Solid phase microextraction of stale flavour volatiles from the headspace of UHT milk

Solid phase microextraction (SPME) offers a solvent-free and less labour-intensive alternative to traditional flavour isolation techniques. In this instance, SPME was optimised for the extraction of 17 stale flavour volatiles (C3-11,13 methyl ketones and C4-10 saturated aldehydes) from the headspace of full-cream ultrahigh-temperature (UHT)-processed milk. A comparison of relative extraction efficiencies was made using three fibre coatings, three extraction times and three extraction temperatures. Linearity of calibration curves, limits of detection and repeatability (coefficients of variation) were also used in determining the optimum extraction conditions. A 2 cm fibre coating of 50130 gm divinylbenzene/Carboxen/polydimethylsiloxane in conjunction with a 15 min extraction at 40 degrees C were chosen as the final optimum conditions. This method can be used as an objective tool for monitoring the flavour quality of UHT milk during storage. (c) 2005 Society of Chemical Industry.

On the optimal ratio of heavy to light chain genes for efficient recombinant antibody production by CHO cells

Monoclonal antibodies (Mab) are heterotetramers consisting of an equimolar ratio of heavy chain (HC) and light chain (LC) polypeptides. Accordingly, most recombinant Mab expression systems utilize an equimolar ratio of heavy chain (he) to light chain (lc) genes encoded on either one or two plasmids. However, there is no evidence to suggest that this gene ratio is optimal for stable or transient production of recombinant Mab. In this study we have determined the optimal ratio of hc:lc genes for production of a recombinant IgG(4) Mab, cB72.3, by Chinese hamster ovary (CHO) cells using both empirical and mathematical modeling approaches. Polyethyleneimine-mediated transient expression of cB72.3 at varying ratios of hc:lc genes encoded on separate plasmids yielded an optimal Mab titer at a hc:lc gene ratio of 3:2; a conclusion confirmed by separate mathematical modeling of the Mab folding and assembly process using transient expression data. On the basis of this information, we hypothesized that utilization of he genes at low hc:lc gene ratios is more efficient. To confirm this, cB72.3 Mab was transiently produced by CHO cells at constant he and varying lc gene dose. Under these conditions, Mab yield was increased with a concomitant increase in lc gene dose. To determine if the above findings also apply to stably transfected CHO cells producing recombinant Mab, we compared the intra- and extracellular ratios of HC and LC polypeptides for three GS-CHO cells lines transfected with a 1:1 ratio of hc:lc genes and selected for stable expression of the same recombinant Mab, cB72.3. Intra- and extracellular HC:LC polypeptide ratios ranged from 1:2 to 1:5, less than that observed on transient expression of the same Mab in parental CHO cells using the same vector. In conclusion, our data suggest that the optimal ratio of hc:lc genes used for transient and stable expression of Mab differ. In the case of the latter, we infer that optimal Mab production by stably transfected cells represents a compromise between HC abundance limiting productivity and the requirement for excess LC to render Mab folding and assembly more efficient.

Salt diffusion and distribution in meat studied by Na-23 nuclear magnetic resonance imaging and relaxometry

This study introduces the use of combined Na-23 magnetic resonance imaging (MRI) and Na-23 NMR relaxometry for the study of meat curing. The diffusion of sodium ions into the meat was measured using Na-23 MRI on a 1 kg meat sample brined in 10% w/w NaCl for 3-100 h. Calculations revealed a diffusion coefficient of 1 x 10(-5) cm(2)/s after 3 h of curing and subsequently decreasing to 8 x 10(-6) cm(2)/s at longer curing times, suggesting that changes occur in the microscopic structure of the meat during curing. The microscopic mobility and distribution of sodium was measured using Na-23 relaxometry. Two sodium populations were observed, and with increasing length of curing time the relaxation times of these changed, reflecting a salt-induced swelling and increase in myofibrillar pore sizes. Accordingly, the present study demonstrated that pore size and thereby salt-induced swelling in meat can be assessed using Na-23 relaxometry.

