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Biomedical subjects

B Akesson

Publications and source records attributed to B Akesson.

At least 55 records · Page 3Linked to original sources

Selenoprotein P in serum as a biochemical marker of selenium status.

Some characteristics of a radioimmunoassay of selenoprotein P (a major selenoprotein in human serum) are described. Polyclonal antibodies generated in rabbits were used and goat anti-rabbit-IgG antiserum was used as a second antibody. Depending on the concentration of selenoprotein P, 1-10 microliters of human serum were used in the assay. The relative standard deviation for the concentration of selenoprotein P was 6.3% between assays and 7.7% within assays. Different animal sera gave no significant interference, indicating that the antibodies did not react with non-human analogues of selenoprotein P. No indication of cross-reactivity could be found concerning extracellular glutathione peroxidase (another selenoprotein in serum). Addition of increasing amounts of normal human serum and partially purified selenoprotein P to the radioimmunoassay resulted in parallel curves. Incubation at 4 degrees C gave somewhat higher binding of labelled selenoprotein P than incubation at room temperature. The epitope, recognized by the antibodies, was apparently stable after storage of serum (in the frozen state for years, and in the cold for months). No significant amount of selenoprotein P could be demonstrated in red blood cells, and analysis of haemolysed whole blood gave expected data. Investigations of selenium status in different study groups indicated that in most cases the concentration of selenoprotein P in serum was positively correlated to that of glutathione peroxidase and serum selenium. In an intervention study, where subjects decreased their selenium intake to 50%, the serum levels of glutathione peroxidase and selenium decreased, but no significant decrease of selenoprotein P could be demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS)

Biomarkers↗

Selenoprotein P and glutathione peroxidase (EC 1.11.1.9) in plasma as indices of selenium status in relation to the intake of fish.

In Sweden fish is considered to be an important source of dietary Se. Therefore Se status was assessed in forty-one middle-aged men with widely varying fish consumption. Glutathione peroxidase (EC 1.11.1.9) and selenoprotein P in plasma were measured by radioimmunoassay. Plasma Se among the men increased slightly with increasing consumption of fish, but no such increases in the concentrations of glutathione peroxidase and selenoprotein P in plasma were observed. Moreover, no correlation was found between plasma Se and glutathione peroxidase or selenoprotein P. Instead, glutathione peroxidase was significantly correlated with selenoprotein P (r 0.73, P < 0.001), indicating that both glutathione peroxidase and selenoprotein P were functional indicators of Se status in this group. The proportion of plasma Se located in glutathione peroxidase decreased with increasing plasma Se. The results suggest that the Se consumed from fish had no apparent effect on the amount of Se incorporated into the functional selenoproteins of plasma. It is concluded that in some cases selenoproteins may be better biological markers of Se status than the total concentration of Se.

Animals↗

Influence of nitric oxide synthase inhibition, nitric oxide and hydroperoxide on insulin release induced by various secretagogues.

