Bone mineral metabolism after total gastrectomy.

Gastric surgery is mostly needed for treatment of gastric malignancy. To investigate the effect of total gastrectomy on bone mineral density (BMD) and bone mineral metabolism we evaluated 18 patients after total gastrectomy. Mean interval since operation was 71 +/- 20 months. BMD results were compared with age- and gender-matched controls (n = 46) and also expressed as T and Z scores. Bone mineral density measured by dual-energy X-ray absorptiometry (DXA) was found to be significantly lower in patients after total gastrectomy compared with healthy controls in the lumbar spine (p = 0.017 for women, p = 0.002 for men), femoral neck (p = 0.004 for women, p = 0.001 for men), Ward's triangle (p = 0.031 for women, p = 0.003 for men), and greater trochanter (p = 0.001 for women, p = 0.001 for men). Z scores for lumbar spine, femoral neck, Ward's triangle, and greater trochanter were -0.83, -1.54, -1.02, and -1.19, respectively. Biochemical measurements correlated poorly with BMD and were found to be of lesser value in diagnosing reduced bone mass as well as in differential diagnosis of etiology of osteopenia. The results of our study show the deleterious effect of total gastrectomy on bone mineral status and suggest an increased fracture risk in these patients.

Does apolipoprotein E genotype relate to BMD and bone markers in postmenopausal women?

OBJECTIVES: Bone mineral density (BMD) and development of osteoporosis are partly determined by genetic factors. The associations between one of suggested candidate, apolipoprotein E (apo E) genotype to bone mineral density (BMD) and bone biochemical markers was studied in 464 subjects recruited from a population-based group of early postmenopausal women (n = 13100). Additionally, the influence of apo E genotype on BMD changes during a 5-year follow-up with or without hormone replacement therapy (HRT) was investigated. METHODS: Participants were randomized into two treatment groups: HRT group: Sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate with or without vitamin D3, 100-300 IU/day + calcium lactate, 500 mg/day (n = 232), and the non-HRT group: Calcium lactate, 500 mg/day alone or in combination with vitamin D3, 100-300 IU/day (n = 232). BMD was measured from the lumbar spine and proximal femur at baseline and after 5 years of treatment (n = 352). In a subgroup (n = 59), the serum concentrations of bone biochemical markers (intact osteocalcin (OC), bone-specific alkaline phosphatase (BAP) and type I collagen carboxy-terminal telopeptide (ICTP)) were measured at baseline and after 1 year of follow-up. RESULTS: At baseline, the BMDs were similar between the five apo E genotype groups (2/3, 2/4, 3/3, 3/4, 4/4). No significant differences in lumbar or femoral neck BMDs of women with the apo E4 allele were found compared with those without it. There was a statistically significant difference in 5-year BMD changes between the HRT and non-HRT groups. After 5 years, the BMD of the femoral neck had remained constant and the mean lumbar spine BMD had increased by 1.5% in the HRT group, whereas both BMDs had decreased by 4-5% in the non-HRT group. However, the apo E genotype did not modify the changes in BMD in either group. Additionally, the baseline concentrations of bone metabolic markers and their 1-year changes showed no genotype-related associations. CONCLUSIONS: The results of our population-based study indicate that apo E genotype does not modify lumbar or femoral neck BMDs or serum bone biochemical markers or their response to HRT in early postmenopausal Caucasian women.

Urinary bone resorption markers in monitoring treatment of symptomatic osteoporosis.

