Micro-techniques for analysis of human adipose tissue fatty acid composition in dietary studies.

BACKGROUND AND AIMS: Adipose tissue (AT) fatty acid (FA) composition is considered to be the gold standard long-term biomarker of dietary fatty acid intake. Typically this measurement is made directly from samples collected via large-needle-biopsy or incision. However, with growing interest in the role of AT in relation to health, ideally the fatty acid composition would be analysed along with other measurements, such as gene expression or histology, on a single AT sample. Here we assess alternative ways of obtaining AT for measuring FA composition, in some cases in conjunction with other measurements. METHODS AND RESULTS: The FA composition of tissue obtained via different methods was compared to that of tissue collected via large-needle or surgical biopsy. Fatty acid composition was not significantly different in AT collected by small-needle mini-biopsy (n = 10), from an RNA 'lipid layer' (obtained during RNA extraction, 2 sites, n = 6 for each), or from cryosectioned tissue prepared for histology (n = 10). We also assessed the usefulness of the composition of plasma NEFA as a surrogate marker of subcutaneous AT (n = 58-80). Most FAs in plasma NEFA correlated strongly with those in AT (P < 0.05). CONCLUSION: It is feasible to measure the FA composition of AT on very small amounts of tissue. Additionally, it is possible to measure FA composition on the lipid rich 'by-product' of AT samples undergoing RNA extraction for gene expression. Samples sectioned for histology are also suitable. This provides further opportunities for multidisciplinary collaborations that may lead to a better application of dietary biomarkers.

Differentiation of patient-specific void urine-derived human induced pluripotent stem cells to fibroblasts and skeletal muscle myocytes.

Cell-based therapy is a major focus for treatment of stress urinary incontinence (SUI). However, derivation of primary cells requires tissue biopsies, which often have adverse effects on patients. A recent study used human induced pluripotent stem cells (iPSC)-derived smooth muscle myocytes for urethral sphincter regeneration in rats. Here, we establish a workflow using iPSC-derived fibroblasts and skeletal myocytes for urethral tissue regeneration: (1) Cells from voided urine of women were reprogrammed into iPSC. (2) The iPSC line U1 and hESC line H9 (control) were differentiated into fibroblasts expressing FSP1, TE7, vinculin, vimentin, αSMA, fibronectin and paxillin. (3) Myogenic differentiation of U1 and H9 was induced by small molecule CHIR99021 and confirmed by protein expression of myogenic factors PAX7, MYOD, MYOG, and MF20. Striated muscle cells enriched by FACS expressed NCAM1, TITIN, DESMIN, TNNT3. (4) Human iPSC-derived fibroblasts and myocytes were engrafted into the periurethral region of RNU rats. Injected cells were labelled with ferric nanoparticles and traced by Prussian Blue stain, human-specific nuclear protein KU80, and human anti-mitochondria antibody. This workflow allows the scalable derivation, culture, and in vivo tracing of patient-specific fibroblasts and myocytes, which can be assessed in rat SUI models to regenerate urethral damages and restore continence.

Validity and application of immunoturbidimetric and enzyme-linked immunosorbent assays for the measurement of adiponectin concentration in ponies.

