Convergent validity, ceiling, and floor effects of the English-ISYQOL against established quality of life questionnaires (SRS-22r and SAQ) and curve angles in adolescents with idiopathic scoliosis.

Scoliosis significantly impacts Quality of Life (QOL). Current quality of life questionnaires for adolescents with idiopathic scoliosis (AIS) have limitations. A new questionnaire for measuring QOL in AIS called the Italian Spine Youth Quality of Life (ISYQOL) has been developed to address these limitations but the English translation has not yet been validated. To determine the ceiling and floor effects, and the convergent validity of the ISYQOL questionnaire against established QOL questionnaires and Cobb angle in AIS. One hundred consecutive females with AIS, (10-18 years old), treated non-operatively. The English translation of the ISYQOL was compared to the following established questionnaires: Scoliosis Research Society-22r and the Spinal Appearance Questionnaire. The participants were 100 females (13.89+/-1.8 years) with 28.75+/-13.9° curve angles. The convergent validity of the ISYQOL score (60.3+/-12.44) was supported by significant correlation with the SRS-22r total score, function, pain, self-image, and mental health scores (r = 0.70, 0.54, 0.57, 0.52 and 0.50, respectively), and with the SAQ general, waist, and expectations domains (r = -0.6. -.52, and -0.56, respectively). Correlation with the Cobb angle was (r = -.37)(see Table 1). No ceiling effect was observed in the ISYQOL. Ceiling effects were observed for the SRS-22r and the SAQ. The ISYQOL demonstrated evidence of convergent validity. This study supports its suitability for QOL research in AIS. ISYQOL appears more likely to detect changes in evaluative studies than the SRS- 22r and the SAQ.

Vitamin E supplementation and oxidative damage to DNA and plasma LDL in type 1 diabetes.

OBJECTIVE: To determine the effect of 400 IU/day of the antioxidant vitamin E on the susceptibility of plasma LDL and lymphocyte DNA to oxidative damage in type 1 diabetes. RESEARCH DESIGN AND METHODS: We studied 42 patients with type 1 diabetes and 31 age- and sex-matched control subjects in a randomized prospective double-blind placebo-controlled trial by using 400 IU/day of oral vitamin E for 8 weeks. Measurements were made of single-strand breaks in lymphocyte DNA at baseline and after hydrogen peroxide-induced stress (comet assay) and of copper-induced LDL oxidization and plasma antioxidant profiles. RESULTS: Plasma LDL and lymphocyte DNA were more resistant to induced oxidative change in the type 1 diabetes group than in control subjects. Vitamin E supplementation reduced LDL oxidizability in the control subjects but not in the type 1 diabetes group and had no effect on oxidative DNA damage in either group. The type 1 diabetes group had a significantly poorer plasma antioxidant profile with lower mean serum concentrations of alpha-tocopherol and most carotenoids than control subjects. CONCLUSIONS: Plasma LDL and lymphocyte DNA appear to be more resistant to oxidative change in type 1 diabetic subjects than in control subjects, and there was no evidence of oxidatively induced DNA or LDL change in type 1 diabetes. This study does not support the hypothesis of oxidative damage in these patients, and a dose of vitamin E (400 IU/day) that reduced LDL oxidative susceptibility in control subjects did not do so in patients with type 1 diabetes.

Measurement of aldehydes in low density lipoprotein by high performance liquid chromatography.

A modified procedure is presented for the HPLC determination of nanomolar concentrations of n-alkanals, hydroxyalkenals, malondialdehyde and furfural in biological fluid. The modifications allow aldehyde profile analysis of small samples of fresh, human, low density lipoprotein (LDL), enabling more detailed studies of LDL fatty acid peroxidation. Aldehydes are reacted with 1,3-cyclohexanedione to produce fluorescent derivatives which are separated by gradient, reversed phase, high performance liquid chromatography (HPLC). Analysis time has been reduced by shortening the sample preparation. Sensitivity has been increased by miniaturization of the derivatisation procedure, reducing required sample size. Recoveries of added aldehydes have been improved. In addition, the method presented allows determination of three further aldehydes, not measured previously by CHD methods: malondialdehyde, formaldehyde and furfural. Recovery and variability data and concentrations of aldehydes found in human LDL are given. The capacity of the method for further development, to enable determination of other aldehydes such as the trans, 2-alkenals, is also demonstrated.

