The EU optimal blood use project.

The EU optimal blood use project (EUOBUP) is co-funded by the European Commission and led by the Scottish National Blood Transfusion Service (SNBTS). Its purpose is to develop, evaluate and disseminate a manual that provides practical guidance and support for those seeking to improve the safety of the clinical transfusion process and the effectiveness of the prescribing of blood components. We define the optimal use of blood components as the safe, clinically effective and efficient use of the scarce resource of donated human blood. The project will build on the experience of a pilot project in optimal use of blood in the national health service in Scotland. This pilot developed training resources in the safe and effective use of blood and delivered training to a large number of practitioners. It has also developed systems to provide hospitals with comparative information on their use of blood components for specific clinical groups of patients to assist them in reviewing practice against that of their peers.

Sensitivity of markers of DNA stability and DNA repair activity to folate supplementation in healthy volunteers.

We have previously reported that supplementation with folic acid (1.2 mg day(-1) for 12 week) elicited a significant improvement in the folate status of 61 healthy volunteers. We have examined effects of this supplement on markers of genomic stability. Little is known about the effect of folate supplementation on DNA stability in a cohort, which is not folate deficient. Preintervention, there was a significant inverse association between uracil misincorporation in lymphocyte DNA and red cell folate (P < 0.05). In contrast, there were no associations between folate status and DNA strand breakage, global DNA methylation or DNA base excision repair (measured as the capacity of the lymphocyte extract to repair 8-oxoGua ex vivo). Folate supplementation elicited a significant reduction in uracil misincorporation (P < 0.05), while DNA strand breakage and global DNA methylation remained unchanged. Increasing folate status significantly decreased the base excision repair capacity in those volunteers with the lowest preintervention folate status (P < 0.05). Uracil misincorporation was more sensitive to changes in folate status than other measures of DNA stability and therefore could be considered a specific and functional marker of folate status, which may also be relevant to cancer risk in healthy people.

The effects of cranberry juice consumption on antioxidant status and biomarkers relating to heart disease and cancer in healthy human volunteers.

BACKGROUND: Consumption of fruit and vegetables is associated with a decreased risk of heart disease and cancer. This has been ascribed in part to antioxidants in these foods inactivating reactive oxygen species involved in initiation or progression of these diseases. Non-nutritive anthocyanins are present in significant amounts in the human diet. However, it is unclear whether they have health benefits in humans. AIM: To determine whether daily consumption of anthocyanin-rich cranberry juice could alter plasma antioxidant activity and biomarkers of oxidative stress. METHODS: 20 healthy female volunteers aged 18-40 y were recruited. Subjects consumed 750 ml/day of either cranberry juice or a placebo drink for 2 weeks. Fasted blood and urine samples were obtained over 4 weeks. The total phenol, anthocyanin and catechin content of the supplements and plasma were measured. Anthocyanin glycosides were identified by tandem mass spectrometry (MS-MS). Vitamin C, homocysteine (tHcy) and reduced glutathione (GSH) were measured by HPLC. Total antioxidant ability was determined using electron spin resonance (ESR) spectrometry and by the FRAP assay. Plasma total cholesterol, high density lipoprotein (HDL), and low density lipoprotein (LDL) cholesterol and triglycerides (TG) were measured. Glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) activities were measured in erythrocytes. Urine was collected for analysis of malondialdehyde (MDA) by HPLC and 8-oxo-deoxyguanosine (8-oxo-dG) by ELISA. Endogenous and induced DNA damage were measured by single cell gel electrophoresis (SCGE) in lymphocytes. RESULTS: Vitamin C, total phenol, anthocyanin and catechin concentrations and FRAP and ESR values were significantly higher in the cranberry juice compared with the placebo. Cyanidin and peonidin glycosides comprised the major anthocyanin metabolites [peonidin galactoside (29.2%) > cyanidin arabinoside (26.1%) > cyanidin galactoside (21.7%) > peonidin arabinoside (17.5%) > peonidin glucoside (4.1%) > cyanidin glucoside (1.4 %)]. Plasma vitamin C increased significantly (P<0.01) in volunteers consuming cranberry juice. No anthocyanins (plasma) or catechins (plasma or urine) were detectable and plasma total phenols, tHcy,TC,TG,HDL and LDL were unchanged. The antioxidant potential of the plasma, GSH-Px, CAT and SOD activities, and MDA were similar for both groups. Supplementation with cranberry juice did not affect 8-oxo-deoxyguanosine in urine or endogenous or H(2)O(2)-induced DNA damage in lymphocytes. CONCLUSIONS: Cranberry juice consumption did not alter blood or cellular antioxidant status or several biomarkers of lipid status pertinent to heart disease. Similarly, cranberry juice had no effect on basal or induced oxidative DNA damage. These results show the importance of distinguishing between the in vitro and in vivo antioxidant activities of dietary anthocyanins in relation to human health.

