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Biomedical subjects

C E West

Publications and source records attributed to C E West.

At least 55 records · Page 3Linked to original sources

The barley scutellar peptide transporter: biochemical characterization and localization to the plasma membrane.

Thiol-affinity labelling was used to identify and characterize components of the peptide transport system in the barley (Hordeum vulgare) scutellar epithelium. SDS-PAGE and 2D-PAGE in conjunction with fluorography were used to study derivatized proteins. Membrane proteins of 42 kDa and 66 kDa were identified using a strategy devised to label substrate protectable protein with the thiol specific reagent [14C] N-ethylmaleimide (NEM). The scutellar plasma membrane is the anticipated site of transporters involved in the mobilization of endosperm storage reserves in the germinating barley grain. The subcellular localization of these proteins to the plasma membrane was demonstrated by thiol-affinity labelling of high purity plasma membrane vesicles isolated from barley scutellar tissue. A peptide transporter, HvPTR1, specific to the barley scutellum has recently been cloned in this laboratory. A 66 kDa protein, comparable to the predicted molecular mass of HvPTR1, was identified by [14C]NEM labelling studies of Xenopus laevis oocytes expressing HvPTR1 cRNA, but not water injected controls. Peptide antiserum raised to HvPTR1 also cross-reacted with a 66 kDa membrane protein in barley scutellar tissue. This confirms that the 66 kDa protein identified here by thiol-affinity labelling studies is the barley scutellum peptide transporter HvPTR1, and demonstrates that this protein is localized to the plasma membrane of scutellar epithelial cells during germination.

Affinity Labels↗

Efficacy of oral iodised oil is associated with anthropometric status in severely iodine-deficient schoolchildren in rural Malawi.

The effect of anthropometric status on the efficacy of an oral supplement of iodised oil (1 ml Lipiodol Ultrafluide, 490 mg I; Laboratoire Guerbet, Aulnay-sous-Bois, France) was examined in 8-10-year-old schoolchildren (n 197) of Ntcheu, a severely I-deficient district of Malawi. The study was a controlled trial using the I concentration of casual urine samples to monitor the I status. The median urinary I concentration increased from 0.15 micromol/l at baseline (51.3 % of children < 0.16 micromol/l, 89.7 % < 0.40 micromol/l, 95.7 % 0.79 micromol/l) to 0.32 micromol/l at 40 weeks (29.1 % of the children < 0.16 micromol/l, 71.0 % < 0.40 micromol/l, 96.1 % < 0.79 micromol/l) while the total goitre prevalence fell from 63 % to 21 %. Variables of efficacy were estimated from a hyperbolic function describing the longitudinal pattern of urinary I excretion after the dose. The I retention and I elimination rate, and the periods of protection from mild (< 0.79 micromol/l) or moderate (< 0.40 micromol/l) I deficiency were obtained for groups of children with differing anthropometric status at baseline. Initial height-for-age and mid upper-arm circumference were not significantly related to efficacy. However, both the I retention and I elimination rate were reduced in children with lower initial weight-for-height. Children with lower skinfold thickness at baseline also had reduced I retention, which resulted in shorter protection periods from recurrent moderate and mild I deficiency. The efficacy of the oral iodised-oil supplement was not related to changes in anthropometric status during follow-up, nor was it related to the consumption of a food supplement of 1610 kJ immediately before the iodised-oil dose. Very low (< 0.16 micromol/l) urinary I concentration, and the presence of goitre at baseline were both associated with higher I retention and elimination rate. Children with goitre at baseline were found to have a prolonged duration of protection against recurrent moderate I deficiency. We conclude that in apparently healthy schoolchildren in I-deficient areas, general anthropometric status has a little influence on the efficacy of oral iodised oil for correcting I deficiency.

Administration, Oral↗

Comparison of three methods for estimating daily individual discretionary salt intake: 24 hour recall, duplicate-portion method, and urinary lithium-labelled household salt excretion.

