Sampling and processing of fresh blood samples within a European multicenter nutritional study: evaluation of biomarker stability during transport and storage.

INTRODUCTION: Analysis of several biological markers improves the quality and physiologic comprehension of data obtained in epidemiological nutritional studies. AIM: To develop a methodology that guarantees the centralized analysis and quality assurance of the most relevant blood parameters from fresh blood samples in adolescents in a European multicenter study. MATERIALS AND METHODS: Stability of selected nutrients and biomarkers (vitamins, fatty acids, iron metabolism and immunological parameters) chosen with respect to time and temperature of sample transport and storage was evaluated as part of the pilot study of the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) project. RESULTS: Routine biochemistry and iron status parameters included in the HELENA Cross-Sectional Study (CSS) protocol could be analyzed within 24 h from fresh blood samples without any stability problems (coefficient of variation (CV)<5%, P<0.05). However, stability tests for lymphocyte subpopulations, vitamin C and fatty acids showed that they are very unstable at room temperature without any treatment. Therefore, a special handling for these samples was developed. Vitamin C was stabilized with metaphosphoric acid and transported under cooled conditions (CV 4.4%, recovery rate >93%, P>0.05). According to the results, a specific methodology and transport system were developed to collect blood samples at schools in 10 European cities and to send them to the centralized laboratory (IEL, Bonn, Germany). To guarantee good clinical practice, the field workers were instructed in a training workshop and a manual of operation was developed. CONCLUSION: The handling and transport system for fresh blood samples developed for the European multicenter study HELENA is adequate for the final part of the HELENA-CSS and will provide, for the first time, reference values for several biological markers in European adolescents.

Vitamin B6 status, deficiency and its consequences--an overview.

BACKGROUND: Vitamin B6 is thought to be a most versatile coenzyme that participates in more than 100 biochemical reactions. It is involved in amino acid and homocysteine metabolism, glucose and lipid metabolism, neurotransmitter production and DNA/RNA synthesis. Vitamin B6 can also be a modulator of gene expression. Nowadays, clinically evident vitamin B6 deficiency is not a common disorder, at least in the general population. Nevertheless, a subclinical, undiagnosed deficiency may be present in some subjects, particularly in the elderly. OBJECTIVE: This review gives a complete overview over the metabolism and interactions of vitamin B6. Further, we show which complications and deficiency symptoms can occur due to a lack of vitamin B6 and possibilities for public health and supplemental interventions. METHODS: The database Medline (www.ncvi.nlm.nih.gov) was searched for terms like "vitamin B6", "pyridoxal", "cancer", "homocysteine", etc. For a complete understanding, we included studies with early findings from the forties as well as recent results from 2006. These studies were summarised and compared in different chapters. RESULTS AND CONCLUSION: In fact, it has been proposed that suboptimal vitamin B6 status is associated with certain diseases that particularly afflict the elderly population: impaired cognitive function, Alzheimer's disease, cardiovascular disease, and different types of cancer. Some of these problems may be related to the elevated homocysteine concentrations associated to vitamin B6 deficiency, but there is also evidence for other mechanisms independent of homocysteine by which a suboptimal vitamin B6 status could increase the risk for these chronic diseases.

Research goals for folate and related B vitamin in Europe.

In the past decade, the understanding of folate bioavailability, metabolism and related health issues has increased, but several problems remain, including the difficulty of delivering the available knowledge to the populations at risk. Owing to the low compliance of taking folic acid supplements, for example, among women of child-bearing age who could lower the risk of having a baby with a neural tube defect, food-based strategies aimed at increasing the intake of folate and other B-group vitamins should be a priority for future research. These should include the development of a combined strategy of supplemental folate (possibly with vitamin B(12)), biofortification using engineered plant-derived foods and micro-organisms and food fortification for increasing folate intakes in the general population. Currently, the most effective population-based strategy to reduce NTDs remains folic acid fortification. However, the possible adverse effect of high intakes of folic acid on neurologic functioning among elderly persons with vitamin B(12) deficiency needs urgent investigation. The results of ongoing randomized controlled studies aimed at reducing the prevalence of hyperhomocysteinemia and related morbidity must be available before food-based total population approaches for treatment of hyperhomocysteinemia can be recommended. Further research is required on quantitative assessment of folate intake and bioavailability, along with a more thorough understanding of physiological, biochemical and genetic processes involved in folate absorption and metabolism.

