Improved methods for the isolation and study of the C18, C20 and C22 monoethylenic fatty acid isomers of biological samples: Hg adducts,HPLC, AgNO3-TLC/FID,and ozonolysis

The monoethylenic isomers of C₁₈, C₂₀ and C₂₂ chain lengths of the depot fat of a nonhominid primate (cynomolgus monkeys,Macaca fascicularis), fed a partially hydrogenated herring oil (IV=76.0) for 30 months, were examined by 2 different approaches. The first isolation method involved preparative gas liquid chromatography and argentation thin layer chromatography (TLC). The second sequence involved a chain-length fractionation system based on the TLC of the methoxy-bromomercuri quence involved a chain-length fractionation system based on the TLC of the methoxy-bromomercuri adducts of the total methyl esters to isolate groups of acids of common degrees of unsaturation, and then high performance liquid chromatography on a reverse-phase column. In both cases, the monoethylenic isomer distribution was determined by ozonolysis in BF₃/MeOH. Comparable results were obtained with the 2 methods. The second approach is recommended for small biological samples, especially for those containing a relatively high proportion of di- and other polyethylenic isomers which might interfere. Presented in part at the AOCS annual meeting, New Orleans, May 1981.     

Viscosity measurements of liquids under pressure by using the quartz crystal resonators

A quartz crystal viscometer has been developed for measuring viscosity in liquids under pressure. It employs an AT-cut quartz crystal resonator of fundamental frequency 3 MHz inserted in a variable-volume vessel designed for working up to 80 MPa. Viscosity is determined by two methods from resonance frequency and bandwidth measurements along up to eight different overtones. The resonance frequency allows an absolute measurement of the viscosity but leads to an accuracy limited to 5% whereas the bandwidth technique which works in a relative way provides an accuracy of 2%. The techniques were tested by carrying out measurements in two pure compounds: heptane and toluene. Measurement results demonstrate the feasibility of the technique in this viscosity range. The apparatus was also used to determine the viscosity of n-decane with dissolved methane. The results obtained with these mixtures reveal the applicability of the apparatus for reservoir fluids study.     

Influence of temperature and surfactant on Escherichia coli inactivation in aqueous suspensions treated by moderate pulsed electric fields

This research employed a conductometric technique to estimate the inactivation kinetics of Escherichia coli cells in aqueous suspensions (1 wt.%) during simultaneous pulsed electric fields (PEF) and thermal treatments. The electric field strength was E = 5 kV/cm, the effective PEF treatment time t_PEF was within 0–0.2 s, the pulse duration t_i was 10^− 3 s, the medium temperature was 30–50 °C, and the time of thermal treatment t_T was within 0–7000 s. The damage of E. coli was accompanied by cell size decrease and release of intracellular components. The synergy between PEF and thermal treatments on E. coli inactivation was clearly present. The non-ionic surfactant Triton X-100 additionally improved its inactivation. The characteristic damage time followed the Arrhenius law within the temperature range 30–50 °C with activation energies W = 94 ± 2 kJ mol^− 1 and W = 103 ± 5 kJ mol^− 1 with and without the presence of surfactant, respectively. Relations between cell aggregation, cell ζ-potentials and presence of surfactant were analysed.     

Isolation and characterization of copolymers of beta-lactoglobulin, alpha-lactalbumin, kappa-casein, and alphas1-casein generated by pressurization and thermal treatment of raw milk

Raw skim milk was submitted to high pressure (300 to 600 MPa) and temperature (4 to 70 degrees C) treatments for 2 or 5 min. The combined effects of pressure and temperature on milk proteins induced structural changes and polymer and copolymer formation characterized by anion-exchange and size-exclusion fast protein liquid chromatography and electrophoretic techniques. Approximately half of the beta-lactoglobulin formed polymers, and the other half formed large copolymers, mainly with kappa-casein, alpha-lactalbumin via intermolecular disulfide bond exchange, and alpha(s1)-casein via physicochemical interactions, in proportions of 1.0:0.7:0.3:0.1, respectively. Minor whey proteins (serum albumin, immunoglobulins, and lactoferrin) also participated in the formation of the copolymers but to a lesser extent. Two populations of the copolymers were found with apparent molecular masses ranging from 440 to 2000 kDa for the first and more than 2000 kDa for the second. On the contrary, for heated milks the aggregation kinetics obtained by combination of high pressure and thermal treatment were very fast, as no intermediates such as dimers and small size oligomers were observed after pressurization, whatever the temperature studied. Lactosylation of proteins as well as proteolysis were very limited. A beta-casein amino-terminal peptide of 22 kDa was specifically recovered in milk samples treated under the more drastic conditions (500 MPa/55 degrees C per 5 min and 600 MPa/70 degrees C per 5 min) and might have been generated by neutral proteases such as elastase released from somatic cells present in milk. No casein was released from the micelle whatever the combination of high pressure and temperature studied.     

