Exocrine pancreatic function in hepatocyte nuclear factor 1ß-maturity-onset diabetes of the young (HNF1B-MODY) is only moderately reduced: compensatory hypersecretion from a hypoplastic pancreas.

OBJECTIVES: To examine the exocrine pancreatic function in carriers of the hepatocyte nuclear factor 1ß gene (HNF1B) mutation by direct testing. METHODS: Patients with HNF1B mutations and control subjects were assessed using rapid endoscopic secretin tests and secretin-stimulated magnetic resonance imaging. Seven patients and 25 controls underwent endoscopy, while eight patients and 20 controls had magnetic resonance imaging. Ductal function was assessed according to peak bicarbonate concentrations and acinar function was assessed according to peak digestive enzyme activities in secretin-stimulated duodenal juice. The association of pancreatic exocrine function and diabetes status with pancreatic gland volume was examined. RESULTS: The mean increase in secretin-stimulated duodenal fluid was smaller in patients than controls (4.0 vs 6.4 ml/min; P = 0.003). We found lower ductal function in patients than controls (median peak bicarbonate concentration: 73 vs 116 mEq/L; P < 0.001) and lower acinar function (median peak lipase activity: 6.4 vs 33.5 kU/ml; P = 0.01; median peak elastase activity: 0.056 vs 0.130 U/ml; P = 0.01). Pancreatic fluid volume outputs correlated significantly with pancreatic gland volumes (r² = 0.71, P = 0.008) in patients. The total fluid output to pancreatic gland volume ratios were higher in patients than controls (4.5 vs 1.3 ml/cm³; P = 0.03), suggesting compensatory hypersecretion in the remaining gland. CONCLUSION: Carriers of the HNF1B mutation have lower exocrine pancreatic function involving both ductal and acinar cells. Compensatory hypersecretion suggests that the small pancreas of HNF1B mutation carriers is attributable to hypoplasia, not atrophy.

Effect of vitamin D supplementation and ultraviolet B exposure on serum 25-hydroxyvitamin D concentrations in healthy volunteers: a randomized, crossover clinical trial.

BACKGROUND: Solar ultraviolet (UV) radiation during the summer and vitamin D supplementation are two major sources of vitamin D for humans at northern latitudes. However, little is known about the relative efficiency of these two vitamin D sources. OBJECTIVES: The main goal was to compare the efficiency of high-dose oral vitamin D3 supplementation (2000 IU per day for 30 days) with a simulated summer UV exposure [10 sunbed sessions to a total dose of 23·8 standard erythema doses (SED)] to improve vitamin D status. METHODS: Healthy volunteers were randomized into two groups: group 1 received vitamin D supplementation followed by 10 whole-body sunbed exposures; group 2 started with 10 sunbed exposures followed by vitamin D supplementation. RESULTS: The oral supplementation with vitamin D3 resulted in a mean (SEM) serum 25-hydroxyvitamin D [25(OH)D] increase of 25·3 (5·4) nmol L(-1) . A similar increase, 19·8 (5·4) nmol L(-1) , was observed after simulated summer UV exposure. At the end of the study, serum 25(OH)D concentrations were similar in both groups. CONCLUSIONS: Twice-weekly whole-body sunbed exposure to a dose of 4·8 SED is equal to 2000 IU daily of oral vitamin D supplementation for 30 days and enough to achieve and maintain serum 25(OH)D concentrations > 75 nmol L(-1) in ~55% of cases. Based on our calculations, this dose corresponds to a cumulative weekly whole-body exposure of 3·4 SED (~ 40 min around midday during the summer at the latitude of Oslo).

Plasma 25 hydroxyvitamin D level and blood gene expression profiles: a cross-sectional study of the Norwegian Women and Cancer Post-genome Cohort.