1. Recent studies have suggested that the generation of nitric oxide (NO) and hydrogen peroxide (H2O2) by islet NO synthase and monoamine oxidase, respectively, may have a regulatory influence on insulin secretory processes. We have investigated the pattern of insulin release from isolated islets of Langerhans in the presence of various pharmacological agents known to perturb the intracellular levels of NO and the oxidation state of SH-groups. 2. The NO synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) dose-dependently increased L-arginine-induced insulin release. D-Arginine did not influence L-arginine-induced insulin secretion. However, D-NAME which reportedly has no inhibitory action on NO synthase, modestly increased L-arginine-induced insulin release, but was less effective than L-NAME. High concentrations (10 mM) of D-arginine as well as L-NAME and D-NAME could enhance basal insulin release. 3. The intracellular NO donor, hydroxylamine, dose-dependently inhibited insulin secretion induced by L-arginine and L-arginine+L-NAME. 4. Glucose-induced insulin release was increased by NO synthase inhibition (L-NAME) and inhibited by the intracellular NO donor, hydroxylamine. Sydnonimine-1 (SIN-1), an extracellular donor of NO and superoxide, induced a modest suppression of glucose-stimulated insulin release. SIN-1 did not influence insulin secretion induced by L-arginine or the adenylate cyclase activator, forskolin. 5. The intracellular 'hydroperoxide donor' tert-butylhydroperoxide in the concentration range of 0.03-3 mM inhibited insulin release stimulated by the nutrient secretagogues glucose and L-arginine. Low concentrations (0.03-30 microM) of tert-butylhydroperoxide, however enhanced insulin secretion induced by the phosphodiesterase inhibitor isobutylmethylxanthine (IBMX). 6. Islet guanosine 3':5'-cyclic monophosphate (cyclic GMP) content was not influenced by 10 mML-arginine or tert-butylhydroperoxide at 3 or 300 micro M but was markedly increased (14 fold) by a high hydroxylamine concentration (300 micro M). In contrast, islet adenosine 3':5'-cyclic monophosphate (cyclicAMP) content was increased (3 fold) by L-arginine (10 mM) and (2 fold) by tert-butylhydroperoxide(300 micro M).7. Our results strongly suggest that NO is a negative modulator of insulin release induced by the nutrient secretagogues L-arginine and glucose. This effect is probably not mediated to any major extent by the guanylate cyclase-cyclic GMP system but may rather be exerted by the S-nitrosylation of critical thiol groups involved in the secretory process. Similarly the inhibitory effect of tert-butylhydroperoxide is likely to be elicited through affecting critical thiol groups. The mechanism underlying the secretion promoting action of tert-butylhydroperoxide on IBMX-induced insulin release is probably linked to intracellular Ca2+-perturbations affecting exocytosis.8. Taken together with previous data the present results suggest that islet production of low physiological levels of free radicals such as NO and H202 may serve as important modulators of insulin secretory processes.

1-Methyl-3-isobutylxanthine↗

Effect of dietary intake of trimethylamine on human metabolism of the industrial catalyst dimethylethylamine.

OBJECTIVES: The aim was to study the effect of trimethylamine (TMA) on the metabolism of the industrial catalyst dimethylethylamine (DMEA) to ascertain whether biological monitoring of industrial exposure to DMEA is compromised and excretion of the malodorous DMEA in sweat and urine is increased by dietary intake of TMA. METHODS: DMEA (0/25 mg) and TMA (0/300/600 mg) were given simultaneously once weekly for six weeks to five healthy volunteers. Plasma was collected before and one hour after the doses, and urine 0-2, 2-4, 4-6, 6-8, and 8-24 hours after the doses. Specimens were analysed by gas chromatography with a nitrogen sensitive detector. RESULTS: Both amines were readily absorbed from the gastrointestinal tract and excreted in urine within 24 hours (DMEA 80%; TMA 86%). Oral intake of TMA increased the DMEA content of plasma and urine dose dependently, although there were large individual differences. Plasma and urinary TMA concentrations also increased, but not dose dependently. Moreover, the findings suggested the formation of endogenous TMA, little dealkylation of DMEA and TMA, and considerable first-pass metabolism. CONCLUSIONS: Although intake of TMA reduced N-oxygenation of DMEA and TMA, total urinary DMEA values (aggregate of DMEA and its oxide DMEAO excretion) were unaffected. Thus, monitoring occupational exposure to DMEA by analysis of biological specimens is not confounded by dietary intake of TMA, provided that total urinary DMEA is monitored. Although the increased urinary and hidrotic excretion of DMEA may contribute to body odour problems, they were primarily due to TMA excretion, which is much the greater.

Administration, Oral↗

Fish consumption and exposure to persistent organochlorine compounds, mercury, selenium and methylamines among Swedish fishermen.

