Platinum/iridium/carbon: a high-resolution shadowing material for TEM,STM and SEM of biological macromolecular structures

Thin Pt/Ir/C coating films (1.5 nm) show a fine granularity and provide a high structural resolution in the transmission electron microscope (TEM) when applied to freeze-dried biological macromolecules. They keep their structure when exposed to atmospheric conditions, without the need of an additional stabilizing carbon layer, in contrast to conventional high-resolution shadowing materials such as Ta/W and Pt/C. However, the correct ratio of the components has turned out to be crucial. When evaporating Pt/Ir/C from the source electrode in an electron-beam-heated evaporator, the ratio of the three elements changes progressively, and, consequently, the properties of such films depend strongly on the mass that has been pre-evaporated. In this paper we present a quantitative analysis of the composition of Pt/Ir/C films by wavelength-dispersive X-ray analysis (WDX) undertaken in association with TEM experiments. We applied Pt/Ir/C shadowing to two regular biological test specimens, the phage T4 type III polyhead and the HPI-layer of Deinococcus radiodurans. It turns out that Pt/Ir/C films containing at least 25% C are three-dimensionally stable on the freeze-dried macromolecular samples. By the dramatically improved resolution power of the latest scanning electron microscopes (SEM) and the invention of the scanning tunnelling microscope (STM), two new surface-sensitive tools for the investigation of biological macromolecular structures became available. The Pt/Ir/C coating has proved to be well suited for STM and SEM imaging of freeze-dried biological structures because of its good electrical conductivity and its direct three-dimensional stability. We compare STM, SEM and TEM images of freeze-dried and Pt/Ir/C-coated polyheads.     

Cryofixation of vascular endothelium

Cryofixation refers to the immobilization of tissue components by the rapid removal of heat from the specimen, so that the structure is interred and stabilized in a natural embedding medium, namely, frozen (amorphous or microcrystalline) tissue water. Cryofixation is now often used as a complement to the more traditional fixation methods, especially when the cell structure is delicate or dynamic and may be inaccurately preserved by the slow selective action of chemical fixatives. Vascular endothelial cells are specialized for transcellular transport and for the regulation of blood flow and composition. The dynamic and labile subcellular organization of these cells, presumably reflecting these functional specializations, makes them ideal candidates for cryofixation. Several different types of endothelial cells were directly frozen at temperatures below 20 degrees Kelvin by pressing them against a liquid-helium-cooled block. These samples were subsequently processed for structural analysis by freeze-substitution. Detailed rationales, designs, and protocols are described for both freezing and freeze-substitution. Electron micrographs of cryofixed arterial and venous capillaries (rete mirabile of the American eel), iliac vein (rabbit), and cultured endothelium from the iliac vein (human) reveal that the organization of the characteristic intracellular membrane system of endothelial vesicles is qualitatively similar to that seen in chemically fixed endothelium, especially with regard to the interconnection of clusters of individual vesicles to form elaborate networks. The luminal and abluminal networks are not in communication, at least not in static images. Quantitatively, however, most directly frozen endothelial cells have far fewer vesicular profiles than comparable glutaraldehydefixed cells. The differences can be explained by presuming that the rapid action of cryofixation (approximately 1 msec) gives a more accurate picture of the vesicular network because it captures the transient structure of labile or dynamic membranes.     

Overheating in Auckland homes: testing and interventions in full-scale and simulated houses

ABSTRACT

New Zealand dwellings have thermal conditions managed with relatively light regulation. No minimum airtightness standards exist and historical increases in required insulation levels aimed to reduce winter heating energy consumption. A consequence of this policy is an increased potential for overheating in summer. There has been a steady increase in the use of heat pumps, risking heating energy savings being outweighed by cooling energy increases. Internal temperatures and humidity were monitored in the living spaces of three unoccupied, transportable houses over all four seasons of the Auckland climate. The houses are located on the same site and are of identical construction, apart from selected interventions which were tested to explore their potential to mitigate overheating. Results indicate that overheating can be extreme and long lasting. High internal temperatures are very closely connected with solar gains. Internal temperatures reached 32°C in autumn. Roof space temperatures reached 51°C in summertime. Interventions resulted in modest improvements and an airtight construction provided a small thermal benefit. A thermal model for the houses was developed using EnergyPlus and compared with actual measurements and the interventions. Early results point to the further need to reduce solar gain, increase roof-space ventilation and increase mass, where feasible.     