B-vitamin status and intake in European adolescents. A review of the literature.

BACKGROUND: National and international recommendations for the intake of B vitamins in adolescents consist of estimates and extrapolations from adult values. Due to increasing growth and therefore relatively high energy and nutrient requirements adolescents are a vulnerable group from the nutritional point of view. In addition, a deficient intake of several B vitamins is strongly connected with the development of cancer, neural tube defects and cardiovascular diseases. OBJECTIVE: The aim of this work is to assess dietary intake and status of B vitamins and homocysteine of European adolescents on the basis of published data. METHODS: The database Medline (www.ncvi.nlm.nih.gov) was searched for terms like "Vitamin B", "homocysteine", "Europe", etc. Studies published between June 1980 and December 2004 were analysed for this review. Results of the intake of B vitamins were compared with the EAR or AI, respectively, as recommended by the U.S. Institute of Medicine. Due to lacking reference values for adolescents results of blood status as well as homocysteine were compared to different thresholds for adults. RESULTS: Considering the limitations of the comparability between the reviewed studies e.g. by different methodologies, sample size, age groups, the average intake of B vitamins surpassed the EAR and AI. Boys were better supplied with B vitamins than girls. The intake decreased with increasing age in both genders. A possible deficiency of folate was noticed and girls in particular seemed to be more at risk. Clear regional tendencies for the vitamin intake could not be observed. Results of vitamin B6, B12, folate in blood, and homocysteine were levelled in-between the thresholds. Though the great standard deviation of folate increased the probability of a deficient supply in parts of the population. CONCLUSIONS: European girls seem to be at risk of folate deficiency. Supplements and fortified food were not taken into consideration by most of the published studies which additionally distorts the real intake. Standardized methods of dietary surveys and reference values for B vitamins as well as homocysteine still must be established. Hence, further investigations are of great relevance. folate increased the probability of a deficient supply in parts of the population. CONCLUSIONS: European girls seem to be at risk of folate deficiency. Supplements and fortified food were not taken into consideration by most of the published studies which additionally distorts the real intake. Standardized methods of dietary surveys and reference values for B vitamins as well as homocysteine still must be established. Hence, further investigations are of great relevance.

Short-term impact of a lactovegetarian diet on adrenocortical activity and adrenal androgens.

The aim of this study was to determine whether definite diet changes affect adrenocortical activity and/or adrenal androgen metabolism. A controlled experimental diet study with four consecutive diet periods (repeated measure design) was carried out in six healthy adult volunteers. Four nearly isoenergetic diets, two normal (N) moderately protein-rich, one protein-rich (P), and one low protein lactovegetarian (L), were fed. At the end of each 5-day diet period a blood sample and two 24-h urine specimens were obtained from each subject. Plasma levels of dehydroepiandrosterone sulfate (DHEAS) were elevated with diet L (6.5 +/- 1.4 vs. 5.3 +/- 1.1 mumol/L; P < 0.05) compared to diet N, whereas other plasma hormones, including cortisol and insulin-like growth factor I did not vary markedly. A marked increase of 60% was seen in the urinary 24-h output of 3 alpha-androstanediol glucuronide with diet P. Urinary 24-h excretion rates for C peptide, free cortisol, DHEAS, and total 17-ketosteroid sulfates were clearly reduced with diet L compared to those with diet N or P. Our results show that a lactovegetarian diet can reduce adrenocortical activity (at least after a short term diet change). In addition, this vegetarian nutrition leads to a particular metabolic situation (elevated plasma DHEAS and reduced urinary DHEAS output) that usually is characteristic of fasting. Peripheral androgen metabolism as reflected by urinary 3 alpha-androstanediol glucuronide appears to be influenced only by high protein intake (diet P). Further research (controlled dietary long term investigation) is required 1) to validate whether the effects of diet on adrenocortical activity represent sustained endocrine changes and 2) to elucidate the underlying mechanism.

