Multiple centrosomal microtubule organising centres and increased microtubule stability are early features of VP-16-induced apoptosis in CCRF-CEM cells

Microtubular reorganisation contributing to apoptotic morphology occurs in normal and neoplastic cells undergoing apoptosis induced by cytotoxic drugs [1-3]. The aim of this study was to correlate the changes in the microtubules (MTs) with behavior of the centrosome in apoptotic cells, and to see whether post-translational changes in tubulin occurred with the emergence of apoptotic MT bands. Apoptosis was induced in the human T-cell leukaemia line (CCRF-CEM) by treatment with 17 microM etoposide over a 4 h period. The time course of changes was assessed using flow cytometry (FCM) and immunocytochemistry in cells labelled for a centrosomal antigen (CSP-alpha) or alpha-tubulins. One hour following treatment we observed multiple centrosomal microtubule organising centres (MTOCs) associated with the nucleus and the transient appearance of a subset of stable MTs detected with an antibody specific for acetylated alpha-tubulin, as the bands of MTs which lobulate the nucleus are formed. The altered properties of the MTs thus reflect changes in function as apoptosis progresses.     

Application of immunological methods for the detection of species adulteration in dairy products

A number of enzyme‐linked immunosorbent assays (ELISAs) have been developed for the detection of milk adulteration in dairy products. Target antigens have been caseins, lactoglobulins, immunoglobulins and other whey proteins. Polyclonal and monoclonal antibodies have been used in a variety of formats including direct, indirect, competitive and sandwich ELISAs. ELISAs have been successfully applied to the detection of cows' milk adulteration of sheep, goat and buffalo milk. Goat milk adulteration of sheep milk has also been detected. A number of ELISAs have also been applied to cheese. It is recommended that ELISA should be used in combination with PCR to ensure compliance with current legislation.     

Visualisation of mechanisms involved in Co2 injection and storage in hydrocarbon reservoirsand water-bearing aquifers

Storage of carbon dioxide (CO₂) in hydrocarbon reservoirs and saline aquifers is considered as one of the promising mitigation strategies to reduce the negative impact of this greenhouse gas. The static and dynamic behaviour of CO₂ in these storage sites which are located at various depths and geographical locations, affects the efficiency of this strategy. Understanding the impact of the conditions of these storage sites on mechanisms involved in CO₂ flow, displacement and trapping is also critical for the purpose of site selection and the design of CO₂ storage projects. In this paper we report the results of a series of CO₂ injection (CO₂I) flow visualisation (micromodel) experiments conducted using high-pressure transparent porous media representing various aquifer and depleted oil reservoirs storage conditions. The impact of pertinent parameters on the interaction between the stored CO₂ and the reservoir fluids were investigated. Both sub-critical and supercritical CO₂ were used to investigate the effect of pressure (depth) of the storage site on CO₂ trapping mechanisms. A faster CO₂ breakthrough (BT) was observed in the micromodel test simulating CO₂I into depleted oil reservoirs, compared to that into aquifers, reducing the sequestration capacity of the depleted oil reservoirs. Compared to the injection of supercritical CO₂, the BT of gaseous CO₂ happened faster, adversely affecting the CO₂ displacement performance. The results of these direct flow visualization experiments significantly improve our understanding of the complex mechanisms and interactions involved in CO₂I and storage in geological formations. This knowledge is essential for identifying storage conditions that would lead to maximising CO₂ storage capacity, for better understanding the ultimate fate of the stored CO₂ and the storage safety.     

Saponin content of soya and some commercial soya products by means of high-performance liquid chromatography of the sapogenins

Quantitative high-performance liquid chromatography of the sapogenins was used to estimate the saponin content of soya beans (Glycine max (L.) Merr.) and commercially prepared soya flours, protein concentrate, protein isolate and soya milks. The level of saponin in whole soya beans as estimated (0.47%) was in agreement with other recent estimates. Saponins were also found in the various soya flours (0.43–0.67%), protein isolate (0.76%), soya milks (0.022–0.026%) but not in the protein concentrate.     