We have studied the clinical usefulness of urinary bone resorption markers in postmenopausal women with symptomatic osteoporosis. The study design is a randomised double-blind placebo controlled study, in which the subjects were daily treated for 24 months either with a hormone analogue (2.5 mg Livial, generic name Tibolone, Organon, Amsterdam, Holland) plus 800 mg calcium (n = 14, age 63+/-5 years, range 52-68 years), or with placebo plus 800 mg calcium (n = 19, age 66+/-7 years, range 50-75 years). The laboratory methods for urinary bone resorption markers were enzyme immunoassays (EIA) for urinary pyridoline (PYD) and deoxypyridoline crosslinks (DPD), and for cross-linked N-telopeptides of Type I Collagen (NTx), and an HPLC assay for urinary hydroxyproline (HOP). All the urine assay results were calculated per mmol creatinine. All the resorption markers decreased during the two-year study period in both groups. The Z scores (discriminating power, i.e. ability of the different tests to distinguish the hormone treated subjects from the placebo treated subjects) for HOP and PYD were rather low: 0.06-1.52 for HOP and 0.68-1.47 for PYD. The differences between the two treatment groups were statistically significant for DPD at 12 and 24 months of treatment (P = 0.0471 and P = 0.0466, respectively), the Z scores ranging 0.45-1.90. NTx showed the most prominent decrease from the beginning of the study especially in the hormone treatment group: the differences between the two treatment groups were statistically highly significant for NTx already at 6 months of treatment (P = 0.0015), and the Z scores remained high ranging 2.11-3.82 throughout the two-year study period. Dual X-ray absorptiometry (DXA) of the lumbar spine and femoral neck did not show statistically significant differences between the two treatment groups throughout the two-year study period. After 2 years there was, however, a significant increase in bone density both in the spine (+ 6.6%, P = 0.0002) and in the femoral neck (+ 3.4%, P = 0.0389) in the women with hormone treatment. In the control group a significant increase (+ 5.1%, P = 0.0012) in the spine, whereas a non-significant decrease (-1.5%, n.s.) in the femoral neck was observed. We suggest that measurement of urinary cross-linked peptides derived from Type I collagen (NTx and DPD) might be a useful biochemical method of observing the positive clinical effect (i.e. reduction in bone resorption) following hormone replacement therapy in postmenopausal fracture patients.

A high-stearic acid diet does not impair glucose tolerance and insulin sensitivity in healthy women.

Results in epidemiological and experimental studies suggest that a diet rich in saturated fat may affect insulin sensitivity. However, no published data are available on the effect of stearic acid in this respect. Therefore, we examined the effects of a high-stearic acid diet and a high-oleic acid diet on glucose metabolism, serum lipids and lipoproteins, and blood coagulation factors in 15 healthy female subjects. Subjects followed the two experimental diets for 4 weeks according to a randomized crossover design. Both experimental diet periods were preceded by consumption of a baseline diet for 2 weeks. The diets provided 36% of energy (E%) as fat. In the experimental diets, 5 E% stearic or oleic acid was substituted for 5 E% of saturated fatty acids in the baseline diet. After the experimental diets, no differences were found in the insulin sensitivity index (mean+/-SEM, 5.4+/-1.9 v 5.2+/-1.6 x 10(-4) min(-1) x microU(-1) x mL(-1), nonsignificant [NS]), glucose effectiveness (0.026+/-0.006 v 0.026+/-0.003 min(-1), NS), or first-phase insulin reaction ([FPIR] 368+/-57 v 374+/-66 mU/L x min, NS). The concentration of serum lipids and lipoproteins and blood coagulation factors did not differ after the diet periods. In conclusion, a diet rich in stearic acid did not deteriorate glucose tolerance or insulin action in young healthy female subjects as compared with a diet rich in oleic acid.

Effects of postmenopausal hormone replacement therapy with and without vitamin D3 on circulating levels of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D.