BACKGROUND: Circulating adiponectin concentrations were lower in ponies with a history of endocrinopathic laminitis and in nonlaminitic ponies that subsequently developed laminitis. The assays used in these studies have been discontinued or are no longer valid. OBJECTIVES: (1) to determine the validity of immunoturbidimetric (IT) and enzyme linkedimmunosorbent (ELISA) assays for equine total and high molecular weight (HMW) [adiponectin] measurement and (2) to investigate the association between [adiponectin] measured using these assays and endocrinopathic laminitis. STUDY DESIGN: Method validation and cohort study. METHODS: Accuracy and precision of IT and ELISA assays for measuring total (TAC) and HMW (HMWAC) [adiponectin] were determined. Using the IT assay, the effects of anti-coagulant and storage temperature were assessed, TAC was measured in previously laminitic (PL) and never laminitic (NL) ponies (n = 6/group). Comparison with a previously validated radioimmunoassay was made in NL ponies (n = 223). Association between TAC and subsequent laminitis development in NL ponies was investigated using univariable logistic regression and ROC curve analysis. RESULTS: The IT assay was precise and demonstrated good agreement with the previously validated radioimmunoassay. TAC was significantly (P<0.01) lower in PL (mean ± s.d. 8.9 ± 2.9 µg/mL) compared to NL (24.2 ± 11.8 µg/mL) ponies and in NL ponies that developed laminitis within 12 months (median 4.8 µg/mL; IQR 2.65-13.4 µg/mL) compared to those that remained nonlaminitic (19.9 µg/mL; 9.95-31.5 µg/mL). TAC was significantly (P = 0.01) associated with laminitis occurrence within 12 months. Use of the area under the ROC curve to distinguish animals that did and did not develop laminitis showed good accuracy (0.76). None of the ELISA methods validated satisfactorily. MAIN LIMITATIONS: Laminitis risk is based on data from ponies in one region. CONCLUSIONS: The IT method is suitable for measurement of equine TAC. TAC is lower in ponies with previous or future laminitis. The ELISA methods are not suitable for measurement of equine HMWAC or TAC.

Magnetic resonance imaging of abdominal adiposity in a large cohort of British children.

OBJECTIVE: To describe abdominal adipose tissue distribution in a large sample of contemporary British children; to determine the influence of gender, stage of maturation and body mass index (BMI) on abdominal adipose tissue distribution; and to compare the ability of BMI and waist circumference to predict abdominal adipose tissue. SUBJECTS AND METHODS: A total of 74 boys (mean age 13.4+/-0.4 years) and 96 girls (mean age 13.5+/-0.5 years) were selected from volunteer children enrolled in the Avon Longitudinal Study of Parents and Children (ALSPAC). Height, weight and waist circumference were measured and BMI calculated. Stage of sexual maturation was available for 113 children using a self-report questionnaire based on Tanner's criteria. Magnetic resonance imaging was used to assess subcutaneous abdominal adipose tissue (SAAT) and intra-abdominal adipose tissue (IAAT) volumes and patterning. RESULTS: Boys had lower levels of IAAT (P=0.036) and SAAT (P=0.003) than girls. IAAT and SAAT were higher in overweight and obese boys and girls when compared with normal weight children (P<0.0001). This pattern was also reflected in waist circumference groups. Boys had higher IAAT/SAAT ratios than girls, indicating proportionately more adipose tissue deposited intra-abdominally (P=0.002). However, both boys and girls deposited less than 10% of their abdominal fat as internal adipose tissue. WC predicted 67.4% of the variance in IAAT (P<0.001), and BMI predicted 84.8% of the variance in SAAT (P<0.001). However, BMI as the best single predictor explained only 8.4% of the variance in the IAAT/SAAT ratio (P<0.001). CONCLUSIONS: At this age and stage of sexual maturation, the amount of IAAT remains relatively small. WC and BMI offer a feasible alternative to the MRI estimation of IAAT and SAAT, respectively, in a population-based sample of boys and girls. International Journal of Obesity (2008) 32, 91-99; doi:10.1038/sj.ijo.0803780; published online 27 November 2007.

Dietary patterns in pregnancy and associations with socio-demographic and lifestyle factors.