BACKGROUND/OBJECTIVES: Vitamin D deficiency has been associated with increased risk of developing several diseases, but much is unknown about the molecular effects involved. Gene expression technology is increasingly being used to elucidate molecular mechanisms related to nutritional factors, and in this study of free-living, middle-aged Norwegian women, we aimed at identifying gene expression pathways in the blood associated with vitamin D status. SUBJECTS/METHODS: Blood samples and questionnaires were collected as a part of the Norwegian Women and Cancer Post-genome Cohort (500 invited subjects, 218 included). Plasma 25 hydroxyvitamin D (25(OH)D) concentrations were measured using high-performance liquid chromatography, and we compared groups with sufficient versus deficient vitamin D status (25(OH)D >50 nmol/l (n=66) versus <37.5 nmol/l (n=83)), to identify differences in gene expression profiles obtained using full-genome microarrays. RESULTS: In a targeted pathway-level analysis, several immunological processes, immune cell functions and major signaling pathways were differentially regulated according to vitamin D status (P<0.01). To a certain degree, results from in vitro studies reported in the literature were reflected in this population setting. CONCLUSIONS: We conclude that vitamin D status measured as 25(OH)D was associated with molecular pathways that may ultimately affect the potential onset of diseases. The use of gene expression analysis in a population setting may give valuable input to the study of effects of nutritional factors.

Two cases with fatal outcome following total lung irradiation for metastatic bone sarcoma.

We report a single institution experience with total lung irradiation in 53 metastatic bone sarcoma patients in the context of two young female patients who died from treatment-induced pulmonary toxicity. A radiation dose of 19.5 Gy in 1.5 Gy daily fractions was given as two opposing fields with a conventional technique. Both patients succumbed within 3 months following radiotherapy. One patient had osteosarcoma whereas the other advanced Ewing's sarcoma; both with widespread metastases to the lungs at primary diagnosis. In retrospect, most likely high dose methotrexate lung toxicity observed in the osteosarcoma patient, and the GI-toxicity following pelvic radiotherapy in Ewing's case, both observed during the initial phase of their multimodal treatment, might indicate an increased individual radiosensitivity. In view of this, a review of our experience in 53 bone sarcoma patients (19 with Ewing's sarcoma and 34 with osteosarcoma) treated at our institution was conducted. We have not previously experienced significant toxicity following total lung irradiation. Among these, 42% (8/19) with Ewing's sarcoma and 9% (3/34) with osteosarcoma are long-term survivors and without clinically significant lung toxicity.

Faecal calprotectin in HIV-infected, HAART-naïve Ugandan children.

OBJECTIVES: Calprotectin is a calcium- and zinc-binding protein and a marker in faeces of gastrointestinal inflammation. Reference values have been established in children older than 4 years. The aim of the present study was to determine the concentration of faecal calprotectin (FC) in human immunodeficiency virus (HIV)-infected, highly active antiretroviral therapy-naïve Ugandan children and compare it with the reference value. METHODS: We tested 193 HIV-infected children ages 0 to 12 years in a hospital-based survey for FC. A standardised interview with sociodemographic information and medical history was used to assess risk factors. A cluster of differentiation 4 (CD4) cell percentage was prevalent in all of the children. RESULTS: The median FC concentrations decreased with increasing age, as in healthy children. The median concentration was 208 mg/kg in infants 0 to 1 year, 171 mg/kg among toddlers 1 to 4 years, and 62 mg/kg for children 4 to 12 years. Children with advanced disease and a low CD4 cell percentage had significantly higher FC concentrations than those with a high CD4 cell percentage. Children older than 4 years with diarrhoea had significantly higher FC concentrations compared with those without diarrhoea. CONCLUSIONS: HIV-infected children older than 4 years had a median FC concentration above the reference value, and gut inflammation in the children with elevated values is likely. Children with more advanced disease had increased FC concentrations regardless of age.

Health status at long-term follow-up in patients treated for extremity localized Ewing Sarcoma or osteosarcoma: a Scandinavian sarcoma group study.

