Optimized methods for extracting circulating small RNAs from long-term stored equine samples.

Circulating miRNAs in body fluids, particularly serum, are promising candidates for future routine biomarker profiling in various pathologic conditions in human and veterinary medicine. However, reliable standardized methods for miRNA extraction from equine serum and fresh or archived whole blood are sorely lacking. We systematically compared various miRNA extraction methods from serum and whole blood after short and long-term storage without addition of RNA stabilizing additives prior to freezing. Time of storage at room temperature prior to freezing did not affect miRNA quality in serum. Furthermore, we showed that miRNA of NGS-sufficient quality can be recovered from blood samples after >10 years of storage at -80 °C. This allows retrospective analyses of miRNAs from archived samples.

Major and trace element concentration of melt inclusions from Zhupanovsky volcano, Kamchatka

The paper presents data on naturally quenched melt inclusions in olivine (Fo 69-84) from Late Pleistocene pyroclastic rocks of Zhupanovsky volcano in the frontal zone of the Eastern Volcanic Belt of Kamchatka. The composition of the melt inclusions provides insight into the latest crystallization stages (~70% crystallization) of the parental melt (~46.4 wt % SiO2, ~2.5 wt % H2O, ~0.3 wt % S), which proceeded at decompression and started at a depth of approximately 10 km from the surface. The crystallization temperature was estimated at 1100 ± 20°C at an oxygen fugacity of deltaFMQ = 0.9-1.7. The melts evolved due to the simultaneous crystallization of olivine, plagioclase, pyroxene, chromite, and magnetite (Ol: Pl: Cpx : (Crt-Mt) ~ 13 : 54 : 24 : 4) along the tholeiite evolutionary trend and became progressively enriched in FeO, SiO2, Na2O, and K2O and depleted in MgO, CaO, and Al2O3. Melt crystallization was associated with the segregation of fluid rich in S-bearing compounds and, to a lesser extent, in H2O and Cl. The primary melt of Zhupanovsky volcano (whose composition was estimated from data on the most primitive melt inclusions) had a composition of low-Si (~45 wt % SiO2) picrobasalt (~14 wt % MgO), as is typical of parental melts in Kamchatka and other island arcs, and was different from MORB. This primary melt could be derived by ~8% melting of mantle peridotite of composition close to the MORB source, under pressures of 1.5 ± 0.2 GPa and temperatures 20-30°C lower than the solidus temperature of 'dry' peridotite (1230-1240°C). Melting was induced by the interaction of the hot peridotite with a hydrous component that was brought to the mantle from the subducted slab and was also responsible for the enrichment of the Zhupanovsky magmas in LREE, LILE, B, Cl, Th, U, and Pb. The hydrous component in the magma source of Zhupanovsky volcano was produced by the partial slab melting under water-saturated conditions at temperatures of 760-810°C and pressures of ~3.5 GPa. As the depth of the subducted slab beneath Kamchatkan volcanoes varies from 100 to 125 km, the composition of the hydrous component drastically changes from relatively low-temperature H2O-rich fluid to higher temperature H2O-bearing melt. The geothermal gradient at the surface of the slab within the depth range of 100-125 km beneath Kamchatka was estimated at 4°C/km.

Food labeling background papers.

Issued jointly with U.S. Department of Agriculture, Federal Trade Commission, Bureau of Consumer Protection.

Development of double isotope derivative dilutions analyses for selected food addititives /

"Contract No. AT(30-1)-2715."

Animal drug amendments of 1966.

Mode of access: Internet.

Report to the Commissioner of the Food and Drug Administration by the FDA Task Force on the use of antibiotics in animal feeds.

Appended: Press briefing by Commissioner Charles C. Edwards and by C. D. Van Houweling.

Adverse Health Effects of Chronic Exposure to Low-Level Cadmium in Foodstuffs and Cigarette Smoke

Cadmium is a cumulative nephrotoxicant that is absorbed into the body from dietary sources and cigarette smoking. The levels of Cd in organs such as liver and kidney cortex increase with age because of the lack of an active biochemical process for its elimination coupled with renal reabsorption. Recent research has provided evidence linking Cd-related kidney dysfunction and decreases in bone mineral density in nonoccupationally exposed populations who showed no signs of nutritional deficiency. This challenges the previous view that the concurrent kidney and bone damage seen in Japanese itai-itai disease patients was the result of Cd toxicity in combination with nutritional deficiencies, notably, of zinc and calcium. Further, such Cd-linked bone and kidney toxicities were observed in people whose dietary Cd intakes were well within the provisional tolerable weekly intake (PTWI) set by the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives of 1 mug/kg body weight/day or 70 mug/day. This evidence points to the much-needed revision of the current PTWI for Cd. Also, evidence for the carcinogenic risk of chronic Cd exposure is accumulating and Cd effects on reproductive outcomes have begun to emerge.