Oxidative insult specifically decreases levels of a mitochondrial transcript.

The effects of oxidative insult, applied with hydrogen peroxide, on gene transcript levels in a human lymphocyte cell line (Molt-17) were investigated using mRNA differential display. Several cDNA fragments corresponding to putatively up- or down-regulated transcripts were isolated. One of these was found to hybridize to two discrete transcripts on Northern blots of Molt-17 cell RNA. The more abundant transcript, that has previously been demonstrated to correspond to the mRNA for mitochondrial ATPase subunits 8 and 6, was unaffected by the hydrogen peroxide treatment. In contrast, levels of the rarer, larger transcript were consistently reduced in a rapid, sustained, and dose-dependent manner following hydrogen peroxide treatment. Prior supplementation of the cells with beta carotene provided some protection against the reduction in levels of this transcript following hydrogen peroxide treatment. In contrast, vitamins C and E had no effect at the concentrations tested. We have now cloned the cDNA corresponding to this stress-responsive transcript and demonstrated that it is an incompletely processed product of the mitochondrial genome encompassing ATPase subunits 8 and 6 plus the adjacent gene for cytochrome c oxidase subunit 3. This decrease in one specific mitochondrial transcript may represent a novel mechanism for differential expression of mitochondrially-encoded genes.

Measurement of cellular repair activities for oxidative DNA damage.

Two related assays capable of determining cell extract repair activities for different oxidative lesions in DNA are described. Both assays measure the incorporation of radiolabeled nucleotides during repair of an oxidatively damaged template in a cell-free system. The assays differ in the type of oxidative damage present in the DNA. In one, singlet oxygen is used to generate predominantly 8-oxo-2'-deoxyguanosine lesions. In the other, hydroxyl radicals are used to generate a broad spectrum of damage including oxidized bases and strand breaks. Assay conditions were adjusted to ensure that radiolabel incorporation was directly proportional to cell extract repair activity. These assays represent sensitive tools for investigating the regulation of repair systems for oxidative DNA damage.

Beta-carotene enhances the recovery of lymphocytes from oxidative DNA damage.

Epidemiological studies have revealed a strong correlation between high intake of fruit and vegetables and low incidence of certain cancers. Micronutrients present in these foods are thought to decrease free radical attack on DNA and hence protect against mutations that cause cancer, but the fine details of the causal mechanism have still to be elucidated. Whether dietary factors can modulate DNA repair--a crucial element in the avoidance of carcinogenesis--is an intriguing question that has not yet been satisfactorily answered. In order to investigate the effects of beta-carotene on oxidative damage and its repair, volunteers were given a single 45 mg dose and lymphocytes taken before and after the supplement were treated in vitro with H2O2. DNA strand breaks and oxidised pyrimidines were measured at intervals, to monitor the removal of oxidative DNA damage. We found inter-individual variations in response. In cases where the baseline plasma beta-carotene concentration was high, or where supplementation increased the plasma concentration, recovery from oxidative damage (i.e. removal of both oxidised pyrimidines and strand breaks) was relatively rapid. However, what seems to be an enhancement of repair might in fact represent an amelioration of the continuing oxidative stress encountered by the lymphocytes under in vitro culture conditions. We found that culture in a 5% oxygen atmosphere enhanced recovery of lymphocytes from H2O2 damage.

High performance liquid chromatography method for the determination of pyridoxal-5-phosphate in human plasma: how appropriate are cut-off values for vitamin B6 deficiency?

