Attenuating mutations in the influenza virus genome which may increase the safety of vaccine production

Influenza virus epidemics occur on an annual basis and cause severe disease in the very young and old. The vaccine administered to high-risk groups is generated by amplifying reassortant viruses, with chronologically relevant viral surface antigens, in eggs. Every 20 years or so, influenza pandemics occur causing widespread fatality in all age groups. These viruses display novel viral surface antigens acquired from a zoonotic source, and vaccination against them poses new issues since production of large amounts of a respiratory virus containing novel surface antigens could be dangerous for those involved in manufacture. To minimise risks, it is advisable to use a virus whose genetic backbone is highly attenuated in man. Traditionally, the A/PR/8/34 strain of virus is used, however, the genetic basis of its attenuation is unclear. Cold-adapted (CA) strains of the influenza virus are all based on the H2N2 subtype, itself a virus with pandemic potential, and again the genetic basis of temperature sensitivity is not yet established. Reverse genetics technology allows us to engineer designer influenza viruses to order. Using this technology, we have been investigating mutations in several different gene segments to effectively attenuate potential vaccine strains allowing the safe production of vaccine to protect against the next pandemic. (C) 2003 Elsevier B.V. All rights reserved.

Changes in milk composition and processing properties during the spring flush period

The period, known to UK farmers and processors as the "spring flush", when the cows' diet changes from dry feed to spring pasture, has long been established as a time of change in milk properties and processing characteristics. Although it is believed to be a time when problems in processing are most likely to occur (e.g. milk that does not form clots or forms weak gels during cheesemaking), there is little evidence in the literature of detailed changes in milk composition and their impact on product manufacture. In this study, a range of physicochemical properties were analysed in milk collected from five commercial dairy herds before, during and after the spring flush period of 2006. In particular, total and ionic calcium contents of milk were studied in relation to other parameters including rennet clotting, acid gel properties, heat coagulation, alcohol stability, micelle size and zeta potential. Total divalent cations were significantly reduced from 35.4 to 33.4 mmol.L-1 during the study, while ionic calcium was reduced from 1.48 to 1.40 mmol.L-1 over the same period. Many parameters varied significantly between the sample dates. However, there was no evidence to suggest that any of the milk samples would have been unsuitable for processing - e.g. there were no samples that did not form clots with chymosin within a reasonable time or formed especially weak rennet or acid gels. A number of statistically significant correlations were found within the data, including ionic calcium concentration and pH; rennet clotting time (RCT) and micelle diameter; and RCT and ethanol stability. Overall, while there were clear variations in milk composition and properties over this period, there was no evidence to support the view that serious processing problems are likely during the change from dry feed to spring pasture.

An investigation into the mechanics of cutting using data from orthogonally cutting Nylon 66

Cutting force data for Nylon 66 has been examined in terms of various different models of cutting. Theory that includes significant work of separation at the tool tip was found to give the best correlation with experimental data over a wide range of rake angles for derived primary shear plane angle. A fracture toughness parameter was used as the measure of the specific work of separation. Variation in toughness with rake angle determined from cutting is postulated to be caused by mixed mode separation at the tool tip. A rule of mixtures using independently determined values of toughness in tension (mode 1) and shear (mode 11) is found to describe well the variation with rake angle. The ratio of modes varies with rake angle and, in turn, with the primary shear plane angle. Previous suggestions that cutting is a means of experimentally determining fracture toughness are now seen to be extended to identify the mode of fracture toughness as well.

Galactooligosaccharides (GOS) inhibit Vibrio cholerae toxin binding to its GM1 receptor