OBJECTIVE: To compare methods for estimating discretionary salt intake, that is, salt added during food preparation and consumption in the home. SETTING: The study was carried out in a rural Guatemalan village. SUBJECTS: Subjects were selected non-randomly, based on their willingness to cooperate. Nine mother-son dyads participated; the sons were aged 6-9 y. INTERVENTIONS: Three approaches for estimating the discretionary salt consumption were used: 24 h recall; collection of duplicate portions of salt; and urinary excretion of lithium during consumption of lithium-labelled household salt. Total salt intake was assessed from the excretion of chloride over 24 h. RESULTS: The mean discretionary salt consumption based on lithium excretion for mothers was 3.9+/-2.0 g/d (mean +/- s.d.) and for children 1.3+/-0.6 g/d. Estimates from the 24 h recalls and from the duplicate portion method were approximately twice and three times those measured with the lithium-marker technique respectively. The salt intake estimated from the recall method was associated with the lithium-marker technique for both mothers and children (Spearman correlation coefficient, 0.76 and 0.70 respectively). The mean daily coefficient of variation in consumption of discretionary salt measured by the three methods, for mothers and boys respectively, were: lithium marker, 51.7 and 43.7%; 24 h recall, 65.8 and 50.7%; and duplicate portion, 51.0 and 62.6%. CONCLUSIONS: We conclude that an interview method for estimating discretionary salt intake may be a reasonable approach for determining the relative rank-order in a population, especially among female food preparers themselves, but may grossly overestimate the actual intake of salt added during food preparation and consumption.

Adult↗

Bioavailability of lutein from vegetables is 5 times higher than that of beta-carotene.

BACKGROUND: To gain more insight into the relation between vegetable consumption and the risk of chronic diseases, it is important to determine the bioavailability of carotenoids from vegetables and the effect of vegetable consumption on selected biomarkers of chronic diseases. OBJECTIVE: To assess the bioavailability of beta-carotene and lutein from vegetables and the effect of increased vegetable consumption on the ex vivo oxidizability of LDL. DESIGN: Over 4 wk, 22 healthy adult subjects consumed a high-vegetable diet (490 g/d), 22 consumed a low-vegetable diet (130 g/d), and 10 consumed a low-vegetable diet supplemented with pure beta-carotene (6 mg/d) and lutein (9 mg/d). RESULTS: Plasma concentrations of vitamin C and carotenoids (ie, alpha-carotene, beta-carotene, lutein, zeaxanthin, and beta-cryptoxanthin) were significantly higher after the high-vegetable diet than after the low-vegetable diet. In addition to an increase in plasma beta-carotene and lutein, the pure carotenoid-supplemented diet induced a significant decrease in plasma lycopene concentration of -0.11 micromol/L (95% CI: -0.21, -0.0061). The responses of plasma beta-carotene and lutein to the high-vegetable diet were 14% and 67%, respectively, of those to the pure carotenoid- supplemented diet. Conversion of beta-carotene to retinol may have attenuated its plasma response compared with that of lutein. There was no significant effect on the resistance of LDL to oxidation ex vivo. CONCLUSIONS: Increased vegetable consumption enhances plasma vitamin C and carotenoid concentrations, but not resistance of LDL to oxidation. The relative bioavailability of lutein from vegetables is higher than that of beta-carotene.

Adolescent↗

beta-carotene does not change markers of enzymatic and nonenzymatic antioxidant activity in human blood.

In vitamin A-replete populations, increased concentrations of serum carotenoids have been associated with a decreased risk of degenerative diseases. The mechanism of action of carotenoids in determining antioxidant activity is largely unknown. The aim of the study was to examine the effect of carotenoid supplementation and spinach intake on erythrocyte enzyme antioxidant activities, serum or plasma nonenzymatic antioxidant concentrations, and concentrations of oxidatively damaged amino acids in plasma. Subjects received for 3 wk a basic diet (n = 10), a basic diet with a carotenoid supplement (n = 12) or with a spinach product (n = 12 per group), i.e., whole-leaf, minced, liquefied or liquefied spinach plus added dietary fiber. After 3 wk of dietary intervention, changes in serum or plasma concentrations of ascorbic acid, alpha-tocopherol, FRAP (ferric reducing ability of plasma) and uric acid and erythrocyte enzyme activities were assessed, and differences among experimental groups were tested. Consumption of spinach resulted in greater (P < 0.01) erythrocyte glutathione reductase activity and lower (P < 0.05) erythrocyte catalase activity and serum alpha-tocopherol concentration compared with the control group. Consumption of the carotenoid supplement led to lower alpha-tocopherol responses (P = 0.02) compared with the basic diet only. Our data suggest that the short-term changes in erythrocyte glutathione reductase activity and serum alpha-tocopherol concentration can be attributed to an increased carotenoid (lutein and zeaxanthin) intake, but beta-carotene is unlikely to be a causative factor. Lower erythrocyte catalase activity after intervention with spinach products may be related to other constituents in spinach such as flavonoids.

Adolescent↗

The food matrix of spinach is a limiting factor in determining the bioavailability of beta-carotene and to a lesser extent of lutein in humans.