OBJECTIVES: Application of a HPLC (high performance liquid chromatography) method, using cyanide derivatisation, to the determination of plasma pyridoxal-5-phosphate (PLP) concentrations as an indicator of vitamin B6 adequacy. SETTING: The study was performed at the Institute of Food Research, Norwich, UK. Blood samples were taken at the Institute, at Health Centres, or in the volunteer's home. SUBJECTS: 51 adolescent, 131 adult, 68 non-institutionalized elderly and 44 aged (>73 y) volunteers were recruited from local authority schools, local Health Centres and General Practitioners. RESULTS: The mean PLP recovery was 92.8%. The intra- and inter-assay coefficients of variation were 2.8% and 5.2% respectively. Mean PLP concentrations for males and females, respectively, were: adolescents (13-14 y), 36.4 and 43.5 nM; adults (20-64 y), 39.2 and 40.0 nM; elderly (68-73 y), 34.8 and 35.3 nM; aged (>73 y), 57.8 and 49.0 nM. Percentages of subjects with PLP concentrations <34.4 nM were over 26% in all population groups. Mean vitamin B6 intakes (microg/g protein intake), as assessed by weighed dietary records, were all above reference nutrient intakes (15 microg/g protein). CONCLUSIONS: An HPLC method, using cyanide derivitisation, has been applied to the determination of plasma PLP. Comparisons of results for local population groups with current cut-off values for plasma PLP, show large numbers of volunteers at risk of vitamin B6 deficiency although this is not reflected by vitamin B6 intakes calculated from food tables. The 34.4 nM cut-off value for value for plasma PLP, indicating deficiency, is questioned.

Dietary fat intake and plasma lipid levels in adolescents.

The relationship between dietary fat intake and fasting plasma lipid levels was assessed in 35 female and 19 male adolescents recruited from two local education authority schools in Norwich, UK. Dietary intakes were assessed using a 7-day weighed dietary record method, coupled with the collection of duplicate diets. Fat and energy intakes calculated using food composition tables were compared with values obtained by direct analysis of duplicate diets. Fasting plasma lipid levels (total, HDL and LDL cholesterol and triglycerides) were measured and compared with total dietary lipids and fatty acid intakes. The average proportion of energy consumed as fat was higher than is currently considered desirable but lower than previous studies have reported for adults. Mean serum total cholesterol values were 4.2 (SEM 0.1) mmol for females and 4.5 (SEM 0.2) mmol for males; this difference was not statistically significant. In male subjects the dietary fatty acid profiles were significantly correlated with several parameters of plasma lipid status which are thought to be risk factors for coronary heart disease, and in particular with the ratio of total:HDL cholesterol.

Increased fruit and vegetable consumption: potential health benefits.

In this European Union project, a Core human study was conducted in Ireland, Northern Ireland, Spain, France and The Netherlands. Oxidative and antioxidant status, vegetable and fruit consumption, and carotenoid intake of volunteers from different countries were compared. Response to increased carotenoid intake was determined. Attention was paid to whether the antioxidant capability of beta-carotene, lutein and lycopene demonstrated in vitro was apparent in relation to increased oxidation resistance of low-density-lipoprotein (LDL). Other (complementary) studies were undertaken and included determination of: protective effects of carotenoid-rich foods against LDL and DNA oxidative damage; carotenoid absorbability; barriers to increased vegetable consumption; and carotenoid content of fruits and vegetables frequently consumed in Europe. Our results demonstrated that carotenoid supplementation did not increase LDL oxidation resistance. However, increased consumption of carotenoid-rich fruits and vegetables did increase LDL oxidation resistance, and higher plasma concentration of total and specific carotenoids (pre-supplementation) was associated with lower DNA damage.

The effectiveness of various iron-supplementation regimens in improving the Fe status of anaemic rats.

Less frequent iron supplementation may be equally as beneficial to Fe-deficient subjects as routine daily supplementation because of the short-term suppressive effect of oral dosing with large amounts of Fe on subsequent Fe absorption. In the present study, the possibility that the administration of an Fe supplement every 2nd or 3rd day may be as effective in improving Fe status as a daily supplement was investigated in anaemic rats. Anaemic rats were given a 4 mg Fe supplement every day, on alternate days or every 3rd day, as a single dose with a midday meal or as a multiple dose with a morning, midday and evening meal. A low-Fe diet (13 mg/kg) was given at all other times. After 7 d, erythrocyte count, packed cell volume, mean cell volume, haemoglobin concentration and total liver Fe were measured and compared with those of meal-fed rats which had not been given any supplemental Fe. Rats which received a supplement every 3rd day, a total supplement of 12 mg, had a similar Fe status to those receiving a daily supplement, a total supplement of 28 mg. Administration of the supplement as a multiple, rather than as a single dose did not improve recovery from the Fe deficiency. It is suggested that less frequent supplementation with a smaller total amount of Fe, should be considered in human subjects. Such a regimen would minimize unpleasant side-effects of oral Fe therapy, decrease the risk of adverse effects of Fe on the absorption of other essential minerals and substantially cut the cost of supplementation programmes.