Effects of folic acid and combinations of folic acid and vitamin B-12 on plasma homocysteine concentrations in healthy, young women.

BACKGROUND: Elevated plasma homocysteine concentrations are considered to be a risk factor for vascular disease and fetal malformations such as neural tube defects. Recent studies have shown that plasma homocysteine can be lowered by folic acid in amounts corresponding to 1-2 times the recommended dietary allowance. Preliminary evidence indicates that vitamin B-12 may be beneficial when included in supplements or in a food-fortification regimen together with folic acid. OBJECTIVE: We aimed to compare the homocysteine-lowering potential of a folic acid supplement with that of 2 supplements containing different doses of vitamin B-12 in addition to folic acid. DESIGN: Female volunteers of childbearing age (n = 150) received a placebo for 4 wk followed by a 4-wk treatment with either 400 microg folic acid, 400 microg folic acid + 6 microg vitamin B-12, or 400 microg folic acid + 400 microg vitamin B-12. RESULTS: Significant reductions (P < 0.001) in plasma homocysteine were observed in all groups receiving vitamin treatment. The effect observed with the combination of folic acid + 400 microg vitamin B-12 (total homocysteine, -18%) was significantly larger than that with a supplement containing folic acid alone (total homocysteine, -11%) (P < 0.05). Folic acid in combination with a low vitamin B-12 dose (6 microg) affected homocysteine as well (-15%). CONCLUSIONS: These results suggest that the addition of vitamin B-12 to folic acid supplements or enriched foods maximizes the reduction of homocysteine and may thus increase the benefits of the proposed measures in the prevention of vascular disease and neural tube defects.

A moderate increase in daily protein intake causing an enhanced endogenous insulin secretion does not alter circulating levels or urinary excretion of dehydroepiandrosterone sulfate.

To study the effect of a moderate increase in insulin secretion produced by an increased daily protein intake on dehydroepiandrosterone sulfate (DHEAS), a balanced randomized crossover trial consisting of three strictly controlled dietary regimens was performed in six healthy male volunteers. The basic diet (B) contained 50 g protein/d; diets P and M (also basic diets) were enriched with either 32 g protein/d (P) or 10 mmol L-methionine/d (M). Methionine was given (as a specific nonprotein source of endogenously derived sulfate) to control for possible confounding effects on DHEAS due to an increased sulfate supply. At the end of each 4-day diet period, blood and 24-hour urine samples were collected. Fasting plasma levels of testosterone, cortisol, insulin-like growth factor-I (IGF-I), and insulin, as well as urinary output of total (hot acid-cleaved) testosterone conjugates and 3alpha-androstanediol glucuronide, did not show significant changes in response to dietary manipulations. Endogenous sulfate availability (as reflected by renal sulfate output per 24 hours) approximately doubled with diets P and M. However, plasma levels (6.3 +/- 1.5, 6.8 +/- 1.8, and 6.9 +/- 2.1 micromol/L for B, P, and M, respectively) and urinary excretion (8.8 +/- 9.8, 9.4 +/- 11.2, 8.0 +/- 8.3 micromol/d) of DHEAS remained unaffected. Considering the clear increments (P < .01) in urinary C-peptide excretion with diet P (20.4 +/- 10.3 nmol/d) versus diets B and M (12.6 +/- 5.1 and 13.2 +/- 3.6 nmol/d), respectively, our results suggest that a moderately strong diet-induced increase in daily insulin secretion does not alter urinary and plasma levels of DHEAS.