Cryopreserved versus freshly isolated lymphocytes in human biomonitoring: endogenous and induced DNA damage, antioxidant status and repair capability.

Lymphocytes are routinely used in human biomonitoring to assess the potential toxic and cytoprotective effects of diet on both DNA damage and repair and, by implication, health. Logistically, samples may require to be cryopreserved and stored. How this affects cells used in human biomonitoring is often not considered. In this study we have evaluated the influence of cryopreservation on endogenous and induced DNA strand breakage, altered bases (oxidized purines, oxidized pyrimidines and misincorporated uracil), antioxidant capacity and DNA repair capability in human peripheral blood lymphocytes. Neither isolation nor freezing increased DNA strand breakage above endogenous levels found in freshly isolated human lymphocytes. Oxidized bases (both pyrimidines and purines) and misincorporated uracil, were similar for fresh and frozen lymphocytes. Fresh and frozen lymphocytes responded almost identically to hydrogen peroxide. Quercetin-mediated cytoprotection against hydrogen peroxide-induced strand breakage was maintained in cryopreserved lymphocytes after short-term (24 h) and longer term (2 months) storage compared with freshly isolated and treated cells. Hydrogen peroxide-induced DNA strand breakage was repaired in fresh lymphocytes. Cryopreserved lymphocytes were unable to repair oxidant-induced DNA strand breaks. Frozen human lymphocytes can therefore be successfully used for most aspects of DNA damage biomonitoring, but not for repair.

Stimulus satiation: effects of repeated exposure to foods on pleasantness and intake.

Frequent and repeated exposure to foods produces stimulus satiation or monotony. To explore further the nature of stimulus satiation, two experiments were conducted. Experiment 1 investigated the influence of initial pleasantness and frequency of intake on monotony. Tests showed that bread and butter was eaten more frequently but was liked less than chocolate. Therefore, normal-weight, healthy males were randomly assigned to either a chocolate condition (CC, N=13) or bread and butter condition (BC, N=16). All subjects received fixed amounts of the assigned food (67g/1473kJ of chocolate or 95g/1355kJ of bread and butter) every day for 22 days. On days 1, 8, 15 and 22 subjects consumed this food ad libitum. Pleasantness of taste and desire to eat chocolate declined significantly over time but no such changes were observed for bread and butter. Experiment 2 examined intake, pleasantness and desire to eat chocolate in 53 subjects over a 15 day period, with 3 conditions: control (CS: N=15), fixed (FS: N=20) and variable (VS: N=18). CS received no chocolate except on test days (days 1, 8 and 15), FS received 67g/1473kJ of chocolate daily and VS received increasing amounts of chocolate from 57g/1251kJ on day 1 to 86g/1888kJ by day 12. Pleasantness and desire to eat chocolate declined over time with this being more pronounced for F and V subjects. However, ad libitum intake increased over time. Both experiments demonstrated significant changes in pleasantness and desire to eat chocolate, but no commensurate decline in intake. Thus, although stimulus satiation occurred for subjective ratings of pleasantness and desire to eat chocolate, intake remained unaffected. This apparent dissociation between pleasantness and intake may reflect different processes underlying liking and wanting.

Stimulation of appetite by alcohol.