Carotenoid bioavailability depends, amongst other factors, on the food matrix and on the type and extent of processing. To examine the effect of variously processed spinach products and of dietary fiber on serum carotenoid concentrations, subjects received, over a 3-wk period, a control diet (n = 10) or a control diet supplemented with carotenoids or one of four spinach products (n = 12 per group): whole leaf spinach with an almost intact food matrix, minced spinach with the matrix partially disrupted, enzymatically liquefied spinach in which the matrix was further disrupted and the liquefied spinach to which dietary fiber (10 g/kg wet weight) was added. Consumption of spinach significantly increased serum concentrations of all-trans-beta-carotene, cis-beta-carotene, (and consequently total beta-carotene), lutein, alpha-carotene and retinol and decreased the serum concentration of lycopene. Serum total beta-carotene responses (changes in serum concentrations from the start to the end of the intervention period) differed significantly between the whole leaf and liquefied spinach groups and between the minced and liquefied spinach groups. The lutein response did not differ among spinach groups. Addition of dietary fiber to the liquefied spinach had no effect on serum carotenoid responses. The relative bioavailability as compared to bioavailability of the carotenoid supplement for whole leaf, minced, liquefied and liquefied spinach plus added dietary fiber for beta-carotene was 5.1, 6.4, 9.5 and 9.3%, respectively, and for lutein 45, 52, 55 and 54%, respectively. We conclude that the bioavailability of lutein from spinach was higher than that of beta-carotene and that enzymatic disruption of the matrix (cell wall structure) enhanced the bioavailability of beta-carotene from whole leaf and minced spinach, but had no effect on lutein bioavailability.

Adolescent↗

Dietary folate from vegetables and citrus fruit decreases plasma homocysteine concentrations in humans in a dietary controlled trial.

Elevated total plasma homocysteine (tHcy) concentrations are considered a risk factor for neural tube defects (NTD) and cardiovascular disease. Supplementation with folic acid decreases the risk of women having children with NTD. In both sexes, it decreases tHcy levels. We investigated the efficacy of natural dietary folate in improving folate and homocysteine status. We performed a 4-wk dietary controlled, parallel design intervention trial with 66 healthy subjects (18-45 y) divided into 3 treatment groups: the dietary folate group, the folic acid group and the placebo group. Each day each group was fed a different diet. The dietary folate group received a diet high in vegetables and citrus fruit (total folate content approximately 560 microgram) plus a placebo tablet. The folic acid group received a diet naturally low in folate (approximately 210 microgram) plus 500 microgram folic acid and placebo tablet on alternate days, i.e., 250 microgram folic acid/d. And the placebo group received the same low-folate diet as the folic acid group plus a placebo tablet. After 4 wk of intervention, folate status improved, and tHcy concentrations decreased in both the dietary folate and the folic acid groups. From the amount of additional folate (350 microgram/d) and folic acid (250 microgram/d) consumed, the relative bioavailability of dietary folate compared to folic acid was calculated to be 60-98%, depending on the endpoint used. In conclusion, increasing the consumption of vegetables and citrus fruit, both good sources of folate, will improve folate status and decrease tHcy concentrations. This may contribute to the prevention of cardiovascular disease and NTD in the general population

Adolescent↗

Anti-malarial drug use among preschool children in an area of seasonal malaria transmission in Kenya.

The aims of this study were to estimate the proportion of asymptomatic Kenyan preschool children using anti-malarial drugs, to identify factors associated with chloroquine use, and to assess the validity of frequency of febrile episodes and drug use reported by mothers or carers. Of 318 children studied, 38% (95% confidence interval [CI] = 30-47%]) tested positive for chloroquine or sulfadoxine. Of chloroquine-positive children, 15% had concentrations exceeding the estimated minimum therapeutically effective values. Among those testing negative for sulfadoxine, chloroquine-positive children were more frequently parasitemic (odds ratio = 2.6, 95% CI = 1.3-5.2), and had lower mean hemoglobin concentrations (6.1 g/L, 95% CI = 2.1-10.1) than chloroquine-negative children. Mothers over-reported the frequency of malaria or fever episodes as usually defined in medical studies, and underreported anti-malarial drug use. We conclude that anti-malarials are frequently given for treatment of malaria or malaria-associated illness, rather than prophylactically or for symptoms unrelated to malaria. Questionnaire surveys cannot replace biochemical markers to obtain information on anti-malarial drug use.

Adult↗

Development of a method for quantitation of retinol and retinyl palmitate in human serum using high-performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry.