It is widely reported that cholera toxin (Ctx) remains a significant cause of gastrointestinal disease globally, particularly in developing countries where access to clean drinking water is at a premium. Vaccines are prohibitively expensive and have shown only short-term protection. Consequently, there is scope for continued development of novel treatment strategies. One example is the use of galactooligosaccharides (GOS) as functional mimics for the cell-surface toxin receptor (GM1). In this study, GOS fractions were fractionated using cation exchange chromatography followed by structural characterization using a combination of hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization mass spectrometry (ESI-MS) such that their molecular weight profiles were known. Each profile was correlated against biological activity measured using a competitive inhibitory GM1-linked ELISA. GOS fractions containing > 5% hexasaccharides (DP6) exhibited > 90% binding, with EC50 values between 29.27 and 56.04 mg/mL. Inhibition by GOS DP6, was dose dependent, with an EC50 value of 5.10 mg/mL (5.15 mu M MW of 990 Da). In removing low molecular weight carbohydrates that do possess prebiotic, nutraceutical, and/or biological properties and concentrating GOS DP5 and/or DP6, Ctx antiadhesive activity per unit of (dry) weight was improved. This could be advantageous in the manufacture of pharmaceutical or nutraceutical formulations for the treatment or prevention of an acute or chronic disease associated with or caused by the adhesion and/or uptake of a Ctx or HLT.

Achievement of dietary fatty acid intakes in long-term controlled intervention studies: approach and methodology

Objective: To describe the calculations and approaches used to design experimental diets of differing saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA) compositions for use in a long-term dietary intervention study, and to evaluate the degree to which the dietary targets were met. Design, setting and subjects: Fifty-one students living in a university hall of residence consumed a reference (SFA) diet for 8 weeks followed by either a moderate MUFA (MM) diet or a high MUFA (HM) diet for 16 weeks. The three diets were designed to differ only in their proportions of SFA and MUFA, while keeping total fat, polyunsaturated fatty acids (PUFA), trans-fatty acids, and the ratio of palmitic to stearic acid, and n-6 to n-3 PUFA, unchanged. Results: Using habitual diet records and a standardised database for food fatty acid compositions, a sequential process of theoretical fat substitutions enabled suitable fat sources for use in the three diets to be identified, and experimental margarines for baking, spreading and the manufacture of snack foods to be designed. The dietary intervention was largely successful in achieving the fatty acid targets of the three diets, although unintended differences between the original target and the analysed fatty acid composition of the experimental margarines resulted in a lower than anticipated MUFA intake on the HM diet, and a lower ratio of palmitic to stearic acid compared with the reference or MM diet. Conclusions: This study has revealed important theoretical considerations that should be taken into account when designing diets of specific fatty acid composition, as well as practical issues of implementation.

Viability of Salmonella enterica subsp. enterica during the preparation and cold storage of Egyptian soft cheeses and ice-cream

Changes occurring in the viability of Salmonella enterica subsp. enterica during the preparation and cold storage of Domiati cheese, Kariesh cheese and ice-cream were examined. A significant decrease in numbers was observed after whey drainage during the manufacture of Domiati cheese, but Salmonella remained viable for 13 weeks in cheeses prepared from milks with between 60 and 100 g/L NaCl; the viability declined in Domiati cheese made from highly salted milk during the later stages of storage. The method of coagulation used in the preparation of Kariesh cheese affected the survival time of the pathogen, and it varied from 2 to 3 weeks in cheeses made with a slow-acid coagulation method to 4-5 weeks for an acid-rennet coagulation method. This difference was attributed to the higher salt-in-moisture levels and lower pH values of Kariesh cheese prepared by the slow-acid coagulation method. A slight decrease in the numbers of Salmonella resulted from ageing ice-cream mix for 24 h at 0degreesC, but a greater reduction was evident after one day of frozen storage at -20degreesC. The pathogen survived further frozen storage for four months without any substantial change in numbers.

Prebiotic capacity of inulin-type fructans

The human gut microbiota plays a significant role in human health through its ability to digest food ingredients and manufacture metabolites. This can be positive or negative for host welfare. Moreover, the microflora plays an active role in host defense whereby colonization resistance affords protection against pathogens. Prebiotics are nondigestible food ingredients that target beneficial components of the gut microflora (mainly colonic), particularly the bifidobacteria. In vitro and in vivo evidence has accumulated to confirm the prebiotic effects of inulin-derived fructans.