Studies of iron:zinc interactions in adult rats and the effect of iron fortification of two commercial infant weaning products on iron and zinc status of weanling rats.

The effect of iron on zinc absorption in the rat, and vice versa, was investigated from single starch:sucrose test meals (containing 65Zn or 59Fe) by whole body counting. Zinc had no effect on iron absorption, but iron reduced zinc absorption when the total ionic species in the meal (iron plus zinc) reached 1.36 mg. Below this level, high iron:zinc molar ratios (10:1) had no effect on zinc absorption, presumably because the transport mechanism for zinc had not reached full capacity. Previous iron intake had no effect on zinc absorption. The relevance of these findings to infant foods was explored by feeding rats exclusively a vegetable or cereal weaning product, with or without additional iron, for 12 d and measuring zinc and iron status. The added iron raised body iron stores and caused a small reduction in zinc status in animals fed the oat, but not the vegetable, diet as measured by plasma and femur zinc concentrations. Since the threshold level of 1.36 mg ionic species would be exceeded when the animal ate 3-4 g of the iron-fortified weaning food at any one time, it appears that the iron:zinc interactive effect was absent in the vegetable and less potent in the oat formulation than in a carbohydrate test meal. Alternatively, it may be the case that the animals had responded over time to reduced zinc availability by increasing whole body zinc retention.(ABSTRACT TRUNCATED AT 250 WORDS)

Nucleotide concentrations in cellular tissues during potassium and magnesium deficiency.

Potassium deficiency reduced the concentrations of ATP and GTP in rat thigh muscle but not in liver. The concentrations of UTP, CTP and the UDP-sugar derivatives with glucose, galactose and N-acetylglucosamine in liver were all increased. Magnesium deficiency, however, had little effect on the nucleotide concentrations in liver.

Changes in cellular and subcellular composition during potassium deficiency.

1. A specific dietary deficiency of potassium in young rats reduced the potassium concentration in thigh muscle by 48%, and in heart and kidney by 18%, but did not significantly affect the concentration in liver or brain. Conversely the sodium concentration rose in liver, heart and thigh muscle, and thigh muscle also accumulated increased amounts of magnesium. Apart from an increase in the water content of many tissues, no consistent changes in the composition of major cell constituents were observed. 2. The loss of potassium and accumulation of sodium and magnesium occurred predominantly in the supernatant fraction of the cell. The supernatant of all tissues studied contained about 80% of the total cellular potassium and sodium, and the potassium was present entirely in the ionic form. 3. Potassium and magnesium are the two most abundant intracellular metals, but their deficiencies have very different effects on the cell. The relationship between them is more complex than the inverse relationship between potassium and sodium.

Determination of total long-chain fatty acids in human plasma and lipoproteins, before and during copper-stimulated oxidation, by high-performance liquid chromatography.

An improved method for the high-performance liquid chromatographic determination (HPLC) of total or free long-chain (> C12) fatty acids in small volumes (10 microL) of human plasma and lipoprotein samples is described. The method is based on the formation of 2-nitrophenyl-hydrazine (2-NPH) derivatives and offers an alternative to gas chromatographic (GC) fatty acid determination. The retention of 2-NPH fatty acid derivatives on the HPLC system differs from the typical pattern produced by GC separation, thus offering a powerful tool for confirmation of peak identification where GC peak resolution is poor. Fatty acids determined include saturates [myristic acid, C14:0; palmitic acid, C16:0; stearic acid, C18:0; eicosanoic acid, C20:0; docosanoic acid, C22:0; and tetracosanoic acid, C24:0], monounsaturates [palmitoleic, C16:1; petroselenic, C18:1n12; oleic, C18:1n9; and erucic, C22:1], and polyunsaturates [linoleic, C18:2; linolenic, C18:3n3; gamma-linolenic acid, C18:3n6; eicosatrienoic, C20:3; arachidonic, C20:4; eicosapentanoic, C20:5; docosahexanoic, C20:6; and docosatetraenoic, C22:4]. Mean recoveries of fatty acids added to LDL samples were 94.1-109.4%, and intraassay coefficients of variation for the major fatty acids in human plasma were 2.7-6.9%. The potential of the method for further development is discussed. Long-chain fatty acid profiles are given for plasma and very low-, low, and high-density lipoprotein (before and during copper-stimulated oxidation) from human blood.