To investigate the effects of alcohol on appetite and food intake, 26 males attended the laboratory on three occasions. On each occasion, they were given a standard breakfast. Visual analog scale ratings of hunger, desire to eat and fullness (appetite ratings) were recorded from before breakfast until their return to the laboratory for lunch. Thirty minutes before lunch, subjects either rested (baseline), were given 330 ml of a no-alcohol lager (264 kJ: no-alcohol condition) or 330 ml of the same lager spiked with 3 units of alcohol (24 g ethyl alcohol; total energy=969 kJ: alcohol condition). Ratings of appetite were taken before and after the preload or baseline rest period and again before and hourly after lunch. The test meal at lunch consisted of a buffet-style array of foods and chilled water. Ad libitum intake at lunch (excluding energy from alcohol) was significantly higher following alcohol (7301+/-442 kJ) compared to both baseline (6365+/-334 kJ) and the no-alcohol conditions (6479+/-289 kJ). Appetite ratings failed to demonstrate any differences between alcohol and the no-alcohol condition. Total energy intake (including energy from alcohol) was enhanced in the alcohol condition by 30%, suggesting that energy from alcohol is not compensated in the short-term and may even have a stimulatory effect on food intake.

Folate deficiency in vitro induces uracil misincorporation and DNA hypomethylation and inhibits DNA excision repair in immortalized normal human colon epithelial cells.

Epidemiological studies have indicated that folic acid protects against a variety of cancers, particularly cancer of the colorectum. Folate is essential for efficient DNA synthesis and repair. Moreover, folate can affect cellular S-adenosylmethionine levels, which regulate DNA methylation and control gene expression. We have investigated the mechanisms through which folate affects DNA stability in immortalized normal human colonocytes (HCEC). DNA strand breakage, uracil misincorporation, and DNA repair, in response to oxidative and alkylation damage, were determined in folate-sufficient and folate-deficient colonocytes by single cell gel electrophoresis. In addition, methyl incorporation into genomic DNA was measured using the bacterial enzyme Sss1 methylase. Cultured human colonocyte DNA contained endogenous strand breaks and uracil. Folate deficiency significantly increased strand breakage and uracil misincorporation in these cells. This negative effect on DNA stability was concentration dependent at levels usually found in human plasma (1-10 ng/ml). DNA methylation was decreased in HCEC grown in the absence of folate. Conversely, hypomethylation was not concentration dependent. Folate deficiency impaired the ability of HCEC to repair oxidative and alkylation damage. These results demonstrate that folic acid modulates DNA repair, DNA strand breakage, and uracil misincorporation in immortalized human colonocytes and that folate deficiency substantially decreases DNA stability in these cells.

Enhancing care for transfusion patients.

Frequent hospital attendance and excessive waiting times for treatments are often part of haematological patient care. Home platelet transfusions reduced hospital visits for patients. Hospital waiting times decreased and the quality of care improved.

Adaptation of pulmonary receptors in the spontaneously breathing anaesthetized rat.

It has been suggested that species with high breathing frequencies have pulmonary stretch receptors which adapt more rapidly than species with low breathing frequencies. This has proved not to be so. Our hypothesis is that this theory is in fact correct if modified so that overall rate of adaptation of afferent vagal activity, i.e. the sum of stretch and rapidly adapting receptors, is considered. A rapidly breathing species, such as the rat, would thus have a greater proportion of rapidly adapting receptors, than a more slowly breathing species. To test this hypothesis, we measured the proportion of rapidly adapting pulmonary mechanoreceptors in spontaneously breathing rats for comparison with existing results from more slowly breathing species. We found there to be one rapidly adapting receptor for every three slowly adapting receptors present. This measurement has not previously been made in spontaneously breathing rats. The ratio of rapidly to slowly adapting pulmonary receptors in the species sequence cat-rabbit-rat is the same as the ratio of their breathing frequencies (3:4:10). We propose that the difference in proportion of slowly to rapidly adapting pulmonary receptors in different species may be related to their eupnoeic breathing frequency.

A novel ultrasonic aerosol generator.

An ultrasonic aerosol generator constructed from a domestic humidifier is described which has been used to produce liquid aerosols for physiological investigations. The instrument was constructed from a Pifco domestic humidifier modified to include an energy guide to direct the oscillations of the transducer through the coupling water, which would normally be aerosolized, onto a small membrane based sample chamber containing the liquid to be aerosolized. The size distribution of the aerosol produced was found to be between 2 and 6 mm, optimum for diffuse intrapulmonary deposition. Up to 4 ml/min of aqueous liquid was used; however the sample chamber could be made small enough to contain economic amounts of expensive material to administer by inhalation. The instrument has proved to be reliable over a period of three years.