外墙彩色砂浆的泛碱问题

在干混砂浆工业中．泛碱造成水泥基材料褪色而影响美观性的现象是一个大家普遍关心的问题．泛碱是由于盐分沉积在矿物建筑材料如混凝土．砂浆或砖墙的表面引起的．目前．泛碱对于装饰砂浆如彩色抹灰砂浆和瓷砖填缝剂而言依然是一个令人头痛的问题．泛碱通常不会造成砂浆明显的破坏．而主要是影响产品的感官质量．泛碱经常在建筑物投入使用后的很短时间内产生．此时正是工人．建筑师和业主对新建筑物外观质量最关心的时候．特别是当彩色砂浆的颜色较深时．表面形成的白斑会引起对比度强烈的不规则斑痕．     

Compact Saloplastic Poly(Acrylic Acid)/Poly(Allylamine) Complexes: Kinetic Control Over Composition,Microstructure, and Mechanical Properties

Durable compact polyelectrolyte complexes (CoPECs) with controlled porosity and mechanical properties are prepared by ultracentrifugation. Because the starting materials, poly(allylamine hydrochloride) (PAH) and poly(acrylic acid sodium salt) (PAA), are weak acids/bases, both composition and morphology are controlled by solution pH. In addition, the nonequilibrium nature of polyelectrolyte complexation can be exploited to provide a range of compositions and porosities under the influence of polyelectrolyte addition order and speed, and concentration. Confocal microscopy shows these “saloplastic” materials to be highly porous, where pore formation is attributed to a combination of deswelling of the polyelectrolyte matrix and expansion of small inhomogenities by osmotic pressure. The porosity (15–70%) and the pore size (<5 μm to >70 μm) of these materials can be tuned by adjusting the PAA to PAH ratio, the salt concentration, and the pH. The modulus of these CoPECs depends on the ratio of the two polyelectrolytes, with stoichiometric complexes being the stiffest due to optimized charge pairing, which correlates with maximized crosslinking density. Mechanical properties, pore sizes, and pore density of these materials make them well suited to three dimensional supports for tissue engineering applications.     

Inkjet Printing of Amphotericin B onto Biodegradable Microneedles Using Piezoelectric Inkjet Printing

The delivery of amphotericin B, a pharmacologic agent with activity against a broad spectrum of fungi as well as against parasitic protozoa, has been complicated by the fact that amphotericin B exhibits poor solubility in aqueous solutions at physiologic pH levels. In this study, piezoelectric inkjet printing was used to modify the surfaces of Gantrez 169 BF microneedles (Ashland, Covington, KY). These amphotericin B-loaded microneedles demonstrated activity against Candida parapsilosis in a radial diffusion assay. The results of this study suggest that a combination of visible light dynamic mask microstereolithography, micromolding, and piezoelectric inkjet printing may be used to prepare amphotericin B-loaded microneedles with antifungal properties. It is envisioned that microneedles containing amphotericin B may be used for transdermal delivery of pharmacologic agents for the treatment of cutaneous fungal infections as well as cutaneous leishmaniasis.     

Contents of dietary fibre components and their relation to associated bioactive components in whole grain wheat samples from the HEALTHGRAIN diversity screen

A large and diverse material collection of whole grain wheat samples (n = 129) was analysed for total dietary fibre (TDF) content and composition, including fructan (11.5–15.5%). Correlations between the dietary fibre components, associated bioactive components (e.g. tocols, sterols, phenolic acids and folates) and agronomic properties previously determined on the same samples were found with multivariate analysis (PCA). Samples from the same countries had similar characteristics. The first PC described variation in components concentrated in the starchy endosperm (e.g. starch, β-glucan and fructan) and the dietary fibre components concentrated in the bran (e.g. TDF, arabinoxylan and cellulose). The second PC described the variation in kernel weight and other bran components such as alkylresorcinols, tocols and sterols. Interestingly, there was no correlation among these different groups of bran components, which reflected their concentration in different bran tissues. The results are of importance for plant breeders who wish to develop varieties with health-promoting effects.