Egil S. Erichsen

Ultrastructural characterization of the delimiting membranes of isolated autophagosomes and amphisomes by freeze-fracture electron microscopy

The delimiting membranes of isolated autophagosomes from rat liver had extremely few transmembrane proteins, as indicated by the paucity of intramembrane particles in freeze-fracture images (about 20 particles/microm2, whereas isolated lysosomes had about 2000 particles/microm2). The autophagosomes also appeared to lack peripheral surface membrane proteins, since attempts to surface-biotinylate intact autophagosomes only yielded biotinylation of proteins from contaminating damaged mitochondria. All the membrane layers of multilamellar autophagosomes were equally particle-poor; the same was true of the autophagosome-forming, sequestering membrane complexes (phagophores). Isolated amphisomes (vacuoles formed by fusion between autophagosomes and endosomes) had more intramembrane particles than the autophagosomes (about 90 particles/microm2), and freeze-fracture images of these organelles frequently showed particle-rich endosomes fusing with particle-poor or particle-free autophagosomes. The appearence of multiple particle clusters suggested that a single autophagic vacuole could undergo multiple fusions with endosomes. Only the outermost membrane of bi- or multilamellar autophagic vacuoles appeared to engage in such fusions.

Nickel and iron in saliva of patients with fixed orthodontic appliances

Saliva was obtained from patients receiving treatment with fixed orthodontic appliances. One saliva sample was taken without appliances, and another at least 3 weeks after placement. In some patients samples were also taken immediately after insertion of the appliance. Nickel and iron were quantified by electrothermal atomic absorption spectroscopy. There was a large scatter in the results. No statistically significant differences were found either in the concentrations or in absolute masses of nickel or iron in samples taken without appliances and in those obtained when the appliances had been in the mouth for at least 3 weeks. For samples taken immediately after placement of the appliance, there was a significant increase in both concentrations and masses of nickel and iron. It thus seems that there is a high initial release of metals, and the effect diminishes with time.

Metal particles and tissue changes adjacent to miniplates A retrieval study

Peri-implant soft tissue and bone from 12 patients undergoing removal of stainless steel miniplates and screws after healing of jaw fractures were studied with regard to histomorphology and metal content. Three patients with titanium plates were also included. Light microscopy and scanning electron microscopy with energy-dispersive X-ray microanalysis were used. Non-osseous tissue adjacent to devices of both materials showed fibrosis, including areas of mild chronic inflammation. The cellular picture was dominated by fibroblasts with small aggregates of lymphocytes and scattered macrophages. A connective tissue collar was found between the bone tissue and the screws of both stainless steel and titanium. Bone formation was also evident adjacent to screws of both materials. Stainless steel or titanium particles 5-50 microns in diameter were found in both soft tissue and bone next to implants of their corresponding bulk material. The amount of metal impregnation varied between individual sections, and fewer particles were found in the bone specimens than in soft tissue. The mild inflammatory changes were not restricted to areas of metal impregnation.

Corrosion of silver soldered orthodontic wires

The amounts of metals liberated from silver soldered stainless steel and cobalt-chromium orthodontic wires were measured. Measurements were taken after 3 days and 24 days immersion in 0.9% sodium chloride solution. High amounts of copper and zinc and some cadmium from the silver solder were found. The silver soldered stainless steel wire corroded more than the cobalt-chromium type, and liberated more nickel and chromium than did the cobalt-chromium wires.

Metal Release from Cobalt-Chromium Partial Dentures in the Mouth

The amounts of cobalt and chromium released to the saliva in individuals wearing cobalt-chromium partial dentures have been measured. Saliva was collected during a 5-min period, both with and without dentures in the mouth, and the two samples compared. More than 80% of the 30 cases showed increased amounts of metals with the denture in the mouth. Newer and larger dentures liberated more than did older and smaller ones. The median increase in chromium was approximately twice that of cobalt.

Autophagosome-associated variant isoforms of cytosolic enzymes

In a search for autophagosome-associated proteins, two-dimensional gel separations of proteins from purified autophagosomes, postnuclear supernatant, cytosol, lysosomes, mitochondria, endosomes and a cytomembrane fraction (mostly endoplasmic reticulum) were compared. Three proteins, with monomeric molecular masses of 43, 35 and 31 kDa, were enriched in total or sedimentable fractions of autophagosomes relative to the corresponding fractions of postnuclear supernatant, suggesting an association with the autophagosomal delimiting membrane. These proteins were also present on lysosomal membranes, but they were absent from mitochondria, and detected only in small amounts in the cytomembrane fraction and in endosomes, indicating that they were not associated with organelles sequestered by autophagy. However, all three proteins were present in the cytosol, suggesting that they were cytosolic proteins binding peripherally to the delimiting membrane of autophagosomes, probably to its innermost surface as indicated by their resistance to treatment of intact autophagosomes with proteinase or protein-stripping agents. Amino acid sequencing identified these proteins as an isoform of argininosuccinate synthase, an N-truncated variant of glyceraldehyde-3-phosphate dehydrogenase, and a sequence variant of short-chain 2-enoyl-CoA hydratase.