OBJECTIVES: This study assessed dietary habits and exposure to selenium, persistent organochlorine compounds, methylmercury, and methylamines among Swedish fishermen. METHODS: Two hundred and fifty randomly selected subjects from a cohort of 2896 fishermen from the Swedish east coast (Baltic Sea) and 8477 fishermen from the west coast (Skagerrak and Kattegatt) were interviewed along with 250 referents. Subgroups of fishermen and referents from different coastal areas were also selected for blood and urine sampling. RESULTS: The interview data showed that fishermen ate almost twice as much fish as the 250 referents from the general population. The blood levels of mercury were twice as high, and the plasma selenium levels were 10-15% higher in the fishermen than in the referents. There was, however, no difference between the fishermen's cohorts with respect to these variables. Fishermen from the east coast ate more fatty fish than fishermen from the west coast, and they also had higher blood levels of persistent organochlorine compounds such as polychlorinated biphenyls and polychlorinated dibenzo p-dioxins and furans (present in fatty fish species in the Baltic Sea) than both the westcoast fishermen and the referents. CONCLUSIONS: A cohort of Swedish eastcoast fishermen might be a suitable study base for epidemiologic studies on the mortality and cancer morbidity associated with dietary exposure to persistent organochlorine compounds.

Animals↗

Variation in selenoprotein P concentration in serum from different European regions.

OBJECTIVE: To study the variation in selenoprotein P levels in serum among healthy subjects from 17 European regions representing nine countries. SUBJECTS: Survey of 414 37-70-year-old subjects (197/217 males/females) from 17 regions. RESULTS: The level of selenoprotein P varied significantly among regions and countries. When the highest concentration of selenoprotein P encountered in samples from Maldegem (Belgium), was set at 100%, the mean concentration for other regions was: 92% Barcelona; 91% Ipswich and London; 89% Malmö and Vosselaan; 86% Lisbon; 85% Netherlands; 83% Umeå; 82% Uppsala; 80% Paris; 79% Heidelberg; 78% Gothenburg and Grenoble; 74% Giessen; 71% Ioannina and 69% Epirus. When the data were pooled for the nine countries the highest selenoprotein P concentration was observed for subjects from Spain. When this concentration was set at 100%, the mean concentration for other countries was: 99% Belgium and United Kingdom, 94% Portugal, 92% Netherlands and Sweden, 86% France, 83% Germany and 76% Greece. The linear correlation between serum selenium and serum selenoprotein P in the whole sample was 0.68, P < 0.001 (n = 414), and it was similar in men and women. Despite this high correlation the mean ratio between selenoprotein P and selenium varied 1.3-fold among regions. CONCLUSIONS: Both the concentration of selenoprotein P as well as its proportion of total serum selenium in adult subjects vary among different European regions. Further studies are necessary to evaluate the physiological or medical implications of these differences.

Adult↗

The thioredoxin and glutaredoxin systems are efficient electron donors to human plasma glutathione peroxidase.

Human plasma glutathione peroxidase (GSH-Px) is a distinct extracellular selenoenzyme that detoxifies hydroperoxides when used with GSH in high (mM) non-physiological concentrations. We have discovered that NADPH and human thioredoxin reductase (TR) by itself or with thioredoxin (Trx) are efficient electron donors to this human plasma peroxidase. Incubation of 0.05 microM TR with 0.25 microM GSH-Px, in a system free from GSH, resulted in reduction of t-butyl hydroperoxide. Addition of Trx, 2.5 and 5 microM, respectively, further increased the rate of the reaction. These data were obtained using an assay measuring the oxidation of NADPH. A direct assay demonstrated the formation of cumyl alcohol from cumene hydroperoxide in this GSH-independent peroxidase reaction. Incubation of 0.25 microM GSH-Px with a low concentration of GSH (10 microM), representing the upper level in plasma, plus excess glutathione reductase and NADPH did not result in any reduction of t-butyl hydroperoxide. However, after addition of 2.5 microM human glutaredoxin, a linear peroxidase reaction started. The results suggest that extracellular TR, Trx, or glutaredoxin are reductants for the selenium-dependent peroxidase rather than GSH.

Electron Transport↗

Purification of selenoprotein P from human plasma.