Chemical, physical, and sensory properties of dairy products enriched with conjugated linoleic acid

Recent studies have illustrated the effects of cis-9, trans-11 conjugated linoleic acid (CLA) on human health. Ruminant-derived meat, milk and dairy products are the predominant sources of cis-9, trans-11 CLA in the human diet. This study evaluated the processing properties, texture, storage characteristics, and organoleptic properties of UHT milk, Caerphilly cheese, and butter produced from a milk enriched to a level of cis-9, trans-11 CLA that has been shown to have biological effects in humans. Forty-nine early-lactation Holstein-British Friesian cows were fed total mixed rations containing 0 (control) or 45 g/kg ( on dry matter basis) of a mixture (1:2 wt/wt) of fish oil and sunflower oil during two consecutive 7-d periods to produce a control and CLA-enhanced milk, respectively. Milk produced from cows fed the control and fish and sunflower oil diets contained 0.54 and 4.68 g of total CLA/100 g of fatty acids, respectively. Enrichment of CLA in raw milk from the fish and sunflower oil diet was also accompanied by substantial increases in trans C18:1 levels, lowered C18: 0, cis-C18:1, and total saturated fatty acid concentrations, and small increases in n-3 polyunsaturated fatty acid content. The CLA-enriched milk was used for the manufacture of UHT milk, butter, and cheese. Both the CLA-enhanced butter and cheese were less firm than control products. Although the sensory profiles of the CLA-enriched milk, butter, and cheese differed from those of the control products with respect to some attributes, the overall impression and flavor did not differ. In conclusion, it is feasible to produce CLA-enriched dairy products with acceptable storage and sensory characteristics.

A study of the effects of adjunct cultures on the aroma compounds of Feta-type cheese

The influence of adjunct brine cultures on the volatile compounds in Feta-type cheeses made from bovine milk was studied. Four batches of brine were produced: one with no added adjuncts, a second containing Lactobacillus paracasei subsp. paracasei, a third containing Lb. paracasei subsp. paracasei plus Debaryomyces hansenii and a fourth with Lb. paracasei subsp. paracasei plus Yarrowia lipolytica. All the cultures were isolated from commercial Feta brines. Aroma compounds were analysed by dynamic headspace analysis, on-line coupled with GC/MS. The most important volatile compounds were quantified in the experimental cheeses; it was concluded that the use of Lb. paracasei subsp. paracasei and D. hansenii as adjuncts in the manufacture of Feta-type cheeses contribute to the formation of a richer pattern of aroma compounds, namely alcohols, aldehydes and esters. The inclusion of Y. lipolytica resulted in the production of undesirable aroma compounds that are not part of the usual volatile profile of high quality Feta cheeses. (C) 2004 Elsevier Ltd. All rights reserved.

Effect of chymosin reduction and salt substitution on the properties of white salted cheese

A whey salts mixture was used as a partial substitute for sodium chloride to provide a modified Na:K ratio (1:3.4) in the manufacture of white salted cheese using ultrafiltration. Reduction of chymosin addition from 20 to 8 mu L kg(-1) of cheese was also investigated. Variation of salt and chymosin levels did not result in any significant differences in composition and physicochemical properties. The rates of proteolysis in terms of water-soluble nitrogen (WSN) and nitrogen soluble in 12% trichloroacetic acid (TCA-SN) were affected by chymosin levels but not by salt treatment. Urea-PAGE electrophoretic analysis of caseins from the cheeses manufactured using three levels of chymosin and two salt types showed that the hydrolysis of alpha(s1)-casein was higher than for beta-caseins but the differences between the cheeses were not significant (P > 0.05). The chymosin level did not have a significant effect (P > 0.05) on hardness and fracturability, suggesting that any variation in hardness due to the initial hydrolysis was being confounded by other variables. Cheeses including the whey salts product were harder and more fracturable (P < 0.01) than the cheese treated with NaCl only. Both hardness and fracturability values decreased (P < 0.05) over the maturation period. The scores for bitterness were low; neither the effects of salt nor chymosin levels were significant (P > 0.05). (c) 2005 Elsevier Ltd. All rights reserved.