BACKGROUND: The purpose of this study was to evaluate late effects and symptom complaints in long-term survivors (>5 years) of Extremity Bone Sarcoma (EBS survivors). The results were compared with findings in age- and gender-matched individuals from the general population (NORMs). PATIENTS AND METHODS: Among 155 EBS survivors approached, 133 (86%) were included, and 110 of them (83%) attended an outpatient examination. Health status was evaluated by a mailed questionnaire concerning demographic and current health issues, and physical examinations at the outpatient clinic. Age- and gender-adjusted normative controls were drawn from participants of the Health Study of Nord-Trøndelag County (HUNT 2). RESULTS: Median age at follow-up was 29 (15-57) years. Median follow-up was 12 (6-22) years. Of EBS survivors 42% had > or =1 somatic disease, 33% had ototoxicity and 13% had reduced renal function. EBS survivors were more likely to have heart disease (odds ratio [OR], 7.9; 95% confidence interval [95% CI], 2.5-25.3; P = 0.001), hypertension (OR, 3.4; 95% CI, 1.1-10.1; P = 0.03) and thyroid disease (OR, 3.0; 95% CI, 1.1-8.3; P = 0.04) compared to NORMs. EBS survivors reported more diarrhoea (29% vs. 19%, P = 0.02), palpitations (23% vs. 13%, P = 0.01) and shortness of breath (11% vs. 5%, P = 0.01) than NORMs. CONCLUSIONS: EBS survivors have poorer health status compared to age- and gender-matched controls. Long-term follow-up of these patients is therefore mandatory.

Limb-sparing surgery preserves more function than amputation: a Scandinavian sarcoma group study of 118 patients.

We evaluated the long-term functional outcome in 118 patients treated for osteosarcoma or Ewing's sarcoma in the extremities a minimum of five years after treatment. We also examined if impaired function influenced their quality of life and ability to work. The function was evaluated according to the Musculoskeletal Tumor Society (MSTS) score and the Toronto Extremity Salvage Score (TESS). Quality of life was assessed by using the Short Form-36 (SF-36). The mean age at follow-up was 31 years (15 to 57) and the mean follow-up was for 13 years (6 to 22). A total of 67 patients (57%) initially had limb-sparing surgery, but four had a secondary amputation. The median MSTS score was 70% (17% to 100%) and the median TESS was 89% (43% to 100%). The amputees had a significantly lower MSTS score than those with limb-sparing surgery (p < 0.001), but there was no difference for the TESS. Tumour localisation above knee level resulted in significantly lower MSTS scores and TESS (p = 0.003 and p = 0.02, respectively). There were no significant differences in quality of life between amputees and those with limb-sparing surgery except in physical functioning. Of the patients 11% (13) did not work or study. In multivariate analysis, amputation, tumour location above the knee and having muscular pain were associated with low physical function. We conclude that most of the bone tumour survivors managed well after adjustment to their physical limitations. A total of 105 are able to work and have an overall good quality of life.

The vitamin D receptor and its ligand 1alpha,25-dihydroxyvitamin D3 in Atlantic salmon (Salmo salar).

Seaward migration of Salmo salar is preceded by preparatory physiological adaptations (parr-smolt transformation) to allow for a switch from freshwater (FW) to seawater (SW), which also means a switch in ambient calcium from hypocalcic (<1 mM Ca(2+)) to the plasma (~1.25 mM Ca(2+)) and to strongly hypercalcic (8-12 mM Ca(2+)). Uptake, storage (skeleton, scales) and excretion of calcium need careful regulation. In fish, the vitamin D endocrine system plays a rather enigmatic role in calcium physiology. Here, we give direct evidence for calcitriol involvement in SW migration. We report the full sequence of the nuclear vitamin D receptor (sVDR0) and two alternatively spliced variants resulting from intron retention (sVDR1 and sVDR2). In FW parr, SW adapting smolts, and in SW adults, plasma concentrations of 25(OH)D(3) and 24,25(OH)(2)D(3) did not change significantly. Plasma calcitriol concentrations were lowest in FW parr, doubled during smoltification and remained elevated in SW adults. Increased calcitriol coincided with a twofold decrease in sVDR mRNA levels in gill, intestine, and kidney of FW smolts and SW adults, when compared with parr. Clearly, there was a negative feedback and dynamic response of the vitamin D endocrine system during parr-smolt transformation. The onset of these dynamic changes in FW parr warrants a further search for the endocrines that initiate these changes. We speculate that the vitamin D system plays a crucial role in calcium and phosphorus handling in Atlantic salmon.