Selenoprotein P was partially purified (> 1000-fold) from human plasma in four chromatographic steps using 75Se-labeled selenoprotein P secreted by HepG2 cells in culture as a marker. The purified preparation was injected into mice and monoclonal antibodies, which precipitated the labeled protein, were generated. Neither of two different monoclonal antibodies had cross-reactivity with plasma from five animal species. Antibodies were coupled to agarose, and selenoprotein P was purified from human plasma by immunoaffinity chromatography followed by chromatography on heparin agarose. With two different matrix-bound monoclonal antibodies, the purification procedure gave two bands on SDS-PAGE with mobilities corresponding to 61 and 55 kDa. Both bands stained for carbohydrate and showed increased electrophoretic mobility after enzymatic deglycosylation. Immunoaffinity chromatography removed approx. one-third of the selenium from plasma or 0.4 mumol Se/l at a total selenium concentration of 1.1 mumol/l, indicating that selenoprotein P constituted this proportion of total plasma selenium in healthy US blood donors.

Amino Acid Sequence↗

Urinary excretion of methylamines in men with varying intake of fish from the Baltic Sea.

Fish contain methylamines, especially trimethylamine N-oxide (TMAO), trimethylamine (TMA), and dimethylamine (DMA). Further, DMA may be formed TMA and TMAO. DMA is a precursor of nitrosodimethylamine (NDMA), which is a potent carcinogen. Levels of DMA, TMA, and TMAO in urine were used as indicators of the dietary exposure and in vivo formation of these amines in 44 men, representing 3 groups with different fish consumption habits. The levels of TMA (median 0.24 mmol/mol creatinine; range 0-2.7) and TMAO (median 38 mmol/mol creatinine; range 8-290) were significantly associated with the weekly intake of fish (r = .47, p = .001, and r = .53, p = .0002, respectively), while no such relation was found for DMA (median 24 mmol/mol creatinine; range 5-46). Further, urinary levels of TMA and TMAO were dependent on recent intake of fish.

Adult↗

Trace element concentrations in hair of subjects from two South Pacific Islands, Atafu (Tokelau) and Kitava (Papua New Guinea).

The concentrations of trace elements in hair were measured in 108 healthy subjects (60 males and 48 females, mean age 46-years) resident on Atafu (Tokelau) and in 83 subjects (63 males and 20 females, mean age 26-years) living on Kitava island (Papua New Guinea). On Kitava, a traditional subsistence lifestyle uninfluenced by Western dietary habits is still maintained, but on Atafu the dietary habits have been modified by the inclusion of such imported foods as rice, flour, sugar and canned meat and fish. The concentrations of zinc and magnesium in hair were significantly higher in the Kitava than in the Atafu subjects, whereas those of copper were similar in both groups, and those of selenium, mercury, lead and cadmium were higher in Atafu subjects. The levels of serum copper, magnesium and selenium concentrations in Kitava subjects were not so low as to indicate any trace element deficiency. The higher hair content of mercury, lead and cadmium found among Atafu subjects might be due to consumption of marine foods contaminated with metals, consumption of canned foods, or frequent cigarette smoking, though it is difficult to single out any specific factor.

Adolescent↗

Radioimmunoassay of glutathione peroxidase in human serum.

Glutathione peroxidase (GSHPx) was purified from human serum and used for immunization of rabbits. Antiserum bound up to 75% of added 125I-GSHPx after precipitation with a second antibody. Human serum, but not sera from eight animal species, inhibited the binding of labelled GSHPx, indicating that the antiserum did not react with GSHPx from these species. GSHPx could be measured in less than 10 microliters of human serum by radioimmunoassay. In sera with widely varying selenium concentrations (0.1-2.9 mumol/l) the amount of GSHPx protein (0.3-6.3 mg/l) was strongly correlated with GSHPx activity (r = 0.94) and it was also correlated with serum selenium concentrations (r = 0.64). This indicates that GSHPx protein may be a valuable biological marker of selenium status. In samples with serum selenium concentrations of 0.8-1.2 mumol/l, the concentration of GSHPx was 3.3 (0.4) mg/l (mean (S.D.)), or 0.04 (0.005) mumol/l. This corresponded to 0.16 (0.02) mumol/l of GSHPx selenium and 16% (2.8)% of total serum selenium. The data suggest that the method can be used to measure the proportion of serum selenium that is located in GSHPx. Following storage of serum at room temperature, both serum GSHPx protein and activity declined, but addition of glutathione protected both GSHPx protein and activity.