Effect of chymosin and salt reduction on the quality of ultrafiltrated white-salted cheese

This study demonstrated that both chymosin and salt-in-moisture (SM) were important factors for proteolysis in the manufacture of ultrafiltrated white-salted cheese, with significant effects on water-soluble nitrogen and nitrogen soluble in trichloroacetic acid. In contrast, the levels of free amino acids were not significantly affected by chymosin and salt treatments. The cheeses made using high levels of chymosin with low SM had lower levels of residual α(s1)- and β-casein at the end of ripening. On texture profile analysis, the hardness and fracturability of the cheeses significantly increased with SM and decreased during ripening. Increases in chymosin significantly contributed to the overall weakening of the structure throughout ripening. Bitter flavour was detected after 12 weeks in the cheese made with the higher chymosin level and lower SM, which could be the result of accumulation of γ-casein fractions. The sensory data indicated that the hedonic responses for low chymosin with low SM cheeses were good and acceptable in flavour, which may be due to the moderate levels of proteolysis products.

Interference filters and coatings for mid-infrared astronomy (8-30µm)

The VISIR instrument for the European Southern Observatory (ESO) Very Large Telescope (VLT) is a thermal-infrared imager and spectrometer currently being developed by the French Service d'Astrophysique of CEA Saclay, and Dutch NFRA ASTRON Dwingeloo consortium. This cryogenic instrument will employ precision infrared bandpass filters in the N-( =7.5-14µm) and Q-( =16-28µm) band mid-IR atmospheric windows to study interstellar and circumstellar environments crucial for star and planetary formation theories. As the filters in these mid-IR wavelength ranges are of interest to many astronomical cryogenic instruments, a worldwide astronomical filter consortium was set up with participation from 12 differing institutes, each requiring instrument specific filter operating environments and optical metrology. This paper describes the design and fabrication methods used to manufacture these astronomical consortium filters, including the rationale for the selection of multilayer coating designs, temperature-dependant optical properties of the filter materials and FTIR spectral measurements showing the changes in passband and blocking performance on cooling to <50K. We also describe the development of a 7-14µm broadband antireflection coating deposited on Ge lenses and KRS-5 grisms for cryogenic operation at 40K

A technique for evaluation of interactive evolutionary systems

Very large scale scheduling and planning tasks cannot be effectively addressed by fully automated schedule optimisation systems, since many key factors which govern 'fitness' in such cases are unformalisable. This raises the question of an interactive (or collaborative) approach, where fitness is assigned by the expert user. Though well-researched in the domains of interactively evolved art and music, this method is as yet rarely used in logistics. This paper concerns a difficulty shared by all interactive evolutionary systems (IESs), but especially those used for logistics or design problems. The difficulty is that objective evaluation of IESs is severely hampered by the need for expert humans in the loop. This makes it effectively impossible to, for example, determine with statistical confidence any ranking among a decent number of configurations for the parameters and strategy choices. We make headway into this difficulty with an Automated Tester (AT) for such systems. The AT replaces the human in experiments, and has parameters controlling its decision-making accuracy (modelling human error) and a built-in notion of a target solution which may typically be at odds with the solution which is optimal in terms of formalisable fitness. Using the AT, plausible evaluations of alternative designs for the IES can be done, allowing for (and examining the effects of) different levels of user error. We describe such an AT for evaluating an IES for very large scale planning.

Impedance mismatch: some differences between the way humans and robots control interaction forces

In over forty years of research robots have made very little progress still largely confined to industrial manufacture and cute toys, yet in the same period computing has followed Moores Law where the capacity double roughly every two years. So why is there no Moores Law for robots? Two areas stand out as worthy of research to speedup progress. The first is to get a greater understanding of how human and animal brains control movement, the second to build a new generation of robots that have greater haptic sense, that is a better ability to adapt to the environment as it is encountered. A remarkable property of the cognitive-motor system in humans and animals is that it is slow. Recognising an object may take 250 mS, a reaction time of 150 mS is considered fast. Yet despite this slow system we are well designed to allow contact with the world in a variety of ways. We can anticipate an encounter, use the change of force as a means of communication and ignore sensory cues when they are not relevant. A better understanding of these process has allowed us to build haptic interfaces to mimic the interaction. Emerging from this understanding are new ways to control the contact between robots, the user and the environment. Rehabilitation robotics has all the elements in the subject to not only enable and change the lives of people with disabilities, but also to facilitate revolution change in classic robotics.