The effects of postmenopausal hormone replacement therapy (HRT) and vitamin D3 on vitamin D metabolites (25OHD and 1,25(OH)2D) were studied in a population-based prospective 1-year study. The serum concentrations of intact parathyroid hormone (PTH), calcium, and phosphate were also studied. A total of 72 women were randomized into four treatment groups: HRT group (sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate), Vit D3 group (vitamin D3 300 IU/day + calcium lactate 500 mg/day), HRT + Vit D3 group (both above) and placebo group (calcium lactate 500 mg/day). Serum samples were taken in March-April, when vitamin D formation from sunlight in Finland is minimal after the dark winter. Serum concentrations of 25OHD increased in the Vit D3 group (33.5%, P < 0. 001) and in the HRT + Vit D3 group (38.2%, P < 0.001) but had not changed significantly in the HRT and placebo groups at the 1-year follow-up examination. Serum concentrations of calcitriol (1, 25(OH)2D) increased, however, only in the HRT group (23.7%, P < 0. 05), and remained unchanged in other groups. Serum concentrations of PTH decreased by 23.2% (P < 0.05) in the placebo group, but did not change significantly in the other three groups. The concentrations of serum calcium increased in the nonhormone groups (P < 0.001), whereas serum phosphate concentrations decreased in the hormone groups (P < 0.05 and 0.001). Our results confirm the positive effect of 1 year of HRT on serum calcitriol. Vitamin D3 supplementation increased 25OHD concentrations, but did not affect calcitriol concentrations even though the initial levels were low. Interestingly, the combination of HRT and vitamin D3 did not increase serum calcitriol concentrations as much as HRT alone.

A modification of the acid diazo coupling method (Malloy-Evelyn) for the determination of serum total bilirubin.

A simple and reliable method for the determination of total bilirubin from human serum is described. In this method, indirect bilirubin is liberated by the tenside in 0.12 mol l-1 HCl (R1), and the total bilirubin is coupled with a 2,5-dichlorobenzene diazonium (DBD) salt to obtain the corresponding azobilirubin having a lambda max of about 520-522 nm. The method can easily be applied to the KONE Delta, a fully automated, discrete random access clinical analyser, and also to less modern instruments. A sample volume of 5 microliters, R1 volume of 180 microliters, and R2 volume of 36 microliters was used on the KONE Delta. After a 5-min incubation at 37 degrees C, measurement at 575 nm was done (main wavelength). The within-run imprecision (CV%) varied from 2.9 to 0.3% within the serum total bilirubin range of 14-290 mumol l (n = 10). The between-run imprecision was from 2.2 to 1.3% within the range 13-97 mumol l-1 (n = 8). The method is linear up to at least 340 mumol l-1 (19.8 mg dl-1), and dilution extends the test limit to 3400 mumol l-1 (198.8 mg dl-1). The linearity of dilution was good over the practical measuring range. The present method had a strong linear correlation with the Boehringer 2,5-dichlorophenyl diazonium (DPD) method on the Hitachi 717 analyser: y(DBD) = 1.018x(DPD)+0.758, r = 0.9955 (n = 61). The stability of R2 (diazo reagent) in the analyser reagent compartment lasts at least 2 weeks.

Vitamin C deficiency and risk of myocardial infarction: prospective population study of men from eastern Finland.

OBJECTIVE: To examine the association between plasma vitamin C concentrations and the risk of acute myocardial infarction. DESIGN: Prospective population study. SETTING: Eastern Finland. SUBJECTS: 1605 randomly selected men aged 42, 48, 54, or 60 who did not have either symptomatic coronary heart disease or ischaemia on exercise testing at entry to the Kuopio ischaemic heart disease risk factor study in between 1984 and 1989. MAIN OUTCOME MEASURES: Number of acute myocardial infarctions; fasting plasma vitamin C concentrations at baseline. RESULTS: 70 of the men had a fatal or non-fatal myocardial infarction between March 1984 and December 1992.91 men had vitamin C deficiency (plasma ascorbate < 11.4 mumol/l, or 2.0 mg/l), of whom 12 (13.2%) had a myocardial infarction; 1514 men were not deficient in vitamin C, of whom 58 (3.8%) had a myocardial infarction. In a Cox proportional hazards model adjusted for age, year of examination, and season of the year examined (August to October v rest of the year) men who had vitamin C deficiency had a relative risk of acute myocardial infarction of 3.5 (95% confidence interval 1.8 to 6.7, P = 0.0002) compared with those who were not deficient. In another model adjusted additionally for the strongest risk factors for myocardial infarction and for dietary intakes of tea fibre, carotene, and saturated fats men with a plasma ascorbate concentration < 11.4 mumol/l had a relative risk of 2.5 (1.3 to 5.2, P = 0.0095) compared with men with higher plasma vitamin C concentrations. CONCLUSIONS: Vitamin C deficiency, as assessed by low plasma ascorbate concentration, is a risk factor for coronary heart disease.