Gingerol content of diploid and tetraploid clones of ginger (Zingiber officinale Roscoe)

Ginger (Zingiber officinale, Roscoe), a monocotyledonous, sterile cultigen, is widely used as a spice, flavoring agent, and herbal medicine. The pungency of fresh ginger is due to a series of homologous phenolic ketones of which [6]-gingerol is the major one. The gingerols are thermally unstable and can be converted to their corresponding shogaols, which are present in dried ginger, Fresh rhizomes of 17 clones of Australian ginger, including commercial cultivars and experimental tetraploid clones, were assayed by HPLC for gingerols and shogaols. [6]-Gingerol was identified as the major pungent phenolic compound in all samples, while [8]- and [10]-gingerol occurred in lower concentrations. One cultivar known as Jamaican contained the highest concentrations of all three gingerols and was the most pungent of the clones analyzed. Gingerols were stable in ethanolic solution over a 5-month period when stored at 4 degrees C. Shogaols were not identified in the extracts prepared from fresh rhizomes at ambient temperature, confirming that these compounds are not native constituents of fresh ginger, In contrast to previous findings, this study did not find significant differences in gingerol concentrations between the tetraploid clones and their parent diploid cultivar.

Bilberry adulteration using the food dye amaranth

Vaccinium myrtillus or bilberry fruit is a commonly used herbal product. The usual method of determining the anthocyanin content is a single-wavelength spectrophotometric assay. Using this method, anthocyanin levels of two extracts were found to be 25% as claimed by the manufacturers. When high-performance liquid chromatography (HPLC) was used, however, one extract was found to contain 9% anthocyanins probably not derived from V. myrtillus but from an adulterant. This adulterant was subsequently identified, using HPLC, mass spectroscopy, and nuclear magnetic resonance, as amaranth, that is, 3-hydroxy-4-[(4-sulfo-1-naphthalenyl)azo]-2,7-naphthalenedisulfonic acid trisodium saltsa synthetic dark red sulfonic acid based naphthylazo dye. As described in this study, if deliberate adulteration occurs in an extract, a single-wavelength spectrophotometric assay is inadequate to accurately determine the levels of compounds such as anthocyanins. Detection of deliberate adulteration in commercial samples thus requires the use of alternative, more sophisticated, methods of analysis such as HPLC with photodiode array detection as a minimum.

Biological response of broiler chickens fed peas (Pisum sativum L.) expressing the bean (Phaseolus vulgaris L.) alpha-amylase inhibitor transgene

The nutritive value of transgenic peas expressing an a-amylase inhibitor (alpha-Ail) was evaluated with broiler chickens. The effects of feeding transgenic peas on the development of visceral organs associated with digestion and nutrient absorption were also examined. The chemical composition of the conventional and the transgenic peas used in this study were similar. In the two feeding trials, that were conducted normal and transgenic peas were incorporated into a maize-soybean diet at concentrations up to 500 g kg(-1). The diets were balanced to contain similar levels of apparent metabolisable energy (AME) and amino acids. In the first trial, the birds were fed the diets from 3 to 17days post-hatching and with levels of transgenic peas at 250 g kg(-1) or greater there was a significant reduction in body weight but an increase in feed intake resulting in deceased feed conversion efficiency. In the second trial, in which the birds were fed diets containing 300 g kg(-1) transgenic peas until 40 days of age, growth performance was significantly reduced. It was also demonstrated that the ileal starch digestibility coefficient (0.80 vs 0.42) was significantly reduced in the birds fed transgenic peas. Determination of AME and ileal digestibility of amino acids in 5-week-old broilers demonstrated a significant reduction in AME (12.12 vs 5.08 MJ kg(-1) DM) in the birds fed the transgenic peas. The AME value recorded for transgenic peas reflected the lower starch digestibility of this line. Real digestion of protein and amino acids was unaffected by treatment. Expression of a-Ail in peas did not appear to affect bird health or the utilisation of dietary protein. However, the significant reduction in ileal digestion of starch in transgenic peas does reduce the utility of this feedstuff in monogastric diets where efficient energy utilisation is required. (c) 2006 Society of Chemical Industry.

Iron (Fe) bioavailability and the distribution of anti-Fe nutrition biochemicals in the unpolished, and bran fraction of five rice gei polished grain riotypes