OBJECTIVE: To obtain distinct dietary patterns in the third trimester of pregnancy using principal components analysis (PCA); to determine associations with socio-demographic and lifestyle factors. DESIGN AND METHODS: A total of 12 053 pregnant women partaking in a population-based cohort study recorded current frequency of food consumption via questionnaire in 1991-1992. Dietary patterns identified using PCA were related to social and demographic characteristics and lifestyle factors. RESULTS: Five dietary patterns were established and labelled to best describe the types of diet being consumed in pregnancy. The 'health conscious' component described a diet based on salad, fruit, rice, pasta, breakfast cereals, fish, eggs, pulses, fruit juices, white meat and non-white bread. The 'traditional' component loaded highly on all types of vegetables, red meat and poultry. The 'processed' component was associated with high-fat processed foods. The 'confectionery' component was characterized by snack foods with high sugar content and the final 'vegetarian' component loaded highly on meat substitutes, pulses, nuts and herbal tea and high negative loadings were seen with red meat and poultry. There were strong associations between various socio-demographic variables and all dietary components; in particular, a 'health conscious' diet was positively associated with increasing education and age and non-white women. There was a negative association with increased parity, single, non-working women, those who smoked and who were overweight pre-pregnancy. Opposite associations were seen with the 'processed' component. CONCLUSIONS: Distinct dietary patterns in pregnancy have been identified. There is clear evidence of social patterning associated with the dietary patterns, these social factors need to be accounted for in future studies using dietary patterns. This study will form the basis for further work investigating pregnancy outcome.

Adjusting for energy intake in dietary pattern investigations using principal components analysis.

OBJECTIVE: The effect of energy adjustment on variables entered into principal component analysis (PCA) to derive dietary patterns has received little attention. DESIGN AND METHODS: As part of regular self-completion questionnaires, used in the Avon Longitudinal Study of Parents and Children, pregnant women were asked to record the frequency of consumption of a variety of food items. A total of 12 053 women completed the questionnaire. Individual dietary types were identified using PCA, before and after adjusting the food variables for energy intake. Associations with estimated nutrient intakes and with birthweight were examined for the two solutions and when energy adjustment was performed at a later stage of the analysis. RESULTS: Slight differences were seen in terms of the components extracted and the factor loadings obtained. The associations with nutrient intakes showed that there was a general reduction in the size of the correlation coefficients for the energy-adjusted components compared to the unadjusted components. There did not appear to be any difference in the size of the effects of the dietary pattern scores on birthweight, whether energy was adjusted for before entry into the PCA or after. CONCLUSIONS: In this sample, it is not necessary to adjust for energy intake before entry into a PCA analysis to determine dietary patterns when using food frequency questionnaire data. Effects of energy intake can be determined at a later stage in the analytical process.This study determines the effect of adjusting for energy on dietary patterns resulting from PCA and the subsequent effect on future outcomes.

Sodium intake in infancy and blood pressure at 7 years: findings from the Avon Longitudinal Study of Parents and Children.

BACKGROUND: Infancy may be a sensitive period regarding effects of sodium intake on future blood pressure (BP). This has only been demonstrated in one randomized trial of low sodium formulae with follow-up in adolescence in one-third of participants. OBJECTIVE: To prospectively assess associations between sodium intake in infancy and BP at 7 years in the Avon Longitudinal Study of Parents and Children (ALSPAC). SUBJECTS: A total of 533 children with sodium data at 4 months and 710 children with sodium at 8 months. RESULTS: 0.4% of participants at 4 months and 73.0% at 8 months exceeded recommended levels for infant sodium intake. After minimal adjustment (child age, sex, energy), sodium intake at 4 months was positively associated with systolic blood pressure (SBP) at 7 years (beta=0.54 mm Hg/mmol; 95% CI: 0.09, 0.98 mm Hg; P=0.02). This changed little following adjustment for confounders but attenuated after adjusting for breastfeeding. This association was not mediated by sodium intake at 7 years. Due to high sodium-potassium correlations, effects of sodium independent of potassium could not be estimated with reasonable precision. Sodium intake neither at 8 months nor at 7 years was associated with SBP at 7 years. CONCLUSION: The association between sodium intake at 4 months and future SBP requires replication in studies that can control for effects of potassium before we can conclude that early infancy is a sensitive period with respect to effects of sodium on future BP. The majority of infants exceeded recommended levels of sodium intake at 8 months, and interventions to reduce sodium in infants' diets should be considered.

Quality of food eaten in English primary schools: school dinners vs packed lunches.