Elemental composition of natural populations of key microbial groups in Atlantic waters

Intracellular carbon (C), nitrogen (N) and phosphorus (P) content of marine phytoplankton and bacterioplankton can vary according to cell requirements or physiological acclimation to growth under nutrient limited conditions. Although such variation in macronutrient content is well known for cultured organisms, there is a dearth of data from natural populations that reside under a range of environmental conditions. Here, we compare C, N and P content of Synechococcus, Prochlorococcus, low nucleic acid (LNA) content bacterioplankton and small plastidic protists inhabiting surface waters of the North and South subtropical gyres and the Equatorial Region of the Atlantic Ocean. While intracellular C:N ratios ranged between 3.5 and 6, i.e. below the Redfield ratio of 6.6, all the C:P and N:P ratios were up to 10 times higher than the corresponding Redfield ratio of 106 and 16, respectively, reaching and in some cases exceeding maximum values reported in the literature. Similar C:P or N:P ratios in areas with different concentrations of inorganic phosphorus suggests that this is not just a response to the prevailing environmental conditions but an indication of the extremely low P content of these oceanic microbes.

An unaccounted fraction of marine biogenic CaCO3 particles.

Biogenic production and sedimentation of calcium carbonate in the ocean, referred to as the carbonate pump, has profound implications for the ocean carbon cycle, and relate both to global climate, ocean acidification and the geological past. In marine pelagic environments coccolithophores, foraminifera and pteropods have been considered the main calcifying organisms. Here, we document the presence of an abundant, previously unaccounted fraction of marine calcium carbonate particles in seawater, presumably formed by bacteria or in relation to extracellular polymeric substances. The particles occur in a variety of different morphologies, in a size range from <1 to >100 µm, and in a typical concentration of 104–105 particles L−1 (size range counted 1–100 µm). Quantitative estimates of annual averages suggests that the pure calcium particles we counted in the 1–100 µm size range account for 2–4 times more CaCO3 than the dominating coccolithophoride Emiliania huxleyi and for 21% of the total concentration of particulate calcium. Due to their high density, we hypothesize that the particles sediment rapidly, and therefore contribute significantly to the export of carbon and alkalinity from surface waters. The biological and environmental factors affecting the formation of these particles and possible impact of this process on global atmospheric CO2 remains to be investigated.

Monodisperse mesoporous silica nanoparticles of distinct topology

Monodisperse and uniform high-quality MCM(Mobil Composition of Matter)-48-type CMSNs (Cubic Mesoporous Silica Nanoparticles) are readily prepared by simply optimizing the molar ratio of ethanol and surfactant in the system TEOS-CTAB-NaOH-H2O-EtOH (TEOS=tetraethyl orthosilicate, CTAB=cetyltrimethylammonium bromide, EtOH=ethanol). In the absence of ethanol only hexagonal mesoporous silica with ellipsoidal and spherical morphology are obtained. The presence of ethanol drives a mesophase transformation from hexagonal to mixed hexagonal/cubic, further to purely cubic, and finally to a mixed cubic/lamellar. This is accompanied by a morphology evolution involving a mixture of ellipses/spheres, regular rods, uniform spheres, and finally a mixture of spheres/flakes. Preserving the three-dimensional (3D) cubic MCM-48 structure, use of a small amount of ethanol is beneficial to the improvement of the monodispersity of the CMSNs. Moreover, the quality of the CMSNs can also be controlled by changing the surfactant concentration or adjusting the stirring rate. All MSNs were characterized using powder X-ray diffraction, transmission electron microscopy, scanning electron microscopy, and N2 physisorption, indicating highly long-range ordered pore arrays, high specific surface areas (max. 1173 m2g-1) as well as high pore volumes (max. 1.14 cm3g-1). The monodispersity of the CMSNs was verified by statistical particle size distribution from SEM (scanning electron microscopy)/TEM (transmission electron microscopy) images and DLS (dynamic light scattering). The mesophase transformation can be rationalized on the basis of an ethanol-driven change of the surfactant packing structure and charge matching at the surfactant/silicate interface. The corresponding morphology evolution can be elucidated by an ethanol-controlled hydrolysis rate of TEOS and degree of condensation of oligomeric silicate species via a nucleation and growth process.

The effect of abrasion on corrosion of dental Co-Cr alloys: An in vitro study

The effect of abrasion on corrosion of two dental Co-Cr alloys was investigated in vitro. The metals were abraded with a toothbrush and with a piece of tooth enamel. Changes in corrosion potentials and metal release due to the abrasion were measured. Abrasion by tooth enamel caused a persistent drop in corrosion potentials. The release of both chromium and, in particular, cobalt was higher than during brushing. The two alloys tested did not significantly differ with regard to mode and rate of corrosion. It was demonstrated that the corrosion behavior of electrochemically passive dental alloys was affected by abrasion of a magnitude normally encountered during clinical service.