Re-examination of the effect of hCG on plasma levels and renal excretion of dehydroepiandrosterone sulfate in healthy males.

To reinvestigate the effect of hCG on circulating and urinary dehydroepiandrosterone sulfate (DHEAS), a hCG stimulation test (5000 IU administered i.m. at 8.30 h on 3 consecutive days) was performed in 6 healthy males (aged 24 to 35 years). Blood specimens and 24-h urine samples were collected immediately before the first and directly after the last hCG administration. Contrary to previous findings in normal men, the present study revealed significant DHEAS responses after testicular stimulation with hCG: plasma DHEAS increased from 7.9 +/- 2.3 to 9.6 +/- 2.2 mumol/L (P < 0.05) and urinary DHEAS from 5.7 +/- 3.6 to 9.3 +/- 5.2 mumol/day (P < 0.05). There was also a marked rise (P < 0.05) in the urinary excretion of total 17-ketosteroid sulfates. Clear increases of unconjugated plasma dehydroepiandrosterone as well as of circulating and renally excreted androstenedione and testosterone definitely confirmed an adequate Leydig cell stimulation. Significant post-hCG changes were additionally observed for plasma and urinary 3 alpha-androstanediol glucuronide (149% and 79% increases, respectively) and for urinary cortisol (21% decrease). Significant correlations were found for the post-hCG percent increases of plasma androstenedione versus plasma DHEAS (r = 0.86) and for the percent increases of plasma testosterone versus urinary DHEAS (r = 0.98), indicating that the extent of gonadal androgen elevations in the circulation of normal men is a determinant of DHEAS increases in blood or urine. These findings provide an explanation for the frequently observed sex differences for DHEAS in adults.(ABSTRACT TRUNCATED AT 250 WORDS)

Measurement of urinary androgen sulfates without previous hydrolysis: a tool to investigate adrenarche. Validation of a commercial radioimmunoassay for dehydroepiandrosterone sulfate.

A commercial 125I radioimmunoassay designed for the quantification of dehydroepiandrosterone sulfate (DHEAS) in blood samples was tested for its direct applicability to 24-hour urine samples from children and adults. Average recoveries in parallelism and spiking experiments were found to be near 100%. Intra- and inter-assay coefficients of variation were below 10%. Urinary DHEAS concentrations determined directly by the radioimmunoassay (x) differed only slightly from corresponding radioimmunoassay values (y) obtained after C18 reversed-phase extraction and LH-20 chromatography (y = 0.85x + 0.12; r = 0.99). Cross-reactivity data from related steroids suggested only a small contribution to the DHEAS titer by other steroids. In 8-year-old children compared to preadrenarchal children (4 years old) a clearly increased median daily urinary DHEAS output could be observed both for absolute excretion data (0.163 versus 0.05 mumol/d, P < 0.01) and for excretion values related to body surface area (0.181 versus 0.071 mumol/d/1.73 m2, P < 0.05). However, this "onset of adrenarche" was no longer statistically significant when urinary creatinine was taken as adjustment parameter for renal androgen sulfate output. After correction with the individual body surface area, rises of urinary DHEAS from childhood to adulthood were nearly in the order of the literature data on age-corresponding serum increases of DHEAS. In conclusion, the direct radioimmunological quantification of DHEAS in 24-hour urine samples with subsequent correction for individual body surface area appears to present a physiologically meaningful way to assess the adrenal gland's secretory activity for this androgen sulfate.

Alcohol consumption and its relation to cardiovascular risk factors in Germany.