Effect of intrafollicular indomethacin injection on gonadotropin surge-induced expression of select extracellular matrix degrading enzymes and their inhibitors in bovine preovulatory follicles

A growing body of evidence supports an obligatory role for intrafollicular prostanoids in the mechanism of ovulation. However, the prostanoid-dependent mediators of the follicular extracellular matrix degradation required for ovulation are unknown. The objectives of this study were to determine the cellular compartment(s) in which the gonadotropin surge-induced regulation of select extracellular matrix degrading enzymes and their cognate inhibitors occurs in bovine preovulatory follicles, and to test whether such regulation is blocked by intrafollicular administration of the prostanoid synthesis and ovulation inhibitor, indomethacin (INDO). Follicular fluid prostaglandin E2 concentrations were elevated in diluent-treated follicles before ovulation (24 h after GnRH injection), but the increase was blocked in INDO-treated follicles. Real-time PCR analysis revealed the specific follicular cell types where gonadotropin surge-induced increases in mRNA abundance for members of the matrix metalloproteinase/tissue inhibitor of metalloproteinase and plasminogen activator families occurred. INDO treatment increased thecal cell mRNA for tissue inhibitor of metalloproteinase-4 and its protein abundance in the apex of preovulatory follicles before ovulation, but suppressed granulosal cell mRNA and activity for tissue plasminogen activator in follicular fluid and the follicle apex. Plasmin activity was also suppressed in the follicular fluid of INDO-treated follicles. Effects of INDO injection on select matrix metalloproteinases were not observed. The results suggest that gonadotropin surge-induced regulation of tissue inhibitor of metalloproteinase-4 and tissue plasminogen activator may be prostanoid dependent, and support a potential role for increased tissue plasminogen activator expression and decreased tissue inhibitor of metalloproteinase-4 expression in the mechanism of ovulation.     

Harmonised procedures for producing new data on the nutritional composition of ethnic foods

Food composition databases provide important data that can be used in a variety of ways to improve the nutritional quality of food and the health of the populations. The multicultural nature of European populations, together with increased travel and the globalisation of the food supply, has led to an increase in the consumption of ethnic foods by both mainstream and ethnic populations. However, the information that is available on the composition of ethnic foods is currently incomplete and fragmentary which both prevents effective health and disease interventions and limits the provision of dietary advice and information. One of the aims of the ethnic foods work package within the European Commission’s FP6 Network of Excellence EuroFIR is the provision of new and reliable data on the nutritional composition of ethnic foods from seven EU Member States and Israel for inclusion in national food composition databases. To achieve this goal, specific standards and mechanisms were developed to sample and analyse foods to harmonise procedures. Given the anticipated usage of the data it was essential that they were representative of the foods consumed in the country or region and that proper sampling procedures of the foods were employed. The most important nutrients were prioritised and analysed by selected accredited laboratories. The analytical methods selected were appropriate to fulfil the requirements of the resulting data. Quality assurance measures (ISO 17025, CRMs, IQC) were in place and the data accurately reflected the composition of chosen foods. This paper describes the procedures for producing validated new data for inclusion in forthcoming publications.     

Techniques for detailed heat transfer measurements in cold supersonic blowdown tunnels using thermochromic liquid crystals

The paper presents methods for measurement of convective heat transfer distributions in a “cold flow” supersonic blowdown wind tunnel. The techniques involve use of the difference between model surface temperature and adiabatic wall temperature as the driving temperature difference for heat transfer and no active heating or cooling of the test gas or model is required. Thermochromic liquid crystals are used for surface temperature indication and results presented from experiments in a Mach 3 flow indicate that measurements of the surface heat transfer distribution under swept shock wave boundary layer interactions can be made.     

Alteration in immune responsiveness during the peripartum period and its ramification on dairy cow and calf health

Substantial evidence indicates that innate and acquired defense mechanisms are lowest from 3 wk precalving to 3 wk postcalving. This lowered responsiveness includes aspects of systemic and mammary gland immunity that may account, at least in part, for the increased incidence of peripartum disease. The physical and metabolic stresses of pregnancy, calving, and lactation may contribute to this decrease in host resistance and the subsequent increase in disease incidence. However, variation among cows in their host resistance mechanisms suggests that genotype and phenotype may possibly be used to identify cows that are able to mount beneficial immune responses over the periparturient period. Our own studies suggest that cows may be categorized as high or low responders based on the peripartum antibody responses to ovalbumin and Escherichia coli J5. Low responders were hyporesponsive to these test antigens and had a higher incidence of peripartum diseases, particularly mastitis. In many species, a functional link exists between the immune and endocrine systems, and, during periods of stress or physical injury, neuropeptides and neuroendocrine hormones function as immunomodulators. Initial investigations of peripartum cows reveal positive relationships between growth hormone kinetics and profiles of antibody response. Whether hormone fluctuations during the periparturient period are responsible for the alterations observed in immune responsiveness remains uncertain.