Increased levels of urinary leukotriene E4 in children with severe atopic eczema/dermatitis syndrome.

BACKGROUND: Leukotrienes are thought to play a role in the pathogenesis of atopic eczema/dermatitis syndrome (AEDS). Urinary leukotriene E4 (U-LTE4) is a marker of whole-body cysteinyl-leukotriene production. AIMS OF THE STUDY: To evaluate the role of leukotrienes in children with AEDS by measuring levels of U-LTE4, and to evaluate whether levels of U-LTE4 may reflect disease activity and allergic sensitization in AEDS. METHODS: U-LTE4 was measured by enzyme-linked immunosorbent assay in 87 children with mild (n=32), moderate (n=34) and severe (n=21) AEDS, as well as in 72 nonatopic healthy controls. Fifty-eight of the children with AEDS were sensitized to common allergens, and 29 were not. RESULTS: Levels of U-LTE4 were higher in children with severe AEDS (140; 66-166 microg/mmol creatinine, median; quartiles) than in controls (52; 30-90, P <0.05), whereas levels of U-LTE4 in moderate and mild disease were similar to controls. U-LTE4 levels were similar in children with or without sensitization to common allergens, but severe AEDS children with sensitization had higher levels of U-LTE4 than those without sensitization. CONCLUSION: The results suggest a role for leukotrienes in the pathogenesis of severe AEDS, and may support a role for leukotriene-antagonists in the treatment of this disorder. Levels of U-LTE4 may reflect the disease severity and sensitization to allergens in AEDS.

Vitamin D status in a rural population of northern Norway with high fish liver consumption.

OBJECTIVE: To assess vitamin D status and the impact of three fish meals consisting of cod liver and fresh cod-liver oil on the plasma level of vitamin D metabolites in an area with high consumption of cod liver and cod-liver oil. DESIGN: Experimental field study. METHODS: Thirty-two volunteers from the Skjervøy (70 degrees N) municipality in northern Norway were recruited to consume three traditional mølje meals, consisting of cod, cod liver, fresh cod-liver oil and hard roe, in one week. The liver and fresh cod-liver oil consumed by the participants were weighed and recorded. Blood samples were collected before the first meal, and subsequently 12 h and 4 days after the last meal. The blood samples were analysed for the vitamin D metabolites 25-hydroxyvitamin D (25(OH)D) and 1,25-dihydroxyvitamin D (1,25(OH)2D). All participants answered a semi-quantitative food-frequency questionnaire, which was used to estimate usual daily nutrient intake. The study was carried out in the last part of March 2001. RESULTS: The median daily vitamin D intake estimated from the questionnaire was 9.9 microg. The proportion of subjects with baseline 25(OH)D level below 50 nmol l(-1) was 15.4% and none were below 37.5 nmol l(-1). Only "mølje consumption" and "time spent in daylight" were significantly associated with baseline log 25(OH)D. The mean total intake of vitamin D in the three servings was 272 microg (standard deviation 94 microg), ranging from 142 to 434 microg. Relative to baseline plasma concentration, the mean level of 25(OH)D decreased slightly in both post-consumption samples (P< or =0.03), while 1,25(OH)2D peaked 12 h after the final meal (P=0.03). CONCLUSION: Three mølje meals provided, on average, an amount of vitamin D equal to 54 times the recommended daily dose. Subjects with food consumption habits that included frequent mølje meals during the winter sustained satisfactory vitamin D levels in their blood, in spite of the long "vitamin D winter" (i.e. absence of ultraviolet-induced vitamin D production in the skin).

Vitamin D status of middle-aged women at 65-71 degrees N in relation to dietary intake and exposure to ultraviolet radiation.