The effect of alcoholic beverages on iron and zinc metabolism in the rat.

1. Male Wistar rats (approximately 200 g) were given distilled water and a semi-synthetic control diet for 6 d. On day 7, 37 kBq 65Zn were administered intramuscularly and the rats were given distilled water, beer, cider, red wine, whisky or ethanol as their only source of fluid. The wine, whisky and ethanol were diluted so that each of the beverages contained a similar ethanol concentration (approximately 30 g/l). Food and fluid intake, growth rate and whole-body 65Zn were measured regularly over 11 d, after which animals were killed and blood haemoglobin (Hb) concentration, liver iron stores and the Zn concentration in testes determined. 2. There were no differences in body-weight gain or food intake between groups but fluid intake for the beer group was considerably higher than that for the other groups. 3. There was a significant effect of the type of alcoholic beverage consumed on whole-body 65Zn retention. Rats given whisky had a smaller daily loss of 65Zn than those given water, beer or cider. The ethanol group also showed a lower rate of 65Zn loss compared with the water group. The observed changes in whole-body 65Zn retention could be explained by an adverse influence of ethanol on Zn absorption from the diet. 4. Blood Hb and testes Zn concentration were similar in all groups but the type of liquid consumed influenced liver Fe levels. The cider group had the lowest liver Fe values and the ethanol group the highest values. 5. It is apparent from the present study that ethanol and alcoholic beverages affect Zn and Fe metabolism, but that the effects of ethanol are moderated by other components of the alcoholic beverages.

Fetal growth, glucose tolerance and plasma insulin concentration in rats given a marginal-zinc diet in the latter stages of pregnancy.

1. Wistar rats were fed on a control semi-synthetic diet throughout pregnancy, or a control diet in the first 2 weeks and a marginal-zinc diet in the 3rd week of pregnancy. On day 20, after an overnight fast, half the animals in each group were given glucose by gavage and the 0-30 min rise in blood glucose measured in tail blood. After 60 min blood was taken by cardiac puncture for glucose and insulin assay. Maternal pancreases were removed and the Zn contents measured. Fetuses from each litter were combined for wet/dry weights, protein and DNA determinations. 2. Plasma insulin concentration was higher, and glucose concentration and pancreatic Zn content lower, in pregnant v. non-pregnant animals of similar age, fed on the same diet. Pancreatic Zn content was lowest in the marginal-Zn group of pregnant rats. Fetuses from mothers fed on the marginal-Zn diet during the last week of pregnancy were slightly heavier than controls and had a significantly higher protein: DNA ratio. The 0-30 min rise in blood glucose was significantly greater in the marginal-Zn animals. 3. In a second experiment, pregnant rats were given similar diets to those used in the first study, but the marginal-Zn diet was given for a shorter period (days 15-19 of pregnancy). On day 19 the rats were meal-fed and on day 20, after an overnight fast, an oral glucose dose was administered. Tail-blood was taken at timed intervals up to 60 min post-dosing for glucose assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Hexose absorption from jejunal loops in situ in zinc-deficient and Zn-supplemented rats.

1. Immature, male Wistar rats were given a low-zinc semi-synthetic diet (2 mg Zn/kg) for 22-28 d. Control groups received a similar diet supplemented with 58 mg Zn/kg either ad lib., or in amounts matched to the consumption of the Zn-deficient group. There was a rapid onset of reduced food consumption and growth retardation in the Zn-depleted animals. 2. Serosal surface area of small intestines taken from Zn-deficient rats was significantly reduced compared with that of control animals. Villi, dissected from samples of proximal jejunum, were markedly smaller than those of control rats and were present in greater numbers per unit area of serosa. 3. Luminal loss of galactose from jejunal loops in situ was significantly greater in the Zn-deficient rats compared with controls when expressed in terms of unit dry weight of intestine and serosal or villous surface area. Since only a small proportion of the total galactose remained in the mucosal tissue and associated extracellular space, this loss could only be accounted for by an increased efficiency of net trans-epithelial transport. Differences in total galactose absorption per unit length of jejunum were not so marked. 4. This intestinal adaptation to Zn-deficiency allows the maintenance of normal, and possibly increased, rates of hexose transfer into the body of animals exhibiting severe growth retardation, reduced food utilization and abnormal glucose metabolism.