A method for the quantitative analysis of the vitamin A compounds all-trans-retinol and all-trans-retinyl palmitate was developed using high-performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (APCI-LC-MS). Unlike previous quantitative mass spectrometric methods for vitamin A, HPLC separations were carried out using a C30 reversed-phase column instead of GC separation. Because no sample hydrolysis or derivatization was necessary, retinyl palmitate was preserved for analysis instead of being hydrolyzed to retinol. Human serum was analyzed following simple hexane extraction without saponification or any additional purification. A comparison of APCI and electrospray ionization showed that only APCI produced a linear response over all four orders of magnitude of retinol and three orders of magnitude of retinyl palmitate concentrations. Selected ion monitoring of the fragment ion of m/z 269 was used for APCI quantitation of both retinol and retinyl palmitate, since it was the base peak and the only abundant ion in the mass spectra of both compounds and the internal standard, retinyl acetate. The ion of m/z 269 corresponded to loss of water, loss of palmitic acid, or elimination of acetic acid from the protonated molecules of retinol, retinyl palmitate and retinyl acetate, respectively. The limit of detection of APCI-LC-MS for all-trans-retinol and all-trans-retinyl palmitate was determined to be approximately 34 fmol/microliter and 36 fmol/microliter (0.670 pmol all-trans-retinol and 0.720 pmol all-trans-retinyl palmitate injected in 20 microliters on-column), respectively. The limit of quantitation was approximately 500 fmol/microliter and 250 fmol/microliter (10 pmol and 5 pmol injected in 20 microliters on-column) for retinol and retinyl palmitate, respectively.

Chromatography, High Pressure Liquid↗

Cloning and functional characterisation of a peptide transporter expressed in the scutellum of barley grain during the early stages of germination.

A peptide transport gene (HvPTR1) expressed in the scutella of germinating barley grain has been cloned by an RT-PCR approach. Sequence analysis of the full length cDNA (2260 bp) revealed an open reading frame encoding a 579 amino acid protein of predicted molecular mass 63 kDa, which displayed 58% identity to the Arabidopsis thaliana peptide transporter AtPTR2-B. Expression of HvPTR1 in Xenopus laevis oocytes conferred a 48-fold increase in alanyl-[14C]phenylalanine uptake relative to water injected oocytes, confirming the function of HvPTR1 as a peptide transporter. HvPTR1 expression was detectable only in the scutellum of the germinating barley grain, with no transcript found in roots, shoots or the embryo axis. Transcript levels increased rapidly from 6 to 24 h imbibition, correlating with the development of peptide transport activity in the barley scutellum. Peptide transport provides a significant source of organic nitrogen to the barley embryo for use in germination and growth processes associated with the early stages of seedling development. The temporal and spatial pattern of HvPTR1 expression is consistent with a central role for HvPTR1 in the transport of peptides in the germinating barley grain.

Amino Acid Sequence↗

Determination of discretionary salt intake in rural Guatemala and Benin to determine the iodine fortification of salt required to control iodine deficiency disorders: studies using lithium-labeled salt.

The use of discretionary salt, which is salt added during cooking and at the table, as a suitable vehicle for iodine intake was assessed by measuring salt consumption using the lithium-marker technique in rural areas of Guatemala and Benin. In both countries, we studied boys aged 6-12 y and their mothers. Subjects used lithium-labeled salt after all unlabeled salt was removed from their households. In Guatemala, 24-h urine samples for 9 mother-son pairs were collected at baseline and on days 7, 8, and 9 during the use of lithium-labeled salt. Total maternal salt intake averaged 5.2 +/- 1.7 g/d (mean +/- SD), of which 77 +/- 24% came from discretionary sources, whereas Guatemalan boys consumed 1.8 +/- 0.6 g salt/d, of which 72 +/- 12% came from discretionary sources. In Benin, urine collection from 13 mother-son pairs took place at baseline and on days 5 and 7. Beninese mothers had a total salt intake of 9.0 +/- 2.9 g/d and their sons had an intake of 5.7 +/- 2.8 g/d; discretionary salt contributed 52 +/- 14% and 50 +/- 13%, respectively, of total salt consumed. Therefore, fortification of household salt appears to be an appropriate method of controlling iodine deficiency in both countries, although fortification of other salt sources could be considered in Benin.

Adult↗

Orange fruit is more effective than are dark-green, leafy vegetables in increasing serum concentrations of retinol and beta-carotene in schoolchildren in Indonesia.