How to visualize the spider mite silk?

Tetranychus urticae (Acari: Tetranychidae) is a phytophagous mite that forms colonies of several thousand individuals. Like spiders, every individual produces abundant silk strands and is able to construct a common web for the entire colony. Despite the importance of this silk for the biology of this worldwide species, only one previous study suggested how to visualize it. To analyze the web structuration, we developed a simple technique to dye T. urticae'silk on both inert and living substrates. Fluorescent brightener 28 (FB) (Sigma F3543) diluted in different solvents at different concentrations regarding the substrate was used to observe single strands of silk. On glass lenses, a 0.5% dimethyl sulfoxide solution was used and on bean leaves, a 0.1% aqueous solution. A difference of silk deposit was observed depending the substrate: rectilinear threads on glass lenses and more sinuous ones on bean leaves. This visualizing technique will help to carry out future studies about the web architecture and silk used by T. urticae. It might also be useful for the study of other silk-spinning arthropods. Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc.     

Organotin levels in seafood and its implications for health risk in high-seafood consumers

Fish and fishery products are considered as the main source of organotin compounds (OTC). Unfortunately, little national contamination data is available to assess food exposure of organotins from French consumers. To provide a more accurate estimate of risks to human health, the butyltin, phenyltin and octyltin compounds sampling in four French coastal areas were measured in 159 composite samples (96 fresh and frozen fish, 28 mollusks, 14 crustaceans, 1 echinoderm, 11 canned foods, 4 smoked fish, 5 prepared seafood-based dishes) by capillary gas chromatography coupled with a microwave induced plasma atomic-emission spectrometer (CGC-MIP-AES). In these samples, butyltins were usually predominant and the range of the contamination levels was generally below those of earlier studies (fish: mean 5.6; min-max 1.1-23 microg/kg; fishery products: mean 6; min-max 0.8-14 microg/kg). Fish, especially tuna, salmon, mackerel, saithe/coalfish and cod were largely the main contributors (38%) to the total organotin exposure. With the supplementary contribution of great scallop, surimi, squid and oysters, the exposure exceeded 50% in all. However, the utmost OTC exposure was lesser than 47% of the provisional tolerable weekly intake [EFSA (European Food Safety Agency). Opinion of the Scientific Panel on Contaminants in the food chain on a request from the Commission to assess the health risks to consumers associated with exposure to organotins in foodstuffs. (Question N EFSA-Q-2003-110). The EFSA Journal, 102, 1-119, 2004. http://www.efsa.eu.int]. Nobody would exceed this limit. Finally, as this study has some limitations and since some other sources and health effects have not been clearly evaluated, it appears rational from public health and environmental viewpoints to continue to reduce the OTC levels in the environment.     

Design and use of a hypermedia system at the University level

Abstract This paper reports on an experiment in which a whole semester course in psychology was replaced by a mixed formula consisting of a CD-ROM complemented by a series of seminars and workshops. The CD-ROM was conceived as a collection of documents (hypertexts, research data, references, videos and activities) linked together with genuine Netscape facilities. Students were invited to search through these documents for information to answer questions (called challenges ) on the topic. A multiple-choice questionnaire accompanied each challenge in order to foster students' self-evaluation. The seminars, held every other week, served both as forums to discuss each of the topics under the guidance of an expert. Careful analysis of students' answers to two questionnaires at the beginning and end of the course and during interviews, showed that such a formula was favourably accepted by a large majority of students, although it lead to more anxiety and work load than a traditional course. Positive effects were also observed on learning.     

Contribution of Protein Flexibility to the Foaming Properties of Casein

The effect of biopolymer flexibility on the foaming properties of casein was investigated. Flexibility was altered by: (1) chemical modification (covalent binding of a monosaccharide on the lysyl residues) or (2) pH change. Electron Spin Resonance was used to measure the reorientational frequency of casein residues labeled with nitroxide radicals. High levels of glycosylation induced increased protein flexibility and improved the foaming capacity. Good agreement was observed between higher values of flexibility and improved surface properties near the isoelectric point.