OBJECTIVE: To analyse the association of alcohol consumption and blood lipids, haemostatic factors, and homocysteine in German adults by gender and age groups. DESIGN: Cross-sectional population-based survey. SETTING: Data from the German National Health Interview and Examination Survey 1998, representative for age, gender, community size, and federal state. SUBJECTS: From a sample of 7124 Germans between 18 and 79 y old, 2420 women and 2365 men were selected. Only individuals who were not currently receiving medical treatment or did not have disorders related to cardiovascular disease were selected for this study. RESULTS: Using analyses of variance, mean blood levels of total cholesterol, HDL cholesterol, HDL/total cholesterol ratio, total glycerides, fibrinogen, antithrombin III, and homocysteine adjusted for age, socioeconomic status, East/West Germany residence, body mass index, tobacco use, sports activity, and coffee consumption, if appropriate are presented by alcohol consumption groups (0, >0-10, >10-20, >20-30 and >30 g/day). The HDL/total cholesterol ratio increased with higher alcohol groups up to 10-20 g/day (+15%) for women and >30 g/day (+18%) for men, showing the strongest rise among men aged 55-79 y. Fibrinogen decreased with higher alcohol groups up to 10-20 g/day for women and 20-30 g/day for men. Among women, homocysteine levels showed a U-shaped curve with a minimum of 8.49 mmol/l at 10-20 g alcohol/day (-8%, reference: nondrinking), whereas an inverse association was observed for men. CONCLUSIONS: Moderate alcohol consumption is associated with favourable levels of several cardiovascular risk factors. The most favourable cardiovascular risk factor profile among women was observed among those drinking 10-20 g alcohol/day. Beneficial effects seem to be more pronounced among older men.

Folate economy in pregnancy.

Pregnant women are prone to becoming folate deficient because there is a significant increase in folate requirement during pregnancy and folate intakes of pregnant women are often insufficient. Reduced folate levels in blood and neutrophilic hypersegmentation reflect a negative folate balance. Possible consequences of a low maternal folate status may be pregnancy complications such as abortion, abortus imminens, abruptio placentae, and congenital malformations. The role of folic acid in the etiology of neural tube defects has been discussed for decades. The importance of an adequate maternal folate status in the prevention of neural tube defects has been demonstrated by observational and controlled intervention trials. However, the mechanism of the protective effect of periconceptional folate supplementation is not completely understood. Metabolic disorders are probably involved in the pathogenesis of neural tube defects so that a relative folate shortage rather than folate deficiency seems to be responsible for the disturbed neural tube development, which can be compensated for by a higher folate intake.

Studies on the distribution of (1-14C) pantothenic acid in rats.

The distribution of (1-14C) pantothenic acid was studied in two different experiments with rats. Single dose kinetics were carried out over a period of 168 hours. In this experiment it could be shown that besides the urinary and faecal output a third route of excretion exists: the respiratory pathway. The tissues with the highest retention capacity were the kidneys, pituitary gland, heart, muscle, liver and the adrenal glands. In a second experiment - performed as steady state study, lasting 35 days - during the investigated period and finally in the steady state the above mentioned tissues showed the highest affinity for pantothenic acid, indicating an intensive metabolism of this vitamin. These results are discussed, especially towards pantothenic acid and acetyl coenzyme A-depending functions.

[Investigation or oral substitution of iron and folic acid (author's transl)]. / Untersuchungen zur oralen Eisen-Folsäure-Substitution.

In a study of apparently healthy volunteers (5 male, 5 female) the bioavailability of iron and folacin was calculated using Tardyferon¿ resp. Tardyferon-Fol¿ as test substances. Blood samples for serum iron and folacin level determinations were obtained before and 1/2, 1, 2, 4, 6 and 8 hours after dosing. Because of the diurnal fluctuation of serum iron and folate levels, baseline levels were obtained at the corresponding 0, 1/2, 1, 2, 4, 6 and 8 hours one week prior to the study. Serum iron and folacin levels above the baseline values and the area under the curve were calculated. The data suggested a good bioavailability of the tested substances. The folacin absorption after a single dose of 0.35 mg produced a maximal serum concentration of mean = 28 ng/ml. Twice the dose resulted in a maximum of serum folacin level of mean = 51 ng/ml. The oral administration of 80 mg Fe2+ resp. 160 mg Fe2+ resulted in a significant increase of the serum iron levels, too.