Flow cytometric method to measure urea-resistant alkaline phosphatase from blood neutrophils for use in the prenatal screening of Down's syndrome.

The histochemical measurement of urea-resistant alkaline phosphatase from maternal blood neutrophils is known to have a high detection rate for the prenatal detection of Down's syndrome pregnancies. However, because the histochemical method is laborious and subjective to use, it has not gained widespread acceptance in prenatal screening programmes. We present a simple and objective method for the measurement of urea-resistant alkaline phosphatase by flow cytometry. The method should allow the design of larger studies aimed at evaluating the role of neutrophil urea-resistant alkaline phosphatase in the prenatal screening for Down's syndrome.

Determination of retinyl palmitate and total vitamin A content in liver and liver-based ready-to-eat foods.

The determination of retinyl palmitate and total vitamin A in liver and liver-based ready-to-eat foods is described. The method is very simple as sample preparation is minimal, and the isocratic elution of the C18 column with pure methanol does not necessitate a sophisticated instrumental set-up. The method is accurate with high recoveries (100.6 +/- 9.3%, mean +/- S.D., n = 23), and precise with within-day and between-day coefficients of variation of less than 5.5% (n = 13) and less than 16% (n = 6), respectively.

Epidemiology of vitamins A, E, D and C in rural villages in Finland: biochemical, nutritional and socioeconomical aspects.

We carried out an extensive health profile analysis in spring-winter 1986 in four Eastern Finnish rural villages as a part of the Healthy Village Study. Altogether, 793 people at working age (20-64 years of age, 427 men and 366 women) participated (80%). Serum lipids (total cholesterol, HDL-cholesterol and triglycerides) and plasma vitamins (vitamin A, D, E and C) were measured as biochemical indicators of health. The dietary habits were reflected in high serum total cholesterol, and in low plasma vitamin C (ascorbic acid, mean 34.4 mumol/l in men, and 51.2 mumol/l in women). The plasma levels of the other vitamins studied were, in general, satisfactory. The mean plasma concentration of vitamin A (retinol) was 2.70 mumol/l in men, and 2.23 mumol/l in women. The gender, high body weight and the use of animal fats had the strongest association to apparent plasma retinol concentrations. The corresponding plasma concentrations of vitamin D (25-hydroxy-D) were 34.1 nmol/l and 35.4 nmol/l, and vitamin E (d-alpha-tocopherol) 22.1 mumol/l and 22.2 mumol/l. Vitamin D deficiency (plasma 25-OHD less than 12.5 nmol/l) was seen in 5% of the subjects. A good vitamin D status was correlated with the use of vitamin supplements, and, surprisingly, with the frequent consumption of alcohol.

Nephrotoxicity of tobramycin. Value of examining various protein and enzyme markers.

The value of various protein and enzyme markers for the assessment of early nephrotoxicity of tobramycin was studied in 18 patients with febrile infection. The renal clearance of creatinine and the excretion of the following protein and enzyme markers were measured on the first and the last day of the 5-8 day treatment period: albumin, N-acetyl-beta-D-glucosaminidase (NAG), beta 2-microglobulin, alpha-amylase, lysozyme and retinol-binding protein (RBP). Diurnal excretions of beta 2-microglobulin, lysozyme and RBP, all markers of tubular dysfunction, were already increased on the first treatment day, compared to similar control patients treated with non-aminoglycoside antibiotics and reference values of our laboratory. The excretion of NAG, an enzyme released from tubular cell lysosomes, was not increased initially, but on the last treatment day it was increased most consistently of all the markers studied. Glomerular filtration rate was halved in 5 of the patients. The results suggest that the initial increase in beta 2-microglobulin, lysozyme and RBP excretion is a result of an early tubular transfer block by tobramycin, whereas the late increase in NAG excretion probably reflects the progress of tubular cell damage during the course of aminoglycoside therapy.

Use of non-collagen markers in osteoporosis studies.