Iron (Fe) bioavailability in unpolished, polished grain and bran fraction of five rice genotypes with a range of Fe contents was measured by in vitro digestion and cultured Caco-2 cells of cooked grain. There was a significant difference in Fe bioavailability among the five rice genotypes tested, in both the unpolished and polished grain. The range of Fe bioavailability variation in polished rice was much wider than that of unpolished, suggesting the importance of using Fe levels and bioavailability in polished rice grain as the basis for selecting high-Fe rice cultivars for both agronomic and breeding purposes. Milling and polishing the grain to produce polished (or white) rice increased Fe bioavailability in all genotypes. Iron bioavailability in polished rice was high in the UBON2 and Nishiki, intermediate in both IR68144 and KDML105, and low in CMU122. All genotypes had low bioavailability of Fe in bran fraction compared to unpolished and polished grain, except in CMU122. CMU122 contained the lowest level of bioavailable Fe in unpolished and polished grain and bran, because of the dark purple pericarp colored grain and associated tannin content. The level of bioavailable Fe was not significantly correlated with grain Fe concentration or grain phytate levels among these five genotypes tested. The negative relationship between Fe bioavailability and the levels of total extractable phenol was only observed in unpolished (r = -0.83**) and bran fraction (r = -0.50*). The present results suggested that total extractable phenol and tannin contents could also contribute to lowering bioavailability of Fe in rice grain, in addition to phytate. (c) 2006 Society of Chemical Industry

Total and ileal digestible tryptophan contents of feedstuffs for broiler chickens

Reliable values of total and digestible tryptophan in components of feed formulation matrices are needed because tryptophan is often the third limiting amino acid in practical poultry diets. However, tryptophan is oxidatively destroyed during acid hydrolysis in routine amino acid analysis and its determination requires a separate analytical procedure. The variability in contents and apparent ileal digestibility for 6-week-old broiler chickens of tryptophan in 74 samples representing 24 feedstuffs are presented in this paper. The average ileal tryptophan digestibility coefficient in wheat was 0.83, in sorghum and triticale 0.75, maize 0.71, soybean meal 0.84, sunflower meal 0.81, canola meal 0.78 and cottonseed meal 0.75. Among the grain legumes, tryptophan in lupins was better digested than that in chickpeas, fababeans and field peas. Among the animal protein meals, the tryptophan digestibility coefficients in fish meal (0.77) and blood meal (0.84) were substantially higher than those in meat meal (0.64), meat-and-bone meal (0.63) and feather meal (0.52). Marked variations in tryptophan digestibility were also observed among samples of fish meal, meat-and-bone meal and meat meal, highlighting significant batch-to-batch differences. For most feedstuffs, considerable variability was observed in the tryptophan concentrations, but such variations were not reflected in digestibility coefficients. (c) 2006 Society of Chemical Industry.

Survival of probiotics encapsulated in chitosan-coated alginate beads in yoghurt from UHT- and conventionally treated milk during storage

Survival of the microencapsulated probiotics, Lactobacillus acidophilus 547, Bifidobacterium bifidum ATCC 1994, and Lactobacillus casei 01, in stirred yoghurt from UHT- and conventionally treated milk during low temperature storage was investigated. The probiotic cells both as free cells and microencapsulated cells (in alginate beads coated with chitosan) were added into 20 g/100 g total solids stirred yoghurt from UHT-treated milk and 16 g/100 g total solids yoghurt from conventionally treated milk after 3.5 h of fermentation. The products were kept at 4 degrees C for 4 weeks. The survival of encapsulated probiotic bacteria was higher than free cells by approximately 1 log cycle. The number of probiotic bacteria was maintained above the recommended therapeutic minimum (10(7) cfu g(-1)) throughout the storage except for R bifidum. The viabilities of probiotic bacteria in yoghurts from both UHT- and conventionally treated milks were not significantly (P > 0.05) different. (c) 2004 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Assembly of Human Papillomavirus Type-16 Virus-Like Particles: Multifactorial Study of Assembly and Competing Aggregation

Pentameric capsomeres of human papillomavirus capsid protein L1 expressed in Escherichia coli self-assemble into virus-like particles (VLPs) in vitro. A multifactorial experimental design was used to explore a wide range of solution conditions to optimize the assembly process. The degree of assembly was measured using an enzyme-linked immunosorbent assay, and a high-throughput turbidity assay was developed to monitor competing aggregation. The presence of zinc ions in the assembly buffer greatly increased the incidence of aggregation and had to be excluded from the experiment for meaningful analysis. Assembly of VLPs was optimal at a pH of about 6.5, calcium and sodium ions had no measurable effect, and dithiothreitol and glutathione inhibited assembly. Tryptophan fluorescence spectroscopy demonstrated that an increase in urea concentration reduced the rate of VLP formation but had no effect on the final concentration of assembled VLPs. This study demonstrates the use of the hanging-drop vapor-diffusion crystallization method to screen for conditions that promote aggregation and the use of tryptophan fluorescence spectroscopy for real-time monitoring of the assembly process.