OBJECTIVE: To compare the food and nutrient intakes of primary school children eating school dinners and packed lunches. SUBJECTS: Six-hundred and twenty-one 7-year-old children participating in the Avon Longitudinal Study of Parents and Children in South West England. METHODS: Diet was assessed by 3-day unweighed food record. RESULTS: The composition of both types of school meals compared unfavourably with dietary guidelines. Intakes of energy, non-starch polysaccharides (NSP), calcium, iron, folate, retinol equivalents, zinc, copper, magnesium, iodine and riboflavin were too low, and intakes of total and saturated fat were too high. However, children who ate school dinners had higher lunchtime intakes of protein, starch, NSP and most vitamins and minerals and lower intakes of sugar (14.2 and 20.9% of energy in school dinners and packed lunches, respectively, P<0.001) and saturated fat (12.0 and 16.2%, P<0.001). Only around half of the recommended amount of fruit and vegetables was eaten by children having either type of school meal. There were also differences in the whole day's nutrient intake according to school meal type. Children eating packed lunches had lower daily intakes of potassium and zinc, and higher intakes of sugar and saturated fat. Differences in nutrient intake were independent of maternal education. CONCLUSIONS: The food and nutrient content of both school dinners and packed lunches needed improvement. However, the standard of food brought from home by children was, if anything, worse than that served at school. Recent moves to improve school dinners will need to be complemented by education about what constitutes a healthy packed lunch.

Qualitative evaluation of a formal bedside clinical teaching programme in an emergency department.

BACKGROUND: Bedside clinical teaching in emergency departments is usually opportunist or ad hoc. A structured bedside clinical teaching programme was implemented, where a consultant and registrar were formally allocated to teaching and learning roles separated from the usual departmental management or clinical roles. Themes emphasised included clinical reasoning, practical clinical knowledge, communication, physical examination, procedural and professional skills. AIM: To evaluate the perceived educational value, effects on patient care and areas for ongoing development. METHODS: The study setting was an urban, tertiary referral, university-affiliated emergency department with prospectively allocated educational shifts of 4 or 5 h duration over a 6-month period. Evaluation was by session and course evaluation questionnaires, with respondents ranking predetermined themes and giving free-text responses. Qualitative presentation of results allowed exploration of the themes identified. RESULTS: Learners ranked history taking and physical examination technique as the most frequently learnt item, but clinical reasoning as the most important theme learnt. Informal discussion and performance critique or constructive feedback were the most frequent teaching methods. The biggest obstacle to learning was learner apprehension. The most frequent positive effect on patient care was faster management, decision making or disposition. Most often, no negative effect on patient care was identified. CONCLUSION: Formal bedside teaching is effective if organised with adequate staffing to quarantine the teacher and learner from routine clinical duties, and concentrating on themes best taught in the patient setting. Clinical reasoning and clinical knowledge were perceived to be most important, with positive effects on patient care through more thorough assessment and faster decision making.

Revascularization in the rabbit hindlimb: dissociation between capillary sprouting and arteriogenesis.

OBJECTIVE: Animal models of hindlimb ischemia are critical to our understanding of peripheral vascular disease and allow us to evaluate therapeutic strategies aimed to improve peripheral collateral circulation. To further elucidate the processes involved in revascularization following ischemia, we evaluated the temporal association between tissue ischemia, vascular endothelial cell growth factor (VEGF) release, angiogenesis (capillary sprouting), arteriogenesis (growth of the larger muscular arteries), and reserve blood flow (functional collateral flow). METHODS: New Zealand White rabbits (male 3-4 kg) were evaluated at specific days (0, 5, 10, 20 or 40) following femoral artery removal for measurement of hindlimb blood flow, skeletal muscle lactate production and VEGF content, capillary density (a marker of angiogenesis), and angiographic score (a marker of arteriogenesis). RESULTS: Maximal capillary sprouting occurred within 5 days of femoral artery removal and was temporally associated with reduced resting hindlimb blood flow, increased lactate release and detectable levels of skeletal muscle VEGF. The growth of larger angiographically visible collateral vessels occurred after 10 days and was not temporally associated with ischemia or skeletal muscle VEGF content, but did coincide with a large functional improvement in the reserve blood flow capacity of the limb. CONCLUSIONS: Following femoral artery removal in the rabbit, the time course of angiogenesis and arteriogenesis were clearly distinct. Tissue ischemia and/or VEGF may stimulate capillary sprouting, but this response does not translate to a significant improvement in collateral flow. The growth and development of the larger collateral vessels was correlated with a large functional improvement in collateral flow, and occurred at a time when VEGF levels were undetectable.