The effect of dietary protein source and guar gum on gastrointestinal growth and enteroglucagon secretion in the rat.

1. Male Wistar rats (approximately 100 g) were given fibre-free semi-synthetic diets containing either casein or albumin (168 g/kg diet) as the protein source with or without guar gum (75 g/kg diet) (casein, albumin, casein guar gum and albumin-guar gum groups). 2. Small intestinal length, weights of caecal tissue and contents and plasma enteroglucagon concentration were significantly increased in guar-gum-fed animals compared with the fibre-free groups. 3. Total caecal weight and plasma enteroglucagon concentration were higher in the albumin-guar gum group compared with the casein-guar gum group. The weights of caecal tissue and contents were significantly increased in rats given the fibre-free albumin diet compared with those consuming a similar diet with casein as the protein source, although daily food intake tended to be lower. 4. It is concluded that the effect of materials classed as dietary fibre may be significantly influenced by the non-polysaccharide component of the diet, and that such interactions may influence both the growth and endocrine activity of the gastrointestinal tract.

The effect of differences in dietary iron intake on 59Fe absorption and duodenal morphology in pregnant rats.

The effect of iron intake on 59Fe absorption throughout pregnancy, and on maternal and fetal Fe status towards the end of pregnancy, was investigated in rats. The influence of pregnancy and dietary Fe on duodenal morphology was also studied. Female rats were fed on a diet containing 17 or 100 mg Fe/kg for 2 weeks before and throughout pregnancy. 59Fe absorption was measured on days 1 or 2, 8 or 9 and 17 or 18 of pregnancy, and maternal and fetal Fe status was determined on days 18 or 19. Pregnancy resulted in a fall in haemoglobin (Hb) concentration. Compared with non-pregnant counterparts, total liver Fe was reduced in the low-Fe group, but not in the high-Fe group, indicating that the fall in Hb in the high-Fe rats was not associated with an Fe-deficient state. 59Fe absorption in rats fed on both diets increased throughout pregnancy, demonstrating that Fe supplementation of the diet, to a level that prevented the development of Fe-deficiency, failed to suppress an increase in absorption. Fetal weight appeared to be an important determinant of the efficiency of Fe absorption in late pregnancy. Poor maternal Fe status was accompanied by a reduction in fetal Fe concentration but results also suggested that fetuses were partly protected from maternal Fe-deficiency. Pregnancy resulted in increased duodenal circumference and villus dimensions, whilst high dietary Fe reduced duodenal growth in both pregnant and non-pregnant animals. The relevance of this finding is discussed. It was concluded that, in rats, pregnancy per se causes an enhancement in Fe absorption and that the degree of enhancement is in part related to fetal mass.

Measurement of non-haem iron absorption in non-anaemic rats using 59Fe: can the Fe content of duodenal mucosal cells cause lumen or mucosal radioisotope dilution, or both, thus resulting in the underestimation of Fe absorption?

Male Wistar rats (188 g) were fed on a semi-synthetic (SS) diet (38 mg iron/kg) ad lib. for 7 d and then meal-fed for 1 d. After a 21 h fast each rat was given one meal (10 g) of high-Fe SS (500 mg Fe/kg; high-Fe group) or control (38 mg Fe/kg; control group) diet. After 16 h 2 ml of an 59Fe-labelled ferrous sulphate solution (18 kBq 59Fe; 120 micrograms Fe) was administrated by gavage and equal numbers of rats from each group were killed 6 or 24 h after dosing. Mucosal uptake of 59Fe from the gut lumen and transfer of 59Fe from mucosa into the carcass were measured. Total Fe content of the duodenum was also determined. Mucosal 59Fe uptake and transfer were markedly lower in the high-Fe group compared with the control group. The Fe content of the duodenum, the major region of Fe absorption, was significantly greater in the high-Fe group than in the controls. A larger amount of Fe may thus have been released into the lumen of the high-Fe rats, via mucosal cell turnover, resulting in a greater lumen dilution of the 59Fe dose in this group compared with the controls. Calculations are presented which demonstrate that such an effect could not possibly account for the observed difference in mucosal 59Fe uptake between groups.(ABSTRACT TRUNCATED AT 250 WORDS)