OBJECTIVE: To determine the vitamin D status of middle-aged women living in the Norwegian arctic and its relationship with vitamin D intake and exposure to ultraviolet (UV) radiation. DESIGN: Cross-sectional study. SUBJECTS AND SETTING: This study is based on measurements of 25-hydroxyvitamin D (25(OH)D) levels in a sub-sample of the Norwegian component of the EPIC biological bank, which consists of blood samples from a random selection of participants in the Norwegian Women and Cancer Study. From November 2001 until June 2002, 309 blood samples were collected from a total of 443 invited middle-aged women (44-59 years) in northern Norway (65-71 degrees N) (crude response rate, 69.8%). Questionnaire data provided information on dietary sources of vitamin D and UV exposure. RESULTS: Median plasma 25(OH)D concentration for the whole group was 55.0 nmol l(-1) (range 8.1-142.8 nmol l(-1)). Vitamin D intake was a significant predictor of 25(OH)D status (P=0.0003). The time of the year when the blood sample was collected significantly predicted plasma 25(OH)D level (P=0.005). Levels of 25(OH)D were positively associated (P=0.0002) with estimated hours per day of exposure to UV-B radiation. Residing in northern Norway during the summer prior to blood sampling was negatively associated with 25(OH)D concentration (P=0.001). The prevalence of moderate hypovitaminosis D was highest in January-February, when a quarter of the participants had 25(OH)D concentrations < or =37.5 nmol l(-1). CONCLUSIONS: Increased ingestion of marine food items that provide vitamin D should be promoted and further studies should be carried out to investigate vitamin D status in arctic populations in relation to both UV exposure and traditional food sources.

High frequency of coeliac disease among patients with autoimmune adrenocortical failure.

BACKGROUND: Coeliac disease (CD) is an autoimmune disease of the small intestine caused by gluten ingestion in genetically predisposed subjects. It can occur isolated or in combination with other autoimmune diseases. Autoimmune Addison's disease is frequently associated with other organ-specific autoimmune diseases. We have investigated the prevalence of CD among a large cohort of patients with autoimmune Addison's disease. METHODS: Seventy-six patients (44 women) with Addison's disease, 52% of whom had polyendocrine failure, were recruited from a registry of organ-specific autoimmune diseases in Norway. All sera were analysed for antibodies against gliadin (AGA), endomysium (EMA) and tissue transglutaminase (tTG). Patients with positive EMA and/or anti-tTG were offered endoscopy. The human leucocyte antigen (HLA) class II genotypes were determined. RESULTS: Five patients had antibodies against both endomysium and tissue transglutaminase. In these five patients, CD was verified by biopsy. One patient had known CD prior to the study. All six patients with CD carried the CD-associated HLA haplotype DR3-DQ2. The total prevalence of CD was 7.9%. CONCLUSION: CD is frequently associated with Addison's disease. The risk of developing CD seems to be higher than can be explained by the common DR3-DQ2 association alone. It is often asymptomatic or associated with unspecific symptoms. Addison patients should be screened for the presence of CD on a regular basis.

Faecal calprotectin levels in infants with infantile colic, healthy infants, children with inflammatory bowel disease, children with recurrent abdominal pain and healthy children.

UNLABELLED: This study investigated faecal calprotectin concentration, a measure of intestinal inflammation, in infants and children with abdominal pain. Faecal calprotectin was measured by an enzyme-linked immunosorbent assay kit in spot stool samples in 76 infants with typical infantile colic, 7 infants with transient lactose intolerance and 27 healthy infants. All infants were 2-10 wk of age. In addition, 19 children with recurrent abdominal pain (RAP; mean age 11.5 y), 17 with inflammatory bowel disease (IBD; mean age 11.1 y; 10 had Crohn's disease and 7 ulcerative colitis) and 24 healthy children (mean age 5.3 y) were studied. In infants with infantile colic the mean faecal calprotectin concentration was not different from that in healthy infants (278 +/- 105 vs 277 +/- 109 mg kg(-1), p = 0.97) or in infants with transient lactose intolerance (300.3 +/- 124 mg kg(-1), p = 0.60). The calprotectin level was similar in boys and girls and fell significantly with age (p = 0.04). Children with IBD had faecal calprotectin levels (293 +/- 218 mg kg(-1)) much higher than healthy children (40 +/- 28 mg kg(-1), p < 0.0001) and children with RAP without identified organic disease (18 +/- 24 mg kg(-1), p < 0.0001). CONCLUSION: Faecal calprotectin may differentiate between functional abdominal pain and IBD in school-aged children. In young infants high faecal calprotectin levels are normal.