Maternal fish intake in late pregnancy and the frequency of low birth weight and intrauterine growth retardation in a cohort of British infants.

OBJECTIVE: To investigate the relation between maternal fish intake in late pregnancy and the frequency of low birth weight and intrauterine growth retardation (IUGR). PARTICIPANTS: 11 585 pregnant women in south west England. METHODS: Information on fish intake was obtained from a food frequency questionnaire sent to the women at 32 weeks' gestation, and used to calculate n-3 fatty acid (n-3FA) intake from fish. IUGR was defined as a birth weight for gestational age and sex below the 10th centile. Confounding variables considered included maternal age, height, weight, education, parity, smoking and drinking in pregnancy, and whether the mother was living with a partner. Only singleton, liveborn infants were included. MAIN RESULTS: Mean daily intakes of fish and n-3FAs were 32.8 g and 0.147 g respectively. In unadjusted analyses there were positive associations between mean birth weight and fish intake or n-3FA intake, but these disappeared on adjustment for potential confounders. The frequency of IUGR decreased with increasing fish intake-the OR (95%CI) of IUGR in those eating no fish was 1.85 (1.44 to 2.38) compared with those in the highest fish intake group. On adjustment this relation was attenuated (adjusted OR 1.37 (1.02 to 1.84)), but the decline in the frequency of IUGR with increasing fish intake remained statistically significant. No relation was observed between mean gestation and fish or n-3FA intake. CONCLUSIONS: These results lend some support to the hypothesis that raising fish or n3-FA intake during pregnancy may increase fetal growth rate. However, they provide no evidence that increasing fish consumption is associated with an increase in mean gestation.

Hyponatremia in ultradistance triathletes.

PURPOSE: Hyponatremia ([plasma sodium] <135 mmol x L(-1)) is a potentially serious complication of ultraendurance sports. However, the etiology of this condition is still uncertain. This observational cohort study aimed to determine prospectively the incidence and etiology of hyponatremia in an ultradistance triathlon. METHODS: The subjects consisted of 605 of the 660 athletes entered in the New Zealand Ironman triathlon (3.8-km swim, 180-km cycle, and 42.2-km run). Subjects were weighed before and after the race. A blood sample was drawn for measurement of plasma sodium concentration after the race. RESULTS: Complete data on pre- and postrace weights and plasma sodium concentrations were available in 330 race finishers. Postrace plasma sodium concentrations were inversely related to changes in body weight (P = 0.0001). Women (N = 38) had significantly lower plasma sodium concentrations (133.7 vs 137.4 mmol x L(-1); P = 0.0001) than men (N = 292) and lost significantly less relative weight (-2.7 vs -4.3%; P = 0.0002). Fifty-eight of 330 race finishers (18%) were hyponatremic; of these only 18 (31%) sought medical care for the symptoms of hyponatremia (symptomatic). Eleven of the 58 hyponatremic athletes had severe hyponatremia ([plasma sodium] < 130 mmol x L(-1)); seven of these 11 severely hyponatremic athletes were symptomatic. The relative body weight change of the 11 severely hyponatremic athletes ranged from 2.4% to +5%; eight (73%) of these athletes either maintained or gained weight during the race. In contrast, relative body weight changes in the 47 athletes with mild hyponatremia ([plasma sodium] 130-134 mmol x L(1)) were more variable, ranging from -9.25% to +2.2%. CONCLUSIONS: Hyponatremia is a common biochemical finding in ultradistance triathletes but is usually asymptomatic. Although mild hyponatremia was associated with variable body weight changes, fluid overload was the cause of most (73%) cases of severe, symptomatic hyponatremia.