The politics of knowledge in central Asia:science between Marx and the market

Through careful historical and ethnographic research and extensive use of local scholarly works, this book provides a persuasive and careful analysis of the production of knowledge in Central Asia. The author demonstrates that classical theories of science and society are inadequate for understanding the science project in Central Asia. Instead, a critical understanding of local science is more appropriate. In the region, the professional and political ethos of Marxism-Leninism was incorporated into the logic of science on the periphery of the Soviet empire. This book reveals that science, organizes and constructed by Soviet rule, was also defined by individual efforts of local scientists. Their work to establish themselves 'between Marx and the market' is therefore creating new political economies of knowledge at the edge of the scientific world system.

Transnational environmental policy:reconstructing ozone

Transnational Environmental Policy analyses a surprising success story in the field of international environmental policy making: the threat to the ozone layer posed by industrial chemicals, and how it has been averted. The book also raises the more general question about the problem-solving capacities of industrialised countries and the world society as a whole. Reiner Grundmann investigates the regulations which have been put in place at an international level, and how the process evolved over twenty years in the US and Germany.

The genius of Erasmus Darwin

The Genius of Erasmus Darwin provides insight into the full extent of Erasmus Darwin's exceptional intellect. He is shown to be a major creative thinker and innovator, one of the minds behind the late eighteenth-century industrial revolution, and one of the first, if not the first, to perceive the living world (including humans) as part of a unified evolutionary scenario. The contributions here provide contextual understandings of Erasmus Darwin's thought, as well as studies of particular works and accounts of the later reception of his writings. In this way it is possible to see why the young Samuel Taylor Coleridge was moved to describe Darwin as 'the first literary character in Europe, and the most original-minded man'. Erasmus Darwin, Charles Darwin's grandfather, was one of the leading intellectuals of eighteenth-century England. He was a man with an extraordinary range of interests and activities: he was a doctor, biologist, inventor, poet, linguist, and botanist. He was also a founding member of the Lunar Society, an intellectual community that included such eminent men as James Watt and Josiah Wedgwood. Contents: Introduction; Setting the scene, Jonathan Powers; Prologue 'Catching up with Erasmus Darwin in the New Century', Desmond King-Hele. Section 1: Medicine: Physicians and physic in 17th and 18th century Lichfield, Dennis Gibbs; Dr Erasmus Darwin MD FRS (1731–1802): England's greatest physician?, Gordon Cook; William Pale (1743–1805) and James Parkinson (1755–1824): two peri-Erasmatic thinkers (and several others), Christopher Gardner-Thorpe; The vertiginous philosophers: Erasmus Darwin and William Charles Wells on vertigo, Nicholas Wade. Section 2: Biology: The Antipodes and Erasmus Darwin: the place of Erasmus Darwin in the heritage of Australian literature and biology, John Pearn; Erasmus Darwin on human reproductive generation: placing heredity within historical and Zoonomian contexts, Philip Wilson; All from fibres: Erasmus Darwin's evolutionary psychobiology, C.U.M. Smith; Two special doctors: Erasmus Darwin and Luigi Galvani, Rafaella Simili. Section 3: Education: But what about the women? The lunar society's attitude to women and science and to the education of girls, Jenny Uglow; The Derbyshire 'Darwinians': the persistence of Erasmus Darwin's influence on a British provincial literary and scientific community, c.1780–1850, Paul Elliot. Section 4: Technology: Designing better steering for carriages (and cars); with a glance at other inventions, Desmond King-Hele; Mama and papa: the ancestors of modern-day speech science, Philip Jackson; Negative and positive images: Erasmus Darwin, Tom Wedgwood and the origins of photography, Alan Barnes; Section 5: Environment: Erasmus Darwin's contributions to the geological sciences, Hugh Torrens; The air man, Desmond King-Hele; Erasmus Darwin, work and health, Tim Carter; Section 6: Literature: The progress of society: Darwin's early drafts for the temple of nature, Martin Priestman; The poet as pathologist: myth and medicine in Erasmus Darwin's epic poetry, Stuart Harris; 'Another and the same': nature and human beings in Erasmus Darwin's doctrines of love and imagination, Maurizio Valsania. Epilogue: 'One great slaughter-house the warring world': living in revolutionary times, David Knight; Coda: Midlands memorabilia, Nick Redman; Appendix: The Creation of the Erasmus Darwin Foundation and Erasmus Darwin House, Tony Barnard; Index.