[Evaluation of the folacin status in children with regard to the cobalamin and iron status. 1. Changes in the differential blood picture as a function of the vitamin and ferritin levels of the serum vs. erythrocytes]. / Untersuchungen zur Bewertung der Folatversorgung bei Kindern unter Berücksichtigung des Cobalamin- und Eisenhaushalts. 1. Mitteilung: Veränderungen im Differentialblutbild in Abhängigkeit von den Vitamin- und Ferritinkonzentrationen in Serum bzw. Erythrozyten.

The biochemical data concerning the evaluation of the folacin status in children are insufficient, as there are no biologically based borderlines available. In a study with 165 children between 1 and 15 years we looked for serum and erythrocytic folate concentrations and correlated the results with morphological alterations in the blood picture (lobe average of the neutrophilic granulocytes). As cobalamin and iron status influences different folacin depending parameters we estimated the vitamin B12- and ferritin concentration in serum as well. In our study neither the iron nor the cobalamin status influenced the blood picture and the blood folate concentrations. That's why all samples could be taken for the final evaluation. It could be demonstrated that folacin serum levels below 4.0 ng/ml respectively erythrocytic concentrations below 250 ng/ml correspond with an increasing lobe average above 3.5 compared with normal values ranging between 2.8-3.5. These biologically based borderlines were comparable between the different age groups. On the basis of these data it is possible to assess the duration and severity of folacin deficiency in children.

[Evaluation of the folacin status in children with regard to the cobalamin and iron status. 2. Incidence and severity of folate deficiency]. / Untersuchungen zur Bewertung der Folatversorgung bei Kindern unter Berücksichtigung des Cobalamin- und Eisenhaushalts. 2. Mitteilung: Häufigkeit und Schweregrad eines Folatmangels.

In a study with 165 children the folate, cobalamin and iron status was investigated by biochemical and haematological methods. On the basis of biologically based borderlines it could be demonstrated, that 15% of the children showed folate deficiency in a progressive state. This means that low serum resp. low erythrocytic folate concentrations correlated with alterations in the blood picture (lobe average of the neutrophilic granulocytes). Especially children in the age of puberty (11-15 years) are to be regarded as vulnerable; 30% of this group show deficiency symptoms whereas in very young children (1-5 years) only in 10% the supply situation is insufficient. The folate status in pupils (6-10 years) is comparable to adults (15% deficient). As cobalamin and iron status may influence folate depending parameters the status of these nutrients was estimated, too. Cobalamin deficiency was not detectable in any case. Though the iron supply situation was not adequate in all cases, interactions with the folate depending parameters could be excluded.

[Assessment of the folate supply in children with regard to cobalamin and iron maintenance. 3: Modification of the supply status by various diseases]. / Untersuchungen zur Bewertung der Folatversorgung bei Kindern unter Berücksichtigung des Cobalamin- und Eisenhaushalts. 3. Mitteilung: Beeinflussung der Versorgungssituation durch verschiedene Erkrankungen.

Assessment and Evaluation of the Folacin Supply in Children with Respect to the Cobalamin- and Iron Status - 3rd Communication. On the basis of biochemical data concerning folate-, cobalamin- and iron status of healthy children (n = 165) we estimated the supply situation in 258 children with different diseases. Patients suffering from celiac disease showed in a high frequency symptoms of a "subclinical-manifest" folate deficiency. In spite of folate therapy in Crohn's disease the supply situation was as bad. The supply situation of the controls and infected children was comparable. Anticonvulsant (folate antagonists) therapy had a significant influence on the folate status demonstrated with radioimmunological technique. In contrast to these results patients suffering from phenylketonuria had exceptionally better folate values than the controls. Obviously the vitamin content of the diet products exceeds the recommendations.

Causes and consequences of hyperhomocyst(e)inemia.