Fat content of the diet among pre-school children in Britain; relationship with food and nutrient intakes.

OBJECTIVE: To investigate the relationship between the percentage of energy from fat and food and nutrient intakes at 18 and 43 months of age. DESIGN: Diet was assessed using a 3 day unweighed food record. The children were divided into quartiles of fat intake as a percentage of energy (QFI), and food and nutrient intakes in the different QFIs were compared. SUBJECTS: A total of 1026 children at 18 months and 863 children at 43 months, taking part in the Avon Longitudinal Study of Pregnancy and Childhood, participated. RESULTS: At 18 months the mean (s.d.) fat intake was 31.2 (2.8) % of energy in the lowest QFI and 43.1 (2.2) % in the highest. Energy intake increased slightly with increasing QFI. The percentage of energy derived from total sugar, and in particular non-milk extrinsic sugar fell as QFI increased, as did the ratio of polyunsaturated to saturated fatty acids. Intakes of retinol equivalents and zinc increased significantly with increasing QFI, while intakes of iron and most water soluble vitamins fell. There was a particularly marked fall in vitamin C intake as fat intake increased, from 11.8 mg/MJ in the lowest QFI, to 6.0 mg/MJ in the highest (P<0.001). Consumption of whole milk increased substantially with percentage energy from fat, from 51 g/MJ in the lowest QFI to 116 g/MJ in the highest (P<0.001), while consumption of fruit and fruit juice fell. The results obtained at 43 months were very similar. CONCLUSIONS: The chances of a suboptimal intake of zinc and retinol were higher at lower fat intakes. However, intakes of the fat-soluble vitamins E and D were unrelated to fat intake and intakes of iron and vitamin C fell as fat intakes increased. Some suggestions for improving diet in this age group have been given. SPONSOR: University of Bristol.

Diet during pregnancy in a population of pregnant women in South West England. ALSPAC Study Team. Avon Longitudinal Study of Pregnancy and Childhood.

OBJECTIVE: To describe the diet of a population of pregnant women. SUBJECTS: Eleven thousand, nine hundred and twenty-three pregnant women resident in the south-west of England. DESIGN: A self-completion unquantified food-frequency questionnaire was sent to the women at 32 weeks gestation. Estimated daily nutrient intakes were calculated from the answers to the questionnaire. RESULTS: On the whole, nutrient intakes of the pregnant women in this survey compared very closely with the reported nutrient intakes for all women aged 16-64 in the last Dietary and Nutritional Survey of British Adults (DNSBA). The exceptions were sugar, calcium, folate and vitamin C where the estimated intakes were somewhat higher, and retinol, where the estimated intake was somewhat lower than the DNSBA. Mean and median estimated nutrient intakes were above the RNIs for the majority of nutrients investigated except for energy, iron, magnesium, potassium and folate. Levels of supplementary vitamin and mineral use were fairly low, the two most commonly taken supplements were iron--taken by 22.5% of the pregnant women before 18 weeks and 43% at 32 weeks, and folate, taken by 9% and 18% of the women, respectively. CONCLUSIONS: These results suggest that the diets of pregnant women in this country are likely to contain adequate amounts of most nutrients, the most likely exceptions being iron, magnesium, potassium and folate. The relatively low intakes of folate and small proportion of women taking folate supplements is of concern, because of the association between inadequate amounts of folate in the diet and neural tube defects.

The effect of maternal smoking status, educational level and age on food and nutrient intakes in preschool children: results from the Avon Longitudinal Study of Parents and Children.