Effect of metal chlorides on the sintering and densification of hydroxyapatite adsorbent

This work is part of a series of studies dealing with the evaluation of the effects of major elements of solid waste, especially metallic oxides, nitrates, sulfates, and chlorides, on the sintering and the densification of calcium hydroxyapatite (Ca-HAP) adsorbent. The effects of chloride salts of potassium (KCl) and zinc (ZnCl2) on sintering and densification of Ca-HAP were studied using surface area reduction and shrinkage measurements. The addition of KCl (2% w/w) activated the sintering process by bringing a swift reduction in surface area and lowering the densification temperature. However, a low final densification was achieved. Increasing the amount of this additive to 10% w/w further lowered the final densification and lowered the densification temperature of hydroxyapatite by 150 degrees C. On the other hand, the addition of 2 wt % of ZnCl2 deactivated the sintering process by slowing down the densification process and raising the densification temperature. However, the reduction of surface area was comparable to that of Ca-HAP. The densification rate contained two or more rate maxima indicating the additives (salts) bring multiple speeds in the densification process.

Effect of carbon/noncarbon addition on hydrogen storage behaviors of magnesium hydride

Various Mg/carbon and Mg/noncarbon composite systems were prepared by mechanical milling and their hydrogen storage behaviors were investigated. It was found that all the carbon additives exhibited prominent advantage over the noncarbon additives, such as BN nanotubes (BNNTs) or asbestos in improving the hydrogen capacity and dehydriding/hydriding kinetics of Mg. And among the various carbon additives, purified single-walled carbon nanotubes (SWNTs) exhibited the most prominent catalytic effect on the hydrogen storage properties of Mg. The hydrogen capacities of all Mg/C composites at 573 K reached more than 6.2 wt.% within 10 min, about 1.5 wt.% higher than that of pure MgH2 at the identical operation conditions. Under certain operation temperatures, H-absorption/desorption rates of Mg/carbon systems were over one order of magnitude higher than that of pure Mg. Furthermore, the starting temperature of the desorption reaction of MgH2 has been lowered to 60 K by adding SWNTs. On the basis of the hydrogen storage behavior and structure/phase investigations, the possible mechanism involved in the property improvement of Mg upon adding carbon materials was discussed. (c) 2005 Elsevier B.V. All rights reserved.

Crisis management and risk communication: Why the food industry must get it right

There is public unease about food-related issues including food additives, food poisoning bacteria and GM ingredients. The public wants evidence of no risks, but all regulators can ever offer is no evidence of risk or evidence of a very small risk. The situation is complex because experts and non-experts can perceive the same risk in vastly different ways. The way in which the food industry manages crises and communicates risks will determine the public acceptance and success of new technologies such as GM foods and nanomaterials. There is a need for the food industry (including regulators and scientific experts) to sharpen up their risk communication skills to ensure that technical innovations are accepted by consumers, and crises such as food recalls do not undermine the public's confidence in the food industry. The AIFST has a key role to play in driving the risk communication process and allaying public unease about food-related issues.

Crystallization and preliminary X-ray analysis of a Kunitz-type inhibitor, textilinin-1 from Pseudonaja textilis textilis

Textilinin-1 (Txln-1), a Kunitz-type serine protease inhibitor, is a 59-amino-acid polypeptide isolated from the venom of the Australian Common Brown snake Pseudonaja textilis textilis. This molecule has been suggested as an alternative to aprotinin, also a Kunitz-type serine protease inhibitor, for use as an anti-bleeding agent in surgical procedures. Txln-1 shares only 47% amino-acid identity to aprotinin; however, six cysteine residues in the two peptides are in conserved locations. It is therefore expected that the overall fold of these molecules is similar but that they have contrasting surface features. Here, the crystallization of recombinant textilinin-1 (rTxln-1) as the free molecule and in complex with bovine trypsin (229 amino acids) is reported. Two organic solvents, phenol and 1,4-butanediol, were used as additives to facilitate the crystallization of free rTxln-1. Crystals of the rTxln-1-bovine trypsin complex diffracted to 2.0 angstrom resolution, while crystals of free rTxln-1 diffracted to 1.63 angstrom resolution.

Vaccine components and constituents: Responding to consumer concerns

Vaccination remains a vital strategy in the prevention of infectious disease. Commercial vaccine formulations contain a range of additives or manufacturing residuals, which may contribute to patient concerns about vaccine safety. Primary health care professionals are well placed to address patient concerns about vaccine safety. We describe the key constituents present in vaccines, discuss issues related to safety and acceptability of these constituents, and provide a table highlighting constituents of commercially available vaccines in Australia.