Antagonistic effects of 24R,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3 on L-type Ca2+ channels and Na+/Ca2+ exchange in enterocytes from Atlantic cod (Gadus morhua).

There is mounting evidence that vitamin D and its metabolites play important roles in regulating plasma calcium concentrations in teleost fish as in other vertebrates. The aims of the present study were to elucidate the possible cellular target mechanisms for the rapid actions of 24R,25(OH)(2)D(3), 25(OH)D(3) and 1,25(OH)(2)D(3) in Atlantic cod enterocytes at physiological doses, and to establish the concentration and thus the physiological range of circulating 24R,25(OH)(2)D(3), 25(OH)D(3) and 1,25(OH)(2)D(3) in the Atlantic cod. The plasma concentrations of 25(OH)D(3), 1,25(OH)(2)D(3) and 24R,25(OH)(2)D(3) were 15.3 +/- 2.7nM, 125.1 +/- 12.3pM and 10.1 +/- 23.5nM respectively. Exposure of enterocytes to 10mM calcium (Ca(2+)) evoked an increase in intracellular Ca(2+) concentrations ([Ca(2+)](i)). This increase was suppressed by 24R,25(OH)(2)D(3) dose-dependently, with an EC(50) of 4.9nM and a maximal inhibition of 60%. 24R,25(OH)(2)D(3) (20nM) abolished an increase in [Ca(2+)](i) (approximately 252%) in the control enterocytes exposed to 10microM S(-)-BAYK-8644, suggesting that the hormone acts by inhibiting Ca(2+) entry through L-type voltage-gated Ca(2+) channels. Administration of 20nM 24R,25(OH)(2)D(3) to enterocytes in the absence of extracellular Ca(2+) increased [Ca(2+)](i) by approximately 20%, indicating a release of Ca(2+) from intracellular stores. Administration of 25(OH)D(3) (20nM) resulted in a biphasic change in the enterocyte [Ca(2+)](i): within 1--5s, it decreased to 87 +/- 12nM below its mean basal [Ca(2+)](i) (334 +/- 13nM), followed by a rapid recovery of [Ca(2+)](i) to a new level, 10% lower than the initial [Ca(2+)](i). The rapid decrease, the recovery rate and the final [Ca(2+)](i) were all affected dose-dependently by 25(OH)D(3), with EC(50) values of 8.5, 17.0 and 18.9nM respectively. Furthermore, the effects of 25(OH)D(3) were sensitive to sodium (Na(+)), bepridil (10microM) and nifedipine (5 microM), suggesting that 25(OH)D(3) regulates the activity of both basolateral membrane-associated Na(+)/Ca(2+) exchangers and brush border membrane-associated L-type Ca(2+) channels. Administration of 25(OH)D(3) (10nM) to enterocytes in the absence of extracellular Ca(2+) increased [Ca(2+)](i) by approximately 18%, indicating a release of Ca(2+) from intracellular stores. 1,25(OH)(2)D(3) also affected enterocyte [Ca(2+)](i) in a biphasic manner: the rapid decrease, the recovery rate, and the mean final [Ca(2+)](i) were all affected dose-dependently, with EC(50) values of 8.3, 24.5 and 7.7nM respectively. The high EC(50) values for 1,25(OH)(2)D(3) compared with circulating concentrations of 1,25(OH)(2)D(3) (130pM) suggest that this effect is pharmacological, rather than of physiological relevance in enterocyte Ca(2+) homeostasis of the Atlantic cod. It is concluded that 24R,25(OH)(2)D(3) has a physiological role in decreasing intestinal Ca(2+) uptake via inactivation of L-type Ca(2+) channels, whereas the physiological role of 25(OH)D(3) is to increase enterocyte Ca(2+) transport via activation of Na(+)/Ca(2+) exchangers, concurrent with activation of L-type Ca(2+) channels.