OBJECTIVE: Studies in adults have demonstrated that smoking status is associated with dietary quality, with smokers tending to have diets that conform less closely to guidelines on healthy eating than nonsmokers. However, there is very little information on the relation between children's dietary quality and parental smoking status. The objective of this study was to investigate the relation between maternal smoking status and nutrient intake in preschool children, allowing for the possible confounding effects of maternal educational level and age at delivery. SUBJECTS: In total, 993 children aged 18 months participating in the Avon Longitudinal Study of Parents and Children. METHODS: Diet was assessed by a 3-day food record. Maternal smoking status and educational level and age at delivery were assessed by questionnaire. RESULTS: In multivariate analysis, the children of smokers had significantly higher intakes of monounsaturated fatty acids and starch, and lower intakes of nonstarch polysaccharides (NSP). They were also less likely to have eaten poultry, buns, cakes and puddings, wholemeal bread and fruit, and more likely to have drunk sugar-sweetened soft drinks. Intakes of NSP and most vitamins and minerals increased significantly with increasing maternal education. Children of more highly educated mothers were less likely to have eaten chocolate, crisps and white bread, and more likely to have consumed wholemeal bread, fruit and fruit juice. CONCLUSIONS: The children of nonsmokers and more highly educated mothers consumed a diet that conformed more closely to current guidelines on healthy eating. These dietary differences may contribute to the excess of ill-health observed in the children of smokers and of less-educated mothers.

Drinks consumed by 18-month-old children: are current recommendations being followed?

OBJECTIVE: To examine the types of drinks consumed by children at 18 months of age, determine any associations with socio-demographic characteristics and investigate the use of a bottle for providing these drinks. DESIGN: A total of 1026 randomly chosen children from the Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC) were studied. METHODS: As part of a dietary diary completed when their children were 18 months of age, mothers were asked to record all drinks consumed over a 24 h period and to report the containers used to give these drinks. Socio-demographic characteristics were obtained via self-completion questionnaire. RESULTS: Dietary records were available for 1026 children at 18 months. A wide variety of drinks were given at this age. In all, 64% of children had received at least one drink in a bottle and 10% solely used a bottle. Bottle users consumed a significantly greater volume of milk drinks compared to those who used a cup or other container. The types of drinks consumed and adherence to the guidelines on weaning were associated with several socio-demographic characteristics. In particular, children with mothers of lower educational level were more likely to receive fizzy drinks and low-calorie soft drinks, more likely to drink from a bottle, and less likely to drink from a cup. CONCLUSION: We have shown that certain groups of mothers with 18-month-old children are not following the current guidelines on weaning. Assuming these guidelines have scientific validity, it is clear that nutritional education needs to be targeted at these mothers to help them feed their children in the optimal way, to protect from nutritional deficiencies, possible later obesity and poor dental health. SPONSORSHIP: University of Bristol.

Financial difficulties, smoking habits, composition of the diet and birthweight in a population of pregnant women in the South West of England. ALSPAC Study Team. Avon Longitudinal Study of Pregnancy and Childhood.

OBJECTIVE: To investigate the relationships between self-assessed financial difficulties, diet in pregnancy and birthweight. DESIGN: Information on diet composition, difficulty in affording food, smoking habits and parity was collected by self-completion questionnaires at 32 weeks of pregnancy. Information on birthweights was obtained from hospital records. SUBJECTS: A geographically defined population 11833 pregnant women resident in the South-West of England. MAIN OUTCOME MEASURES: Nutrient intakes, food choices and birthweight. RESULTS: Women with greater difficulty in affording food had lower intakes of protein, fibre, vitamin C, niacin, pyridoxine, iron, zinc, magnesium and potassium than did women with little or no difficulty. They were more likely to use cooking and spreading fats with a high saturates content, and less likely to eat fish, fruit, vegetables and salad. In a multivariate analysis including parity and smoking status, financial difficulty was found to have no significant relationship with birthweight. CONCLUSION: The quality of diet in pregnancy falls with increasing difficulty in affording food. However, this does not appear to affect mean birthweight of infants born after 32 weeks gestation.

Several epitopes of p85 glycoprotein (CDw44) are dependent on intact disulphide bonds. Isolation of cDNA clones requires a polyclonal antibody raised against the reduced protein.