Clinical and research laboratories routinely measure various hormonal and nonhormonal parameters of calcium and phosphorus metabolism, and markers of bone turnover. Such measurements may help clinical decision-making relating to metabolic bone disease and osteoporosis. Molecular biological and cell-culture techniques are being used in basic biochemical research on bone-cell metabolism. Results may aid understanding of normal and abnormal regulation of the bone-cell metabolism, and thus provide further insights relating to the diagnosis and prevention of osteoporosis.

Plasma catecholamines: laboratory aspects.

In this review the methods used for analysis of plasma catecholamines in clinical chemical laboratories are discussed. The physiology of catecholamines as well as their measuring indications are discussed, together with concise evaluation of the methods most commonly used, namely indirect radioenzymatic assays or direct determinations by high-performance liquid chromatography combined with either electrochemical or fluorometric detection. The main advantage of radioenzymatic assay is its sensitivity and thus the need for only a small sample. Liquid chromatographic methods in general are less tedious, relatively rapid, and cheap, and omit the use of radionuclides. Both of these methods, however, are subject to a number of analytical errors, which can only be avoided by proper development of methods and skilled use of these methods. Little routine work is done using either radioimmunoassay or gas-chromatography.

Alkaline phosphatase isoenzymes in serum determined by high-performance anion-exchange liquid chromatography with detection by enzyme reaction.

This rapid, reproducible method for separating and determining individual alkaline phosphatase (EC 3.1.3.1) isoenzymes in serum is based on high-performance liquid chromatography with a weak anion-exchange column (SynChropak AX 300). The isoenzymes so resolved are detected by using an on-line enzyme reaction followed by spectrophotometric monitoring at 405 nm of the 4-nitrophenol formed. Complete diagnostic profiles of the various isoenzymes present in normal and pathological sera are obtained within 20 min. The mean (and SD) normal concentrations of the bone B1 and intestinal isoenzymes in serum of adults were 3.7 (4.3) and 4.5 (3.9) U/L, respectively (n = 14), and of the bone isoenzyme B2 and liver isoenzymes L1 and L2, 5.8 (8.6), 33.0 (10.6), and 12.0 (4.8) U/L, respectively (n = 17). Concentrations of the B2 and L1 isoenzymes in adults over age 40 years differed significantly from those in adults younger than 40 years, that of bone isoenzyme being lower (P less than 0.05) and that of the liver isoenzyme being higher (P less than 0.001) in the younger adults.

25-Hydroxyvitamin D and vitamin D in human milk: effects of supplementation and season.

Breast-milk 25-hydroxyvitamin D (25-[OH]D) and vitamin D were measured in mothers supplemented with 2000 or 1000 IU (50 or 25 micrograms) of vitamin D/d or with no supplementation. Fore- and hindmilk samples were collected at two stages of lactation (8 and 15 or 20 wk after delivery) and at different seasons. Season affected the levels of 25-(OH)D and vitamin D. The 25-(OH)D levels were higher in hind- than in foremilk. Supplementation had no effect on vitamin D levels. Milk 25-(OH)D levels of mothers receiving either 1000 or 2000 IU (25 or 50 micrograms) vitamin D/d were significantly higher than those of unsupplemented mothers in February and April. In theory, supplementation with 2000 IU (50 micrograms) vitamin D should have increased the calculated antirachitic activity of the milk in winter to the levels of unsupplemented mothers in September; however, responses varied widely among individuals.

Plasma haemoglobin determination using chlorpromazine as a non-carcinogenic reagent.

Plasma haemoglobin was assayed with the non-carcinogenic reagent phenothiazine. This method is sensitive and allows the measurement of plasma haemoglobin concentrations in the range 4-500 mg/l with a within-run CV of 2.1%, and a between-run CV of 4.3%. A spectrophotometric scanning method (x) based on the determination of haemoglobin as haemiglobin cyanide using the Soret band at 419 nm correlated well with the phenothiazine method (y): y = 1.07x + 15.8, r = 0.995, n = 31. It was found that the absorbances in the phenothiazine method were markedly dependent on the concentration of phosphoric acid.