High prevalence of asymptomatic coeliac disease in Norway: a study of blood donors.

OBJECTIVE: The prevalence of symptomatic coeliac disease in Norway is 1:675. Coeliac disease has previously been reported in presumably healthy people. Our aim was to determine the prevalence of latent coeliac disease in apparently healthy (i.e. asymptomatic) Norwegian individuals. METHODS: Blood donor sera were tested for gluten antibodies (IgA, IgG). Positive samples (IgA AGA > 0.35, IgG AGA > 0.90) were further tested for endomysium antibodies (IgA EMA). EMA positive individuals were offered gastroenterological investigation. RESULTS: Of 2096 sera, 83 fulfilled the criteria for EMA testing (M/F = 55/28). Eight individuals were EMA positive. On biopsy, seven out of eight had villous atrophy (six subtotal, one partial). None of the patients had significant symptoms. Biochemical data showed iron deficiency (two), hypocalcaemia (one), and low serum zinc (five). All patients were treated with a gluten-free diet and followed up. CONCLUSION: The study indicates a prevalence of 1:340 among asymptomatic and presumably healthy people. This is in keeping with studies from other countries. Lack of symptoms does not exclude secondary deficiency conditions.

Fat-soluble vitamin levels in familial hypercholesterolemia.

OBJECTIVE: To examine serum levels of retinol, 25-hydroxyvitamin D, and alpha-tocopherol and their potential determinants in familial hypercholesterolemia (FH). SUBJECTS: Study 1: 151 boys and girls with FH aged 7 to 16 years who were following a lipid-reduced diet but not taking lipid-lowering drugs. Study 2: 87 boys and girls with FH, of whom 24 were taking bile acid-binding resins in addition to the diet, and 30 age- and sex-matched control subjects. DESIGN: Cross-sectional survey. SETTING: Lipid referral clinic. RESULTS: None of the subjects had suboptimal retinol or 25-hydroxyvitamin D levels. Only one girl had a low alpha-tocopherol level and alpha-tocopherol/lipid ratio. In multiple regression analysis, pubertal onset and gender were associated with retinol and 25-hydroxyvitamin D levels. The triglyceride level was positively related to level of retinol, and body mass index was inversely related to 25-hydroxyvitamin D level. Vitamin supplementation was positively related to 25-hydroxyvitamin D level and the alpha-tocopherol/lipid ratio. This ratio was lower in subjects whose total cholesterol level was above the median (8.0 mmol/L (310 mg/dl)) than in subjects whose cholesterol level was below the median (p = 0.01). In study 2, the alpha-tocopherol/lipid ratio in control subjects (median, 5.1 mumol/mmol) was higher than in subjects with FH who were not taking resins (median, 3.3 mumol/mmol; p < 0.05) but similar to the ratio in treated subjects (median, 5.4 mumol/mmol). CONCLUSIONS: Pubertal onset, gender, lipid levels, vitamin supplementation, and body mass index are significant predictors of fat-soluble vitamin levels in children with FH. Though children following a lipid-lowering diet have normal serum levels of fat-soluble vitamins, the alpha-tocopherol level does not appear to increase proportionately to the increase in cholesterol level. Treatment with resins may restore a normal alpha-tocopherol/lipid ratio.

Low dose colestipol in adolescents with familial hypercholesterolaemia.

The effects of orange flavoured colestipol granules, 10 g/day, in 37 boys and 29 girls aged 10-16 years with familial hypercholesterolaemia were examined first in an eight week double blind, placebo controlled protocol, then in open treatment for 44-52 weeks. All patients were on a low fat diet. Low density lipoprotein cholesterol levels were reduced by 19.5% by colestipol v 1.0% by placebo. Levels of serum folate, vitamin E, and carotenoids were reduced in the colestipol group, but not the vitamin E/cholesterol and carotenoid/cholesterol ratios or serum concentrations of vitamins A and D. After one year of colestipol, two thirds of the participants remained in the study, of whom half took > or = 80% of the prescribed dose. Those who took > or = 80% of the dose had a greater decrease in serum 25-hydroxyvitamin D levels than those who took < 80%. No adverse effects on weight gain or linear growth velocity were observed. Although low dose colestipol effectively reduces low density lipoprotein cholesterol levels, only a minority of adolescents adhered to the new formulation for one year. Folate and possibly vitamin D supplementation is recommended.