The objectives of this study were to quantify the effectiveness of dietary retinol sources, orange fruit, and dark-green, leafy vegetables in improving vitamin A status, and to test whether orange fruit is a better source of vitamin A and carotenoids than are leafy vegetables. Anemic schoolchildren aged 7-11 y (n = 238) in West Java, Indonesia, were randomly allocated to 1 of 4 groups to consume 2 complete meals/d, 6 d/wk, for 9 wk: 1) 556 retinol equivalents (RE)/d from retinol-rich food (n = 48); 2) 509 RE/d from fruit (n = 49); 3) 684 RE/d from dark-green, leafy vegetables and carrots (n = 45); and 4) 44 RE/d from low-retinol, low-carotene food (n = 46). Mean changes in serum retinol concentrations of the retinol-rich, fruit, vegetable, and low-retinol, low-carotene groups were 0.23 (95% CI: 0.18, 0.28), 0.12 (0.06, 0.18), 0.07 (0.03,0.11), and 0.00 (-0.06, 0.05) micromol/L, respectively. Mean changes in serum beta-carotene concentrations in the vegetable and fruit groups were 0.14 (0.12, 0.17) and 0.52 (0.43, 0.60) micromol/L, respectively. Until now, it has been assumed that 6 microg dietary beta-carotene is equivalent to 1 RE. On the basis of this study, however, the equivalent of 1 RE would be 12 microg beta-carotene (95% CI: 6 microg, 29 microg) for fruit and 26 microg beta-carotene (95% CI: 13 microg, 76 microg) for leafy vegetables and carrots. Thus, the apparent mean vitamin A activity of carotenoids in fruit and in leafy vegetables and carrots was 50% (95% CI: 21%, 100%) and 23% (95% CI: 8%, 46%) of that assumed, respectively. This has important implications for choosing strategies for controlling vitamin A deficiency. Research should be directed toward ways of improving bioavailability and bioconversion of dietary carotenoids, focusing on factors such as intestinal parasites, absorption inhibitors, and food matrixes.

Anthropometry↗

Bioavailability and bioconversion of carotenoids.

Factors that influence the bioavailability of carotenoids and their bioconversion to retinol are species of carotenoids, molecular linkage, amount of carotenoids consumed in a meal, matrix in which the carotenoid is incorporated, effectors of absorption and bioconversion, nutrient status of the host, genetic factors, host related factors, and mathematical interactions. In this paper, current knowledge of these factors is examined. Although data are not sufficiently comparable to allow an extensive systematic comparison of results, a number of conclusions can be drawn from the information available.

Absorption↗

Potential of vegetable processing to increase the delivery of carotenoids to man.

It has been suggested that carotenoids contribute to the inverse association between vegetable consumption and risk for coronary heart disease and cancer. However, there are indications that the bioavailability of beta-carotene from vegetables is less than previously thought. Therefore, it is of interest to explore whether the bioavailability of carotenoids from vegetables can be improved. We have reviewed data on the effect of vegetable processing on carotenoid bioavailability. Based on this analysis, we conclude that processing of vegetables by mechanical homogenization or heat treatment has the potential of increasing the bioavailability of carotenoids. Addition of fat during processing may also have an effect.

Absorption↗

Quantification of the "SLAMENGHI" factors for carotenoid bioavailability and bioconversion.

The bioavailability of carotenoids is the fraction of ingested carotenoids utilized for normal physiological functions or storage, while bioconversion is the proportion of bioavailable carotenoid converted to retinol (vitamin A). Factors influencing carotenoid bioavailability and bioconversion have been incorporated in the mnemonic "SLAMENGHI": species of carotenoid; molecular linkage; amount of carotenoids consumed in a meal; matrix in which the carotenoid is incorporated; effectors of absorption and bioconversion; nutrient status of the host; genetic factors; host-related factors; and mathematical interactions. In this paper, an attempt is made to quantify the impact of the various SLAMENGHI factors on carotenoid bioavailability and bioconversion although many of the data available are not strictly comparable.

Absorption↗

Malaria is associated with reduced serum retinol levels in rural Zambian children.

Vitamin A deficiency increases the risk of illness, while infections impair vitamin A status. Malaria is highly prevalent in rural Zambia. We describe the relationship between malaria and vitamin A status. We examined dietary vitamin A intake, malaria parasitaemia and serum concentrations of retinol, C-reactive protein (CRP) and alpha 1-acid glycoprotein (AGP) in 210 children under the age of 2. Vitamin A intake was low. Serum retinol was negatively correlated with malaria parasite count and to serum levels of CRP and AGP. Increased malaria parasite density resulted in raised CRP and AGP levels, which were negatively associated with serum retinol. We conclude that improvement of dietary vitamin A intake and prevention of infectious diseases, especially malaria, could alleviate vitamin A deficiency in this population.

Animals↗