Increased plasma concentrations of the sulfur-containing amino acid homocyst(e)ine are designated as hyperhomocyst(e)inemia. Various definitions have been used to derive cut-off levels for hyperhomocyst(e)inemia. The classification by Kang is now generally used distinguishing moderate, intermediate and severe hyperhomocyst(e)inemia. A variety of causes are discussed for the etiology of the disease which can be grouped into genetic and nongenetic factors. Severe hyperhomocyst(e)inemia is accompanied by homocystinuria and several other symptoms occurring early in life. Treatment is mandatory for normal development and prevention of premature atherosclerosis. Even less severe forms of hyperhomocyst(e)inemia imply a substantially elevated risk for vascular diseases. Etiology and severity of defect(s) leading to hyperhomocyst(e)inemia are the basis for treatment. In genetic defects, supplementation with the cofactor(s) of the affected enzyme is used to enhance enzyme activity. Alternative routes in the pathway may also be enhanced. Nongenetic hyperhomocyst(e)inemia often requires correction of suboptimal vitamin concentrations. Nutritive doses of the vitamins may be sufficient for treatment of less severe forms as well as for prevention of hyperhomocyst(e)inemia.

[Biochemical and hematological measures for determination of folate status in humans. 1. Relation between serum folate and segmentation of neutrophil granulocytes]. / Biochemische und haematologische Massstäbe zur Beurteilung des Folatstatus beim Menschen. 1. Mitteilung: Beziehung zwischen Serumfolat und Segmentation der neutrophilen Granulozyten.

In a study on 130 student volunteers the segmentation of neutrophil granulocytes was measured. The serum folate concentration was used as biochemical reference. Both parameters indicate the state of folate supply or the degree of folate deficiency, respectively. Considering the haematologic response as an objective criterion only 71% of the examines showed an adequate folate status. 11% were found "deficient" and 18% were in a marginal state of folate supply. This result was confirmed by repeating the experiment. Further studies are to include folate concentration of erythrocytes and urine concentration of formiminoglutamic acid.

[Biochemical and hematological measures for determination of folate status in humans. 2. Comparative determination of folate in blood and in erythrocytes]. / Biochemische und haematologische Massstäbe zur Beurteilung des Folatstatus beim Menschen. 2. Mitteilung: Vergleichende Messungen von Folat im Serum und in Erythrozyten.

Criteria to judge the folate supply have been evaluated in previous studies. The present study is examining parameters to estimate the degree of folate deficiency. Serum folate levels and erythrocyte folate concentrations were measured in 116 volunteers (patients). The erythrocyte folate is to a large extend independent of exterior supply and is lowered only in severe chronic deficiency. Low levels of erythrocyte folate indicate longstanding undersupply. Our experiments show that folate concentration of greater than 250 ng per ml erythrocytes indicate adequate supply. Concentrations below this limit are characteristic for chronic folate deficiency. To confirm this statement further studies will take into account the urine concentration of formiminoglutamic acid.

[Biochemical and hematological parameters for the estimation of folate status in man. 3. Importance of formiminoglutamic acid excretion in the urine in comparison to other measurements]. / Biochemische und hämatologische Massstäbe zur Beurteilung des Folatstatus beim Menschen. 3. Mitteilung. Bedeutung der Formiminoglutaminsäureausscheidung im Urin im Vergleich zu anderen Messgrössen.

In a study on 250 industrial workers the amount of the formiminoglutamic acid excretion in urine was measured by an enzymatic spectrophotometric method. For the estimation of the folacine status the vitamine concentration in serum and erythrozytes was determined as well as the segmentation of the neutrophilic granulocytes. As shown in previous studies these parameters indicate the degree of folate supply. A correlation between the FIGLU excretion in urine and other folate dependent parameters could not be demonstrated. Criterial to judge the folate supply are the radioimmunological determination of folacine concentration in serum and erythrocytes as well as the estimation of the segmentation of the neutrophilic granulocytes.