Animals↗

Gas chromatographic determination of primary and secondary low-molecular-mass aliphatic amines in urine using derivatization with isobutyl chloroformate.

A simple routine method for the gas chromatographic determination of methylamine, dimethylamine, ethylamine and methylethylamine in urine is presented. The method is based on a two-phase derivatization procedure with isobutyl chloroformate as reagent. The reaction is quantitative in 10 min. We found no artifact formation of either choline or trimethylamine (dietary amine compounds) or of dimethylethylamine or triethylamine (catalyst amines in the industrial setting). The chromatographic behaviour of the amine carbamates was excellent. The recoveries of methylamine, dimethylamine, ethylamine and methylethylamine in spiked urine samples were 82, 89, 100 and 96%, respectively, and the precision (the relative standard deviation) was 3.6, 1.8, 3.3 and 2.0%, respectively. The method was linear for the studied amine carbamates up to 250 mg/l. The endogenous amine concentrations in urine samples from ten normal subjects were: methylamine, 0.9 mg/l (mean; range 0.3-1.5); dimethylamine, 14.7 mg/l (mean; range 4.6-27.6); ethylamine, 0.8 mg/l (mean; range 0.2-2.3); methylethylamine, less than 0.02 mg/l.

Amines↗

Dietary assessment methods evaluated in the Malmö food study.

A study to evaluate the relative validity of two dietary assessment methods, to be used in a prospective cohort study on diet and cancer, was conducted in a random sample of the Malmö population during 1984-1985. The study compared (i) a long, self-administered 'food-use questionnaire' (about 250 food items), asking for frequency of consumption of food and beverages and for usual portion size estimated with the help of a booklet of food pictures, and (ii) a combined method involving a shorter food frequency questionnaire (about 130 food items) and a 2-week diet diary, with (iii) a reference method requiring 18 days of weighed food records. The results indicated that subject compliance was quite good for both methods and, in particular, that the diet diary was well accepted. Both methods produced fairly good correlations with the reference method, of the order of 0.40 to 0.60 for most nutrients after adjustment for energy. Both methods tended to overestimate intake of fat, protein and carbohydrates as well as of vitamins and minerals.

Aged↗

Fatty acid composition of serum phosphatidylcholine in healthy subjects consuming varying amounts of fish.

Fatty acid composition of serum phosphatidylcholine was studied in subjects living in the same area and selected for high consumption of fat fish, high consumption of lean fish, moderate or no consumption of fish. The proportion of eicosapentaenoic acid (20:5) in serum phosphatidylcholine was fourfold higher, and that of docosahexaenoic acid (22:6) twofold higher among high consumers of fish compared to non-consumers, and these proportions were also higher in fat fish consumers than in high consumers of lean fish, and in moderate consumers compared to non-consumers. In the whole group the intake of fat fish was correlated to the proportions of 20:5, 22:6 and total n-3 fatty acids in serum phosphatidylcholine. The estimated intake of total n-3 fatty acids from fish was highly correlated to the proportions of these fatty acids in serum phosphatidylcholine (r = 0.87). Plasma triglyceride concentration was lower among high fat fish consumers than in other groups, but no differences between groups were observed for concentration of cholesterol, LDL cholesterol, and HDL cholesterol. It is concluded that subjects who consume high amounts of fat fish in their normal diet attain approximately the same proportions of n-3 fatty acids in serum phosphatidylcholine as reported for subjects taking fish oil supplements, and that fatty acid composition of serum lipids can be used as a marker of intake of fat fish.

Adult↗

Fish as a source of exposure to mercury and selenium.