Enzymatic production of oligosaccharides in centrifugal fields

The aims of this work have been to identify an enzymatic reaction system suitable to investigate and develop the high-speed centrifuge as a novel reaction system for performing such reactions. The production of galacto-oligosaccharides by the trans-galactosyl activity of the enzyme β-galactosidase on lactose monohydrate was identified as a model enzymatic system to elucidate the principles of this type of process. Galacto-oligosaccharides have attracted considerable commercial interest as food additives which have been shown to be beneficial to the health of the human gastrointestinal tract. The development of a single unit operation capable of controlling the biosynthesis of galacto-oligosaccharides whilst simultaneously separating the enzyme from the reaction products would reduce downstream processing costs. This thesis shows for the first time that by using a combination of (a) immobilised or insolubilised β-galactosidase , (b) a rate-zonal centrifugation technique, and (c) various applied centrifugal fields, that a high-speed centrifuge could be used to control the formation of galacto-oligosaccharides whilst removing the enzyme from the reaction products. By layering a suspension of insolubilised β-galactosidase on top of a lactose monohydrate density gradient and centrifuging, the applied centrifugal fields generated produced sedimentation of the enzyme particles through the substrate. The higher sedimentation rate of the enzyme compared to those of the reaction products allowed for separation to take place. Complete sedimentation, or pelleting of the enzyme permits the possible recovery and re-use. Insolubilisation of the enzyme allowed it to be sedimented through the substrate gradient using much lower applied centrifugal fields than that required to sediment free soluble enzyme and this allowed for less expensive centrifugation equipment to be used. Using free soluble and insolubilised β-galactosidase stirred-batch reactions were performed to investigate the kinetics of lactose monohydrate hydrolysis and galacto-oligosaccharide formation. Based on these results a preliminary mathematical model based on Michaelis-Menten kinetics was produced. It was found that the enzyme insolubilisation process using a chemical cross-linking agent did not affect the process of galacto-oligosaccharide formation. Centrifugation experiments were performed and it was found that by varying the applied centrifugal fields that the yield of galacto-oligosaccharides could be controlled. The higher the applied centrifugal fields the lower the yield of galacto-oligosaccharides. By increasing the applied centrifugal fields the 'contact time' between the sedimenting enzyme and the substrate was reduced, which produced lower yields. A novel technique involving pulsing the insolubilised enzyme through the substrate gradient was developed and this was found to produce higher yields of galacto-oligosaccharide compared to using a single enzyme loading equivalent to the total combined activity of the pulses. Comparison of the galacto-oligosaccharide yields between stirred-batch and centrifugation reactions showed that the applied centrifugal fields did not adversely affect the transgalactosyl activity of the insolubilised enzyme.

Synthesis and evaluation of novel polymer modifiers for biodegradable polymers

The effects of ester plasticizers and copolymers on the mechanical properties of the natural biodegradable polymers, poly(3-hydroxybutyrate) [PHB] and poly(lactic acid) [PLA] have been studied after subjecting to melt processing conditions. Ester plasticizers were synthesized from citric, tartaric and maleic acids using various alcohols. A variety of PLA copolymers have also been prepared from poly(ethylene glycol) derivatives using stannous octanoate catalysed ring opening polymerisations of DL-lactide. A novel PLA star copolymer was also prepared from an ethoxylated pentaerythritol. The structures of these copolymers were determined by NMR spectroscopy. The plasticizing effect of the synthesised additives at various concentrations was determined. While certain additives were capable of improving the mechanical properties of PLA, none were effective in PHB. Moreover, it was found that certain combinations of additives exhibited synergistic effects. Possible mechanisms are discussed. Biotic and abiotic degradation studies showed that the plasticizers (esters and copolymers) did not inhibit the biodegradability of PHB or PLA in compost at 60°C. Simple toxicity tests carried out on compost extract and its ability to support the growth of cress seeds was established. PLA was found to be susceptible to limited thermal degradation under melt processing conditions. Conventional phenolic antioxidants showed no significant effect on this process, suggesting that degradation was not predominantly a free radical process. PLA also underwent photo-oxidative degradation with UV light and the process could be accelerated in the presence of a photoactivator such as iron (III) diisononyl dithiocarbamate. The mechanisms for the above processes are discussed. Finally, selected compounds were prepared on a pilot plant scale. Extruded and blown films were prepared containing these additives with conventional polymer processing equipment. The mechanical properties were similar to those obtained with laboratory produced compression moulded films.

The effect of complexes on combustion

The twin goals of low and efficient fuel use and minimum emissions are increasingly being addressed by research in both the motor and the catalyst industries of the world. This study was designed to attempt to investigate these goals. For diesel engine vehicles, this can be achieved by improving the efficiency of the fuel combustion in the combustion chamber. By having a suitable oxidation catalyst in the fuel one would expect the efficiency of the fuel combustion to be increased and fewer partial oxidation products to be formed. Also by placing a catalyst converter in the exhaust system partial oxidation products may be converted to more desirable final products. Finally, in this research the net catalytic effect of using an additive treated fuel and a blank ceramic monolith to trap the metal in the exhaust gases for potential use as catalytic converter was investigated. Suitable metal additives must yield a stable solution in the fuel tank. That is, they should not react with the air, water and rust that are always present. The research was targeted on the synthesis of hydrocarbon-soluble complexes that might exhibit unusually slow rates of ligand substitution. For materials containing metal ions, these properties are best met by using multi-dentate ligands that form neutral complexes. Metal complexes have been synthesised using acetylacetone derivatives, schiff base ligands and macrocyclic polyamine ligands, as potential pro-oxidant additives. Their thermal stabilities were also investigated using a differential thermal analysis instrument. The complexes were then investigated as potential additives for use in diesel fuel. The tests were conducted under controlled conditions using a diesel combustion bomb simulating the combustion process in the D.I. diesel engine, a test bed engine, and a vehicle engine.