Monoclonal antibodies 50B4 and 50E6 recognize two distinct epitopes of human p85 glycoprotein (CDw44). Both epitopes are destroyed by reduction of the purified glycoprotein as demonstrated by inhibition of cellular radioimmunoassay and Western blot analysis. Endoglycosidase F treated p85 glycoprotein, with an apparent molecular weight of 73,000, is still reactive with both monoclonal antibodies. Thus both epitopes are conformational determinants of the polypeptide chain. A rabbit antibody produced against purified native p85 glycoprotein also reacted only with the non-reduced form of p85. Repeated immunizations with SDS-dissociated and reduced p85 yielded a polyclonal antibody reactive by Western blot analysis with reduced and non-reduced forms of p85 glycoprotein. When a HOON leukemia cell line cDNA expression library was screened with this polyclonal antibody, two cDNA clones were isolated which reacted specifically with the antiserum and not with the control non-immune serum. Preliminary characterization of these clones indicates that they are p85-related.

The accuracy of oral predictive and infrared emission detection tympanic thermometers in an emergency department setting.

OBJECTIVE: To assess the accuracy of an oral predictive thermometer and an infrared emission detection (IRED) tympanic thermometer in detecting fever in an adult emergency department (ED) population, using an oral glass mercury thermometer as the criterion standard. METHODS: This was a single-center, nonrandomized trial performed in the ED of a metropolitan tertiary referral hospital with a convenience sample of 500 subjects. The temperature of each subject was taken by an oral predictive thermometer, an IRED tympanic thermometer set to "oral" equivalent, and an oral glass mercury thermometer (used as the criterion standard). A fever was defined as a temperature of 37.8 degrees C or higher. The subject's age, sex, triage category, and diagnostic group were also recorded. Sensitivity, specificity, positive and negative likelihood ratios, positive and negative predictive values, and corresponding 95% confidence intervals were calculated. Logistic regression was used to identify predictors of fever. RESULTS: The sensitivities and specificities for detection of fever of the predictive and the IRED tympanic thermometers were similar (sensitivity 85.7%/88.1% and specificity 98.7%/95.8%, respectively). The predictive thermometer had a better positive predictive value (85.7%) compared with the IRED tympanic thermometer (66.1%). The positive and negative likelihood ratios for the predictive oral thermometer were 65 and 0.14, respectively, and for the IRED tympanic thermometer 21 and 0.12, respectively, indicating that the predictive thermometer will "miss" 1 in about 7 fevers and the IRED tympanic thermometer will "miss" 1 in about 8 fevers. CONCLUSIONS: Although quick and convenient, oral predictive and IRED tympanic thermometers give readings that cannot always be relied on in the detection of fever. If we are to continue using electronic thermometers in the ED setting, we need to recognize their limitations and maintain the importance of our clinical judgment.

Relactation.

Relactation may be useful in the developing world either if the child has been ill and unable to feed for a time or the mother is ill or has died. Relactation appears to be easier with a younger infant and in women who have lactated previously. However, with appropriate care, support and motivation even some women who have never been pregnant or who have been pregnant but never lactated may be able to start lactation.

PIP: The need for relactation can arise if a child has been ill and unable to nurse for a time, when a relative or adoptive parent wants to assume breast feeding of a child after a maternal death, or if an infant is found to be unable to tolerate formula milk. The degree of breast and nipple stimulation is considered most important to the chance of inducing a good milk supply. Several studies have found that the willingness to suckle decreases as the age of the infant increases. The methods most commonly used by women in developed countries to prepare the breast are hand expression and a hand pump. Women in developing countries have been enabled to relactate after initiation of a high-protein, high-calorie diet. The use of drugs or hormonal preparations has not been adequately researched. In general, relactation seems to be most successful with younger infants, in cases of incomplete postpartum breast involution, and in women who have lactated previously. Most women report the relactation process is stressful and a support group of some type is recommended.