In a total of 395 subjects with varying fish consumption habits, mercury levels in whole blood (B-Hg), and selenium levels in plasma (P-Se) were studied. Also, in subcohorts, mercury levels in blood cells (Ery-Hg; n = 79), plasma (P-Hg; n = 158) and urine (U-Hg; n = 125) were analysed. There were statistically significant associations between fish intake on the one hand, and B-Hg, Ery-Hg and P-Hg, on the other, but not so with U-Hg. In subjects who never had fish, the average B-Hg was 1.8 ng/g, in subjects who had at least two fish meals each week, 6.7 ng/g. Ery-Hg, and to a less extent P-Hg, were associated with levels of marine n-3 polyunsaturated fatty acids (PUFA) in serum phosphatidylcholine. P-Hg and U-Hg were associated with numbers of teeth with amalgam fillings. P-Se also correlated with fish intake. In subjects who never had fish, P-Se averaged 80 micrograms/l, in subjects who had at least two fish meals per week, 91 micrograms/l. There was an association between PUFA and P-Se. Further, there were statistically significant associations between P-Se on the one hand, and B-Hg, Ery-Hg and P-Hg on the other. The data clearly demonstrate the importance of fish for the exposure to methylmercury and selenium in the Swedish diet, and the impact of amalgam as a source of exposure to inorganic mercury.

Adult↗

Trace element status in healthy subjects switching from a mixed to a lactovegetarian diet for 12 mo.

The consequences of a change from a mixed to a lactovegetarian diet for 12 mo on trace element concentrations in plasma, hair, urine, and feces were studied in 16 women and 4 men. After the diet shift, intakes of zinc and magnesium did not change but that of selenium decreased by 40%. Three months after the diet shift, plasma and hair concentrations of zinc, copper, and selenium had decreased but those of magnesium had increased and the concentrations of mercury, lead, and cadmium in hair were lower. Also, the excretion of zinc, copper, and magnesium in urine, and that of selenium in urine and feces had decreased. Only small changes occurred during the remaining lactovegetarian-diet period. Three years later trace element concentrations had reverted towards baseline concentrations; copper values were similar to baseline concentrations but data for magnesium were slightly higher, and more complex patterns were observed for zinc and selenium. It is concluded that a shift to a lactovegetarian diet changes trace element status.

Adult↗

Dietary intake and nutritional status in patients with systemic sclerosis.

Oesophageal dysmotility and abnormalities of intestinal function are important manifestations in systemic sclerosis and may have a significant effect on nutrient absorption and nutritional status. In this study 30 patients with systemic sclerosis with symptoms from the gastrointestinal tract were compared with matched healthy control subjects with respect to nutrient intake (four day record), anthropometric measurements, and biochemical nutritional status. The intake of energy (8.1 and 8.4 MJ/day) and its distribution among nutrients did not differ between patients and control subjects, but the lower intake of dietary fibre among patients with systemic sclerosis suggests that they avoided food with a coarse structure, such as coarse bread. The intake of vegetables and fruit also tended to be lower among patients with systemic sclerosis. Half of the patients had a subnormal arm muscle circumference, and two patients also had a subnormal triceps skinfold thickness, indicating severe malnutrition. The concentration of ascorbic acid, alpha-tocopherol, carotene, selenium, and also the proportion of linoleic acid (18:2) in serum phosphatidylcholine was lower in patients than in control subjects.

Adult↗

Fatty acid composition of adipose tissue and serum micronutrients in relation to common rheumatic complaints in Swedish adults 50-70 years old.

Biochemical markers of nutritional status, such as adipose tissue fatty acid composition and circulating levels of micronutrients, may reflect dietary habits, but may also be related to inflammatory activity. We examined the relationship between common rheumatic complaints such as osteo-arthrosis, arthralgia, tendinitis, low-back pain, and neck pain, and the composition of adipose tissue fatty acids, serum concentrations of retinol, ascorbic acid, alpha-tocopherol and selenium in 502, out of 900 randomly selected individuals 50-70 years old. Compared with control subjects the rheumatic group as a whole (p less than 0.01) as well as subgroups like those with arthralgia (p less than 0.01), subacromial shoulder pain (p less than 0.03), and low-back pain (p less than 0.05), had decreased proportions of adipose tissue arachidonic acid. The abnormalities correlated with the duration of the symptom-free period preceding adipose tissue sampling and could not be explained by the evaluation of possible confounding factors. Regarding other fatty acids and micronutrients there were no differences between symptomatic subjects and controls. One explanation for the low proportion of adipose tissue arachidonic acid may be increased eicosanoid production.

Adipose Tissue↗