum at different hydration levels.

This study focused on the water distribution in human stratum corneum and on the swelling of the corneocytes. For this purpose stratum corneum was hydrated to various levels and used either for Fourier transform infrared spectroscopy or for cryo-scanning electron microscopy. The images were analyzed with respect to water localization and cell shape. The Fourier transform infrared spectra were measured to study the water-lipid interactions. The results show that water only slightly changes the lipid transitions in the stratum corneum even at a hydration level of 300% wt/wt compared to stratum corneum and that water is inhomogeneously distributed in the stratum corneum. No gradual increase in water level was observed in depth. At 57%-87% wt/wt water content the hydration level in the central part of stratum corneum is higher than in the superficial and deeper cell layers. Water domains are mainly present within the corneocytes and not in the intercellular regions. At a very high hydration level (300% wt/wt), the corneocytes are strongly swollen except for the deepest cell layers adjacent to the viable epidermis. The corneocytes in these layers are not swollen. At 300% wt/wt hydration level water domains are also present in intercellular regions. Between 17% wt/wt and 300% wt/wt the cell thickness increases linearly with the hydration level suggesting that swelling of cells mainly occurs in the direction perpendicular to the skin surface. At an increased hydration level, the corneocyte envelope more efficiently surrounds the cell content compensating for the increased cell volume. The changes in stratum corneum morphology with increasing water level have also been observed in dermatomed skin.

SEM Observations on the Marine Nematode Dracognomus simplex (Gerlach, 1954) Allen and Noffsinger, 1978 (Draconematidae: Prochaetosomatinae).

The free-living marine nematode Dracognomus simplex (Gerlach, 1954) Allen &Noffsinger, 1978 was studied by scanning electron microscopy (SEM). The morphology of males and females is described and illustrated in detail. In addition to the typical and modified adhesion tubes, a new type of posterior adhesion tube was discovered. A neotype is proposed for Dracognomus simplex, and D. simplex sensu Decraemer &Gourbault, 1986 is renamed as Dracognomus americanum n. sp. Additionally, a key toward the Dracognomus species is proposed.

Structural differences between two types of basidiomycete septal pore caps.

The septal pore cap (SPC) of Trichosporon sporotrichoides CBS 8245 is vesicular-tubular, connected with flat-tubular endoplasmic reticulum (ER), and stains densely with zinc/iodine/osmium tetroxide, as does the ER. The SPC of Schizophyllum commune CBS 340.81 is more complex, about 600 nm in diameter, with perforations of 80-120 nm diameter, and stains less densely with zinc/iodine/osmium tetroxide than the ER. In high-pressure frozen and freeze-substituted hyphae of T. sporotrichoides the ER is present parallel to the dolipore septa, and electron-dense material occurs opposite the septal pore channel; the SPC rarely showed smooth vesicular-tubular membranes, suggesting that this is an ephemeral function of the SPC. The SPC of S. commune has a smooth outer and inner membrane, which enclose a matrix with a palisade-like substructure. A thin layer of electron-dense material covers the inner surface of the SPC of S. commune, from which beaded filamentous structures connect the SPC and the pore-occluding material. These filamentous structures may maintain the intracellular position of the SPC and possibly play a role in plugging the septal pore channel. The septal pore swellings of T. sporotrichoides contain more 1,6-beta-glucan than the septum, and intracellular glucans are also present near the septal pore channel. This cytosolic 1,6-beta-glucan in T. sporotrichoides may serve as a matrix to keep the tubular membranous structures of the SPC together. In contrast, 1,6-beta-glucan is not observed in the SPC and in the pore-occluding material of S. commune, and hyphal septa of this species show less labelling of 1,6-beta-glucan than the septal swelling. The evolutionary transition from simple to more complex types of SPCs may have resulted in a requirement for different components to maintain the morphological integrity and cell biological function.