Simultaneous determination of retinol, alpha-tocopherol, and 25-hydroxyvitamin D in human serum by high-performance liquid chromatography.

Determination of the serum or plasma levels of retinol (vitamin A), 25-hydroxyvitamin D, and alpha-tocopherol (vitamin E) are the most frequently used parameters to evaluate status of vitamin A, D, and E, and also to assess the gastrointestinal absorption of the vitamins. We present a simple and sensitive method for simultaneous determination of these vitamins in 0.5 ml human serum or plasma. The vitamins were extracted from serum by methanol/iso-propanol (80/20, v/v) and n-hexane. The n-hexane phase was evaporated and injected to a reversed-phase (C-18) high-performance liquid chromatography system. Elution was performed with methanol/water (85:15, v/v) for 25-hydroxyvitamin D and retinol, and after that by methanol/water (98:2, v/v) for alpha-tocopherol. The eluate was monitored by a UV detector at 265 nm for detection of the vitamins. Baseline separation was obtained for all vitamins, and the system also permitted separate determinations of the D2 and D3 forms of 25-hydroxyvitamin D. The limit of detection and interassay variation for determination in 0.5 ml serum were 6.0 nmol/L and 6.2% for 25-hydroxyvitamin D2, 6 nmol/l and 6.1% for 25-hydroxyvitamin D3, 0.035 mumol/L and 5.0% for retinol, and 1.2 mumol/l and 5.5% for alpha-tocopherol.

Effects of 1,25-dihydroxyvitamin D3 and retinoic acid on the proliferation and cell cycle phase distribution of neuroblastoma SK-N-SH cells.

The hormone 1,25-(OH)2D3 has been shown to modulate cell proliferation and induce differentiation in several normal and malignant cell lines. In this work, we examined the effect of the hormone on the neuroblastoma SK-N-SH cell line. The steroid did not influence cell growth and cell cycle distribution, while retinoic acid inhibited proliferation and induced an accumulation of the cells in the G0/G1 phase of the cell cycle. 1,25-(OH)2D3 did not alter cell morphology. The activities of the 1-alpha- and 24-hydroxylases were low and not regulated by the hormone. The level of the total 1,25-(OH)2D3 receptor was low. We conclude that the lack of effect of 1,25-(OH)2D3 on the SK-N-SH cell line is related to the low level of the 1,25-(OH)2D3 receptor.

In vitro toxicity of purified gluten peptides tested by organ culture.

Various subfractions of Frazer fraction III were separated by high-pressure liquid chromatography, and their toxicity in vitro (organ culture) was tested in comparison with alpha-gliadin using duodenal biopsies from 25 patients with active celiac disease and subtotal villous atrophy, 2 patients with partial villous atrophy, and 10 nonceliac controls. One dominating fraction, designated Frazer III-2-VIII, was purified by several steps of rechromatography. It was markedly toxic to duodenal explants from patients with active celiac disease. The mean enterocyte height after culture was 15.9 microns compared with 25.6 microns in gluten-free medium. This difference was statistically significant in all cases except one, in which the lowest concentration (110 micrograms) was used. The in vitro toxicity of Frazer III-2-VIII was comparable with the toxicity of alpha-gliadin in twofold to fivefold higher concentrations. No toxicity could be detected in nonceliac explants (mean enterocyte height, 25.7 vs. 24.9 microns in gluten-free medium). The N-terminal amino acid sequence was (Gln)-Ile-Gln-Val-Phe-Pro-Ser-Gly-Gln-Val-Gln-(Trp)-Pro-Gln-Gln-(Gln)-Gl n-Pro- Phe-Pro. This sequence was not homologous to previously reported sequences of toxic gluten peptides. By use of the SwissProt and GenEMBL databases, it was concluded that the peptide Frazer III-2-VIII is part of the gamma-gliadin fraction.